Mass spectrometry method for identifying and / or verifying a sample

Abstract

The present invention relates to a method for identifying and / or validating a sample, the method comprising: converting at least one chemical compound into the sample; determining the level of the at least one compound after addition to the sample; and comparing the level of the at least one compound to a reference level and identifying the sample based on the comparison with the reference sample and the effect of the sample on the level of the at least one compound. The present invention further relates to the use of at least one compound in the method for identifying and / or validating a sample. Furthermore, the present invention relates to the use of a kit comprising at least one compound in the method of the present invention. In addition, the present invention relates to a composition comprising at least one compound. Furthermore, the present invention relates to a kit comprising the composition provided herein, for example, for calibrating the method provided herein, and / or an apparatus used in the method provided herein.

Description

Technical Field

[0001] The present invention relates to a method for identifying and / or Confirmation of a sample, the method comprising: Such contacting at least one chemical Target compound with the sample; Addition ; The aforesaid determining the level of at least one compound after addition to the sample; Of the aforesaid ; The aforesaid comparing the level of at least one compound with a reference level To do; and, the aforesaid by comparison with a reference sample and The aforesaid identifying the sample based on the effect of the sample on the level of at least one compound. The present invention The aforesaid relates to the use of at least one compound in a method for identifying and / or The aforesaid of a sample. Further, the present invention relates to the Furthermore use of a kit comprising at least one compound. In addition, the present invention relates to a Confirmation composition comprising at least one compound. Further, the present invention relates to a kit The aforesaid comprising a composition provided by, for example, Relevant for calibrating the method provided by, for example, The aforesaid . In the method of the present invention The aforesaid Further, the present invention relates to a kit The present disclosure comprising a composition provided by, for example, And / or the apparatus used in the method provided by the present disclosure for calibrating the method provided by, for example, The present disclosure To

Background Art

[0002] Identifying or Confirmation of a sample is tedious, expensive, and time-consuming. This is particularly true for samples having a complex composition, such as food products (e.g., wine), pharmaceutical compositions, or body fluids. Such samples can contain a large number of metabolites, small molecule chemicals, lipids, peptides, nucleic acids, and / or proteins. Conventional methods are labor-intensive and time-consuming:

[0003] For example, International Publication No. WO 2016 / 196181 (A1) relates to metabolomics and lipidomics and concerns the analysis of metabolites and lipids in simple Mixture and and complex mixtures. More particularly, International Publication No. WO 2016 / 196181 concerns a De -trap Data independent acquisition obtained (data independent acquisition) using 、 a mass spectrometry system having quadrupole and ion mobility separation capabilities. International Publication No. WO 2016 / 196181 discloses For the identification and quantification of metabolites and lipids connecting a quadrupole; an ion mobility spectrometer; and a mass spectrometer for Sa sample identification. Four If necessary, further including HPLC separation

[0004] In addition, International Publication No. WO 2016 / 181299 describes a top-down method for generating a plurality of precursor peptides or protein ions from a sample solution containing at least one peptide or protein. Using an ion source 、 Analysis For

[0005] Furthermore, Mann (Nature Reviews Molecular Cell Biology volume 17, page 678 (2016)) discloses strategies for the unbiased detection of all substances present in a sample. This type of analysis is often limited to a few specialized research institutes. To detect the complete spectrum of a substance for analysis, depletion , digestion, chromatographic separation and / or high-resolution mass spectrometry, etc., time And consuming Required sample preparation techniques To perform are required. For example, in order to identify possible false Among them, the true samples Is Many and Using the same quantify proteins or metabolites By doing so a quantitatively precise and highly reproducible data set To is Do required Is .

[0006] Make it difficult to identify composite samples Required Due to One of is an interference effect caused by the components / substances of the sample, for example, the matrix And Ion suppression and Increase effects.

