26 results about "Neochlorogenic acid" patented technology

This invention belongs to the field of traditional Chinese medicine preparation analysis, specifically disclosing a one-test-multiple-evaluation method for determining the content of seven components in Jinteng Qingbi granules and its application. Using sinomenine as an internal standard, a one-test-multiple-evaluation method is used to establish relative correction factors between sinomenine and gallic acid, neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B, and isochlorogenic acid C in Jinteng Qingbi granules. The contents of gallic acid, neochlorogenic acid, sinomenine, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B, and isochlorogenic acid C in Jinteng Qingbi granules are calculated using these correction factors. This invention's method is highly practical, simple to operate, cost-effective, and provides accurate and reliable test results, overcoming the shortcomings of existing methods for quality testing of Jinteng Qingbi granules and providing a new approach for quality control and intrinsic quality evaluation of Jinteng Qingbi granules.

The invention discloses construction and application of an HPLC (High Performance Liquid Chromatography) fingerprint spectrum for integrally controlling a multi-component material basis of a traumatic injury bone-growing capsule, and belongs to the technical field of quality control of traditional Chinese medicine preparations. According to the method, high performance liquid chromatography and mass spectrometry are combined, and neochlorogenic acid, protocatechuic aldehyde, cryptochlorogenic acid, vanillic acid, rosmarinic acid, alkannic acid, berberine hydrochloride and salvianolic acid B are taken as reference substances; the fingerprint spectrum is successfully constructed by detecting different batches of traumatic injury bone-growing capsules. The method is simple and easy to implement, high in precision, good in reproducibility, accurate and reliable, at least eight components can be confirmed at a time under the same chromatographic condition, the detection efficiency is greatly improved, and time and cost are saved; quality monitoring can be carried out on a plurality of characteristic components of the traumatic injury bone-growing capsule accurately, rapidly and simultaneously, and a powerful guarantee is provided for controlling the quality and ensuring the clinical curative effect of the traumatic injury bone-growing capsule.

The invention relates to a traditional Chinese medicine composition which comprises more than 1.09 permillage of ephedrine hydrochloride, more than 0.86 permillage of pseudoephedrine hydrochloride, more than 0.17 permillage of peiminine, more than 0.03 permillage of peimine, more than 0.17 permillage of chrysin-7-O-glucuronide, more than 0.002 permillage of glycyrrhetinic acid, more than 11.87 permillage of amygdalin and more than 0.01 permillage of neotuberostemonine. The oral liquid is prepared from more than 2.16 per mill of glycyrrhizic acid, more than 1.97 per mill of oroxylin A, more than 1.97 per mill of oroxylin B, more than 1.23 per mill of liquiritin, more than 0.59 per mill of apiose liquiritin, more than 0.55 per mill of baicalin, more than 0.23 per mill of chlorogenic acid, more than 0.18 per mill of praeruptorin A, more than 0.15 per mill of wild melanosin, more than 0.14 per mill of methyephedrine hydrochloride, more than 0.13 per mill of isoliquiritin, more than 0.13 per mill of neochlorogenic acid and more than 0.13 per mill of cryptochlorogenic acid. The composition is prepared from more than 0.10% of peimine, more than 0.08% of baicalein, more than 0.05% of praeruptorin B, more than 0.04% of praeruptorin C, more than 0.011% of xanthoxylin and more than 0.0006% of formononetin.

The application relates to the field of traditional Chinese medicine quality control, and particularly discloses a quality control method for Shuanghuanglian inhalation solution. The method can simultaneously determine the fingerprint spectrum and ingredient content of the Shuanghuanglian inhalation solution. The fingerprint spectrum detection method disclosed by the application adopts UPLC determination, has 17 characteristic peaks, and attributes 8 characteristic peaks. The contents of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B, isochlorogenic acid A, isochlorogenic acid C and forsythoside are simultaneously determined. Compared with the prior art, the method for determining the fingerprint spectrum and content of the Shuanghuanglian inhalation solution is improved, and the quality of the Shuanghuanglian inhalation solution can be comprehensively monitored conveniently, quickly and accurately.

