A method of cultivating, preparing, and using a matrix-blank biomass

Abstract

The invention described herein relates to selectively-bred, matrix-blank organisms selected to: a) not express a bioactive compound or group of compounds; yet b) express, at least substantially qualitatively, most other non-bioactive compounds present in an analytically active comparator organism, wherein the organisms are used to generate a matrix-blank reference material with a chemical profile that resembles that of the comparator organism less the active compound or group of compounds, as well as methods of making the organisms and uses thereof. The invention described herein further relates to the matrix-blank reference materials, methods of making, and uses thereof.The invention described herein relates to selectively-bred, matrix-blank organisms selected to: a) not express a bioactive compound or group of compounds; yet b) express, at least substantially qualitatively, most other non-bioactive compounds present in an analytically active comparator organism, wherein the organisms are used to generate a matrix-blank reference material with a chemical profile that resembles that of the comparator organism less the active compound or group of compounds, as well as methods of making the organisms and uses thereof. The invention described herein further relates to the matrix-blank reference materials, methods of making, and uses thereof.

Description

TECHNICAL FIELD OF THE INVENTION

[0001] The invention described herein relates to novel matrix-blank organisms, the methods used to grow and produce these organisms, and the processes to create a novel bioactive-free organism to be used to generate an analytical matrix blank. Methods to produce extracts free of an active compound or group of compounds obtained therefrom and their uses are also disclosed. In an exemplary embodiment, the novel matrix-blank organism is a Cannabis plant devoid of one or more cannabinoids to produce a representative Cannabis matrix blank. BACKGROUND TO THE INVENTION

[0002] Accurate testing of active compounds in biological organisms is critical for both compliance and quality assurance within an industry. In the context of analytical testing, particularly for quantifying active compounds in complex matrices, a matrix blank devoid of that active compound is essential. Similar to practices in pharmaceutical analysis, this approach helps rule out potential interference from the matrix itself in the analysis.

[0003] Advanced analytical techniques, such as gas chromatography (GC) and liquid chromatography (LC) coupled to mass spectrometry, benefit from matrix-matched calibrations. Leghissa et al. (2018b) discuss the use of GC with triple quadrupole mass spectrometry for analyzing cannabinoids, highlighting the method’s applicability to plant materials and Cannabis products. In another study, Leghissa et al. (2018c) explore the use of gas chromatography with vacuum ultraviolet spectroscopy (GC-VUV) for the rapid detection of cannabinoids. Additionally, Aizpurua-Olaizola et al. (2016) and Dalltige et al. (2003) demonstrate the efficacy of two-dimensional gas chromatography (GC * GC) for analyzing complex cannabinoid mixtures.

[0004] The Cannabis industry has established a unique standard for analytical testing to ensure product safety. This is particularly notable given the Cannabis plant’s ability to produce over 500 bioactive compounds, with cannabinoids being the primary focus. A cannabinoid-free version of Cannabis is crucial for precise measurement of bioactive molecules, while avoiding matrix-induced artifacts as Cannabis is known to produce hundreds of metabolites other than cannabinoids.

[0005] Plants of the genus Cannabis have been used as a drug for centuries as a botanical drug, although the precise basis for the plant’s activity is not known. Both tetrahydrocannabinol (THC) and cannabidiol (CBD), two of the plant’s most abundant cannabinoids, are known to have distinct pharmacological activities. It is noted that botanists disagree regarding whether the genus Cannabis includes only one species or multiple species. For purposes of this disclosure, reference to Cannabis is intended to refer to any species or subspecies of Cannabis, whether the species is sativa, indica, ruderalis or is unspecified.

[0006] For extracts, it is unclear whether the efficacy of a botanical drug extract is attributable to the identified “active(s)” or “markers” and / or other components present in an extract which may provide an unidentified additive or synergistic effect or in fact be directly responsible for the activity.

[0007] In the biosynthesis of cannabinoids within Cannabis plants, a series of enzymatic reactions are involved. The initial step in the formation of pentyl cannabinoids is the enzymatic reaction where geranylpyrophosphate (GPP), a terpenoid, combines with olivetolic acid (OA), a phenolic compound. This reaction results in the creation of cannabigerolic acid (CBGA) and is facilitated by the enzyme geranylpyrophosphate:olivetolate geranyltransferase (GOT).

[0008] The production of GPP involves two distinct pathways: the mevalonate pathway (MVA) in the cytoplasm and the deoxyxylulose pathway (DOX) in the plastid compartments. Research indicates that GPP used in cannabinoids primarily comes from the DOX pathway present in glandular trichome plastids. On the other hand, olivetolic acid (OA) is synthesized through a polyketide-type mechanism involving the condensation of n-hexanoyl-CoA and three malonyl-CoA molecules.

[0009] Further in the pathway, cannabigerolic acid (CBGA) acts as a direct precursor for other cannabinoids like tetrahydrocannabinolic acid (THCA), cannabidiolic acid (CBDA), and cannabichromenic acid (CBCA). These transformations from CBGA to other cannabinoids are also enzyme-driven processes, with specific synthases identified for each conversion.

[0010] In cases where GPP combines with divarinic acid (DA) instead of OA, cannabinoids with propyl side chains are formed, leading to the production of cannabigerovarinic acid (CBGVA). The enzymatic conversion of CBGVA by the cannabinoid synthases results in the formation of the propyl homologues of CBDVA, THCVA, and CBCVA.

[0011] Lastly, the decarboxylation of acidic cannabinoids, a process influenced by factors like UV light, heat, and enzymatic action, leads to the formation of their neutral analog, such as, but not limited to THC, CBD, CBG, CBC, CBDV, CBCV, THCV, and CBGV.

[0012] The exact mechanism of biosynthesis of other chain length cannabinoids, such as oricin and phorol cannabinoids, is unknown. However, it has been hypothesized that these cannabinoids are created through the condensation of other precursors than n-hexanoyl-CoA and malonyl-CoA which leads to the different carbon chain length observed in these cannabinoids. It has been shown that the downstream pathway following the creation of CBGOA or CBGPA is consistent between the various chain length cannabinoids.

[0013] A more likely explanation for the absence of cannabinoids in these plants could be an obstruction in certain biochemical pathways essential for generating precursor molecules leading to CBGA. The unaltered chemical profile of terpenes in these cannabinoid-free plants implies that the disruption does not occur in the initial stages of the MVA and DOX pathways responsible for Isopentenyl Pyrophosphate (IPP) production. This suggests that the interruption might occur downstream of the DOX and MVA biosynthetic pathways.

[0014] Impeding the MVA pathway would not directly impact cannabinoid synthesis, but it could result in reduced levels of sesquiterpenes, sterols, and triterpenes. On the other hand, a disruption in the DOX pathway would hinder the formation of the terpenoid components essential for cannabinoids and could also adversely affect the production of monoterpenes, diterpenes, triterpenes, and tetraterpenes.

[0015] The specific phenotype observed could be due to a malfunction in the enzyme geranyl diphosphate:olivetolate geranyltransferase (GOT), which is vital for combining resorcinolic acids (OA and DA) with GPP to produce cannabigerolic acid. An ineffective GOT enzyme might lead to an accumulation of OA and / or DA. However, this was not confirmed through HPLC analysis for cannabinoid content, which would have detected these acids in both their carboxylated and decarboxylated forms, as per De Meijer et al., 2003.

[0016] Thus, it appears most plausible that the absence of cannabinoids is caused by a disruption in the polyketide pathway that produces OA and DA. The precise nature of this cannabinoid knockout phenomenon remains uncertain, particularly considering that a functional synthase would override a non-functional variant. This raises further questions about the suppressed cannabinoid synthesis in heterozygous genotypes, as noted by De Meijer et al., 2003.

[0017] There is a need for a matrix blank botanical specimen, engineered for comparative analytical purposes. Predicated upon novel cultivation and processing, this exemplary botanical genotype, referred to herein as the “knock-out organism,” is characterized by its absence of conventional organism-specific compounds due to a mutation in the biosynthetic pathway of that compound, as exemplified by the production of cannabinoid-free Cannabis in the case of cannabis, through selectively breeding non-functional cannabinoid biosynthetic genes. Such a specimen is instrumental in corroborating accurate quantification of various bioactive compounds intrinsic to the Cannabis plant and its affiliated extracts.

[0018] Additionally, there is a need for an innovative organism and resulting biomass or extracts designed to facilitate the identification of specific human metabolites post exposure to the organism lacking the drugs to be analyzed. This allows for drug analysis laboratories to have true comparative material to quantify active compounds more accurately. The implications of this innovation extend to the enhanced accuracy in evaluating diverse biological matrices, encompassing but not limited to saliva, blood, hair, breath, and urine from users who have consumed cannabinoid-free Cannabis based products. SUMMARY OF INVENTION

[0019] Embodiments of the invention disclosed herein relate to biological matrix-blank reference materials derived from a selectively-bred organism, the selectively-bred organism being characterized by an absence of one or more bioactive compounds expressed in the comparator organism. For example, in the case of the organism being Cannabis, the selectively, bred Cannabis is characterized by the absence one or more of the following cannabinoids: tetrahydrocannabiphorol (9-THCP), tetrahydrocannabivarin (9-THCV), delta-8-tetrahydrocannabinol (8-THC), delta-9-tetrahydrocannabinol (9-THC), tetrahydrocannabinolic acid (9-THCA), tetrahydrocannabiphorolic acid (9-THCP A), tetrahydrocannabivarinic acid (9-THCVA), cannabichromene (CBC), cannabichromenic acid (CBCA), cannabichromeorcin (CBCO), cannabichromevarin (CBCV), cannabichromevarinic acid (CBCVA), cannabidiol (CBD), cannabidiolic acid (CBDA), cannabidibutol (CBDB), cannabielsoin (CBE), cannabidiol monomethyl ether (CBDM), cannabidiol acid monomethyl ether (CBDAM), cannabidiorcin (CBDO), cannabidiphorolic acid (CBDPA), cannabidivarin (CBDV), cannabidivarinic acid (CBDVA), cannabigerol (CBG), cannabigerolic acid (CBGA), cannabigerol acid monomethyl ether (CBGAM), cannabigerol monomethyl ether (CBGM), cannabigerovarin (CBGV), cannabigerovarinic acid (CBGVA), cannabigerphorolic acid (CBGPA), cannabicyclol (CBL), cannabicyclolic acid (CBLA), cannabinol (CBN), cannabinolic acid (CBNA), cannabicitran (CBT).

[0020] In some embodiments, the selectively-bred Cannabis is further characterized by the presence of trichomes of clear, white, and / or amber coloration. In some embodiments, the selectively-bred Cannabis is the Cannabis variety designated as ‘ZCP.’

[0021] In some embodiments, the matrix-blank organisms or reference materials disclosed herein include flavonoids including cannflavin A, cannflavin B, isocannflavin B, cannflavin C, or other Cannabis-specific flavonoids. In some embodiments, the matrix-blank organisms or reference materials disclosed herein include terpenes. In some embodiments, matrix-blank organisms or reference materials disclosed herein are further characterized by one or more of the presence of both monoterpenes and sesquiterpenes; the absence of either monoterpenes or sesquiterpenes; the absence of both monoterpenes and sesquiterpenes, the presence of di terpenes, the absence of di terpenes, the presence of tri terpenes, the absence of tri terpenes, the presence of tetraterpenes, and / or the absence of tetraterpenes.

[0022] In some embodiments, the matrix-blank reference materials disclosed herein comprise a homogenized bulk biomass lot devoid of a profile of entourage or contaminant compounds or organisms. In some embodiments, the matrix-blank organisms or reference materials disclosed herein are further characterized by at least one of the presence of a known compound or organism spiked in at a known concentration for analytical reference; and / or the addition of one or more compounds selected from the group consisting of flavonoids, alkaloids, terpenoids, carotenoids, cannabinoids, alcohols, ketones, ethers, amines, amides, esters, aldehydes, sterols, pesticides, herbicides, fungicides, hydrocarbons, synthetic cannabinoids, synthetic psychedelics, synthetic opioids, isoxazoles, halogenated compounds, radioactive elements, heavy metals, Aspergillus, Salmonella, Shiga-Toxin Producing Escherichia coli, non-pathogenic Escherichia coli, Pseudomonas, Listeria, Hepatitis, norovirus, Clostridium, Campylobacter, Taxoplasma, Psilocybe species genes, Cryptosporidium, Saccharomyces, hops-latent viroid, beat-curly top virus, tobacco mosaic virus, and radiolabeled nucleic acids. In some embodiments, the spiked amount of the compound is at a regulatory body’s limit of detection and / or limit quantification for that analyte. In some embodiments, the spiked amount of compound is greater than the regulatory body’s limit detection and / or limit of quantification for that given analyte. In some embodiments the spiked amount of compound is less than the regulatory body limit detection and / or limit of quantification for that given analyte.

[0023] In some embodiments, the matrix-blank reference materials disclosed herein for use in biological consumption to create a bioactive-free matrix-blank material from biological tissue or excrement, further characterized by the addition of one or more compounds selected from the group consisting of flavonoids, terpenoids, carotenoids, cannabinoids, alcohols, ketones, pesticides, herbicides, fungicides, solvents, synthetic cannabinoids, synthetic psychedelics, synthetic opioids, isoxazoles, halogenated compounds, radioactive elements, Aspergillus, Salmonella, Escherichia coli, and / or heavy metals.

[0024] For example, in some embodiments, cannabinoid-free Cannabis is sprayed with certain pesticides, fungicides, or herbicides specific to the contaminated matrix blank material desired, such as spraying pyrethrins as a pest management strategy when creating a pesticide-contaminated mixture of biomass to use in determining a laboratories ability to detect that given pesticide.

[0100] In some embodiments, the matrix-blank reference materials disclosed herein can be consumed by humans or animals to create a comparative analytical chemistry matrix. In some embodiments, the matrix-blank reference materials disclosed herein employed for analytical chemistry purposes as a matrix blank to compare to unknown Cannabis samples.

[0025] In some embodiments, the matrix-blank organisms or reference materials disclosed herein are utilized for proficiency testing or regulatory testing enforcement to evaluate the ability of a laboratory to detect and quantify pathogens or adulterants in Cannabis. In some embodiments, the matrix-blank reference materials disclosed herein are used for proficiency testing or regulatory testing enforcement to evaluate the ability of a laboratory to detect and quantify pathogens or adulterants in Cannabis.

[0026] In some embodiments, a method for cultivating Cannabis to produce pesticide-, herbicide-, fungicide-, heavy metal-, mycotoxin-, endotoxin-, and microbial-free Cannabis biomass, as described in Example 1 is provided. In some embodiments, a method for sterile post-harvest processing of Cannabis to prevent environmental contamination, as described in Example 2 is provided.

[0027] In some embodiments, a method for packaging the matrix-blank reference materials disclosed herein in sterile containers meeting or exceeding EPA standards in a clean environment, ensuring no contamination during packaging, transport, or storage are provided. In some embodiments, the packaging is performed under ISO 17034 accreditation as a certified reference material. In some embodiments, the matrix-blank reference materials disclosed herein are provide as certified reference materials.

[0028] In some embodiments, the tissues of the matrix-blank organisms disclosed herein are used for comparison with unknown or known bioactive-containing samples of the same organism.

[0029] In some embodiments, the organism is selected from one of the following organisms: Lion’s Mane mushroom species such as Hericium erinaceus, Hericium coralloides, and Hericium americanum: Reishi mushroom species including Ganoderma lucidum, Ganoderma tsugae, and Ganoderma applanalum: Chaga mushroom, specifically Inonotus obliquus', Cordy cep mushrooms such as Cordyceps militaris, Cordyceps gunni, and Cordyceps dipterigena; Amanita mushroom species such as Amanita citrina, Amanita muscaria, Amanita gemmata, Amanita. Pantherina, Amanita porphyria, Amanita regalis, and Amanita strobiliformis; various psilocybin-containing mushrooms such as Psilocybe tampanensis, Psilocybe cubensis, Psilocybe semilanceala, Psilocybe oaxacana, Psilocybe natalensis, Psilocybe baeocystis, Psilocybe zapatecorm, Psilocybe azurescens, Psilocybe mexicana, Panaeoulus cyanescens, Panaeoulus cinctulus Psilocybe cyanescens, Panaeolus acidus, Panaeolus acuminatus, Panaeolus affinis, Panaeolus africanus, Panaeolus albellus, Panaeolus albidocinereus, Panaeolus albovelutinus, Panaeolus aids, Panaeolus alveolatus, Panaeolus annulatus, Panaeolus anomalus, Panaeolus antillarum, Panaeolus atomatus, Panaeolus atrobalteatus Panaeolus axfordii, Panaeolus bisporus, Panaeolus cambodginiensis, Panaeolus fimicola, Panaeolus moellerianus, Panaeolus olivaceus, Panaeolus fimicoloides, Panaeolus fimiputris, Panaeolus foenisecii, Panaeolus fontinalis, Panaeolus fraxinophilus, Panaeolus georgii, Panaeolus gomphodes, Panaeolus goossensiae, Panaeolus griseofibrillosus, Panaeolus guttulatus, Panaeolus hippophilus, Panaeolus hygrophanus, Panaeolus hypomelas, Panaeolus incanus, Panaeolus indicus, Panaeolus intermedins, Panaeolus lentisporus, Panaeolus lerchenfeldii, Panaeolus leucophanes, Panaeolus lignicola, Panaeolus linnaeanus, and Panaeolus longiguus, Lophophora williamsii, Lophophora diffusa, Lophophora fricii, Trichocereus pachanoi, and Trichocereus peruvianus, Mimosa hostilis / tenuiflora and Acacia confusa, Banisteriopsis caapi, Passiflora incarnata, and Peganum harmala.

[0030] In some embodiments, the selectively-bred organism is selected using a genetic marker complementary to one or more biosynthetic pathway genes that select against the biosynthesis of an active component. In some embodiments, the bioactive compound is selected based on the compound being unique to the organism, for example, cannabinoids in Cannabis, Cordycepin in Cordyceps, and Psilocybin in Psilocybe, and the like.

[0031] In some embodiments, the matrix-blank reference material comprises biomass or an extract of the organism. In some embodiments, the matrix-blank organism is used to create an extract utilizing either a solventless method or a solvent selected from a range including, but not limited to, ethanol, methanol, carbon dioxide, propane, butane, heptane, hexane, naphtha, limonene, pinene, or analogous polar or non-polar solvent system.

[0032] In some embodiments, the matrix-blank reference material is free from one or more molecules of the following classes: herbicides, fungicides, elements, isotopically labels, bioactive molecules, biologicals, and hydrocarbons.