[0007] In mass spectrometry, the detection interference effect is Often causes ion suppression or enhancement phenomena. Such phenomena , Ma in the tricks Of volatile components not only the presence of, Non-volatile compounds that can change the droplet formation (or volatilization) efficiency of analytes, generated in the gas phase but also the amount of analyte ions reaching the detector Minute induced by To T.M. Annesley, Clin.Chem. 49(2003)1041). Various phenomena potentially induced by matrix components include detectability, selectivity, repeatability accuracy, accuracy, response Increase ( (signal vs concentration) ( and Linearity of quantitation limit In terms of method Of has a dramatic No impact on performance. Furthermore, spectral Increase changes caused by ion suppression or Of phenomena make it difficult to There are also build a spectral library. In addition, the absence of fragment ions in the full-scan spectrum significantly reduces the library information. To construct

[0008] Ion suppression and Increase affect both the identification and determination of analytes. When it prevents the detection of existing analytes, it not only causes false-negative diagnoses, but also, for example, when the internal standard (I.S.) level is more suppressed than the analyte level, it can cause false-positive diagnoses (F. Gosetti et al. / J.Chromatogr.A 1217(2010)3929~3937). For example, in the analysis of clenbuterol in urine, the percentage of ion suppression ranges from 37% at an analyte concentration level of 93 g / L Is to Is 45 g / L -1 at a concentration of 、 -1 concentrationIn 6 Up to 9% Width is was found (T.M. Annesley, Clin. Chem. 49 (2003) 1041).

[0009] The choice of positive ion mode (PI) or negative ion mode (NI) can also affect the signal. When the analyte can be ionized in both PI and NI modes and there is no selectivity problem, the use of the NI mode is recommended. For example, when using SI (NI mode) for pesticides (herbicides, insecticides, and fungicides), with only a few exceptions, signal suppression is not significant compared to that observed in the PI mode (J.M. Marin, E. Gracia-Lor, J.V. Sancho, F.J. Lopez, F. Hernandez, J. Chromatogr. A 1216 (2009) 1410). At and selectivity Of problem Occurs is not an issue, a more selective Highly sensitive, more Ma matrix effect Low due to is observed, and the use of the NI mode is recommended. For example, In the environment and wastewater 37 Species of for pesticides (herbicides, insecticides, and fungicides) Determination by E when using SI (NI mode), with only a few exceptions, signal suppression is not significant compared to that observed in the PI mode (J.M. Marin, E. Gracia-Lor, J.V. Sancho, F.J. Lopez, F. Hernandez, J. Chromatogr. A 1216 (2009) 1410). Its (J.M. Marin, E. Gracia-Lor, J.V. Sancho, F.J. Lopez, F. Hernandez, J. Chromatogr. A 1216 (2009) 1410).

[0010] Furey (Talanta. 2013 Oct 15;115:104 - 22. doi:10.1016 / j.talanta.2013.03.048. Epub 2013 Apr 16) Also disclosed the matrix effect in mass spectrometry analysis Influence to analytical chemists A major concern as a And certain

[0011] Said any conventional mass spectrometry method One of is direct injection mass spectrometry (DIMS). In DIMS, a diluted sample, e.g., a urine sample, undergoes no prior Of chromatographic separation To doInstead, it is directly injected into an ESI-equipped mass spectrometer. DIMS exhibits significantly high ionization suppression (Dunn & Ellis, 2005; Dettmer, Aronov, & Hammock, 2007; Want et al., 2010; Dunn et al., 2011). Compared with a standard spiked in an aqueous saline solution, Internal standard spiked into rat urine Including, the o Despite removing salts from urine using on-line SPE extraction, 60% ion suppression Observation Is caused was made (Dettmer, Aronov, & Hammock, 2007). Such Obstacle due to Its use is multiple a number of samples for fingerprint or putative identification Of when investigating for Swift is limited to high-throughput screening purposes (Mikami, Aoki, & Kimura, 2012; Zhang et al., 2012a). (Lit: DOI 10.1002 / mas.21455 MASS SPECTROMETRIC BASED APPROACHES IN URINE METABOLOMICS AND BIOMARKER DISCOVERY)

[0012] A further strategy for identifying samples is disclosed by Khamis (Mass Spectrom Rev. 2017 Mar;36(2):115 - 134.doi:10.1002 / mas.21455.Epub 2015 Apr 16). This method involves a complex Target multi-step Of approach. Alternative Target approaches are provided by Causon et al. (2019) Analytica Chimica Acta 1052, pp.179 - 189. Liquid A fingerprint method for wine using Combined with mass spectrometry liquid chromatography has been proposed.