The invention discloses application of neochlorogenic acid in preparation of a medicine for preventing and / or treating hand-foot-and-mouth disease caused by A16 type coxsackie virus (CVA16) infection, and belongs to the technical field of biological medicine. Researches prove that the neochlorogenic acid has the effect of resisting CVA16 infection on HEK293T cells and H4 cells, after the two cells treated by the neochlorogenic acid are infected by the CVA16, the viral genome RNA abundance is obviously reduced, and the cell lesion death condition is obviously relieved; meanwhile, a cytotoxicity test shows that the neochlorogenic acid has no obvious cytotoxicity to HEK293T cells and only has weak cytotoxicity to H4 cells. Therefore, in a reasonable dosage range, the neochlorogenic acid, which is a low-toxicity natural small molecule compound, can be used for preparing the anti-CVA16 medicine for preventing and / or treating the hand-foot-and-mouth disease caused by CVA16 infection.

The present application relates to a kind of Huahong composition effective part extract and its preparation method, the Huahong composition effective part extract includes the following components: syringic acid, new chlorogenic acid, deacetyl carlina acid methyl ester, geniposidic acid, N-acetyl-L-tyrosine, fraxidin, 4-pyridoxinic acid, 7,8-dihydroxy-4-methyl coumarin, chlorogenic acid, ferulic acid;The total content of above components is 80%-95% of effective part extract.The Huahong composition effective part extract of the present application is from the effective part of Huahong composition.

Provided herein are polyphenol-enriched lettuce powders comprising particles of a polyphenol-enriched lettuce or extract, wherein the polyphenol-enriched lettuce comprises from 40-280 mg / g of polyphenols. The polyphenols include any combination of a chlorogenic acid, a neochlorogenic acid, a chicoric acid, a quercetin, a quercetin derivative, and an anthocyanin. The disclosure also provides compositions and methods of using the polyphenol-enriched lettuce powders to promote wound healing and treat inflammation and skin disorders.

The invention belongs to the technical field of medicinal material quality control, and discloses a honeysuckle flower QAMS (quantitative analysis of multi-components by single marker) quality control method, which comprises the following steps: preparing a single reference solution and a mixed reference solution of chlorogenic acid, neochlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B, isochlorogenic acid A and isochlorogenic acid C, and preparing a test solution; after high performance liquid chromatography detection, the relative correction factors of other components in the honeysuckle are calculated by taking chlorogenic acid as an internal reference substance through chromatographic peak positioning, and the contents of the components in the honeysuckle are obtained by utilizing the relative correction components, so that the synchronous determination of the multiple components in the honeysuckle is realized. The method has the advantages of cost saving, simplicity in operation, high detection accuracy and good stability, has great significance in quality control and clinical curative effect guarantee of the honeysuckle medicinal material, and provides a basis for establishing a quality evaluation standard of the honeysuckle medicinal material.

The present application relates to the technical field of traditional Chinese medicine detection, and provides a genuine three-leafed knot thin-layer identification kit and method, comprising: a chrysophanol control solution, a neochlorogenic acid control solution and a rapid extraction solvent, the rapid extraction solvent contains a low-carbon alcohol, which is used for extracting flavonoid glycoside components in three-leafed knot stem and leaf samples, and the kit is used for identifying the origin of three-leafed knot by detecting the content of chrysophanol and the presence of neochlorogenic acid. The present application uses a chrysophanol and neochlorogenic acid double-index mutual exclusion identification logic, combined with a rapid extraction solvent system without drying, to realize on-site, rapid and accurate identification of the genuineness of three-leafed knot, and effectively solves the problem of origin confusion.

The application discloses application of a medicine composition in preparation of a medicine for treating rheumatoid arthritis (RA), and belongs to the field of RA treating medicines. The medicine composition is composed of swertiamarin (SW) and neochlorogenic acid (NCA) and is screened from two active ingredients in Danteng Yimu Decoction. With the help of animal model and cell model experiments, the action mechanism of the medicine composition in treating RA is explored, the related symptoms and indexes of collagen-induced arthritis (CIA) rats can be effectively improved, joint swelling can be relieved, arthritis inflammation index can be reduced, inflammatory factor release can be inhibited, joint bone quality can be protected, arthritis inflammation infiltration and cartilage destruction can be reduced, and the medicine safety is good. The composition can effectively inhibit the inflammatory factor release and migration ability of rheumatoid arthritis fibroblast-like synoviocytes (RA-FLS). Animal experiments and cell experiments prove that the combination of neochlorogenic acid and swertiamarin (SW+NCA) can be effectively applied to clinical treatment of RA and has a wide application prospect.