[0033] In some embodiments, the matrix-blank reference material is combined with one or more molecules selected from one or more of the following bioactive constituents including, but not limited to, alkaloids, monoterpenes, diterpenes, triterpenes, tetraterpenes, sterols, amino acids; contaminant constituents such as but not limited to lead, mercury, cadmium, arsenic, chromium, nickel, copper, zinc, gold, uranium, radon, silver, synthetic drugs of abuse, myclobutanil, bifenthrin, avermectin, imazalil, permethrin, spiromesifen, spirotetramat, chlorfenapyr, pyrethrins, imidacloprid, carbaryl, malathion, chlorothalonil, mancozeb, iprodione, propiconazole, pyraclostrobin, vinclozolin, copper-based fungicides, aflatoxins, ochratoxins, endotoxins, polycyclic aromatic hydrocarbons, plasticizers, dioxins and pcbs, nitrates and nitrites, acrylamide, and biological contaminants (live or dead cells of Salmonella spp, Escherichia coli, Listeria monocytogenes. Staphylococcus aureus, Botritiys spp, Aspergillus spp, Clostridium botulinum, Bacillus cereus, Campylobacter spp., yeasts, viroids, molds), as well as extracted or synthesized DNA / RNA of these species.

[0034] In some embodiments, the matrix-blank reference material is combined with one or more molecules selected from the group consisting of bioactive constituents, contaminant constituents, biological contaminants, and extracted or synthesized DNA / RNA of the aforementioned species.

[0035] In some embodiments, the tissues of the matrix blank organisms disclosed herein are utilized for analytical chemistry analyses such as, matrix-matched calibrations, validations, and quality controls as a bioactive-free matrix blank. In some embodiments, the tissues of the matrix blank organisms disclosed herein are used for medical studies to determine the efficacy, toxicity, or potential benefits of a given organism or compound produced by that organism. In some embodiments, the tissues of the matrix blank organisms disclosed herein are used for proficiency testing of analytical labs to determine the accuracy, precision, and ability to detect and quantify bioactive or contaminant compounds.

[0036] In some embodiments, the matrix-blank reference materials disclosed herein are utilized as control samples in scientific research, including but not limited to pharmacological, toxicological, and botanical studies. In some embodiments, the matrix-blank reference materials disclosed herein are employed in the development and calibration of analytical instruments and techniques for detecting and quantifying bioactive compounds in an organism’s biomass. In some embodiments, the matrix-blank reference materials disclosed herein are used in educational settings for demonstration and training purposes in fields related to botany, chemistry, and pharmacology.

[0037] In some embodiments, the matrix-blank reference materials disclosed herein are used in the development of new varieties of plants through genetic modification or selective breeding to introduce or enhance desirable traits while maintaining the absence of bioactive compounds. In some embodiments, the matrix-blank reference materials disclosed herein are used as a base material for the production of non-psychoactive health supplements, cosmetics, or food products.

[0038] In some embodiments, the matrix-blank reference materials disclosed herein are combined with one or more molecules selected from the group consisting of bioactive constituents, contaminant constituents, biological contaminants, and extracted or synthesized DNA / RNA of the aforementioned species.

[0039] In some embodiments, the tissues of the matrix-blank organisms are utilized for analytical chemistry analyses such as, matrix-matched calibrations, validations, and quality controls as a bioactive-free matrix blank. In some embodiments, the tissues of the matrix-blank organisms are used for medical studies to determine the efficacy, toxicity, or potential benefits of a given organism or compound produced by that organism. In some embodiments, the tissues of the matrix-blank organisms are used for proficiency testing of analytical labs to determine the accuracy, precision, and ability to detect and quantify bioactive or contaminant compounds.

[0040] In some embodiments, the matrix-blank disclosed herein is utilized as a control sample in scientific research, including but not limited to pharmacological, toxicological, and botanical studies. In some embodiments, the matrix-blank organism disclosed herein is employed in the development and calibration of analytical instruments and techniques for detecting and quantifying bioactive compounds in an organism’s biomass. In some embodiments, the matrix-blank disclosed herein is used in educational settings for demonstration and training purposes in fields related to botany, chemistry, and pharmacology. In some embodiments, the matrix-blank disclosed herein is utilized in the development of new varieties of plants through genetic modification or selective breeding to introduce or enhance desirable traits while maintaining the absence of bioactive compounds.

[0041] In some embodiments, the biological matrix-blanks disclosed herein are used as a base material for the production of non-psychoactive health supplements, cosmetics, or food products. In some embodiments, the biological matrix-blanks disclosed herein are further characterized by an altered nutritional profile, including but not limited to enhanced levels of vitamins, minerals, or dietary fibers.

[0042] In some embodiments, the biological matrix-blanks disclosed herein are used as a substrate or medium in agricultural research, particularly for studying plant growth, nutrient uptake, or disease resistance in the absence of bioactive compounds. In some embodiments, the biological matrix-blanks disclosed herein are used in the production of non-psychoactive, environmentally friendly biofuels or bioplastics. In some embodiments, the biological matrixblanks disclosed herein are used as in horticulture as a grafting stock for other plant species, leveraging its unique genetic attributes. In some embodiments, the biological matrix-blanks disclosed herein are used as in environmental studies to assess the impact of bioactive compounds on ecosystems, particularly in soil and water analyses.

[0043] In some embodiments, a method for using a matrix-blank of a selectively bred Cannabis plant in the manufacturing of textiles and fibers is provided, wherein the absence of cannabinoids does not affect the physical properties of the fibers. In some embodiments, a method for using a matrix-blank material of a selectively-bred Cannabis plant in the production of paper and cardboard materials is provided, where the lack of cannabinoids contributes to a more environmentally sustainable and ergonomically friendly processing of the biomass due to the lack of adhesive resin found in cannabinoid-containing plants.

[0044] In some embodiments, a method for using a matrix-blank of a selectively-bred Cannabis plant in animal feed is provided, wherein the absence of cannabinoids ensures safety and compliance with animal health regulations while still providing the entire Cannabis plant’s nutritional value in the mixture absent of cannabinoids or other bioactive compounds. In some embodiments, a method for using a matrix-blank material of a selectively-bred Cannabis plant as a base for non-psychoactive, cannabinoid-free cosmetic products, including but not limited to creams, lotions, transdermal products, and ointments, is provided.

[0045] In some embodiments, a method for using genetic markers associated with a cannabinoid-free phenotype of a Cannabis plant in a breeding program to develop new varieties of cannabinoid-free plants is provided.

[0046] In some embodiments, a process for the extraction of non-cannabinoid compounds from a bioactive-free matrix of a Cannabis plant, including but not limited to terpenes, lignanamides, esters, ketones, alcohols, flavonoids, alkaloids, and fatty acids, is provided for use in various industries.

[0047] In some embodiments, a method for utilizing a cannabinoid-free Cannabis plant material in environmental remediation and phytoremediation processes is provided to exploit its ability to absorb and break down pollutants in soil or water without the risk of cannabinoid accumulation. In some embodiments, a method for using a matrix-blank of a selectively-bred Cannabis plant in the production of biofuels is provided, where the absence of cannabinoids facilitates a more efficient conversion process. In some embodiments, a method for using a cannabinoid-free Cannabis plant or a biological matrix-blank therefrom, as a research tool in botanical and agricultural studies is provided, providing a unique model for understanding plant biology without the interference of cannabinoid pathways.

[0048] In some embodiments, a method for manufacturing textiles and fibers utilizing a biological matrix-blank derived from any selectively bred, bioactive-free organism is provided, wherein the absence of specific bioactive compounds does not affect the physical properties of the fibers. In some embodiments, a method for producing paper and cardboard materials using a bioactive-free matrix from any selectively bred, bioactive-free organism is provided, wherein the lack of specific bioactive compounds contributes to more environmentally sustainable processing.

[0049] In some embodiments, a method for producing animal feed using a biological matrix-blank from any selectively bred, bioactive-free organism is provided, ensuring safety and compliance with animal health regulations due to the absence of specific bioactive compounds. In some embodiments, a method for creating non-psychoactive, bioactive compound-free cosmetic products, such as creams, lotions, and ointments, using a bioactive-free matrix derived from any selectively bred, bioactive-free organism is provided.

[0050] In some embodiments, a method is provided for developing new varieties of bioactive-free organisms using genetic markers associated with a bioactive-free phenotype in any selectively bred, bioactive-free organism.

[0051] In some embodiments, a process for extracting non-bioactive compounds, such as terpenes, flavonoids, and fatty acids, from the bioactive-free matrix of any selectively bred, bioactive-free organism, is provided for use in various industries.

[0052] In some embodiments, a method for utilizing the bioactive-free biomass of any selectively bred, bioactive-free organism in environmental remediation and phytoremediation processes is provided, leveraging its ability to absorb and break down pollutants in soil or water.

[0053] In some embodiments, a method for producing biofuels using a bioactive-free matrix from any selectively bred, bioactive-free organism, is provided where the absence of specific bioactive compounds facilitates a more efficient conversion process.

[0054] In some embodiments, a method for using a bioactive-free, selectively bred organism as a research tool in botanical and agricultural studies is provided, providing a unique model for understanding plant biology without the interference of specific bioactive pathways.

[0055] In some embodiments, a method is provided for processing a matrix-blank organism wherein the absence of one or more target bioactives in the biomass input provides enhanced and more efficient processing of one or more than one of the following materials: paper, fiber, textile, biofuels, bioplastics, health supplements, cosmetic products, food products, animal feed, or animal bedding.

[0056] In some embodiments, a method is provided for using a matrix-blank organism wherein the absence of one or more target bioactives in the biomass input provides enhanced properties compared to the bioactive-containing organism when used in one or more than of the following applications: bioremediation, phytoremediation, soil erosion cover crop, agricultural crop studies, nutrient use efficiency studies, non-cannabinoid compound production, matrix blank biomass, spiked matrix biomass, disease resistance studies, pest resistance studies, compound-specific medicinal effect studies, nuclear material bioremediation, or chemical contaminant bioremediation.

[0100] In some embodiments, a seed from a Cannabis plant designated ‘ZCP’ wherein a representative sample of seed of said plant has been deposited under. In some embodiments, a Cannabis plant, or plant part, tissue, or cell thereof produced by growing the seed from the Cannabis plant designated ‘ZCP’, or a descendant thereof is provided; wherein flower produced from the plant, or plant part, tissue, or cell thereof comprises a biochemical profile as described herein. In some embodiments, the Cannabis plant part is selected from the group consisting of: a stem, a trichome, a leaf, and a flower. In some embodiments, the Cannabis plant descended from the plant, or plant part, tissue, cell, or seed above is provided, wherein the plant is a clonal descendent.

[0101] In some embodiments, a method of breeding a Cannabis plant, or plant part, tissue, or cell thereof is provided , wherein the plant, plant part, tissue, or cell is produced by growing a seed or clone from: a Cannabis plant designated wherein a representative sample of seed of said plant has been deposited under; or a descendant of the Cannabis plant designated ‘ZCP,’ wherein the plant comprises a cannabinoid profile and / or a terpene profile as described herein, and wherein the method comprises providing the plant as at least one parent in a breeding program and selecting progeny displaying a cannabinoid profile as described herein and / or a terpene profile as described herein. BRIEF DESCRIPTION OF THE DRAWINGS

[0057] Figure 1 shows chromatogram analysis of ‘ZCP’ via HPLC-PDA.

[0058] Figure 2 shows chromatogram analysis of reference materials spiked in to an extract of ‘ZCP’ via HPLC-PDA. DETAILS OF INVENTION:

[0059] Embodiments of the invention disclosed herein provide biological matrix-blank organisms (also referred to herein as “matrix-blank organism” or “bioactive-free organism”) and compositions derived therefrom (each such composition being referred to herein as a “biological matrix-blank reference material” or “matrix-blank material” or “matrix-blank biomass” or “matrix-blank extract”). Organisms are provided in which expression and / or accumulation of one or more key biochemical components (the “deleted analyte” or “deleted analytes”) has been modified, while substantially preserving the expression and accumulation of the remaining biochemical components of the organism (the “background analytes”). Thus, a matrix-blank organism has a substantially normal biochemical profile as compared with the organism from which it was derived (the “reference organism” or “comparator organism”), while having a finite and limited number of deleted analytes that are present in the reference organism. In practice, the matrix-blank organism provides a source of preparation of matrixblank reference materials in any desired form useful for any of the analyses and / or comparisons described herein, such as, but not limited to, biomass and extracts, as well as others that would be evident to those of skill in the art.

[0060] Embodiments of the invention can be any organism including but not limited to plants, fungi, microorganisms, and research animals. While the invention is primarily described in terms of plants, embodiments of the invention are not limited to plants. Likewise, while a specific matrix-blank Cannabis plant is described herein, the invention specifically contemplates other matrix-blank Cannabis varieties as well as varieties of other matrix-blank plants, fungi, microorganisms, and the like.

[0061] In practice, a matrix-blank reference material can be used to provide a baseline, a reference, or a control for a number of different analyses. Since the biochemical profile of the matrix-blank reference material is substantially the same as the biochemical profile of the same type of reference material from the reference organism, except for the deleted analytes, use of the matrix-blank reference material in any of these different analyses permits more accurate and informative results of such analyses. The two matrices being substantially the same, except for the deleted analytes, permits a level of fidelity and quality control on any analytical test that could not be achieved in any other way. This is partly because the tremendous number of variants of some classes of compounds, including but not limited to cannabinoids, for example, can lead to undesirable imprecision in many kinds of testing or analyses.

[0062] For example, if a specific test is intended to detect and quantify the presence of THC, other cannabinoids or terpenes can be mistakenly classified as THC if the quality or precision of the testing is not suitable for precise differentiation between THC and other compounds. Use of a matrix-reference material to establish a baseline of non-THC components in the reference plant permits a vastly more precise detection of actual THC by avoiding false positive identification of background analytes as being THC.

[0063] As used herein, the term “comparator plant” or “reference plant” refers to a plant variety expressing at least one medicinally active compound used to develop a new plant that has been selectively bred to not express the at least one medicinally active compound.

[0064] As used herein, the terms “biological matrix-blank organism” or “matrix-blank organism” or “bioactive-free organism” (or “matrix-blank plant,” or “matrix-blank fungi,” and the like) refer to an organism that has been selectively bred to not express at least one medicinally active or bioactive compound but to express at least one other compound expressed in a comparator organism.

[0065] As used herein, the terms “biological matrix-blank reference material” (or “matrixblank reference material) and “biological matrix-blank extract,” (or “matrix-blank extract”) and “biological matrix-blank biomass” (or “matrix-blank biomass”) are used to refer to a reference material, such as an extract or biomass from an organism that expresses all or substantially all of the compounds of a comparator organism but does not express at least one medicinally active compound.

[0066] As used herein, the term “comparative analytical chemistry matrix” refers to the use of a bioactive-free organism to create a non-organism matrix material. For example, in the case of Cannabis, “comparative analytical chemistry matrix” refers to the use of cannabinoid-free Cannabis to create a non-plant matrix material. In some embodiments, this includes infusing cannabinoid-free Cannabis, or an extract thereof, into a finished product such as, but not limited to a chocolate, cooking oil, tincture, sublingual spray, tablet, topical, candy, or transdermal patch, in order to accurately distinguish cannabinoids from the other metabolites of Cannabis present in cannabinoid-free Cannabis and the matrix material itself. In other embodiments, the cannabinoid-free Cannabis is consumed by a human participant / animal subj ect or used to create one or more biological tissue or excreted samples that contains the other metabolites of Cannabis without having cannabinoids to create a biological material specific Cannabis matrix blank. These biological samples include but are not limited to blood, hair, salvia, urine, fecal matter, and other biological tissues, fluids, and excrement.

[0067] As used herein, the term “medicinally active” refers to a compound that has demonstrated biological effects beneficial for health, such as, for treating, preventing, or managing diseases or medical conditions. In some embodiments, “medicinally active” compounds have pharmacological effects on the body, such as reducing pain, inflammation, or symptoms of diseases. Medicinally active compounds are sometimes referred to as “active” compounds or ingredients, which are responsible for the therapeutic effects of a plant.

[0068] As used herein a “medicinal plant” refers to a plant used or consumed primarily for the purpose of treating or managing health conditions and alleviate symptoms of diseases. As an example, medicinal Cannabis plants typically contain cannabinoids and terpenes, such as cannabidiol (CBD) and / or beta caryophyllene, which have potential therapeutic benefits.

[0069] As used herein a “recreational plant” refers to a plant used or consumed primarily for the purpose of enjoyment, relaxation, or social interaction. As an example, recreational Cannabis is typically consumed for its psychoactive effects primarily from THC. Consumers experience euphoric effects, altered perception, and / pr mood enhancement.

[0070] As used herein, the terms “contaminant chemical” and “contaminant organism” refer to a material incorporated into a matrix blank organism to mimic a sample that would fail testing, for example, regulatory testing for a given set of regulations. Examples of contaminant chemicals and organisms include, but are not limited to: mycotoxins, neurotoxins, drugs of abuse, tryptamines, hericenones, erinacines, ganoderic acids, lucidenic acid, inotodiol, lanosterol, betulinic acid, betulin, mescaline, hordenine, anhalonidine, and tyramine; contaminant constituents including but not limited to Per- and polyfluoroalkyl substances (PFAS), lead, mercury, cadmium, arsenic, chromium, nickel, copper, zinc, gold, uranium, radon, silver, myclobutanil, bifenthrin, avermectin, imazalil, permethrin, spiromesifen, spirotetramat, chlorfenapyr, pyrethrins, imidacloprid, carbaryl, malathion, chlorothalonil, mancozeb, iprodione, propiconazole, pyraclostrobin, vinclozolin, copper-based fungicides, aflatoxins, ochratoxins, endotoxins, polycyclic aromatic hydrocarbons, plasticizers, dioxins and pcbs, nitrates and nitrites, acrylamide); biological contaminant organisms including but not limited to live or dead cells of Salmonella spp, Escherichia coli, Shiga Toxin Producing E. Coli (STEC), Listeria monocytogenes. Staphylococcus aureus, Botritiys spp, Aspergillus spp, Clostridium botulinum, Bacillus cereus, Campylobacter spp, yeasts, viroids, molds, as well as extracted or synthesized DNA / RNA of these species.

[0071] As used herein, the term “spiked” refers to the process of adding a known quantity of a substance, for example, a specific compound or contaminant, to a sample or a matrix for the purpose of analytical testing, calibration, verification, and / or validation.

[0072] As used herein, the term “biomass” refers to the total mass of organic material produced by an organism during its growth cycle. In terms of a plant, for example, this can include leaves, stems, flowers, preflowers, trichomes, and roots. Biomass can be measured in terms of dry weight or fresh weight.