Summary of the Invention

Problems to be Solved by the Invention

[0013] In this way Identifying and / or Confirmation The method of doing so is time-consuming and requires a lot of labor. Therefore, the technical problem underlying the present invention is to provide means and methods for identifying and / or Food samples (e.g., Wa goods, especially To Identifying and / or Confirmation for Rapid and reliable samples).

Means for Solving the Problem

[0014] Such The technical problem is solved by providing Claims its features. In the form described Thereby.

[0015] Therefore, the present invention relates to a method for identifying and / or Confirmation samples, the method comprising: (i) Having variable ionization degrees adding at least one compound to the sample; And ; (ii) determining the level of the at least one compound in the mass spectrum of the at least one compound after adding the sample; Obtained to the sample To after addition In of the at least one compound; (iii) Step comparing the level of the at least one compound determined in (ii) with a reference level; Of the compound determined after addition of the reference sample of the compound ; And ; (iv) identifying the sample based on the comparison in step (iii) and the effect of the sample on the level of the at least one compound. 、 and 、 ; 、 ; The present invention relates to a method comprising the above steps.

Embodiments for Carrying Out the Invention

[0016] The present invention solves the Identified above technical problems set. The following The present disclosure and Described laterAs described in the examples, Surprisingly, to the sample Less at least one compound Added, the compound with respect to the level of The aforesaid Based on the effect of the sample The aforesaid the sample is identified Can to be Actual proven. As shown below The present disclosure As shown in the following, the method of the present invention enables easy, rapid In of Structure constituent substances Part or all of without the need to determine the ( For example Wa in Etc.) easy, rapid 、 and reliable identification of the sample. This Using the method 、 the sample Confirmation was performed may also be. For example, The present disclosure the method provided in enables easy, rapid and reliable determination of whether a sample (e.g., a wine sample) is original or fake. To Also, this The method There is the sample is Original determine whether it is diluted compared to the sample Also is possible. The principle underlying the present invention is to determine the level of at least one compound added to the sample. Therefore, the method of the present invention includes adding at least one compound to the sample. Of Described later In the examples, the following preferred compounds are added to the sample: sulfaguanidine, sodium naproxen, sulfadimethoxine, ciprofloxacin, tetracycline hydrochloride, verapamil hydrochloride, terfenazine, leucine enkephalin acetate hydrate and reserpine. The constituent substances of the sample Of the ionization degree of at least one compound added to the sample , That is , can affect the level on the mass spectrometer By means of the . For example, the constituent components or substances of the sample The aforesaid at least 1 one compound By doing so have an ion suppression or ion Increase effect.​​The aforesaid of at least one compound The ionization degree is at its level can be determined by a bell.

[0017] The prior art shows that ion suppression affects the detection ability, accuracy, and precision of mass spectrometry Relative to adversely. And disclosed . This In the prior art adverse effect is utilized to Sample is at least one compound Of level Given based on the effect Such to identify and / or Confirmation sample Innovative was the idea of the inventors. For example, In the sample substances The aforesaid for at least one compound By doing so ion suppression effect Or is an ion Increase with an effect There may be a possibility , or The aforesaid of at least one compound Ionization degree is substantially changed No no There may be a possibility. The present disclosure The method provided is Utilizes these effects on the level of the at least one compound, the aforesaid by determining at least one compound level 、 to identify and / or Confirmation sample 。 Therefore, the method of the present invention Of the at least one compound after addition to the sample In at least one compound Of determines the unique pattern of the level. Thus obtained at least one compound Of Based on the unique pattern of the level 、 the sample is identified. Such Way method Examples of is The following present disclosure and described later described in In the examples.

[0018] In the prior art, samples are identified based on the substances contained in the sample and their levels. In contrast, The present disclosure in the method provided Of at least one compound after addition to the sample The aforesaid at least one compoundOf Level 、 and The aforesaid at least one compound Of based on the effect of the sample on the level The aforesaid identify the sample. Accordingly, the present invention Within the range of , the sample Of The levels of the components need not be determined for this, and are irrelevant to the identification / Confirmation of the sample. In other words, the sample to be identified / Confirmation is The present disclosure the mass spectrometry used In the present disclosure, to add at least one compound in the matrix used Equivalent .