The invention discloses an establishment method and an application method of a fingerprint spectrum of a honeysuckle and fructus forsythiae mixture, and belongs to the technical field of detection of effective components of traditional Chinese medicine, and the establishment method comprises the following steps: S1, determining chromatographic conditions; s2, preparing a test solution and a mixed reference solution; s3, collecting chromatograms of the test sample and the mixed reference substance; and S4, matching common peaks, calculating the relative retention time and the relative peak area of each common peak, comparing with the chromatogram of the reference substance, identifying the common peaks, and establishing the fingerprint spectrum of the honeysuckle and fructus forsythiae mixture. According to the establishment method of the fingerprint spectrum of the honeysuckle and fructus forsythiae mixture, reference and basis are provided for quality control of the honeysuckle and fructus forsythiae mixture, and the establishment method has the advantages of being simple, convenient, stable, high in precision, good in reproducibility and the like. The fingerprint spectrum has 11 effectively separated common peaks, identifies the characteristic peaks of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, liquiritin and forsythiaside A, can reflect the condition of the effective components of the honeysuckle and fructus forsythiae mixture, and effectively characterizes the quality of the honeysuckle and fructus forsythiae mixture.

The invention discloses a quality control method for a standard sample of a five-ingredient disinfection drink. The quality control method comprises the following steps: (1) establishing a thin layer chromatography to identify honeysuckle, dandelion, herba violae and wild chrysanthemum in the standard sample; (2) establishing a fingerprint spectrum with relatively good separation degree and reproducibility by adopting a high performance liquid chromatography; (3) quantitatively researching 12 index components including neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid and the like through high performance liquid chromatography, and establishing a thin-layer identification method for honeysuckle, dandelion, philippine violet herb and wild chrysanthemum flower in the standard sample of the five-flavor disinfection drink; a method for determining the content of 12 index components such as neochlorogenic acid, chlorogenic acid and cryptochlorogenic acid is established, a reference sample characteristic spectrum method is established, and the similarity of 15 batches of five-ingredient disinfection drink reference samples reaches 0.90 or above.

The present application belongs to the technical field of quality detection of traditional Chinese medicinal materials, and particularly relates to a method for quantitatively analyzing effective components of Xiaochaihu medicinal materials. The method comprises the following steps: a, preparation of a control solution: precisely take new chlorogenic acid, chlorogenic acid, cryptochlorogenic acid, rutin, 3,5-O-dicaffeoylquinic acid, quercitrin and 4,5-O-dicaffeoylquinic acid control samples, and add methanol to prepare a mixed control solution; b, preparation of a test solution: crush Xiaochaihu medicinal materials, add a methanol solution for extraction, and filter to obtain the test solution; and c, determination: precisely take the control solution and the test solution, inject them into an ultra-high performance liquid chromatograph, and determine the content by using an external standard method. The method can simultaneously detect and analyze the content of seven chemical components in Xiaochaihu medicinal materials, has good linearity, repeatability, stability and recovery rate, and can effectively control the quality of Xiaochaihu medicinal materials.

The invention discloses preparation of a folium cortex eucommiae extract and application of the folium cortex eucommiae extract in improvement of age-related macular degeneration, and belongs to the technical field of traditional Chinese medicine extraction. The technical problem to be solved is how to prepare the folium cortex eucommiae extract which is higher in content of chlorogenic acid, cryptochlorogenic acid, neochlorogenic acid, aucubin and geniposidic acid and has a better anti-age-related macular degeneration effect. According to the technical scheme, the folium cortex eucommiae extract is characterized in that the content of chlorogenic acid is 50-100 mg / g, the content of cryptochlorogenic acid is 40-60 mg / g, the content of neochlorogenic acid is 35-55 mg / g, the content of aucubin is 56-79 mg / g, and the content of geniposidic acid is 65-105 mg / g. The folium cortex eucommiae extract obtained by the method can better retain effective components in the folium cortex eucommiae extract, and a better anti-age-related macular degeneration effect is embodied.