[0073] As used herein, the term “extract” refers to a composition derived from the processing of plant or fungal biomass, including but not limited to roots, stems, leaves, flowers, fruiting bodies, spores, or mycelium, through physical, chemical, or biological methodologies to isolate, concentrate, or purify one or more constituents including but not limited to primary or secondary metabolites, lipids, proteins, tryptamines, cannabinoids, alkaloids, phenolic compounds, flavonoids, carbohydrates, nucleic acids, or derivatives thereof. The term encompasses products obtained via solvent extraction (e.g., ethanol, methanol, acetone, supercritical CO2), mechanical processing (e.g., pressing, grinding, maceration), chromatographic fractionation, distillation, enzymatic or microbial biotransformation, or advanced technologies such as cold plasma or microwave-assisted extraction. Specific examples include cannabinoid-rich, flavonoid-rich or terpene-rich compositions derived from Cannabis sativa I.., Cannabis indica. or hybrids thereof using supercritical CO2, mechanical processing, or hydrocarbon extractions, and psilocybin or psilocin compositions from psilocybin-producing fungi such as Psilocybe cubensis obtained through aqueous, alcohol-based, methanol-based, or solid-phase extraction methods. These extracts may exist in crude, semi-purified, or purified forms and are suitable for applications as intermediates, active pharmaceutical ingredients, or final formulations. Matrix-blank plants

[0074] As an exemplary biological matrix-blank organism, in some embodiments, a biological matrix-blank plant is provided which has been selected to not express at least one medicinally active or bioactive compound, but to express at least one other compound expressed in a comparator plant. In some embodiments, the at least one other compound is one or more non-medicinally active compounds. In some embodiments, the amount of at least one non-medicinally active compound expressed by the biological matrix-blank plant is at least greater than 50% (weight / weight) of the total compounds in the plant. In some embodiments, the amount of at least one non-medicinally active compound expressed by the biological matrix-blank plant is greater than 60% (weight / weight) non-medicinally active compounds, or greater than 70% (weight / weight) non-medicinally active compounds, or greater than 80% (weight / weight) non-medicinally active compounds, or greater than 90% (w / w) non-medicinally active compounds, or the greater than 95% (weight / weight) non-medicinally active compounds.

[0075] In some embodiments, the biological matrix-blank plant is a Cannabis plant. Cannabis plants synthesize a diverse spectrum of compounds including cannabinoids, which are identified as primary active components, as well as “entourage” compounds, molecular entities that exhibit minimal interaction with cannabinoid receptors. Prevailing scientific discourse suggests entourage compounds modulate cannabinoid activity, potentially amplifying their pharmacological effectiveness. It is therefore advantageous to engineer a botanical specimen devoid of primary cannabinoids but containing these entourage compounds. Preferably, the biological matrix-blank plant retains other significant compounds in proportions that mirror, both qualitatively and quantitatively, the comparator plant. In some embodiments, the biological matrix-blank plant is one that chemotypically aligns with medicinal plants employed in the formulation of pharmaceuticals, nutraceuticals, or functional foods.

[0076] In some embodiments, the biological matrix-blank plant is a Cannabis plant and the at least one medicinally active compound is at least one cannabinoid, such as tetrahydrocannabinol (THC) or cannabidiol (CBD), and the like. In some embodiments, the biological matrix-blank plant is a THC-free Cannabis plant. In some embodiments, the biological matrix-blank plant is a CBD-free Cannabis plant. In some embodiments, the biological matrix-blank plant is a THC-free and CBD-free Cannabis plant. In some embodiments, the biological matrix-blank plant is a cannabinoid-free Cannabis plant.

[0077] In some embodiments, the at least one other compound present in the comparator plant includes one or more entourage compounds selected from terpenes, lignans, sterols, and flavonoids. In some embodiments, the biological matrix-blank plant does not express at least one of CBG, THC, or CBD but expresses one or more terpenes, lignans, sterols, or flavonoids.

[0078] In some embodiments, the biological matrix-blank plant expresses one or more compounds selected from monoterpenes, diterpenes, carotenoids, alkaloids, triterpenes, flavonoids, sterols, and lignans. In some embodiments, the biological matrix-blank plant lacks expression of one or more compounds selected from monoterpenes, diterpenes, tetraterpenes, alkaloids, triterpenes, flavonoids, sterols, and / or lignans.

[0079] It is imperative to acknowledge the diversity and range of Cannabis species, encompassing both wild and cultivated varieties. These cultivated varieties are distinct in their purposes and genetic makeup. They include fiber and grain-producing plants characterized by low THC content, varieties bred for recreational and medicinal applications with high THC levels, and medicinal plants specifically selected for their cannabinoid profile. The latter category may exhibit a predominance of one or more cannabinoids and, optionally, a specific profile of accompanying compounds, commonly referred to as “entourage compounds.”

[0080] In some embodiments, a biological matrix-blank plant with targeted characteristics, including the absence of one or more medicinally or bioactive molecules, is developed using a selective breeding approach. In some embodiments, the selective breeding approach focuses on reducing the bioaccumulation of one or more cannabinoids with and without adversely impacting the synthesis and presence of compounds present in medicinal and recreational organisms. The methodology employed in achieving this selective breeding and its implications are detailed in the subsequent sections of this application. In some embodiments, the biological matrix-blank plant is a Cannabis plant containing a monogenic or multigenic mutation that blocks the biosynthesis of one or more cannabinoids. In some embodiments, the biological matrix-blank plant comprises a cannabinoid knockout factor governing a reaction in the pathways towards the phenolic moieties olivetolic and divarinic acid. Identification of matrix-blank plants

[0081] In some embodiments, a method is provided for producing a biological matrixblank plant that does not express at least one medicinally active compound yet expresses at least substantially qualitatively, most other non-medicinally active compounds present in a bioactive-containing organism wherein the method includes: a) selecting an organism that does not express at least one medicinally active compound; b) selecting a bioactive-containing organism; and c) crossing the organism which does not express at least one medicinally active compound with the bioactive-containing plant to obtain an Fl progeny and self-crossing the F1 progeny to obtain an F2 progeny which is selected for the characteristics sought. Subsequent steps of breeding and selection, to achieve a desired result in terms of reduced or eliminated expression of the medicinally active compound, including but not limited to backcrosses, selfcrosses, sibling crosses, and the like, are known to those of skill in the art and need not be described herein. Guidance regarding such plant breeding techniques is available, for example, in Singh, D.P., et al.. Plant Breeding and Cultivar Development, Academic Press, 2021; and Loud, J., Cannabis Breeding: The Art and Science of Crafting Distinctive Cultivars, James Loud Publishing, 2024, which are incorporated herein by reference.

[0082] The invention is further described, by way of example only, with reference to novel Cannabis plants (and not specific varieties), which do not express cannabinoids but which otherwise, resemble, chemotypically, medicinal Cannabis plants.

[0083] The cultivation of Cannabis in numerous countries is often limited to fiber hemp cultivars containing specified “low” levels of THC (typically below 0.1 or 0.3% wt. / wt. of dry floral tissue) to adhere to legal restrictions. To comply, various breeding programs have emerged with the goal of meeting these stringent limits.

[0084] A survey of European commercial fiber cultivars identified that the cultivars developed at the Ukrainian Institute of Fibre Crops (formerly, Federal Research Institute of Bast Crops) exhibit the lowest THC contents and total cannabinoid contents. Dating back to 1973, the cannabinoid breeding program at this institute primarily employed selective breeding within cultivars possessing high agronomic value. Their approach involves eliminating plants with relatively high contents before flowering and resulting in a gradual reduction of THC content and total cannabinoid content.

[0085] Researchers Gorshkova et al. conducted an evaluation of glandular trichome densities on various plants' bracteoles. They discovered that plants with stalked trichomes tended to possess higher cannabinoid contents, which positively correlated with the density of these trichomes. Conversely, plants with only sessile trichomes consistently exhibited low contents unrelated to their sessile trichome densities. Moreover, the study identified cannabinoid-free plants lacking glandular trichomes.

[0086] Subsequently, Ukrainian plant breeders reported the existence of cannabinoid-free breeding materials multiple times. Pacifico et al. analyzed individual plants from the Ukrainian cultivar USO 31, revealing that approximately one third contained no cannabinoids. Similarly, they found that a minority of plants (<10%) in a French fiber cultivar, Epsilon 68, were cannabinoid-free.

[0087] These cannabinoid-free plants differ phenotypically and chemotypically from those engineered artificially and those isolated in nature. They potentially manifest because of two distinct physiological conditions: (1) a disruption in the formation of glandular trichomes, which are pivotal for cannabinoid synthesis, as suggested by Sirikantaramas et al. and (2) blockage of biochemical pathways crucial for the formation of biochemical precursors preceding CBGA.

[0088] In 1991, field-grown cannabinoid-free plants resulting from the Gorshkova et al. program exhibited a lack of glandular trichomes on their bracts and bracteoles. Furthermore, these plants did not emit the typical Cannabis fragrance, indicating a potential absence of volatile mono- and sesquiterpenes. Consequently, these cannabinoid-free plants might have been deemed unsuitable for breeding a typical entourage compound-bearing, cannabinoid-free plant.

[0089] The second condition, affecting metabolites beyond cannabinoids, might hinder the common precursor basic pathways for various end products. For instance, the synthesis of cannabinoids and certain terpenes, sterols, and triterpenes are distinct, occurring in different cellular compartments.

[0090] In some embodiments, the matrix-blank Cannabis plants disclosed herein include stalked glandular trichomes. In some embodiments, the trichomes are present at a density comparable to those present in comparator medicinal and recreational cannabinoid-producing plants. In some embodiments, the biological matrix-blank plants have small, grey, dull trichomes of various shapes. The trichomes can be headless; pinhead and / or shriveled trichomes, which may be flat, convex or concave, while others are comparable to drug-type cannabinoid producing plants. In some embodiments, the matrix-blank Cannabis plants are selectively bred for capitate stalked glandular trichomes or bulbous stalked trichomes. In some embodiments, other trichome types are present.

[0091] In some embodiments, the biological matrix-blank plants exhibit branching characteristic of a drug producing phenotype as opposed to a fiber producing phenotype. In some embodiments, the biological matrix-blank plant exhibits vigor, characterized in that the total above ground dry weight is comparable to drug producing phenotypes.

[0092] In some embodiments, the matrix-blank plants disclosed herein are selectively bred to be dioecious to increase biomass yield and eliminate undesired seed production. In some embodiments, the plants autoflower or photoperiod flowering versions of cannabinoid-free Cannabis. Photoperiod insensitivity and dioecy introduction provides a competitive advantage over the plant known in the art.

[0093] In other aspects of this invention, while the methodology described above is specific to the Cannabis species, the methodology described herein of selectively breeding a blank organism to generate a matrix blank material for analytical testing and comparative studies can be applied to other organisms.

[0094] While this invention is exemplified by a cannabinoid-deficient Cannabis plant, one skilled in the art would understand the broader implications of this methodology to be applicable to all natural product organisms including, but not limited to, Psilocybe spp, Panaeolus spp, Hericium spp, Ganoderma spp, Inonotus spp, Lophophora spp, Trichocereus spp, Banisteriopsis caapi. Amanita spp, Salvia spp, Panax ginseng, Withania somnifera, Camellia sinensis, Cordyceps spp., and other species of medicinally and recreationally used organisms. Biological matrix-blank reference material

[0095] In some embodiments, the biological matrix-blank organism disclosed herein is used to generate a biological matrix-blank reference material having a chemical profile that resembles that of the comparator organism less the at least one medicinally active compound.

[0096] As an exemplary embodiment, in the case of a Cannabis plant, the biological matrix-blank reference material exhibits a profile of entourage compounds that is quantitatively similar to that of the biological matrix-blank plant as shown in FIG. 1 and FIG. 2. This similarity is much greater than prior-art attempts to create control materials for testing. Such attempts typically created the control materials by attempting to wash out (or otherwise remove) the analytes to be tested. However, such approaches also typically wash out other biochemical components of the material, resulting in an analytical “blank” that lacked the substantial similarity to the comparator material. In an alternative approach, prior-art attempts to create control materials would involve assembling the major components of a “blank” minus the analytes to be tested, without any effective attempt to replicate the presence of minor components. Since the minor components can interact in various ways and can be, in some testing formats, mistaken for the analytes to be tested, the absence of such minor components had the effect of the “blank” not being an accurate control material.

[0097] In some embodiments, the biological matrix-blank reference material is used for comparison with unknown or known bioactive-containing samples of the same organism. In some embodiments, the biological matrix-blank reference material is used as a standard reference material to be used as a matrix subtraction for analytical chemistry method development and / or method validation.

[0098] In some embodiments, a biological matrix-blank reference material comprising a biomass or an extract from a biological matrix-blank organism, such as a plant or fungus, is provided for use as a reference material. In some embodiments, the biomass is a homogenized bulk biomass. In some embodiments, the homogenized bulk biomass includes a homogenized biomass of stems, leaves, seeds, and / or buds. In some embodiments, the homogenized bulk biomass includes a homogenized biomass of mycelium, spores, and / or fruiting bodies. The biological matrix-blank reference material mirrors the biological matrix-blank plant or fungus in that it lacks at least one medicinally active or bioactive compound but includes at least one other compound expressed in a comparator plant. The biological matrix-blank reference material disclosed herein being derived from a biological matrix-blank organism does not require removal of medicinally active or other organism-produced compounds through means such as extraction, chemical conversion, genetic modification or genetic manipulation.

[0099] The matrix-blank reference materials disclosed herein represent a significant improvement over current material used for matrix blank subtraction, as the removal of noncannabinoid metabolites in efforts to create current cannabinoid-free Cannabis has led to nonrepresentative matrix blanks being used leading to inaccurate quantification due to the inability to properly subtract the background. Furthermore, the matrix-blank reference materials and associated methods disclosed herein provide the first chemically representative matrix-blank reference material that contains all primary and secondary metabolites produced by the comparator organism without containing the bioactive compound of interest, such as, in the case of cannabinoids being absent in ‘ZCP.’ Additionally, many previous attempts at using a cannabinoid-free matrix for Cannabis has led researchers to use other plant biomass; however, due to the lack of other non-cannabinoid Cannabis-specific metabolites, it is limited in use due to not being comparable to the unprocessed Cannabis material being tested. The embodiments herein allow for the first full spectrum representative matrix-blank reference material in Cannabis that is comparable to the chemical compositions of recreational marijuana or hemp plants.

[00100] In some embodiments, a method of producing a biological matrix-blank reference material is provided wherein the method includes selecting a biological matrix-blank organism that does not express at least one medicinally active compound but express at least one non-medicinally active compound present in a comparator plant, generating a reference material therefrom, wherein the reference material has a chemical profile that resembles the chemical profile of the comparator organism minus the at least one medicinally active compound not expressed. Producing organisms, such as plants, and mixtures of biomass derived from the organisms in this embodiment provides additional avenues of using the biomass or products derived from the matrix-blank organism including but not limited to enabling studies to identify bioactive effects of individual components of a given botanical drug substance, as well as providing specialized Cannabis with non-naturally occurring ratios of metabolites that provide medicinal effects.

[00101] In some embodiments, the biological matrix-blank reference materials disclosed herein are prepared by any method generally known in the art, for example, but not limited to, by maceration, percolation, vaporization, chromatography, distillation, recrystallisation and extraction with organic polar and non-polar solvents such as, but not limited to, methanol, ethanol, propanol, butanol, pentanol, norflurane, butane, propane, acetone, acetonitrile, benzene, toluene, chloroform, ethyl ether, xylenes, pyridines, methylene chloride, dimethyl sulfoxide, isobutanol, nitrobenzene, cyclohexane, chlorobenezene, hexanes, heptanes, pentanes, and other alcohols. Such extracts may be prepared also by utilizing polar solvents such as water and supercritical or subcritical liquid carbon dioxide. In some embodiments, the extracts can be obtained by the methods and processes described in international patent application numbers WO02 / 089945 and WO 2004 / 016277, each of which is hereby incorporated by reference in its entirety. Additionally, in some embodiments, the biomass and extract derived therefrom are handled as disclosed in sterile conditions to eliminate the possibility of post-harvest or post-processing contamination so that the material is suitable to be used as an analytical matrix blank material.

[00102] In some embodiments, the biological matrix-blank reference materials disclosed herein include an extract or a mixture of plant biomass from a selectively-bred Cannabis plant characterized by the absence of at least one of the following cannabinoids: Tetrahydrocannabiphorol (9-THCP), Tetrahydrocannabivarin (9-THCV), Delta-8-Tetrahydrocannabinol (8-THC), Delta-9-Tetrahydrocannabinol (9-THC), Tetrahydrocannabinolic Acid (9-THCA), Tetrahydrocannabiphorolic Acid (9-THCPA), Tetrahydrocannabivarinic Acid (9-THCVA), Cannabichromene (CBC), Cannabichromenic Acid (CBCA), Cannabichromeorcin (CBCO), Cannabichromevarin (CBCV), Cannabichromevarinic Acid (CBCVA), Cannabidiol (CBD), Cannabidiolic Acid (CBDA), Cannabidibutol (CBDB), Cannabielsoin (CBE), Cannabidiol monomethyl ether (CBDM), Cannabidiol Acid monomethyl ether (CBDAM), Cannabidiorcin (CBDO), Cannabidiphorolic Acid (CBDPA), Cannabidivarin (CBDV), Cannabidivarinic Acid (CBDVA), Cannabigerol (CBG), Cannabigerolic Acid (CBGA), Cannabigerol acid monomethyl ether (CBGAM), Cannabigerol monomethyl ether (CBGM), Cannabigerovarin (CBGV), Cannabigerovarinic Acid (CBGVA), Cannabigerphorolic Acid (CBGPA), Cannabicyclol (CBL), Cannabicyclolic Acid (CBLA), Cannabinol (CBN), Cannabinolic Acid (CBNA), and Cannabicitran (CBT). In some embodiments, the selectively-bred Cannabis plant is characterized by trichomes of clear, white, and / or amber coloration. In some embodiments, the selectively-bred Cannabis plant is derived from the Cannabis variety, ‘LCP.’ In some embodiments, the selectively-bred Cannabis plant is derived from the Cannabis variety, ‘ZCP.’ In some embodiments, the selectively-bred Cannabis plant is derived from the Cannabis variety, ‘ZCSB.’ In some embodiments, the selectively-bred Cannabis plant is derived from the Cannabis variety, ‘ZCCC.’ In some embodiments, the selectively-bred Cannabis plant is derived from the Cannabis variety, ‘ZCBC.’ In some embodiments, the selectively-bred Cannabis plant is derived from the Cannabis variety, ‘CK15.’ In some embodiments, the selectively-bred Cannabis plant is derived from the Cannabis variety, ‘CP21.’ In some embodiments, the selectively-bred Cannabis plant is derived from the Cannabis variety, ‘ZCO.’