[0019] Accordingly In the following disclosure added to the sample and then Quality analyzed by quantitative analysis Of a predetermined compound Set of (based on at least one compound) Developed is Generally stated for the sake of Described completeness To . Such The analysis level of at least one compound is affected by the substances contained in the sample (matrix). The aforesaid Quantify the effect of the sample on at least one compound level and compare it with a reference sample, i.e., Standard a standard (e.g., Healthy sample, a calibration Sample, or a reference sample Etc ) to enable By doing so the identification of the sample Is is No possible.

[0020] The prior art Of approach attempts to reduce matrix / ion suppression and Target the effects caused by complex To sample compositions. Such prior art Increase approach of is underlying technology time-consuming for analysis (sample total preparation and chromatography of run RO execution of )of including time), the complexity of the analyzer 、 and cost negative has an impact exert . The matrix and ion of suppression or increase effect reduction or exclude typical not means as is to separate specific interfering components from the matrix of by chromatographic methods removal, extended of desalination and buffer / solvent 、 change of and introduce , others into the ion source of . etc. are listed .

[0021] The present invention reverses the conventional impact and uses these negative to reduce / is. That is, quantify the level of one or more libraries of at least one compound qualitative in the complex to sample composition 、 and correlate with the identification of the native of sample increase . In this case, the ion suppression or transformation unique pattern of the complex sample with respect to one or more libraries of at least one compound is used as the fingerprint of the sample itself. As described below make this , a library of at least one compound of , of is also referred to as a composition containing at least one compound cause . The method provided herein is characterized by, for example increase having minimal sample preparation and run time. Further This disclosure , to , In this disclosure there is a dose-response curve (compound vs. sample concentration This disclosure or so far sample vs. compound concentration) limited . The examples described to in many show different 、 or 、 sample-to-compound concentrations) There is also an advantage that accurate quantification of the effect becomes possible . This disclosure in the examples described are differentof the origin Wine can be clearly identified, for example, by using a library of compounds directly with direct injection mass spectrometry or another mass spectrometry method by in a run time of less than 5 minutes. is doing .

[0022] Furthermore, as described later the examples show that a water sample can be identified by the method provided This disclosure therein. See Example 2. In particular, the examples is doing. show that six different water in samples were identified by obtaining the mass spectrum of at least one compound / of . As shown below confirmation , pure water has a different effect on at least one compound This disclosure compared to ion-containing water. 、 Different ion-containing water of samples have been shown to have different effects done on at least one compound. Thus, Also, the method provided of therein can also identify / done, water samples. Thus, to the method provided This disclosure therein can be applied to samples analyzed by mass spectrometry of . confirmation Furthermore, the examples This disclosure show the identification of different alcoholic any possible size beverages as described

[0023] . For example, in the method provided drinkable therein can identify and / or able to gin and whiskey also samples. See Example 3. Example 3 uses additional compounds compared to Example 1 This disclosure . by of confirmation can be done. Actually . is doing .

[0024] Furthermore, Example in provides a method This disclosure for using in, a known compound to to identify a sample by doing . Refer to Example 4 can be which is described 。 therein. That is , This disclosure the method provided in, by using one compound in 、 can be used to identify and / or confirmation characterize a sample. The addition of additional compounds can further improve the confirmation accuracy robustness and 、 precision resolution of the identification. Refer to Example 3 。 . Therefore, the examples provided This disclosure in This disclosure are indicates that the subject can be realized .

[0025] In addition, Example in shows that it is also possible to identify a sample the above when the sample already contains at least one compound 、 or the above at least one compound's a part of which constituent. is doing For example, refer to Example 5 also below. In other words, it is possible to identify and is doing. characterize a sample when at least one compound is present in the sample. For example, confirmation atrazine and quinine even were present in the sample to be identified. but Refer to Example 5 。 . The further addition of atrazine and / or quinine 、 increased the levels of these compounds and made it possible to determine the specific effect of the sample on at least one compound level. Therefore, it is possible to identify and confirmation characterize a sample the above even when the sample already contains at least one compound 、 or the above at least one compound's a part of which constituent.