The present application relates to the technical field of cultivation, in particular to a preparation method of astringent horehound extract, a feed additive and application thereof. The present application takes astringent horehound as raw material, the plant can be planted in large area, and fresh grass yield per mu can reach about 10 tons, and dry grass can reach more than 1 ton. The natural source feed additive active component (i.e. astringent horehound extract) is obtained by ethanol extraction combined with column chromatography. The content of chlorogenic acid in the component can reach more than 12.9%, and the total content of neochlorogenic acid, chlorogenic acid and cryptochlorogenic acid can reach more than 35%. The active component as a feed additive shows good disease resistance in the feeding of Tibetan sheep, Ippy rabbits and small crayfish, and the yield of the animals is greatly improved, and the effect is better than that of the same dose of chlorogenic acid adding group. In addition, the astringent horehound extract prepared by the preparation method can significantly improve the meat quality and increase the yield.

The present application relates to the technical field of traditional Chinese medicine detection, and specifically provides a characteristic spectrum construction method of Herba Centipedae and its preparation. A specific elution procedure is obtained through a large number of experiments by using octadecylsilane bonded silica gel as a filler and a mobile phase including methanol and an acid-containing aqueous solution for gradient elution, 13 or more common characteristic peaks are obtained, and effective separation of characteristic peaks including neochlorogenic acid, caffeic acid, chlorogenic acid, cryptochlorogenic acid, 1,4-dicaffeoylquinic acid, 1,5-dicaffeoylquinic acid, big pothos glycoside and thymotin lactone ketone is achieved, the peak shape is good, the baseline is smooth, the determined components can be completely separated, the detection time is short, and the method provides a basis for quality detection and control of Herba Centipedae and its preparation, can fully reflect the integrity and characteristics of Herba Centipedae and its preparation, and is simple to operate and applicable to quality control of different preparation types of Herba Centipedae.

This invention belongs to the field of traditional Chinese medicine processing and natural product chemistry, specifically relating to a method for the targeted enrichment of chlorogenic acid components in honeysuckle. The method involves placing honeysuckle powder under constant temperature heating conditions for dry heat treatment. Different dry heat treatment conditions are set according to different target concentrators to achieve the targeted enrichment of various chlorogenic acid components, including neochlorogenic acid, chlorogenic acid, isochlorogenic acid B, and isochlorogenic acid C. This method achieves the targeted and efficient enrichment of effective components during the processing of honeysuckle tablets, and the process is simple, convenient to operate, and easy to scale up for industrial production.

The invention discloses a UHPLC-MS / MS (Ultra High Performance Liquid Chromatography-Mass Spectrometry / Mass Spectrometry)-based multi-component content determination method for a compound radix scutellariae throat-clearing capsule, which is characterized in that a multi-component simultaneous quantitative analysis method is established by adopting a UHPLC-MS / MS technology, and the quality of the compound radix scutellariae throat-clearing capsule is evaluated from the quantitative aspect. 21 components including p-hydroxybenzaldehyde, vanillin, caffeic acid, syringaldehyde, citric acid, tanshinol, chrysin, baicalein, genistein, quercetin, chlorogenic acid, neochlorogenic acid, rosmarinic acid, harpagide, puerarin, apigenin-7-O-beta-D-glucoside, baicalin, oleanolic acid, scutellarin, glycyrrhetinic acid and harpagoside are selected as quantitative indexes; and determining the content of the compound radix scutellariae throat-clearing capsule. According to the method, the contents of 21 components in the compound radix scutellariae throat-clearing capsule can be accurately measured at the same time within 14 min, and the method has good precision and stability.

A steviol glycoside solubility enhancer may include a monocaffeoylquinic (MCQ) component that includes at least one compound selected from the group consisting of chlorogenic acid, neochlorogenic acid, cryptochlorogenic acid, and salts thereof; and a dicaffeoylquinic (DCQ) component that includes at least one compound selected from the group consisting of 1,3-dicaffeoylquinic acid, 1,4-dicaffeoylquinic acid, 1,5-dicaffeoylquinic acid, 3,4-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid, 4,5-dicaffeoylquinic acid, and salts thereof. In one suitable implementation, the MCQ component and the DCQ component together comprise more than 50% (wt), preferably more than 60% (wt), more than 70% (wt), more than 80% (wt), more than 90% (wt), or more than 95% (wt) of the steviol glycoside solubility enhancer.