[00103] In some embodiments, the plant material selected as the source of a matrix-blank reference material is derived through breeding. For example, as an exemplary embodiment, the presence of cannabinoids is selected against over a period of five or more generations of inbreeding and chemical selection. In some embodiments, the selection is further bred to introduce various subsets of entourage compounds to be more similar and comparable to commercially available Cannabis varieties. This allows for a more accurate comparison in analytical testing to have a more representative matrix material to compare against, which currently does not exist on the market. Furthermore, the presence of Cannabis-specific flavonoids (e.g., cannflavin A, cannflavin B, and cannflavin C) are indicative of the biomass being Cannabis of which current extracted matrix blank materials are devoid. Thus, biological matrix-blank plants provide an enhanced and more representative matrix material than extracted Cannabis or other matrices that are devoid of cannabinoids.

[00104] In some embodiments, the biological matrix-blank reference materials disclosed herein naturally include flavonoids such as, cannflavin A, cannflavin B, cannflavin C, or other Cannabis-specific flavonoids expressed by the biological matrix-blank plant. In some embodiments, the biological matrix-blank reference materials naturally include terpenes expressed by the biological matrix-blank plant. In some embodiments, the biological matrixblank reference materials disclosed herein are characterized by the presence of one or more of monoterpenes and sesquiterpenes; the absence of either monoterpenes or sesquiterpenes; the absence of both monoterpenes and sesquiterpenes; the presence diterpenes; the absence of diterpenes; the presence of triterpenes; and / or the absence of triterpenes.

[00105] In some embodiments, a method of obtaining a certified biological matrix-blank reference material is provided, wherein the method includes cultivating a biological matrixblank organism under optimized conditions to produce a pesticide-, herbicide-, fungicide-, heavy metal-, mycotoxin-, endotoxin-, and microbial-free biological matrix-blank organism, as described in Example 1, from which a biomass or extract is obtained.

[00106] In some embodiments, a method for sterile post-harvest processing of the biological matrix-blank organism is provided, wherein the method includes post-harvest processing that has been optimized to prevent environmental contamination, as described in Example 2.

[00107] In some embodiments, a method for packaging a reference material from a biological matrix-blank organism in sterile containers meeting or exceeding EPA standards in a clean environment, ensuring no contamination during packaging, transport, or storage is provided, wherein the packaging is performed under ISO 17034 accreditation as a certified reference material.

[00108] In some embodiments, the biological matrix-blank reference material is devoid of a profile of entourage or contaminant compounds or organisms, further characterized by at least one of: the presence of a known compound or organism spiked in at a known concentration for analytical reference; and / or the addition of one or more compounds selected from the group consisting of flavonoids, alkaloids, terpenoids, carotenoids, cannabinoids, alcohols, ketones, ethers, amines, amides, esters, aldehydes, sterols, pesticides, herbicides, fungicides, hydrocarbons, synthetic cannabinoids, synthetic psychedelics, synthetic opioids, isoxazoles, halogenated compounds, radioactive elements, heavy metals, Aspergillus spp., Salmonella spp., Escherichia coli, Pseudomonas spp., Listeria, hepatitis, norovirus, Clostridium, Campylobacter, Taxoplasma, Psilocybe spp., Cryptosporidium, Saccharomyces spp., hops-latent viroid, beat-curly top virus, tobacco mosaic virus, and radiolabeled nucleic acids.

[00109] Some embodiments provide a selectively bred matrix-blank organism wherein the tissues of the organism are used for comparison with unknown or known bioactive-containing samples of the same organism.

[00110] In some embodiments, the matrix-blank organism can be selected from one of the following organisms: Lion's Mane mushroom species such as Hericium erinaceus, Hericium coralloides, and Hericium americanum', Reishi mushroom species including Ganoderma lucidum, Ganoderma tsugae, and Ganoderma applanation, Chaga mushroom, specifically Inonotus obliquus', Cordycep mushrooms such as Cordyceps militaris, Cordyceps gunni, and Cordyceps dipterigena; Amanita mushroom species such as Amanita citrina, Amanita muscaria, Amanita gemmata, Amanita. Pantherina, Amanita porphyria, Amanita regalis, and Amanita strobiliformis; various psilocybin-containing mushrooms such as Psilocybe tampanensis, Psilocybe cubensis, Psilocybe semilanceata, Psilocybe oaxacana, Psilocybe natalensis, Psilocybe baeocystis, Psilocybe zapatecorm, Psilocybe azurescens, Psilocybe mexicana, Panaeoulus cyanescens, Panaeoulus cinctulus Psilocybe cyanescens, Panaeolus acidus, Panaeolus acuminatus, Panaeolus affinis, Panaeolus africanus, Panaeolus albellus, Panaeolus albidocinereus, Panaeolus albovelutinus, Panaeolus aids, Panaeolus alveolatus, Panaeolus annulatus, Panaeolus anomalus, Panaeolus antillarum, Panaeolus atomatus, Panaeolus atrobalteatus Panaeolus axfordii, Panaeolus bisporus, Panaeolus cambodginiensis, Panaeolus fimicola, Panaeolus moellerianus, Panaeolus olivaceus, Panaeolus fimicoloides, Panaeolus fimiputris, Panaeolus foenisecii, Panaeolus fontinalis, Panaeolus fraxinophilus, Panaeolus georgii, Panaeolus gomphodes, Panaeolus goossensiae, Panaeolus griseofibrillosus, Panaeolus guttulatus, Panaeolus hippophilus, Panaeolus hygrophanus, Panaeolus hypomelas, Panaeolus incanus, Panaeolus indicus, Panaeolus intermedins, Panaeolus lentisporus, Panaeolus lerchenfeldii, Panaeolus leucophanes, Panaeolus lignicola, Panaeolus linnaeanus, and Panaeolus longiguus. Medicinal cactus species such as Lophophora williamsii, Lophophora diffusa, Lophophora fricii, Trichocereus pachanoi, and Trichocereusperuvianus and other medicinal plant species such as Mimosa hostilis / tenuiflora, Acacia confusa, Banisteriopsis caapi, Passiflora incarnata, and Peganum harmala.

[00111] In some embodiments, the matrix-blank organism can be selected using a genetic marker complementary to one or more biosynthetic pathway genes that select against the biosynthesis of the active component.

[00112] In some embodiments, the matrix-blank reference material is used to create an extract utilizing either a solventless extraction method including but not limited to ice water extraction, heat and pressure based extraction, or size based extraction or a solvent based extraction method selected from a one or more solvents including ethanol, methanol, carbon dioxide, propane, butane, heptane, hexane, naphtha, limonene, pinene, or analogous polar or non-polar solvent system.

[00113] In some embodiments the matrix-blank or reference material includes one or more molecules spiked in at a known concentration selected from the following classes: pesticides, herbicides, Per- and polyfluoroalkyl substances (PFAS), fungicides, mycotoxins, elements, isotopically labels, bioactive molecules, biologicals, and hydrocarbons. In some embodiments, the matrix-blank or reference material spiked with one or more compounds from the previously mentioned classes of compounds allows for proficiency testing of laboratories using this spiked material, as well as producing known bioactive concentration materials specific to treating a specific condition.

[00114] In some embodiments, the matrix-blank reference material biomass or extract is combined with one or more molecules selected from one or more of the following bioactive constituents including but not limited to tryptamines, hericenones, erinacines, ganoderic acids, lucidenic acid, inotodiol, lanosterol, betulinic acid, betulin, mescaline, hordenine, anhalonidine, and tyramine; contaminant constituents including but not limited to Per- and polyfluoroalkyl substances (PFAS), lead, mercury, cadmium, arsenic, chromium, nickel, copper, zinc, gold, uranium, radon, silver, myclobutanil, bifenthrin, avermectin, imazalil, permethrin, spiromesifen, spirotetramat, chlorfenapyr, pyrethrins, imidacloprid, carbaryl, malathion, chlorothalonil, mancozeb, iprodione, propiconazole, pyraclostrobin, vinclozolin, copper-based fungicides, aflatoxins, ochratoxins, endotoxins, polycyclic aromatic hydrocarbons, residual solvents, plasticizers, dioxins and pcbs, nitrates and nitrites, acrylamide); biological contaminants including but not limited to live or dead cells of Salmonella spp, Escherichia coli, Shiga Toxin Producing E. Coli (STEC), Listeria monocytogenes. Staphylococcus aureus, Botritiys spp, Aspergillus spp, Clostridium botulinum, Bacillus cereus, Campylobacter spp, yeasts, viroids, molds, as well as extracted or synthesized DNA / RNA of these species.

[00115] In some embodiments, the matrix-blank reference material is combined with one or more molecules selected from the group consisting of bioactive constituents, contaminant constituents, biological contaminants, and extracted or synthesized DNA / RNA of the aforementioned species.

[00116] In some embodiments, the biological matrix-blank reference material is devoid of a profile of entourage compounds, for use in biological consumption to create a matrix-blank material from biological tissue or excrement, further characterized by one or more of: the addition of one or more compounds selected from the group consisting of flavonoids, terpenoids, tetraterpenes, triterpenes, diterpenes, alcohols, ketones, pesticides, herbicides, fungicides, solvents, Aspergillus, Salmonella, Escherichia coli, microplastics, and heavy metals; the use of the spiked plant biomass for biological consumption by humans or animals to create a comparative analytical chemistry matrix. Some embodiments include the addition of alkaloids, phenolic compounds, isoxazoles, sterols, vitamins, and / or minerals. Use of biological matrix-blanks

[00117] In some embodiments, the biological matrix-blank reference materials disclosed herein are used for proficiency testing of analytical labs to determine the accuracy, precision, and ability to detect and quantify bioactive or contaminant compounds. In some embodiments, the biological matrix-blank reference materials disclosed herein are spiked with one or more contaminants, such as a microbe or a pesticide, and used for proficiency testing or regulatory testing enforcement to evaluate the ability of a testing facility or laboratory to detect and quantify pathogens or adulterants in a test organism, such as Cannabis.

[00118] In some embodiments, the biological matrix-blank reference materials disclosed herein are used for analytical chemistry analyses such as, matrix-matched calibrations, validations, and quality controls as a bioactive-free matrix blank. In some embodiments, the biological matrix-blank reference materials disclosed herein are employed for analytical chemistry purposes to compare to unknown organism samples, such as Cannabis samples. In some embodiments, the biological matrix-blank reference materials disclosed herein are used for medical studies to determine the efficacy, toxicity, or potential benefits of a given organism or compound produced by that organism.

[00119] In some embodiments, the biological matrix-blank reference materials disclosed herein are used as control samples in scientific research, including but not limited to pharmacological, toxicological, and botanical studies. In some embodiments, the biological matrix-blank reference materials disclosed herein are used in the development and calibration of analytical instruments and techniques for detecting and quantifying bioactive compounds in an organism’s biomass. In some embodiments, the biological matrix-blank reference materials disclosed herein are used to test the hypothesis that at least one active compound is responsible for an extract’s perceived medicinal value. In some embodiments, the biological matrix-blank reference materials disclosed herein are used as a placebo. In some embodiments, a method of testing a hypothesis that one or more compounds present in a reference material are responsible for the pharmacological activity of that reference material is provided wherein the method includes: i) selecting a biological matrix-blank organism as described herein; ii) obtaining a reference material therefrom; and iii) running comparative tests against a reference material obtained from a comparator organism.

[00120] In some embodiments, the biological matrix-blank reference materials disclosed herein are used in educational settings for demonstration and training purposes in fields related to botany, chemistry, and pharmacology.

[00121] In some embodiments, the biological matrix-blank reference materials disclosed herein are used in the development of new varieties of organisms through genetic modification or selective breeding to introduce or enhance desirable traits while maintaining the absence of one or more bioactive compounds.

[00122] In some embodiments, a method of producing a designer biological organism extract used for analytical testing is provided, which method includes: i) selecting an extract obtainable from a biological matrix-blank organism as taught herein; and ii) combining the extract of (i) with one or more medicinally active components. The term “designer organism extract,” as used herein refers to an organism extract which includes one or more medicinally active components which do not naturally occur in the biological matrix-blank organism of part i. While sometimes exemplified by botanical extracts herein, this invention can additionally be further characterized by designer fungal extracts.

[00123] In some embodiments, the medicinally active components are purified naturally occurring compounds, synthetic compounds or a combination thereof. In some embodiments, the medicinally active components are present in a plant extract. In some embodiments, the plant extract is an extract from a “drug producing” plant of the same species as the biological matrix-blank plant of part i). Typically, this drug producing plant will not be the comparator plant to the matrix-blank plant of part i). Biological sample matrix

[00124] In some embodiments, the organisms and reference materials described herein are used to create other matrices that allow for research into the metabolite profiles of these organisms when exposed to the organism without one or more of the bioactive-components. In some embodiments, a method of making such a matrix includes administering to a subject who has not previously ingested the bioactive compounds, a specific amount of a matrix-blank organism or a matrix-blank reference material; and collecting blood, hair, saliva, urine, serum, or other bodily excretions or tissues from the subject after the consumption of the matrix-blank material. In some embodiments, the subject is a human. In some embodiments, the subject is an animal. In some embodiments, the new matrix is used to avoid false positives in analyte testing. In some embodiments, the new matrix material is created through infusing metabolites or metabolite-derived compounds from a cannabinoid-free Cannabis into a human or animal sample or a synthetic matrix created to represent a human or animal tissue or excrement.

[00125] In some embodiments, this method enables researchers and drug testing laboratories to more accurately analyze these matrices in cases where there is a desire to identify and quantify the presence of a given bioactive molecule or metabolite of that molecule in human or animal subjects. For example, in the states where Cannabis use is restricted, urine analysis for 11-HO-THC is commonly performed by employers, courts, and police departments. Having a true matrix-blank sample of urine that contains cannabinoid-free Cannabis metabolites allows for a more accurate determination and detection of the metabolites produced by cannabinoid-containing Cannabis through comparison against the cannabinoid-free infused urine matrix blank. This additionally can be used for validation of new and existing testing technologies that claim to be able to detect cannabinoids and their derived molecules. In some embodiments, this includes other bioactive-producing organisms, including, but not limited to, the use of non-psilocybin Psilocybe mushrooms or matrix-blank reference materials thereof to create urine, blood, salvia, or hair matrix blanks that contain other metabolites of psilocybin-containing mushroom species without containing metabolites of the controlled substances, psilocybin or psilocin. Upregulated cannflavin production

[00126] The potential broader chemical implications of engineering cannabinoid-free plants while expressing selected entourage compounds have significant implications for pharmaceutical Cannabis breeding. While cannabinoids, especially THC, are acknowledged as the primary active components, terpenoids are suggested to modify or enhance their physiological effects, offering greater medicinal benefits than isolated cannabinoids alone, as indicated by McPartland and Russo. Williamson and Whalley summarized that noncannabinoid constituents such as monoterpenes, sesquiterpenes, alcohols, esters, ketones, and flavonoids modulate cannabinoid effects and possess individual medicinal properties. Speroni et al. reported anti-inflammatory effects from an extract obtained from a cannabinoid-free chemotype. In some embodiments, cannabinoid-free Cannabis is harvested at a predetermined developmental stage or only certain tissues are harvested to maximize the production of a compound or certain set of compounds. An exemplary embodiment includes harvesting noncannabinoid Cannabis, such as ‘ZCP’ disclosed herein, at the sprout stage to maximize cannflavin content, or harvesting only the flower and leaves to produce a terpene rich cannabinoid-free mixture of biomass. ‘ZCP’ is unique in that cannflavin production reaches its maximum at final flower / senescence. This is contrary to other published results that currently exist. Additionally, the cannabinoid-free plants disclosed herein provide an advantage to other cannflavin producing Cannabis, as the plant continually produces cannflavins through vegetative growth and flowering. Competitor plants are limited to time-specific harvesting techniques to obtain upregulated amounts of cannflavin A and B. Furthermore, ‘ZCP’ is known to produce a maximum concentration of cannflavins at maturity of the flowering stage, which provides a significant improvement in yield over cannflavin-producing plants previously described that must be harvested at the sprout stage.

[00127] Cannflavins are a group of flavonoids found in Cannabis that have been studied for their potential therapeutic properties. For example, cannflavin A is known for antiinflammatory properties and may have analgesic effects. Cannflavin B also exhibits antiinflammatory effects and may have neuroprotective properties. In some embodiments, the biological matrix blank plants still produce flavonoids and are selected for upregulation of flavonoids, including, but not limited to, cannflavin A, cannflavin B, isocannflavin B and cannflavin C. The resulting biological matrix-blank reference materials having high levels of cannflavins have medicinal applications such as anti-inflammatory properties through their proposed role of inhibiting the production of pro-inflammatory prostanoids and leukotrienes. In some embodiments, the cannflavin-rich cannabinoid-free plant are combined with omega 6 and omega 3 fatty acid triglycerides from the seed oil produced by the cannabinoid-free Cannabis, to enhance the anti-inflammatory properties of the cannflavin-rich biomass mixture. In some embodiments, the resulting biological matrix-blank reference material would contain other non-cannflavin Cannabis-specific phenolic compounds including but not limited to upregulated quantities of cannabispiranes and canniprenes. In some embodiments, the cannabinoid-free Cannabis produces elevated concentrations of lignanamides, including but not limited to, Cannabisin E, Grossamide, (2,3-trans)-3-(3-hydroxy-5-methoxyphenyl)-N-(4-hy droxyphenethyl)-7- { (E)-3 - [(4-hy droxyphenethyl)amino] -3 -oxoprop-1 -enyl} -2,3 -dihydro- benzo[b][l,4]dioxine-2-carboxamide, Cannabisin F, N-trans-Caffeoyltyramine, Cannabisin A, N-trans-Feryroyltyramine, Cannabisin C, 3,3'-Demethyl-Grossamide, and / or Cannabisin D which are known to provide antioxidant and acetylcholinesterase inhibitory activities. Consumable products resembling cannabinoid containing products

[00128] In some embodiments, the biological matrix-blank reference materials disclosed herein are used as a base material for the production of non-psychoactive smokable products, health supplements, cosmetics, or food products. In some embodiments, the biological matrixblank reference materials are characterized by an altered nutritional profile, including but not limited to enhanced levels of vitamins, minerals, or dietary fibers.

[00129] In some embodiments, the biological matrix-blank reference materials disclosed herein are lacking in one or more cannabinoids and rendered non-psychoactive, while retaining desirable aromatic and flavor profile components. In some embodiments, the biological matrixblank reference materials disclosed herein are devoid of medicinally active cannabinoids, such as, but not limited to, CBG, THC and / or CBD. Nevertheless, the biological matrix-blank reference materials still naturally containing non-medicinally active compounds maintains the aromatic compounds and flavor profile characteristic of Cannabis, making it suitable for smoking, vaporizing, or infusion into herbal mixtures. As described herein, the biological matrix-blank reference materials naturally contain terpenes and flavonoids, offering unique flavors and scents, as well as non-cannabinoid biological effects in humans. Accordingly, in some embodiments, the biological matrix-blank reference materials described herein are used as a base material to produce non-psychoactive Cannabis products to individuals seeking the sensory experience of smoking or vaporizing Cannabis without the psychoactive effects associated with traditional Cannabis consumption.