[0026] The method of the present invention and This disclosure the use of the kits and compositions provided thereby offer at least the following advantages over the methods of the prior art. At least one compound is enables the identification of samples, This disclosure so the method provided thereby is faster than the methods of the prior art. Thus, the analysis time can be reduced from about 30 minutes to less than about 3 minutes. For example, neither isotope-labeled standards nor high-resolution mass spectrometry (MS) analyzers are and required. is made In addition, The method provided in this disclosure is a sample containing at least one compound can be directly injected into the ionization chamber, and the reaction of at least one compound can be directly measured, many so sample preparation, which requires much effort, is not needed. For example, This disclosure the method provided thereby does not require a liquid chromatography step. Thus, sample preparation can be significantly reduced or omitted. can be In addition, This disclosure the method provided thereby allows the use of various buffers and solvents. Thus, This disclosure the method provided thereby has few restrictions with respect to the of sample of buffer and solvent to composition. This disclosure The method provided thereby allows for further development applications. For example, To select the compound that most interacts with the substance of the sample at an appropriate concentration, at least one compound added to a sample can be readily obtained by screening a transformation compound library. Furthermore, This disclosure the method provided thereby is applicable to all types of samples and sample mixtures. Thus, This disclosure the method, use, kits and compositions provided thereby solve the underlying technical problems and provide rapid, cost-effective and reliable identification and / or confirmation analysis of samples.

[0027] The present invention relates to the following and the abovedisclosure of relates to the embodiments and items provided in. Hereinafter, the embodiments of the present invention will be described in more detail. These descriptions are This disclosure relates to the methods, uses, compositions and kits provided in.

[0028] This disclosure When used in, "adding at least one compound to the sample" means adding at least one compound to the sample for Alternatively, this term means adding the sample to at least one compound for add this can also mean. two or more When adding a compound to a sample, these before adding the compound to the sample these pre-mix the compound may be done , for example, such mixture is one is contained in the composition may be In such a case, a composition containing at least one compound may be added to the sample also Alternatively, This disclosure in the method provided in these compounds add separately to the sample also is okay.

[0029] This disclosure When used in, "at least one compound" used or included in an embodiment of the present invention is for example chemical qualitative or biochemical qualitative represents a compound 。 Preferably, "at least one compound" may be at least one chemical qualitative compound. In certain embodiments, such the compound has variable ionization degree may be ( That is, ion suppression effect may be sensitive to ), the ion of which is that of the MS detection system in use mass ratio charge ratio in a may be . In particular, the aboveAt least one compound is chemical qualitative or biochemical qualitative compound, particularly, a low molecular weight of chemical qualitative or biochemical qualitative compound, and also is good. For example, at least one compound is a peptide, protein, oligonucleotide, DNA strand or This disclosure a preferred compound provided by also is good. This disclosure When used in, "at least one compound" or "at least one chemical qualitative compound" is This disclosure used interchangeably. In certain embodiments, at least one compound quality has According to the quality, there may be a variable ionization degree in a mass spectrometer. As a result, the sample (or the substance contained in the sample) may affect the level of the at least one compound. Polymer chains such as peptides, proteins, oligonucleotides or DNA about , its charged amino acids and nucleotides increase or decrease the amounts such as by its sensitivity to ion suppression vary can be achieved. Proteins may be analyzed by top-down (whole protein of ionization) or bottom-up (peptide analysis after digestion) approaches. Furthermore, Not only peptides but also proteins are is known to be sensitive to subject to ion suppression. In other words, the sample of has doing, potentially the possibility of affecting at least one compound increase levels. the above disclosure and the examples described later As described above and below this the underlying invention concept is to identify and / or confirmation characterize a sample based on the effect of the sample on at least one compound level, and determine the effect of the sample on at least one compound with a mass spectrometer. In other words, after addition to the sample less at least one compound of level by determining identifies and / or confirmation characterizescan be done is possible not to be