This invention discloses a method for determining the quality of *Hemiberlesia lingua* stems and leaves. The method includes fingerprint spectroscopy establishment and similarity analysis, content determination, cluster analysis, principal component analysis, and partial least squares discriminant analysis. The method of this invention involves UPLC fingerprint analysis and similarity analysis of 15 batches of *Hemiberlesia lingua* stems and leaves from different origins. A total of 28 common peaks were found, and 7 of these common peaks were identified using reference standards: peak 4 (neochlorogenic acid), peak 8 (chlorogenic acid), peak 9 (caffeic acid), peak 11 (cryptochlorogenic acid), peak 22 (isoquercetin), peak 24 (astragalin), and peak 28 (isochlorogenic acid C). This invention's method is highly sensitive, specific, stable, and feasible, providing a reference for the quality evaluation and control of *Hemiberlesia lingua* stems and leaves.

The present application relates to the technical field of traditional Chinese medicine analysis, and particularly relates to a mulberry fruit paste fingerprint spectrum establishment method and fingerprint spectrum. The mulberry fruit paste fingerprint spectrum establishment method comprises the following steps: taking mulberry fruit paste as raw material to prepare a test sample solution, taking chlorogenic acid, neochlorogenic acid, cryptochlorogenic acid, 5-hydroxymethyl furfural, rutin and isoquercitrin to prepare reference sample solutions; injecting the test sample solution and the reference sample solutions into an ultra-high performance liquid chromatograph for determination, recording chromatograms, collecting chromatograms of test sample solutions of multiple batches of mulberry fruit paste, importing a traditional Chinese medicine chromatographic fingerprint spectrum similarity evaluation system, generating a control spectrum, and taking peak area and retention time as parameters to calculate similarity. The method can simultaneously detect multiple components in mulberry fruit paste, and can be effectively used to evaluate product quality stability of mulberry fruit paste.

The present application relates to the technical field of honey detection, and particularly relates to a vitexin, a preparation method thereof and a method for identifying vitex honey. The vitexin has a structure shown in formula 1: The present application finds a new compound, vitexin, in vitex honey through research on the vitex honey. The present application also finds that when the to-be-tested honey contains vitexin (p-hydroxybenzoic acid 1-O-gentio-bioside-5-hydroxy-3,7-diene-[2,6] cyclopentanopyranyl-3-methyl ester), or contains vitexin and new chlorogenic acid, 4-hydroxybenzoic acid, chlorogenic acid, caffeic acid, p-coumaric acid, red flower phaseolus acid, trans, trans-epiabscisic acid, cis, trans-epiabscisic acid, 5-methoxy pinocembrin, pinocembrin, kaempferol, pinocembrin, benzyl caffeate, kojic acid, galangin at the same time, the to-be-tested honey can be determined as vitex honey. The present application also provides a new method for constructing a vitex honey authenticity and quality evaluation system.

The invention discloses a purification method of neochlorogenic acid. The purification method comprises the following steps: dissolving a stevia rebaudiana chlorogenic acid product, and adjusting the pH value to obtain a first to-be-treated solution; carrying out an isomerization reaction, and adjusting the pH of the system to obtain a second to-be-treated solution; adsorbing the solution in a macroporous adsorption resin system, independently analyzing the first resin column after jacking the acid liquor, dealcoholizing the effluent, drying to obtain a stevia polyphenol product, analyzing the rest resin columns in series, adjusting the pH value of the effluent, and drying to obtain a crude product; dissolving the crude product, adding a seed crystal for crystallization, filtering to obtain a first filter cake and a first filtrate, and removing a solvent from the first filtrate for dissolving the raw material; mixing the first filter cake and purified water, adjusting the pH value, heating to dissolve, adjusting the pH value again, slowly cooling to crystallize, filtering to obtain a second filter cake and a second filtrate, combining the second filtrate into a second desorption solution, and drying the second filter cake to obtain neochlorogenic acid. The high-purity neochlorogenic acid is obtained through isomerization, resin separation and fractional crystallization, efficiency is high, conditions are mild, side reactions are few, the resource utilization rate is high, and the method is suitable for large-scale production.