[00130] In some embodiments, cannabinoid-free Cannabis plant parts are combined with other cannabinoid-containing Cannabis plant parts to produce a mixture of plants parts with a lower the cannabinoid content than the cannabinoid-producing Cannabis. In some embodiments, this mixture creates a combined biomass containing less than 0.3% total THC (Total THC = THCA*.877 + THC) by dry weight. In some embodiments, this mixture is combined to produce a biomass with less intoxicating effects as the cannabinoid-containing biomass alone. In some embodiments, the mixture of cannabinoid-free Cannabis and cannabinoid-containing Cannabis is to add a specific non-cannabinoid compound from cannabinoid-free Cannabis to the cannabinoid-containing Cannabis, such as incorporating cannflavin A from ‘ZCP’ into a low-cannflavin producing lineage biomass to produce mixtures of biomass with greater anti-inflammatory properties than the low-cannflavin parental line. In some embodiments, the cannabinoid-free material is spiked with terpenes and / or cannabinoids to produce non-naturally occurring and non-intoxicating chemical profiles of Cannabis, such as THC-free biomass with CBD and CBT, or CBG-free biomass with terpinolene and THC.

[00131] In some embodiments, the resulting products resembling cannabinoid-containing Cannabis provide potential health benefits associated with terpenes and other non-cannabinoid compounds including but not limited flavonoids, alcohols, esters, ketones, and other secondary metabolites. In some embodiments, the resulting biomass produced would contain elevated flavonoids, including but not limited to cannflavin A, B, and / or C that would be extracted and used as an active pharmaceutical ingredient in a formulated Cannabis-derived product, or the elevated non-cannabinoid bioactive compound containing plant parts itself are used as a botanical drug substance within clinical trials or medical applications.

[00132] In some embodiments, the biological matrix blank reference materials described herein are processed and used in various forms, including, for example, but not limited to, dried smokable flower, ingredients in herbal blends, extracts, and vaporization products for nonpsychoactive experiences. In some embodiments, the biological matrix-blank reference materials described herein are used in the manufacture of substitute products for smoking cessation. In some embodiments, the biological matrix-blank reference materials described herein are spiked with Cannabis-derived or non-Cannabis-derived compounds to create a biomass mixture that would encourage smoking cessation more so than the matrix-blank material alone. Other applications

[00133] In some embodiments, the biological matrix-blank reference materials described herein are used as substrates or mediums in agricultural research. In some embodiments, the biological matrix-blank reference materials are used for studying plant growth, nutrient uptake, or disease resistance in the absence of bioactive compounds. In some embodiments, the biological matrix-blank reference materials described herein are used as research tools in botanical and agricultural studies, providing a unique model for understanding plant biology without the interference of cannabinoid pathways. In some embodiments, a method for using a biological matrix-blank reference material as described herein as a research tool in botanical and agricultural studies is provided. In some embodiments, the method provides a unique model for understanding plant biology without the interference of specific bioactive pathways. For example, in some embodiments, the biological matrix blank organisms are used as a comparator plant to understand the biological effects of cannabinoids, terpenoids, or flavonoids as a pest deterrent against specific species of herbivores to identify unique chemical profiles to repel environmental-specific pests. In some embodiments, the biological matrix blank organisms are used as a comparator plant to understand the biological effects of cannabinoids, terpenoids, or flavonoids as anti-microbial compounds against specific species of fungi or bacteria to identify unique chemical profiles to gain resistance to environmental-specific microbes. In some embodiments, the biological matrix blank reference materials are spiked with one or more cannabinoids to determine the level of the one or more cannabinoids necessary to produce anti-fungal, anti-viral, anti-inflammatory, anti-biotic, anti-microbial, antianxiety or other medicinal or agricultural properties. The one or more compound at those amounts are then bred into susceptible Cannabis varieties.

[00134] In some embodiments, the biological matrix-blank organisms and reference materials described herein are used in environmental studies to assess the impact of bioactive compounds on ecosystems, particularly in soil and water analyses. In some embodiments, the biological matrix-blank organisms and reference materials described herein are used in environmental remediation and phytoremediation processes, exploiting its ability to absorb and break down pollutants in soil or water without the risk of cannabinoid accumulation. For example, in some embodiments, a method for using a biological matrix-blank plant in environmental remediation and phytoremediation processes is provided. In some embodiments, the method leverages the ability of the biological matrix-blank plant to absorb and break down pollutants in soil or water.

[00135] For example, Cannabis is known to perform bioremediation; however, this is limited in application due to the presence of controlled substances, namely cannabinoids, produced by the plant. Cannabinoid-free Cannabis used for bioremediation or environmental remediation is not limited by local, state or federal laws that cannabinoid-containing Cannabis is regulated under as the plant contains no illicit or prohibited compounds. In some embodiments, the cannabinoid-free Cannabis shows an increased bioremediation capacity compared to cannabinoid-containing Cannabis due to increased biosynthetic capacity of other pathways useful for remediating environmental contaminants due to the absence of cannabinoid biosynthesis. In some embodiments, the environmental contaminants being bioremediated include petrochemicals such as oil and gas spills. In some embodiments the environmental contaminants being bioremediated include radioactive materials such as those leaked from nuclear power plant disasters. In some embodiments, the environmental contaminants being bioremediated include microplastics and PFAS chemicals. In some embodiments, the environmental contaminants being bioremediated include metals and minerals from the soil. In some embodiments, the environmental contaminants being bioremediated include chemical spills of toxic manufacturing compounds such as vinyl chloride, benzene, styrene or other environmentally detrimental compounds.

[00136] In some embodiments, the biological matrix-blank organisms or reference materials described herein are used in the production of biofuels, where the absence of cannabinoids facilitates a more efficient conversion process. In some embodiments, the biological matrixblank reference materials described herein are used in the production of non-psychoactive, environmentally friendly biofuels or bioplastics. In some embodiments, a method for producing biofuels using a biological matrix-blank organism as described herein is provided, wherein the absence of specific bioactive compounds facilitates a more efficient conversion process. In some embodiments, the biological matrix-blank reference materials described herein are used in the production of biofuels and bioplastics wherein the absence of cannabinoids allows for a greater yield of biofuels or bioplastics per gram of biomass processed. In some embodiments, the biological matrix-blank reference materials described herein are used in the production of biofuels and bioplastics wherein the absence of cannabinoids produces higher quality biofuels and / or stronger tensile strength bioplastics. In some embodiments, the biological matrix-blank reference materials described herein are used in the production of biofuels and bioplastics wherein the absence of cannabinoids allows for a more environmentally sustainable manufacturing process through eliminating the processes used to remove or remediate the cannabinoids from cannabinoid-containing biomass.

[00137] In some embodiments, the biological matrix-blank organisms or reference materials described herein are used in horticulture as a grafting stock for other plant species, leveraging its unique genetic attributes. For example, in some embodiments, due to the agronomic properties and diverse growing conditions cannabinoid-free Cannabis can tolerate, cannabinoid-free Cannabis is used to cultivate grafts of other plant species and varieties that are unable to grow in those conditions naturally. In some embodiments, the cannabinoid-free Cannabis is used as root stock in high salinity environments where certain ornamental or crops are unable to survive due to environmental or pest-related selective pressures. In some embodiments, the cannabinoid-free Cannabis is planted as a biological method to prevent erosive forces where it provides an advantage to other cover crops due to its larger and deeper root mass than traditional cover crops. In some embodiments, cannabinoid-free Cannabis is used as a companion crop to provide pest resistance to a crop that is pest susceptible.

[00138] In some embodiments, the biological matrix-blank material provides economical and technical advantages compared to cannabinoid-containing Cannabis in the manufacturing of raw materials, biofuels, building materials, paper products, animal feed, cosmetic products, and food products, wherein the cannabinoids cause complications in machinery due to the physical properties of the resin. In some embodiments, current best practices for producing derived products from hemp or Cannabis are applied to ‘ZCP,’ which enables the production of these materials without being limited to certain countries’ laws against the scheduled compounds Cannabis is known to produce. In some embodiments, the biological matrix-blank materials described herein are used in the manufacturing of textiles and fibers, wherein the absence of cannabinoids does not affect the physical properties of the fibers and provide advantages of limiting or removing cannabinoid resin content in the raw material that is known to pose issues for industrial manufacturing. In some embodiments, a method for manufacturing textiles and fibers using a biological matrix-blank material as described herein is provided, wherein the absence of specific bioactive compounds improves the physical properties of the fibers as well as allows for easier processing of the raw material due to the lack of adhesive resin found in cannabinoid-containing plants. In some embodiments, the biological matrixblank materials described herein are used in the production of paper and cardboard materials, where the lack of cannabinoids contributes to a more environmentally sustainable processing due to the lack of necessity to remove cannabinoids that is required for processing all cannabinoid-containing hemp, which involves potentially toxic solvents and CO2-generating processes. In some embodiments, a method for producing paper and cardboard materials using a biological matrix-blank material as described herein is provided, wherein the lack of specific bioactive compounds contributes to more environmentally sustainable processing due to the lack of necessity to remove cannabinoids that is required for processing all cannabinoid-containing hemp, which involves potentially toxic solvents and CO2-generating processes. In some embodiments, a method for producing paper and cardboard materials using a biological matrix-blank material as described herein is provided, wherein the lack of specific bioactive compounds decreases the cost of production of these materials through eliminating the need of removing these regulated bioactive compounds. In addition to more environmentally sustainable processing, cannabinoid-free Cannabis provides economic and ergonomic advantages over cannabinoid-containing Cannabis in the manufacturing process through limiting time cleaning cannabinoid-resin from equipment, as well eliminating costs associated with extraction and disposal of cannabinoid waste products produced in the production of paper, cardboard, or other hemp-derived materials. Additionally, cannabinoid-content has posed issues for automating industrial Cannabis processing due to the properties of the cannabinoid resin causing it to accumulate on machinery that limits the throughput of these processing facilities. Due to the unique lack of cannabinoid resins in ‘ZCP,’ it provides ergonomical advantages compared to commercially available industrial hemp cultivars that contain cannabinoid resin. Additionally, with the absence of cannabinoids, all input ingredients and intermediary products would be allowed within the law to be processed anywhere; which provides a solution to regulatory and legal challenges currently faced in the industrial hemp industry and production of industrial hemp derived products.

[00139] In some embodiments, the biological matrix-blank materials described herein are used in animal feed, wherein the absence of cannabinoids ensures safety and compliance with animal health regulations and best practices for animal safety and pharmacology. In some embodiments, a method for producing animal feed using a biological matrix-blank material as described herein is provided. In some embodiments, the method ensures safety and compliance with animal health regulations due to the absence of specific bioactive compounds, such as THC. In some embodiments, the absence of certain bioactive compounds in the biological matrix-blank material prevents adverse reactions for animals known to negatively interact with cannabinoid-containing products. In such embodiment, the biological matrix-blank material is used to provide necessary nutritional content as well as optionally including selected bioactive compounds such as terpenes or flavonoids that are beneficial for a given condition or environment that the animal resides. For example, the incorporation ‘ZCP’ material containing cannflavins in animal feed produces anti-inflammatory effects in inflamed animals without risking adverse reactions to the presence of cannabinoids in the animal feed. In other embodiments, the ‘ZCP’ material is selected based on a combination of terpenes known to cause calming effects in animals, that enables specialty purpose cannabinoid-free Cannabis derived animal feed products.

[00140] In some embodiments, the biological matrix-blank materials described herein are used as a base for non-psychoactive, cannabinoid-free cosmetic products, including but not limited to creams, lotions, and ointments. In some embodiments, a method for creating nonpsychoactive, bioactive compound-free cosmetic products, such as creams, lotions, and ointments, using a biological matrix-blank material as described herein is provided.

[00141] For example, in the case of Cannabis, in some embodiments, the biological matrixblank materials described herein are used as a base for non-psychoactive, cannabinoid-free food products, including but not limited to chocolates, tinctures, gummies, pills, transdermal patches, topicals, drink mixes, and other finished products. In some embodiments, a method for creating non-psychoactive, bioactive compound-free food products, including but not limited to chocolates, tinctures, gummies, pills, drink mixes, topicals, and other finished products is provided. In some embodiments, the non-psychoactive, bioactive compound-free products derived from cannabinoid-free Cannabis are used as product-type specific analytical matrix blank materials, as a placebo formulation for research into the effects of cannabinoids or other non-cannabinoid bioactive metabolites, as a drug delivery system for non-cannabinoid bioactive compounds, or as a method of consuming other cannabis-derived compounds without consuming cannabinoids. In some embodiments, these cannabinoid-free Cannabis products are spiked with a known contaminant compound or organism to allow for the proficiency testing of analytical laboratories. In some embodiments, the biological matrix blank material is incorporated into a kit containing spiked and non-spiked matrix blank material that evaluate the ability of an analytical method to detect and quantify concentrations of target analytes at or below a regulatory body’s specifications for that compound.

[00142] In some embodiments, a process for the extraction of non-cannabinoid compounds, including but not limited to hydrocarbon for primarily terpene extractions; water for primarily flavonoid extraction, and / or chloroform for fatty acid targeted extraction, from the biological matrix-blank materials described herein is provided for use in various industries. In some embodiments, crude extracts are produced via extraction methods described here or known in the art, which are then processed and packaged to be a crude extract matrix-blank, or the extract is further refined to produce high purity isolated compounds that are used as active pharmaceutical or nutraceutical ingredients, or used for the production of reference standards of isolated compounds in the analytical testing industry. In some embodiments, the isolated compound used for an API would include one or more of the following compounds: alkaloids, flavonoids, terpenes, isoxazoles, phenolic compounds, or other medicinal bioactive compounds.

[00143] While the described invention is specific to the Cannabis species, it will be appreciated to one skilled in the art that this methodology disclosed herein could be applied to other organisms for which a bio-active free matrix blank material would be useful to achieve accurate analytical testing and comparative studies. Examples of such organisms ta include, but are not limited to: Lion's Mane mushroom species such as Hericium erinaceus, Hericium coralloides, and Hericium americanum: Reishi mushroom species including Ganoderma lucidum, Ganoderma tsugae, and Ganoderma applanatum', Chaga mushroom, specifically Inonotus obliquus, Cordycep mushrooms such as Cordyceps militaris, Cordyceps gunni, and Cordyceps dipterigena; Amanita mushroom species such as Amanita citrina, Amanita muscaria, Amanita gemmata, Amanita. Pantherina, Amanita porphyria, Amanita regalis, and Amanita strobiliformis; various psilocybin-containing mushrooms such as Psilocybe tampanensis, Psilocybe cubensis, Psilocybe semilanceata, Psilocybe oaxacana, Psilocybe natalensis, Psilocybe baeocystis, Psilocybe zapatecorm, Psilocybe azurescens, Psilocybe mexicana, Panaeoulus cyanescens, Panaeoulus cinctulus Psilocybe cyanescens, Panaeolus acidus, Panaeolus acuminatus, Panaeolus affinis, Panaeolus africanus, Panaeolus albellus, Panaeolus albidocinereus, Panaeolus albovelutinus, Panaeolus aids, Panaeolus alveolatus, Panaeolus annulatus, Panaeolus anomalus, Panaeolus antillarum, Panaeolus atomatus, Panaeolus atrobalteatus Panaeolus axfordii, Panaeolus bisporus, Panaeolus cambodginiensis, Panaeolus fimicola, Panaeolus moellerianus, Panaeolus olivaceus, Panaeolus fimicoloides, Panaeolus fimiputris, Panaeolus foenisecii, Panaeolus fontinalis, Panaeolus fraxinophilus, Panaeolus georgii, Panaeolus gomphodes, Panaeolus goossensiae, Panaeolus griseofibrillosus, Panaeolus guttulatus, Panaeolus hippophilus, Panaeolus hygrophanus, Panaeolus hypomelas, Panaeolus incanus, Panaeolus indicus, Panaeolus intermedins, Panaeolus lentisporus, Panaeolus lerchenfeldii, Panaeolus leucophanes, Panaeolus lignicola, Panaeolus linnaeanus, and Panaeolus longiguus. Additionally, the process applies to psychedelic cacti like Lophophora williamsii, Lophophora diffusa, Lophophora fricii, Trichocereus pachanoi, and Trichocereus peruvianus', dimethyltryptamine containing species such as but not limited to Mimosa hostilis / tenuiflora and Acacia confusa, beta-carboline containing species such as but not limited to Banisteriopsis caapi, Passiflora incarnata, and Peganum harmala.

[00144] Furthermore, the application extends to other high-value natural products like Salvia divinorum (Salvia), Panax ginseng (Ginseng), Withania somnifera (Ashwagandha), Camellia sinensis (Tea plant), and coffee plants such as Coffea arabica and Coffea canephora.

[00145] The primary objective of embodiments disclosed herein is to create variants of these plants and fungi that retain their traditional uses, entourage biochemical profiles, nutritional value, and / or ornamental qualities without producing the psychoactive, toxin, or other specific bioactive compounds they are typically known for. The methods disclosed herein are applicable to all matrix-blank organisms that could be produced, which has been shown in the exemplary matrix-blank organism, Cannabis, through the creation and use of the specific selectively bred variety, ‘ZCP,’ and its progeny that have been specialized for various applications disclosed herein. This technology holds potential for broad applications in industries such as pharmaceuticals, nutraceuticals, and agriculture, as well as applications in analytical testing methodologies that require a matrix-blank material for accurate quantification of the bioactive molecules.

[00146] A further benefit of the matrix-blank organisms disclosed herein is that they can be used to create matrix-blank materials containing other bioactive molecules in quantities and purities, which could not be achieved naturally. Such matrix-blank materials provide benefits arising from the presence of one or more selected entourage compounds. For example, cannabinoids, which can be introduced to the cannabinoid free extracts, include one or more natural cannabinoids, synthetic cannabinoids, or degradation cannabinoids (modified natural cannabinoids). This produces a “designer” plant extract that can be used in analytical method validation and proficiency testing or for the production of customized cannabinoid-ratio materials that can not be achieved naturally.

[00147] In one embodiment, the ‘ZCP’ biomass or extract derived thereof is spiked with >1% by dry weight of CBD and CBC while remaining absent of THC and CBG. Due to the nature of cannabinoid biosynthesis, creating plants without CBGA / CBG but having other cannabinoids is impossible due to CBG being a precursor to the terminal cannabinoids. Thus, through these methods of using the plant materials of this invention, the creation of non-naturally occurring cannabinoid profiles through infusing naturally derived, synthetic, or modified cannabinoids into the Cannabis biomass and extracts derived thereof is possible using the matrix-blank organisms and materials and methods disclosed herein. In some embodiments, this is achieved through solubilizing the cannabinoid in a known volume of liquid solvent, such as ethanol, methanol, or other solvents in which cannabinoids are soluble, that is then applied to the dry Cannabis based on the target dry weight percentage to infuse known amounts of a cannabinoid onto the dried matrix blank material. In other embodiments, a similar infusion process can be used to incorporate one or more compounds from the following classes: chemical contaminants, environmental contaminants, bioactive compounds derived from Cannabis, bioactive compounds not found in Cannabis, or non-bioactive compounds.

[00148] Furthermore, this disclosure enables the use of these methods in organisms other than Cannabis, such as in other medicinal plants or fungi. For example, psilocybin-free Psilocybe cubensis mushrooms can be used to create novel ratios of tryptamines that would not be found in nature, but may provide enhanced or improved medicinal properties. Similarly to CBGA being a precursor to THC A, Norbaeocystin and Baeocystin are precursors to Psilocybin, which prevents a psilocybin-containing mushroom from being baeocystin-free in nature. However, through utilizing the method disclosed above, a psilocybin-free fungal matrix-blank can be used to produce norbaeocystin and psilocybin containing biomass that is free of baeocystin, as well as create other non-natural combinations of tryptamines spiked into the natural background of metabolites found in the organism to allow for reproducible and targeted medicinal formulations of natural products to achieve spiked biomass or extracts derived thereof.

[00149] Other examples of this technology include producing mescaline-free cacti through disruption of the mescaline biosynthesis pathway, ganoderic acid free reishi mushrooms through disruption of the ganoderic biosynthetic pathway, cordycepin-free cordyceps through disruption of the cordycepin biosynthetic pathway, alkaloid-free ibogaine through disruption of alkaloid biosynthetic genes, and hericenone-free and / or erinacine-free lions mane material through the disruption of the triterpenoid biosynthetic pathway. This disruption occurs through numerous selectively bred genotypes including, but not limited to, truncated genes in the biosynthetic pathway, loss-of-function genes in the biosynthetic pathway, upstream and downstream mutations causing differential regulation of the biosynthetic pathway, and mutations in regulatory factors associated with the expression of these biosynthetic genes. Additionally, these same mutations can be introduced through non-selective breeding methods as well, such as CRISPR-CAS, Cre-Lox, and other known gene modification techniques to introduce non-naturally occurring knockouts of these organisms to be used in the disclosed applications of the bioactive-free matrix blank material.

[00150] The benefits of naturally produced cannabinoid-free matrix blank material over extracted Cannabis or similar cannabinoid-free plant material lies in the fact that with over 500 unique bioactive compounds in Cannabis, any extraction is likely to remove other compounds that would prevent the matrix blank from being comparable to non-extracted material. This is important due to the ability to accurately detect and quantify all other biological and synthetic molecules from the cannabinoids. Furthermore, this provides an ideal matrix for proficiency testing and quality assurance testing that allows for the spiking of additives, contaminants or bioactive molecules that allow for the proficiency tester to accurately provide known material to testing laboratories to evaluate their ability to detect and quantify these compounds. This spiking of the sample can be extended to live and / or dead organisms as well in the case of microbial testing proficiency testing, or even viroids or viruses to determine a lab’s ability to detect pathogens of Cannabis. This can also be done by spiking known amounts of synthetic and extracted DNA and / or RNA from that given organism, virus, or viroid.

[00151] In some examples, local, state, or federal governments or other non-governmental organizations use cannabinoid-free Cannabis and spiked cannabinoid-free Cannabis to create a quality assurance system to perform laboratory assessments and determine if a laboratory is accurately quantifying the analytes and contaminants required by the regulations. In other embodiments, local, state, or federal governments or other non-governmental organizations use cannabinoid-free Cannabis and spiked cannabinoid-free Cannabis as a laboratory evaluation kit containing unmodified ‘ZCP’ material in addition to spiked ‘ZCP’ material specific to the industry regulations to evaluate a laboratory’s ability as a consideration of licensing or accreditation.

[00152] Similar logic can be applied in many other species and genera of plants and fungi, to identify, selectively breed, and produce organisms devoid of medically or recreationally relevant compounds and their respective degradants. For example, to produce psilocybin-free Psilocybe species, one would select or knockout the genes in the psilocybin biosynthesis pathway that would prevent the accumulation of tryptamines including norbaeocystin, baeocystin, psilocybin, psilocin, aeruginascin, and 4-hydroxy-trimethyltryptamine. In another example, the hericenone diterpene biosynthetic pathway is selected against to produce hericenone-free Hericium spp. Similar post-harvest processing and packaging steps are applied for these organisms to produce a research-grade or certified reference material grade bioactive-free matrix blank biomass material or extracts thereof that are used according to this disclosure.

[00153] In one embodiment, the matrix-blank organism is spiked with pathogenic or contaminant organisms or metabolites indicative of those organisms to create a proficiency test to test a laboratory’s target species specificity of their assay. In some embodiments, two mixtures of the matrix-blank organism are prepared: one spiked with shiga-toxin producing E. Coli and one spiked with non-pathogenic E. Coli, and sent to a laboratory as an unknown sample to determine the accuracy and specificity of microbial testing specifically in regards to testing products containing the same organism as the matrix-blank organism. In another embodiment, the matrix blank organism are spiked with one or more species of pathogenic aspergillus species, which enables a laboratory to validate the specificity, accuracy and matrix compatibility of microbial testing methods in new organisms where the matrix of that organism has not been tested. EXAMPLES Example 1 Identification & Selective Breeding of Non-Cannabinoid Cannabis Plant

[00154] Briefly, a low-cannabinoid plant LCP) was selectively inbred and outcrossed to develop the parental line non-cannabinoid plant (‘ZCP’) that was used both for the preparation of analytical matrix-blank materials, as well as for breeding to develop other specialized matrix materials. The parental lines showed a combination of monoecy and partial dioecy (otherwise known as hermaphroditism) which prevented in-breeding in some cases. In cases of femalefemale crosses, silver thiosulfate was used to induce masculinity and produced feminized pollen. ‘ZCP’ was selected for its limited hermaphroditic traits, unique ability to produce Cannabis-specific cannflavins, other flavonoids, and some terpenes, without producing any of the cannabinoids shown on Table 1.

[00155] Table 1. Cannabinoids Cannabinoid Name Abbreviation Tetrahydrocannabiphorol 9-THC P Tetrad y dr ocan nabi varin 9-THCV Delta~8~Tetrahydrocannabmol 8-THC Delta-9-Tetrahydrocannabinol 9-THC Tetrahy drocannabinolic Acid 9-THCA Tetrahydrocannabiphorolic Acid 9-THC PA Tetrahydrocannabivarinic Acid 9-THCVA C annahi ch rom en e CBC Cannabichromenic Acid CBCA. Cannabi chrom eorcin CBCO Cannabichromevarin CBCV Cannabichromevarinic Acid CBCVA Cannabidiol CBD Cannabidiol ic Aci d CBDA Cannabi dibutol CBDB Cannabielsoin Ft Cannabidiol monomethyl ether CBDM Cannabidiol Acid monomethyl ether CBDAM Cannabidi orcin CBDO Cannabi di ph orol i c Aci d CBDPA Cannabidivarin CBDV Cannabidivarinic Acid CBDVA Cannabi gerol CBG Cannabigerolic Acid CBGA Cannabigerol acid monomethyl ether CBGAM Camrabigerol nmnomethyl ether CBGM Cannabigerovarin CBGV Cannabi gerovarinic Aci d CBGVA Cannabigerphorolic Acid CBGPA Cannabi cy cl ol CBL C ann ab i cy cl oli c Aci d CBLA Cannabinol CBN Cannabinolic Acid CBNA Cannabicitran CB1"

[00156] All crosses described in the examples use the following nomenclature with the pollen acceptor listed first and the pollen donor listed second: (Female x Male) (Female x Reversed Female)

[00157] (i) Breeding for Yield and True Dioecy

[00158] ‘ZCP’ was selectively bred to it siblings and other distantly related Cannabis plants to provide additional phenotypic and chemotypic traits that allow for improved growth qualities, yield, resistances, and other novel compounds. For example, ‘ZCP’ was crossed to a plant, CK15, that was selected for its yield and dioecy phenotypes. The resulting offspring were in-bred against other siblings and backcrossed to the original ‘ZCP’ parental line, which allowed for various levels of introgression of these traits. The backcrossed population provided a higher percentage of cannabinoid-free plants (-50% of offspring retaining zero cannabinoids); whereas the inbred siblings allowed for a greater introgression and recombination of genes related to yield and dioecy, while still producing 25% of the offspring as zero-cannabinoid producing plants. The newly obtained plants (ZCSB1 & ZCBC1) increased yield by over 20% compared to the original ‘ZCP’ parental line, as well as showed multiple dioecious phenotypes that were selected for further breeding.

[00159] (ii) Breeding for Agronomic Traits

[00160] ‘ZCP’ additionally was bred to other lines for more agronomic qualities as well. In particular, this plant was crossed with a high CBD lineage, CP21, that contained herbivory-induced plant volatiles, such as guaiol, a-farnesene and germacrene D. These compounds are known to be produced in response to pest or herbivore pressure to allow for less pesticide application during the cultivation cycle. Using the Fl offspring of that cross, F2 populations were bred through sibling crosses as well as backcrosses to the ‘ZCP’ parent and the herbivory-induced volatile terpene containing plants were selected for future breeding.

[00161] Similar to the process described above, backcrossing achieved a higher proportion of the cannabinoid-free phenotype with less biologically protective terpenes included, whereas the sibling breeding strategy was employed to increase the likelihood of introgressing and producing more of the biologically protective compounds in a cannabinoid-free phenotype. The resulting plants (ZCSB2 & ZCBC2) contained significantly more terpenes in general compared to the original ‘ZCP’ parental line, as well as increased mold / mildew resistance that was exhibited in the CP21 line.

[00162] (iii) Breeding for Hybrid Vigor

[00163] A unique breeding strategy, termed cousin breeding, was employed to increase genetic diversity and heterozygosity of the ‘ZCP’ lineage while maintaining the cannabinoid-free chemotype, but allowing for the introgression of additional traits. Four F2 lots were created through the selective outcrossing of the ‘ZCP’ parental line to CK15 as well as CP21, producing Fl lots named ZCO1 and ZCO2 respectively. Each Fl seed lot was sibling bred as well as back crossed to the ‘ZCP’ parental line, creating F2 sibling bred lots of each outcross. ZCSB1 is the F2 seed lot derived from the sibling breeding of the ZCO1 offspring that originated from the ZCPxCK15 cross, and then ZCSB2 refers to the F2 seed lot derived from the sibling breeding from the ZCPxCP21 cross, which both F2 lots contained 25% of the individuals having a cannabinoid-free phenotype. A similar strategy was used in the backcrossing of Fl offspring to the ‘ZCP’ parental line, which lead to the production of ZCB1 F2 seed lot derived from the outcross to CK15, as well as the seed lot ZCB2, which contained 50% of the individuals having a cannabinoid-free phenotype. These four F2 seed lots,.ZCSB 1, ZCSB2, ZCBC1, & ZCBC2 were then used in a breeding program to create plants that exhibit greater hybrid vigor than the parental lines while maintaining the cannabinoid-free chemotype of the ‘ZCP’ parental line. After selecting the cannabinoid-free individuals from the four F2 lots, a reciprocal breeding strategy was employed that produced 100% cannabinoid-free plants. Offspring were selected based on the total yield of biomass, faster growth rates, presence of biologically protective terpenes, larger trichome heads for increased secondary metabolite carrying capacity, and having a dioecious phenotype. This reciprocal breeding strategy employed traditional inbreeding methods to produce 100% cannabinoid-free F3 lots of each of the sibling bred and back crossed lots individually allowing for the selection and isolation of traits in these inbred lines, as well as employing the “cousin cross” strategy disclosed here where an Fl-like plant with increased genetic diversity and heterozygosity was produced through crossing the ZCSB1 x ZCSB2.

[00164] The feminized crosses ZCSB1 xZCSB2 and its reciprocal cross (ZCSB2 xZCSBl) produced the highest vigor of all plants due to the novel three-way crossing strategy employed; wherein, a related outcrosses of ‘ZCP’ was created, which were inbred individually and then crossed with each other, allowing for three parental lines genotypes to appear in the offspring of this strategy. Plants derived from this three-way breeding strategy have been terms ZCCC1 for ZCSB1 x ZCSB2 and ZCCC2 for ZCSB2 x ZCSB1. Additionally the F3 crosses between the back crossed lots, crosses within a sibling bred lot, or the crosses between the sibling bred lots and the back crossed lots, produced segregating inbred offspring that allowed for us to select for various subsets of the phenotypes traits of the parental line CK15 and CP21 in the cannabinoid-free offspring.

[00165] (iv) Selection of Ideal Matrix Blank Material

[00166] While the parental ‘ZCP’ line is adequate for matrix blank material, various improvements of the offspring can result from the breeding efforts described herein. Plants exhibiting higher yields but lacking in terpene make ideal candidates to replace the ‘ZCP’ parental line as the matrix blank for spiked additions of cannabinoids, terpenes, flavonoids, heavy metals, pesticides, mycotoxins, microbial contaminants, viruses, residual solvents, foreign materials, synthetic drugs, Per and Polyfluorinated Substances (PFAS), and other laboratory tests. Spiked additions of the aforementioned compound classes can serve as method validation and proficiency testing materials.

[00167] (v) Breeding for Cannflavin Production

[00168] Additional development of the ‘ZCP’ lineage through selectively breeding for increased cannflavin production when compared to the parental lines. In particular, ‘ZCSB1 & ZCBCF were crossed with a known high cannflavin A (CP21) producing plant that contained greater than 0.05% (weight / weight) cannflavin A in outdoor cultivation trials. The hybrid Fl offspring was then subsequently inbred through a combination of sibling breeding and backcrossing to the ‘ZCP’ lineage to remove the cannabinoid biosynthetic capacity while retaining or improving the flavonoid production ability of CP21. Through selection of the F2 seed lots, a plant, ZCF1, was produced that had the cannabinoid-free trait of the ‘ZCP’ lineage parental line, while also showing increased production of cannflavin A at nearly double the concentration of the CP21 cannabinoid-containing parental line. ZCF1 was shown by HPLC analysis to produce greater than 0.1% cannflavin A by dry weight of the biomass material. Example 2 Cultivation & Extraction of ‘ZCP’

[00169] Indoor Cultivation of ‘ZCP’ for Regulatory Testing & Proficiency Testing

[00170] The cultivation of a matrix blank material requires a nearly sterile environment free of pesticides, herbicides, fungicides, molds, mildews, pests, and other pathogens that can cause the material to be unsuitable for this application. Thus, all raw ingredients, including but not limited to, soil, fertilizers, pots, and water were screened prior to being used on the ‘ZCP’ plant or its offspring. Seeds were sprouted in HEPA-filtered positive pressure environments to limit any contamination entering the growth space, and all work performed on the plants was done with growers in new Tyvek suits to limit any unintentional introduction of environmental pesticides, pests, diseases or other compounds. These growth conditions were used for most widely applicable versions of ‘ZPC’ that can be used as a matrix blank for all regulatory testing employed by states with recreational or medical Cannabis programs currently as well as for spiked validation studies and services that are routinely employed during proficiency testing of Cannabis laboratories, where known concentrations of compounds or organisms were added to the ‘ZCP’ material that was then analyzed by a laboratory undergoing accuracy and precision assessments. Regulatory testing includes assaying for various analytes including, for example, but not limited to, Abamectin, Acephate, Acequinocyl, Acetamiprid, Aldicarb, Allethrin, Atrazine, Azadirachtin, Azoxystrobin, Benzovindiflupyr, Bifenazate, Bifenthrin, Boscalid, Buprofezin, Captan, Carbaryl, Carbofuran, Chlorantraniliprole, Chlordane, Chlorfenapyr, Chlormequat chloride, Chlorpyrifos, Clofentezine, Clothianidin, Coumaphos, Cyantraniliprole, Cyfluthrin, k-Cyhalothrin, Cypermethrin, Cyprodinil, Daminozide, Deltamethrin, Diazinon, Dichlorvos (DDVP), Dimethoate, Dimethomorph, Dinotefuran, Diuron, Dodemorph, Endosulfan sulfate, Endosulfan-a, Endosulfan-B, Ethoprophos, Etofenprox, Etoxazole, Etridiazole, Fenhexamid, Fenoxycarb, Fenpyroximate, Fensulfothion, Fenthion, Fenvalerate, Fipronil, Flonicamid, Fludioxonil, Fluopyram, Glyphosate, Hexythiazox, Imazalil, Imidacloprid, Iprodione, Kinoprene, Kresoxim-methyl, Malathion, Metalaxyl, Methiocarb, Methomyl, Methoprene, Mevinphos, MGK-264, Myclobutanil, Naled, Novaluron, Oxamyl, Paclobutrazol, Parathion-methyl, Pentachloronitrobenzene (Quintozene)*, Permethrin, Phenothrin, Phosmet, Piperonyl Butoxide, Pirimicarb, Prallethrin, Propiconazole, Propoxur, Pyraclostrobin, Pyrethrins, Pyridaben, Pyriproxyfen, Resmethrin, Spinetoram, Spinosad, Spirodiclofen, Spiromesifen, Spirotetramat, Spiroxamine, Tebuconazole, Tebufenozide, Teflubenzuron, Tetrachlorvinphos, Tetramethrin, Thiabendazole, Thiacloprid, Thiamethoxam, Thiophanate-methyl, Trifloxystrobin, Aflatoxin Bl, Aflatoxin B2, Aflatoxin Gl, Aflatoxin G2, Ochratoxin A, Deoxynivalenol, Fumonisin Bl, Fumonisin B2, Fumonisin B3, HT-2 toxin, T-2 toxin, Zearalenone, Alpha, beta, gamma amatoxins, Propane, 2-Methylpropane (Isobutane), n-Butane, 2-Methylbutane (Isopentane), 2-2-Dimethylpropane (Neopentane), n-Pentane, 2,2-Dimethylbutane (Neohexane), 2,3-Dimethylbutane, 2-Methylpentane (Isohexane), 3-Methylpentane, n-Hexane, Cyclohexane, 2,2-Dimethylpentane (Neoheptane), 2,3-Dimethylpentane, 2,4-Dimethylpentane, 3,3-Dimethylpentane, 2,2,3-Trimethylbutane (Triptane), 2-Methylhexane (Isoheptane), 3-Methylhexane, 3-Ethylpentane, n-Heptane, Cycloheptane, Benzene, Toluene, Cumene, 1,3-Dimethylbenzene (m-Xylene), 1,4-Dimethylbenzene (p-Xylene), 1,2-Dimethylbenzene (oXylene), Ethylbenzene, Methanol, Ethanol, 1-Propanol, 2-Propanol (Isopropyl Alcohol), 1-Butanol, 2-Butanol, 1-Pentanol, Acetone, 2-Butanone, Tetrahydrofuran, Ethyl Ether, Ethylene Glycol, 2-Ethoxyethanol, 1,2-Dimethoxyethane, 1,4-Dioxane, Ethylene Oxide, Ethyl Acetate, Isopropyl Acetate, Chloroform, Dichloromethane (Methylene Chloride), Trichloroethylene, 1,2-Dichloroethane, 1,1-Dichloroethene, 1,2-Dichloroethene, Sulfolane, Dimethyl Sulfoxide, Acetonitrile, Pyridine, N,N-Dimethylacetamide, N,N-Dimethylformamide, Boron, Chromium, Cobalt, Copper, Lithium, Manganese, Molybdenum, Nickel, Selenium, Silver, Sulfur, Titanium, Tungsten, Zinc, Arsenic, Cadmium, Lead, Mercury, Aspergillus spp, (A. fumigatus, A. flavus, A. niger, A. terreus), Bile-Tolerant Gram-Negative Bacteria, Campylobacter spp, Candida albicans, Coliforms, Escherichia coli, Listeria monocytogenes. Pseudomonas aeruginosa, Salmonella spp, Shiga Toxin-Producing Escherichia coli (STEC), Staphylococcus aureus, Total Aerobic Bacteria, Total Enterobacteriaceae, Total Yeast and Mold, Yersinia spp, Hop Latent Viroid (HLVd), Lettuce Chlorosis Virus (LCV), Cannabis Cryptic Virus (CCV) Tobacco Mosaic Virus, Tocopheryl Acetate, a-Tocopherol, 5-Tocopherol, P-Tocopherol / y-Tocopherol, A8THC, A8THCV, A8-iso-THC, A9THC, A9THC Acetate, A10THC, THCa, THCV, THCVa, Exo-THC, 9R-HHC, 9S-HHC, CBD, CBDa, CBDV, CBDVa, CBC, CBCa, CBG, CBGa, CBL, CBN, CBNA, JWH-018, JWH-073, JWH-200, 5F-ADB, 5F-PB-22, 5F-AKB48, XLR-11, UR-144, PB-22, and the like.