The invention provides a method for determining the content of multiple components in an Aidi preparation, and belongs to the technical field of medicines. The Aidi preparation is prepared from traditional Chinese medicinal materials including ginseng, cantharides, astragalus membranaceus and acanthopanax. The content determination method comprises the following steps: simultaneously determining cantharidin, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, ginsenoside Rc, ginsenoside Rd, isofraxidin, syringin, formononetin, astragaloside, chlorogenic acid, eleutheroside E, calycosin-7-glucoside, astragaloside II, neochlorogenic acid, cryptochlorogenic acid and eleutheroside E1 in the Aidi preparation by adopting liquid chromatography-mass spectrometry; the composition is prepared from effective components such as ginsenoside F3, ginsenoside Rg2, azelaic acid and the like. Compared with the prior art, the method has the advantages that the contents of the 20 effective components in the Aidi preparation are simultaneously measured under the same chromatographic condition through liquid chromatography-mass spectrometry, the method is convenient and rapid, the requirement for product quality detection is greatly improved, the method is more suitable for quality control of drugs in the future, and the drug quality and the clinical drug effect are guaranteed.

This invention provides a method for determining the content of *Zanthoxylum bungeanum* seeds using HPLC, comprising: A) dissolving and extracting the sample raw material using a solvent to obtain a test solution; B) determining the test solution using high-performance liquid chromatography (HPLC) to obtain an HPLC chromatogram of *Zanthoxylum bungeanum* seeds; the HPLC conditions are as follows: a C18 column; mobile phase A is acetonitrile solution, mobile phase B is 0.1% phosphoric acid aqueous solution, and gradient elution is performed; specifically, the gradient elution is as follows: 0–20 min, phase A: 10% → 11%, phase B: 90% → 89%; 20–25 min, phase A: 11% → 18%, phase B: 89% → 82%; 25–40 min, phase A: 18% → 40%, phase B: 82% → 60%. This invention employs high-performance liquid chromatography (HPLC) with acetonitrile-0.1% phosphoric acid solution as the mobile phase for gradient elution. Using chlorogenic acid, neochlorogenic acid, and cryptochlorogenic acid as reference standards, an HPLC method for determining the content of *Zanthoxylum bungeanum* seeds was established, providing more scientific technical means for controlling the medicinal quality of *Zanthoxylum bungeanum* seeds.

The invention relates to the technical field of traditional Chinese veterinary medicine analysis, in particular to a construction method and application of a liver and gall particle HPLC characteristic chromatogram, and the method comprises the following steps: 1) taking liver and gall particles to prepare a test solution; 2) respectively taking oriental wormwood and isatis root to prepare a reference medicinal material solution; 3) taking neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, guanosine, adenosine, uridine and (R, S)-epigoitrin reference substances to prepare a reference substance solution; (4) carrying out liquid chromatography detection on the obtained solution to respectively obtain chromatograms of the test solution, the reference substance solution and the reference medicinal material solution; and (5) introducing the chromatograms of the plurality of batches of test sample solutions into a traditional Chinese medicine chromatographic characteristic chromatogram similarity evaluation system to generate a standard characteristic chromatogram, comparing the chromatograms of the reference substance solutions and the reference medicinal material solutions to identify the attribution of the characteristic peaks in the chromatograms, thereby constructing the HPLC characteristic chromatogram of the liver and gall particles. Compared with the prior art, the method has the advantages that quality evaluation of authenticity, quality and quality of products is realized, source tracing can be carried out on traditional Chinese medicinal materials such as radix isatidis and herba artemisiae scopariae, meanwhile, the content of (R, S)-epigoitrin, adenosine, uridine and guanosine in radix isatidis and the content of neochlorogenic acid, chlorogenic acid and cryptochlorogenic acid in herba artemisiae scopariae are quantitatively analyzed, and finally, the quality evaluation of the products has a basis. Therefore, the quality control of the liver and gall particles is comprehensively realized.