[00171] Cultivation of a Cannabis Matrix Material for Cannabinoid Analysis

[00172] While the above example was suitable for analysis of cannabinoids as well as other natural ingredients and / or contaminants, it is important to note that not all of those procedures must be followed to produce a matrix material solely to be used for Cannabinoid analyses. The main differences are that the application of pesticides, fungicides, herbicides, and other beneficial organisms can be allowed to produce a material that is cannabinoid-free but may contain other commonly used compounds that are routinely seen by Cannabis testing laboratories. Thus, the strict approach to sterility and avoidance of any outside contaminants, while being very useful and essential for a universal biological matrix-blank material, it is not necessary in all cases to produce specific use case biological matrix-blanks. Under less rigorous conditions, in which testing is focused solely on certain analytes produced by the plant rather than extraneous contaminants, the plant can be grown indoors or outdoors without the need for a positive pressure environment. This allows for a laboratory to ensure that no additives, such as a pesticides, being added to the plant during cultivation are being detected as a cannabinoid and provides a cheaper alternative to cultivation of ‘ZCP’.

[00173] Extraction of‘ZCP’ Material

[00174] Given the minimal secondary metabolite concentration in ‘ZCP’, the derivation of an extract analogous to commercial Cannabis extract products poses intricate challenges. A multiplicity of extraction modalities was harnessed, including but not limited to: aqueous extractions, hydrocarbon extractions encompassing butane and propane, temperature-variant ethanol extractions, heat and pressure (rosin) and / or supercritical / subcritical CO2 extractions. The intent behind employing this diverse range of methodologies was to yield botanical extracts that faithfully replicate matrix blank materials, suitable for the analytical evaluation of both bioactive constituents and potential deleterious additives that the Cannabis plant might have been exposed to during its growth cycle. Additionally, the extractive process can be realized through other methodologies conventionally known in the art, including maceration, percolation, vaporization, chromatographic techniques, distillation, recrystallisation, and solvent extractions, such as but not limited to, including but not limited to ethanol, norflurane, butane, propane, propanol, methanol, acetone, acetonitrile, benzene, toluene, butanol, chloroform, ethyl ether, xylenes, pyridines, methylene chloride, dimethyl sulfoxide, isobutanol, nitrobenzene, cyclohexane, chlorobenezene, hexanes, heptanes, pentanes, and / or other alcohols. Significantly, extracts derived from plants nurtured via technique (i) are conspicuously devoid of cannabinoids, pesticides, fungicides, mycotoxins, endotoxins, microbial contaminants, viruses, synthetic drugs, residual solvents, Per and Polyfluorinated Substances (PFAS), and heavy metals. This intrinsic purity endows them with suitability across a spectrum of regulatory, research, and development endeavors that could not be achieved by ‘LCP’ material or extracted cannabinoid-containing matrix-blank material. Due to the myriad of primary and secondary metabolites that are naturally produced by Cannabis, the cannabinoid-free Cannabis allows for the creation of a cannabinoid-free extract that still contains these metabolites that were previously degraded or lost during the separation of cannabinoids and other metabolites found in the plant or extract thereof.

[00175] In order to characterize the chemotypes of the parental and produced offspring plants and the resulting extracts, a chemical analysis of both the cannabinoid content, and selected other chemicals, was undertaken as set out in Example 3: Example 3 Analysis of Cannabinoid and Cannflavin Content

[00176] Mature floral clusters, leaf tissue, and stem biomass were sampled from every individual plant considered in the breeding experiments. Samples were air-dried at roomtemperature and milled to a powder using stainless steel ball bearings. Between 0.2 and 0.5 grams of powder aliquots were solvent extracted in 10mL of HPLC-grade acetone using ultrasonication for a total of 30 minutes at a water temperature no greater than 35°C. Sample extracts were syringe-filtered with 0.22 pm PTFE filters, followed by either a 2-fold dilution for leaf and stem tissue extracts or a 5-fold dilution for floral extracts.

[00177] High-Performance Liquid Chromatography (HPLC) with Ultra-Violet (UV) Photo Diode-Array (PDA) detection was the technique utilized to measure concentrations of cannabinoids and cannflavins. All samples were analyzed using Shimadzu 2030C 3D series HPLC system equipped with an Ascentis Express C18 2.7pm x 150mm x 3mm column held at 24°C. The mobile phases consisted of HPLC grade acetonitrile and HPLC grade water formulated with 8% (v / v) Methanol, 0.035% (v / v) Formic Acid, 1.8mM ammonium formate. The mobile phase flow rate was 0.45mL / min. UV profiles were recorded at 230, 258, 270, 280, and 340nm depending on the lambda maximum for each cannabinoid or cannaflavin. The injection volume for each calibration and sample extract solution was 2 pL.

[00178] Cannabinoid and cannflavin peak areas were converted into concentrations using unique linear calibration equations for 32 cannabinoids and 2 cannflavins at on-column concentrations ranging from 0.1 ug / mL to 800 ug / mL. A solvent containing no analytes was applied to all standards and samples for consistent baseline identification. The total cannabinoid content was calculated and the weight proportions of the individual cannabinoids in the cannabinoid fraction were used to characterize the cannabinoid composition in the original material. Example 4 Handling & Packing Matrix Blank Material

[00179] Each consignment of the ‘ZCP’ material, upon release, must incontrovertibly certify the absence of seeds and further ensure its homogenization into a finely-ground Cannabis biomass or Cannabis extract matrix. The matrix can comprise one or more constituents selected from the group consisting of: resin, seeds, stems, roots, leaves, petioles, flowers, pre-flowers, and trichomes. The stipulation of such components was paramount to simulate an authentic matrix blank of Cannabis. This was predicated on the premise that laboratory samples, against which the ‘ZCP’ material was benchmarked, might encompass one or more of the aforementioned botanical elements.

[00180] In relation to matrix blanks, certain unique and non-obvious criteria were meticulously observed during the packaging process of the said material for its intended application as a matrix blank. Specifically, the environmental prerequisites for packaging closely mirror those requisites for cultivation, with distinct exceptions. Firstly, to mitigate the risk of post-harvest adulteration, the subject material was manipulated exclusively by personnel equipped with sterile gloves, Tyvek® suits, and protective facial coverings. Furthermore, the material was sequestered in containers that met or exceeded EPA performance-based standards for volatile organic analysis, ensuring the containers were unequivocally devoid of pesticides, fungicides, herbicides, PFAS, solvents, and any other potential adulterants that might compromise the pristine quality of the ‘ZCP’ material. During manipulation of the material, careful attention was given to assuring that the surrounding environment remained uncontaminated, particularly given the proclivity of various agents like pesticides, fungicides, microbials, and viruses to become airborne, which, if mishandled, to taint the intended results. Additionally, throughout cultivation and post-harvest processing, it is critical to ensure all scissors, homogenization grinders, blades, chambers, and other components used in the grow are made of medical grade stainless steel, or ceramic in composition to avoid the introduction of metal contaminants in the matrix-blank organism. Example 5: Other Bioactive and Analyte-free Matrix Blank Organisms

[00181] The methodology disclosed herein is applied to Psilocybe cubensis as an example of many other medicinal fungi that are amenable to this approach. Variant individuals are bred and selected that retain their traditional uses, entourage biochemical profiles, nutritional value, and / or ornamental qualities without producing the psychoactive, toxic, or other specific bioactive compounds they are typically known for. These compounds are collectively referred to as “target bioactives,” or “organism-specific bioactives,” or “medicinally-relevant bioactives,” or “bioactives of interest.”

[00182] The methods disclosed herein are applicable to producing a multiplicity of matrixblank fungi, as has been shown in the exemplary bioactive-free matrix-blank organism, Cannabis. This is achieved through the creation and use of the specific selectively bred varieties and their progeny which are specialized for the various applications disclosed herein. Such selections are used in industries such as pharmaceuticals, nutraceuticals, and agriculture, as well as applications in analytical testing methodologies that require a matrix-blank material for accurate quantification of the bioactive molecules.

[00183] The bioactive-free segregants resulting from backcrosses with a high bioactivecontent variety have biochemical profiles that appear to be typical as compared with the parent varieties, except as to the target bioactives which are selected for reduction and / or removal through such breeding.

[00184] The presumption regarding the absence of target bioactives in these fungi is an obstruction in biochemical pathways essential for generating precursor molecules leading to the target bioactives. The unaltered chemical profile of these bioactive-free fungi implies that the disruption does not occur in the initial stages of the pathways. This suggests that the interruption occurs downstream.

[00185] Impeding certain bioactive pathways does not necessarily impact bioactive synthesis, depending upon whether the pathway is impeded far upstream of the final bioactive, or is impeded closer to the final bioactive. In many cases, a disrupted pathway can result in reduced levels of precursors. On the other hand, a disruption in a downstream portion of the pathway hinders the formation of more direct precursors. Where the biosynthetic pathways are well known, analysis of precursor accumulation indicates which point(s) in the pathway are impeded.

[00186] The specific phenotype observed is presumed to be due to a malfunction in an upstream enzyme which is involved in catalyzing the modification of to produce another precursor of the bioactive. An ineffective enzyme thus leads to an accumulation of the upstream precursor and an absence of any other precursors that are downstream of the inactive or absent enzyme.

[00187] Thus, the absence of target bioactives is caused by a disruption in the pathway that produces them. The precise nature of this knockout phenomenon is uncertain, but the absence of the target bioactive produces the desired result. This confirms that breeding and selection that are driven by biochemical phenotype need not always involve dissection and full documentation of the exact point of disruption of the biosynthetic pathway. A fungus that has a substantially normal morphology and that is biochemically very similar to the original parental line from which the breeding and selection was initiated, but which lacks one or more target bioactives, is effectively produced and is functionally very useful, even without such detailed confirmation of the genetic and / or biosynthetic disruptions that cause the absence of the target bioactive.

[00188] A further use of the fungi produced by this approach is that they can be used to create fungal-extracts matrix-blank materials containing other bioactive molecules than target bioactives in quantities and purities, which could not be achieved naturally. Such fungal-extracts matrix-blank materials provide the benefits arising from the presence of one or more selected other compounds. This produces a “designer” fungus extract that can be used in analytical method validation and proficiency testing or for the production of customized bioactive-ratio materials that cannot be achieved naturally.

[00189] In some uses, the Zero Fungus biomass or extract derived thereof is spiked with >0.1% by dry weight of a target bioactive while remaining absent of other target bioactives. Due to the nature of bioactive biosynthesis, creating fungi without certain bioactives but having other bioactives is normally impossible if a given bioactive is a precursor to the terminal bioactives. Thus, through these methods of using the fungal materials of this invention, it is possible to create non-naturally occurring bioactive profiles through infusing naturally derived, synthetic, or modified bioactives into the fungal biomass. Extracts derived therefrom are possible using the matrix-blank organisms and materials and methods disclosed herein. In some uses, this is achieved through solubilizing the bioactives in a known volume of liquid solvent, such as ethanol, methanol, or other solvents in which the target bioactives are soluble. In other uses, a similar infusion process can be used to incorporate one or more compounds from the following classes: chemical contaminants, environmental contaminants, bioactive compounds derived from the fungus, bioactive compounds not found in the fungus, or non-bioactive compounds.

[00190] Thus, for example, it is possible to use psilocybin-free Psilocybe cubensis mushrooms to create novel ratios of tryptamines that are not found in nature, but may provide enhanced or improved medicinal properties. Norbaeocystin and baeocystin are precursors to psilocybin, which prevents a psilocybin-containing mushroom from being baeocystin-free in nature. However, through utilizing the method disclosed herein, one is able to utilize the a psilocybin-free fungal matrix-blank to produce norbaeocystin and psilocybin containing biomass that is free of baeocystin, as well as create other non-natural combinations of tryptamines spiked into the natural background of metabolites found in the organism to allow for reproducible and targeted medicinal formulations of natural products to achieved in the spiked biomass or extracts derived thereof.

[00191] Similar logic is applied in many other species and genera of fungi, to identify, selectively breed, and produce organisms devoid of medically or recreationally relevant compounds and their respective degradants. For example, a psilocybin-free Psilocybe species is produced by selecting or knocking out the genes in the psilocybin biosynthesis pathway that prevent the accumulation of tryptamines including norbaeocystin, baeocystin, psilocybin, psilocin, aeruginascin, and 4-hydroxy-trimethyl tryptamine. In another example, the hericenone diterpene biosynthetic pathway is selected against to produce hericenone-free Hericium species. Similar post-harvest processing and packaging steps are applied for these organisms to produce a research-grade or certified reference material grade bioactive-free matrix blank biomass material or extracts thereof that are used according to this disclosure.

[00192] In another use of this technology, the matrix-blank organism is spiked with pathogenic or contaminant organisms or metabolites indicative of those organisms to create a proficiency test to assess the specificity of a laboratory’s target species assay. In some embodiments, two mixtures of the matrix-blank organism are prepared: one spiked with shiga-toxin producing E. Coli and one spiked with non-pathogenic E. Coli, and are sent to a laboratory as an unknown sample to determine the accuracy and specificity of microbial testing protocols specifically in regards to testing products containing the same organism as the matrix-blank organism. In another embodiment, the matrix blank organism is spiked with one or more than one species of pathogenic Aspergillus species, which using this spiked matrixblank organism enables a laboratory to validate microbial methods in new organisms where the matrix has not been tested. EXAMPLE 6

[00193] The methodology disclosed herein is applied to Lophophora williamsii as an example of many other medicinal cacti that are amenable to this approach.

[00194] This technology is used to produce mescaline-free cacti through disruption of the mescaline biosynthesis pathway, This disruption can occurs through numerous selectively bred genotypes including, but not limited to, truncated genes in the biosynthetic pathway, loss-of-function genes in the biosynthetic pathway, upstream and downstream mutations causing differential regulation of the biosynthetic pathway, and mutations in regulatory factors associated with the expression of these biosynthetic genes. Alternatively, these same mutations are introduced through non-selective breeding methods as well, such as CRISPR-CAS, Cre-Lox, and other known gene modification techniques to introduce non-naturally occurring knockouts of these organisms to be used in the disclosed applications of the bioactive-free matrix blank material.

[00195] The methods disclosed herein are applicable to all producing a multiplicity of matrix-blank cacti, as has been shown in the exemplary bioactive-free matrix-blank organism, Cannabis. This is achieved through the creation and use of the specific selectively bred varieties and their progeny which are specialized for the various applications disclosed herein. Such selections are used in industries such as pharmaceuticals, nutraceuticals, and agriculture, as well as applications in analytical testing methodologies that require a matrix-blank material for accurate quantification of the bioactive molecules.

[00196] The bioactive-free segregants resulting from backcrosses with a high bioactivecontent variety have biochemical profiles that appear to be typical as compared with the parent varieties, except as to the target bioactives which are selected for reduction and / or removal through such breeding.

[00197] The presumption regarding the absence of target bioactives in these cacti is an obstruction in biochemical pathways essential for generating precursor molecules leading to the target bioactives. The unaltered chemical profile of these bioactive-free cacti implies that the disruption does not occur in the initial stages of the pathways. This suggests that the interruption occurs downstream.

[00198] Impeding certain bioactive pathways does not necessarily impact bioactive synthesis, depending upon whether the pathway is impeded far upstream of the final bioactive, or is impeded closer to the final bioactive. In many cases, a disrupted pathway can result in reduced levels of precursors. On the other hand, a disruption in a downstream portion of the pathway hinders the formation of more direct precursors. Where the biosynthetic pathways are well known, analysis of precursor accumulation indicates which point(s) in the pathway are impeded.

[00199] The specific phenotype observed is presumed to be due to a malfunction in an upstream enzyme which is involved in catalyzing the modification of to produce another precursor of the bioactive. An ineffective enzyme thus leads to an accumulation of the upstream precursor and an absence of any other precursors that are downstream of the inactive or absent enzyme.

[00200] Thus, the absence of target bioactives is caused by a disruption in the pathway that produces them. The precise nature of this knockout phenomenon is uncertain, but the absence of the target bioactive produces the desired result. This confirms that breeding and selection that are driven by biochemical phenotype need not always involve dissection and full documentation of the exact point of disruption of the biosynthetic pathway. A cactus that has a substantially normal morphology and that is biochemically very similar to the original parental line from which the breeding and selection was initiated, but which lacks one or more target bioactives, is effectively produced and is functionally very useful, even without such detailed confirmation of the genetic and / or biosynthetic disruptions that cause the absence of the target bioactive.

[00201] A further use of the cacti produced by this approach is that they can be used to create cacti-extracts matrix-blank materials containing other bioactive molecules than target bioactives in quantities and purities, which could not be achieved naturally. Such cacti-extracts matrix-blank materials provide the benefits arising from the presence of one or more selected other compounds. This produces a “designer” cactus extract that can be used in analytical method validation and proficiency testing or for the production of customized bioactive-ratio materials that cannot be achieved naturally. In some embodiments the zero-mescaline cacti would be derived from an inbred line of one of the following species: San Pedro (Echinopsis pachanoi), Peruvian Torch (Echinopsis peruviana), Bolivian Torch (Echinopsis lageniformis), Peyote (Lophophora williamsii), Lophophora diffusa, Echinopsis scopulicola, Echinopsis santaensis, Echinopsis macrogona, Trichocereus bridgesii, Trichocereus terscheckii, Trichocereus cuzcoensis, and / or Trichocereus validus. In some embodiments the zero-mescaline cacti would be derived from an interspecific hybrid line of one or more than one of the following species: San Pedro (Echinopsis pachanoi), Peruvian Torch (Echinopsis peruviana), Bolivian Torch (Echinopsis lageniformis), Peyote (Lophophora williamsii), Lophophora diffusa, Echinopsis scopulicola, Echinopsis santaensis, Echinopsis macrogona, Trichocereus bridgesii, Trichocereus terscheckii, Trichocereus cuzcoensis, and / or Trichocereus validus.

[00202] In some uses, the Zero-mescaline Cactus biomass or extract derived thereof is spiked with >0.1% by dry weight of a target bioactive while remaining absent of other target bioactives. Due to the nature of bioactive biosynthesis, creating cacti without certain bioactives but having other bioactives is normally impossible if a given bioactive is a precursor to the terminal bioactives. Thus, through these methods of using the fungal materials of this invention, it is possible to create non-naturally occurring bioactive profiles through infusing naturally derived, synthetic, or modified bioactives into the fungal biomass. Extracts derived therefrom are possible using the matrix-blank organisms and materials and methods disclosed herein. In some uses, this is achieved through solubilizing the bioactives in a known volume of liquid solvent, such as ethanol, methanol, or other solvents in which the target bioactives are soluble. In other uses, a similar infusion process can be used to incorporate one or more compounds from the following classes: chemical contaminants, environmental contaminants, bioactive compounds derived from the cactus, bioactive compounds not found in the cactus, or non-bioactive compounds.

[00203] In another use of this technology, the matrix-blank organism is spiked with pathogenic or contaminant organisms or metabolites indicative of those organisms to create a proficiency test to assess the specificity of a laboratory’s target species assay. In some embodiments, two mixtures of the matrix-blank organism are prepared: one spiked with shiga-toxin producing E. Coli and one spiked with non-pathogenic E. Coli, and are sent to a laboratory as an unknown sample to determine the accuracy and specificity of microbial testing specifically in regards to testing products containing the same organism as the matrix-blank organism. In another embodiment, the matrix blank organism is spiked with one or more than one species of pathogenic Aspergillus species, which using this spiked matrix-blank organism enables a laboratory to validate microbial methods in new organisms where the matrix has not been tested. EXAMPLE 7

[00204] The technology disclosed herein is used to make zero-bioactive organisms including ganoderic acid free Ganoderma spp mushrooms through disruption of the ganoderic biosynthetic pathway, cordycepin-free cordyceps spp through disruption of the cordycepin biosynthetic pathway, alkaloid-free ibogaine through disruption of alkaloid biosynthetic genes, isoxazole-free amanita spp through the disruption of the ibotenic acid biosynthetic pathway, and hericenone-free and / or erinacine-free lions mane material through the disruption of the triterpenoid biosynthetic pathway. Similar observations and presumptions are applied, and similar conclusions are reached, as those discussed in Examples 5 and 6.

[00205] Other aspects of the invention will be clear to the skilled artisan and need not be repeated here. Each reference cited herein is incorporated by reference in its entirety for the relevant teaching contained therein.

[00206] The terms and expressions that have been employed are used as terms of description and not of limitation, and there is no intention in the use of such terms and expressions of excluding any equivalents of the features shown and described or portions thereof, it being recognized that various modifications are possible within the scope of the invention. REFERENCES Pacifico D., et al. Molecular Breeding (2006) vol. 17; pp. 257-268. Novak, J et al. Flavour and Fragrance Journal 2001; vol. 16, pp. 259-262. Vanhoenacker, G et al. Natural Product Letters, vol. 16, No. 1, pp. 57-63. Wilkinson, J.D. et al. Journal of Pharmacy and Pharmacology 2003; vol. 55, pp. 1687-1694. Small, E. et al. Economic Botany 2003; vol. 57. No. 4; pp. 545-558. Hillig, K. W., “A chemotaxonomic analysis of terpenoid variation in Cannabis, ” Biochem Systematics Ecology 2004; 32:875-891. Kriese, U. et al., “Oil content, tocopherol composition and fatty acid patterns of the seed of 51 Cannabis spp L genotypes,” Euphytica 2004; 137:339-351. Novak, J. et al., “Essential oils of different cultivars of Cannabis spp L and their antimicrobial activity,” Flavour and Fragrance J 2001; 16:259-262. Pacifico, D. et al., “Genetics and Marker-assisted Selection of the Chemotype in Cannabis spp L,” Molecular Breeding Apr. 1,2006; 17(3):257-268. Small, E. et al., “Tetrahydrocannabinol levels in Hemp (Cannabis spp) germplasm resources,” Economic Botany 1 Jan. 2003; 57(4):545-558. Vanhoenacker, G. et al., “Chemotaxonomic features associated with flavonoids of cannabinoid-free Cannabis in relation to hops”, Natural Product Letts 2002: 16(1):57-63. Virovets, V.G. et al., “Selektion auf niedrigen Gehalt der Cannabinoid und hohe Produktivitat im Schaffungsprogramm von Hanfsorten (Cannabis spp L.), die kiene narkotische Aktivitat besitzen,” Tagungsband Zum / Proceedings of the Symposium Biorohstoff Hanf Nova-Institut Fur Politische und OKologische Innovation; Jan. 1, 1997:135-153. Wilkinson, J.D. et al., “Medicinal Cannabis', is delta-9-tetrahydrocannabinol necessary for all its effects?” J Pharmacy Pharmacol 2003; 55:1687-1694. The United Kingdom Parliament, Select Committee on Science and Technology Ninth Report (1998) at http: / / www.parliament.the-stationery-office.co.uk / pa / Idl99798 / ldselect / ldsctech / 151 / 15101.htm. The United Kingdom Parliament, Select Committee on Science and Technology Second Report (Mar. 14, 2001) at http: / / www.publications.parliament.uk / pa / ld200001 / ldselect / ldsctech / 50 / 5001.ht. De Meijer, Fibre hemp cultivars: A survey of origin, ancestry, availability and brief agronomic characteristics. J IntHemp Assoc. 1995;2(2):66-73. De Zeeuw et al., Cannabinoids with a propyl side chain in Cannabis', occurrence and chromatographic behavior. Science. Feb. 18, 1972;175(23):778-9. Fellermeier et al., Biosynthesis of cannabinoids. Incorporation experiments with (13)C-labeled glucoses. Eur J Biochem. Mar. 2001;268(6):1596-604. Fellermeier et al., Prenylation of olivetolate by a hemp transferase yields cannabigerolic acid, the precursor of tetrahydrocannabinol. FEBS Lett. May 8, 1998;427(2):283-5. McPartland et al., Cannabis and Cannabis extracts: greater than the sum of their parts?. J Cannabis Therapeutics. 2001;1:103-32. Morimoto et al., Enzymological Evidence for Cannabichromenic Acid Biosynthesis. J Nat Prod. Aug. 1997;60(8):854-7. Morimoto et al., Purification and characterization of cannabichromenic acid synthase from Cannabis spp. Phytochemistry, Nov. 1998;49(6): 1525-9. Rahaijo et al., Cloning and over-expression of a cDNA encoding a polyketide synthase from Cannabis spp. Plant Physiol Biochem. Apr. 2004;42(4):291-7. Rahaijo et al., Olivetol as product of a polyketide synthase in Cannabis spp L. Plant Science. Feb. 2004;166(2):381-5. Taura et al., First direct evidence for the mechanism of .DELTA. 1-tetrahydrocannabinolic acid biosynthesis. J Am Chern Soc. Sep. 1995; 117(38):9766-7. Taura et al., Purification and characterization of cannabidiolic-acid synthase from Cannabis spp L.. Biochemical analysis of a novel enzyme that catalyzes the oxidocyclization of cannabigerolic acid to cannabidiolic acid. J Biol Chern. Jul. 19, 1996;271(29): 17411-6. Virovets, Interview. J Int Hemp Assoc. 1998;5:32-4. Virovets, Selection for Non-Psychoactive Hemp Varieties (Cannabis spp L.) in the CIS (former USSR). J Int Hemp Assoc. 1996;3:13-5. Samuelsson, Drugs of natural origin, 4th ed. Swedish Pharmaceutical Press. 1999; 551. De Meijer et al., The inheritance of chemical phenotype in Cannabis spp L. (II): Cannabigerol predominant plants. Euphytica. 2005;145(l-2):189-98. Evans, Pharmacognosy 15thed. 2002;585. Gaoni et al., Cannabichromene, a new active principle in hashish. Chern Comm 1966; 1:20-1. Gorshkova et al., Methods of evaluating hemp plants for content of cannabinoid compounds. Referativnyi Zhurnal. 1988;12.65.322. Abstract only. Sirikantaramas et al., Tetrahydrocannabinolic acid synthase, the enzyme controlling marijuana psychoactivity, is secreted into the storage cavity of the glandular trichomes. Plant Cell Physiol. Sep. 2005;46(9): 1578-82. Epub Jul. 15, 2005. Speroni et al., Antiinflammatory effects of Cannabis spp L. extracts containing nonpsychoactive cannabinoids. Borrelli et al. eds. Proceedings 3rdInternational Symposium on Natural Drugs, Indena, Naples. 2003;107-14. Virovets et al., Narcotic activity of Cannabis spp L. and prospects of its selection for decreased content of cannabinoids. Agricultural Biology 1. 1991;35-49. Vree et al., Identification of the methyl and propyl homologues of CBD, THC and CBN in hashish by a new method of combined gas chromatography-mass spectrometry. Acta Pharma Suecica. 1971;8:683-4. Williamson et al., Cannabis as a medicine: evidence for synergy. In Medicinal uses of Cannabis, 26th LOF Symposium, Leiden 2002. LaVigne, J.E., Hecksei, R., Keresztes, A. et al. Cannabis sativa terpenes are cannabimimetic and selectively enhance cannabinoid activity. Sci Rep 11,   8232   (2021). https: / / doi.org / 10.1038 / s41598-021-87740-8

Claims

1. A biological matrix-blank reference material derived from a selectively-bred organism, wherein the selectively-bred organism has been selected to not express at least one bioactive compound, but to express at least one other compound expressed in a comparator plant, and wherein the matrix-blank reference material has a chemical profile that resembles that of the comparator organism less the at least one bioactive compound.

2. The biological matrix-blank reference material of claim 1, wherein the selectively-bred organism is selected from one of the following organisms: Lion's Mane mushroom species such as Hericium erinaceus, Hericium coralloides, and Hericium americanum: Reishi mushroom species including Ganoderma lucidum, Ganoderma tsugae, and Ganoderma applanatum', Chaga mushroom, specifically Inonotus obliquus, Cordycep mushrooms such as Cordyceps militaris, Cordyceps gunni, and Cordyceps dipterigena; Amanita mushroom species such as Amanita citrina, Amanita muscaria, Amanita gemmata, Amanita. Pantherina, Amanita porphyria, Amanita regalis, and Amanita strobiliformis; various psilocybin-containing mushrooms such as Psilocybe tampanensis, Psilocybe cubensis, Psilocybe semilanceata, Psilocybe oaxacana, Psilocybe natalensis, Psilocybe baeocystis, Psilocybe zapatecorm, Psilocybe azurescens, Psilocybe mexicana, Panaeoulus cyanescens, Panaeoulus cinctulus Psilocybe cyanescens, Panaeolus acidus, Panaeolus acuminatus, Panaeolus affmis, Panaeolus africanus, Panaeolus albellus, Panaeolus albidocinereus, Panaeolus albovelutinus, Panaeolus aids, Panaeolus alveolatus, Panaeolus annulatus, Panaeolus anomalus, Panaeolus antillarum, Panaeolus atomatus, Panaeolus atrobalteatus Panaeolus axfordii, Panaeolus bisporus, Panaeolus cambodginiensis, Panaeolus fimicola, Panaeolus moellerianus, Panaeolus olivaceus, Panaeolus fimicoloides, Panaeolus fimiputris, Panaeolus foenisecii, Panaeolus fontinalis, Panaeolus fraxinophilus, Panaeolus georgii, Panaeolus gomphodes, Panaeolus goossensiae, Panaeolus griseofibrillosus, Panaeolus guttulatus, Panaeolus hippophilus, Panaeolus hygrophanus, Panaeolus hypomelas, Panaeolus incanus, Panaeolus indicus, Panaeolus intermedins, Panaeolus lentisporus, Panaeolus lerchenfeldii, Panaeolus leucophanes, Panaeolus lignicola, Panaeolus linnaeanus, and Panaeolus longiguus, Lophophora williamsii, Lophophora diffusa, Lophophora fricii, Trichocereus pachanoi, and Trichocereus peruvianus, Mimosa hostilis / tenuiflora and Acacia confusa, Banisteriopsis caapi, Passiflora incarnata, and Peganum harmala.

3. The biological matrix-blank reference material of claim 1, wherein biological matrix-blank reference material is used for comparison with unknown or known bioactivecontaining samples of the same organism.

4. The biological matrix-blank reference material of claim 1, wherein the reference material comprises an extract or biomass of the selectively-bred organism.

5. The biological matrix-blank reference material of claim 4, wherein the biomass is a homogenized bulk biomass.

6. The matrix-blank reference material of claim 1, wherein the selectively-bred organism is a Cannabis plant.

7. The matrix-blank reference material of claim 1, wherein the selectively-bred Cannabis plant is characterized by an absence one or more of the following cannabinoids: tetrahydrocannabiphorol (9-THCP), tetrahydrocannabivarin (9-THCV), delta-8-tetrahydrocannabinol (8-THC), delta-9-tetrahydrocannabinol (9-THC), tetrahydrocannabinolic acid (9-THCA), tetrahydrocannabiphorolic acid (9-THCP A), tetrahydrocannabivarinic acid (9-THCVA), cannabichromene (CBC), cannabichromenic acid (CBCA), cannabichromeorcin (CBCO), cannabichromevarin (CBCV), cannabichromevarinic acid (CBCVA), cannabidiol (CBD), cannabidiolic acid (CBDA), cannabidibutol (CBDB), cannabielsoin (CBE), cannabidiol monomethyl ether (CBDM), cannabidiol acid monomethyl ether (CBDAM), cannabidiorcin (CBDO), cannabidiphorolic acid (CBDPA), cannabidivarin (CBDV), cannabidivarinic acid (CBDVA), cannabigerol (CBG), cannabigerolic acid (CBGA), cannabigerol acid monomethyl ether (CBGAM), cannabigerol monomethyl ether (CBGM), cannabigerovarin (CBGV), cannabigerovarinic acid (CBGVA), cannabigerphorolic acid (CBGPA), cannabicyclol (CBL), cannabicyclolic acid (CBLA), cannabinol (CBN), cannabinolic acid (CBNA), cannabicitran (CBT).

8. The matrix-blank reference material of claim 7, wherein the selectively-bred Cannabis plant is further characterized by trichomes of clear, white, and / or amber coloration.

9. The biological matrix-blank reference material of claim 8, wherein the plant is the Cannabis variety designated as ‘ZCP.’10. The biological matrix-blank reference material of claim 7, wherein the reference material comprises one or more flavonoids selected from cannflavin A, cannflavin B, isocannflavin B, and cannflavin C.

11. The biological matrix-blank reference material of claim 7, wherein the reference material comprises terpenes, further characterized by one or more of:the presence of both monoterpenes and sesquiterpenes;the absence of either monoterpenes or sesquiterpenes;the absence of both monoterpenes and sesquiterpenes;the presence of diterpenes;the absence of diterpenes;the presence of triterpenes;the absence of tri terpenes;the presence of tetraterpenes; orthe absence of tetraterpenes.

12. The biological matrix-blank reference material of claim 1, wherein the reference material is devoid of a profile of entourage or contaminant compounds or organisms.

13. The biological matrix-blank reference material of claim 12, further characterized by at least one of:the presence of a known compound or organism spiked in at a known concentration for analytical reference; orthe addition of one or more compounds selected from the group consisting of flavonoids, alkaloids, terpenoids, carotenoids, cannabinoids, alcohols, ketones, ethers, amines, amides, esters, aldehydes, sterols, pesticides, herbicides, fungicides, hydrocarbons, synthetic cannabinoids, synthetic psychedelics, synthetic opioids, isoxazoles, halogenated compounds, radioactive elements, heavy metals, Aspergillus, Salmonella, Shiga-Toxin Producing Escherichia coli, non-pathogenic Escherichia coli, Pseudomonas, Listeria, Hepatitis, norovirus, Clostridium, Campylobacter, Taxoplasma, Psilocybe species genes, Cryptosporidium, Saccharomyces, hops-latent viroid, beat-curly top virus, tobacco mosaic virus, and radiolabeled nucleic acids.

14. The biological matrix-blank reference material of claim 1, for use in biological consumption to create a bioactive-free matrix-blank material from biological tissue or excrement.

15. The biological matrix-blank reference material of claim 1, wherein the referencematerial is further characterized by the addition of one or more compounds selected from the group consisting of flavonoids, terpenoids, carotenoids, cannabinoids, alcohols, ketones, pesticides, herbicides, fungicides, solvents, synthetic cannabinoids, synthetic psychedelics, synthetic opioids, isoxazoles, halogenated compounds, radioactive elements, Aspergillus, Salmonella, Escherichia coli, and heavy metals16. The biological matrix-blank reference material of claim 1 for biological consumption by humans or animals to create a comparative analytical chemistry matrix.

17. The biological matrix-blank reference material of claim 1, wherein the reference material is for proficiency testing or regulatory testing enforcement to evaluate the ability of a laboratory to detect and quantify pathogens or adulterants in Cannabis.

18. A method for cultivating Cannabis to produce pesticide-, herbicide-, fungicide, heavy metal-, mycotoxin-, endotoxin-, and microbial-free Cannabis biomass, as described in Example 1.

19. A method for sterile post-harvest processing of Cannabis to prevent environmental contamination, as described in Example 2.

20. A method for packaging a biological matrix-blank reference material in sterile containers meeting or exceeding EPA standards in a clean environment, ensuring no contamination during packaging, transport, or storage, wherein the packaging is performed under ISO 17034 accreditation.

Images