[0030] size to determine whether the sample (or the substance contained in the sample) is at least at one compound level doing to affect or not determine to determine means can be recognized by those skilled in the art recognition possible For example, the level of at least one compound of can be determined by adding at least one compound to the sample without doing In other words, the level of at least one compound is determined in the absence of the sample. In a further step, the sample can be added to at least one compound. In other words, the level of at least one compound is determined in the presence of the sample. The comparison of the level of at least one compound in the presence of the sample and in the absence of the sample reveals whether the sample is at least at one compound level and to the presence or absence of the sample doing to affect or not. Therefore size the sample is at least at one compound level doing to affect so that The at least one compound has a mass spectrometer variable ionization degree by is provided

[0031] The variable ionization degree is in positive mode may be determined by or in negative mode to determine also is fine

[0032] The terms "ionization" or "ionization degree" or "ionization efficiency" or "ionization efficiency" can be used interchangeably This disclosure in this context. These terms represent the generation of gas-phase ions suitable for detection and determination in a mass spectrometer or a mass filter of a mass spectrometer. Therefore, the content of a specific compound decomposition can be determined by determining the level of a specific compound (in particular, the ions of a specific compound) in a mass spectrometer. The terms " ionization degree " or " variable ionization degree " mean that at least one of at least one compound variable ionization efficiency is (caused by the substance contained in the sample) subject to the matrix effectis sensitive to In other words, the term " variable ionization degree " or " variable ionization efficiency " refers to a sample (or a substance contained in a sample) is (1) less At least one compound done Has ion suppression effect can be , (2) less At least one compound done ion increase Effective can be or 、 (3) less At least one compound ionization degree Substantially change obtain Therefore, at least one compound variable ionization degree is a compound that is present in the sample in both the presence and absence of the sample. variable ionization efficiency Thus, in certain embodiments, at least one compound has a function in the presence and absence of a sample. variable ionization degree Thus, at least one compound variable ionization degree has an effect on the level of at least one compound in the sample. size At least one compound in the presence and absence of the sample of By determining levels and comparing these levels with each other 、the effect on the level of at least one compound As described above, the sample of The effect on levels is based on whether the sample contains at least one compound. ionization degree The term "at least one compound ionization degree "It does not substantially change described with Or its grammatical equivalent utilize The shape indicates that the sample reacts with at least one compound. is Measurable ion suppression effect increase It means that the sample has no measurable effect on a particular compound. Thus, the sample has no measurable effect on a particular compound. Thus, at least one compound level was not altered by the sample.

[0033] The term "ion suppression effect" or "ion suppression" refers to the phenomenon where the ionization efficiency of at least one compound is 、 reduced by the presence of substances contained in the sample. For example, in the following cases is , ion suppression or ion of the increase possibility is greater than. of the target analyte / substance in the sample While , in a sample containing a complex matrix for when present in only trace amounts, when performing only minimal sample clean-up, when an acid or alkali buffer or ion-pairing agent is present in the LC effluent, when using a short non-resolving chromatography run, and / or 、 when no chromatographic separation is performed.

[0034] Ion suppression and increase occur early in the ionization process in the ion source and depend on the compounds used in the method of the present invention. This can be due to polar and non-retained matrix components (e.g., substances contained in the sample) or overloading of the LC column. i On suppression and increase the phenomenon of has been as, various mechanisms proposed come to be: its example and as follows: (a) competition between the sample and the matrix components of the analyte ions co-eluting in the spray solution for access to the droplet surface for gas-phase release, (b) interference of the matrix that competes for the available charge, (c) A matrix that binds to the analyte or co-precipitates the analyte, (d) Analyte ions that can be neutralized by gas-phase of acid / base reactions, (e) Mobile phase of additives, and (f) Instrument design are mentioned.

[0035] The term "ion increase "Effect" or "ion" increase " is a phenomenon in which the ionization efficiency of at least one compound 、 is increased by the presence of substances contained in the sample. The ion suppression effect and the ion increase effect can be determined by the level of a specific compound. In certain aspects of the present invention, the at least one compound is specific to the sample substances or the ion suppression and / or ion increase effect is sensitive to of the selected substances. In other words, the level of at least one one compound is preferably altered by the substances contained in the sample. In particular, As at least one compound having a variable ionization degree or ionization efficiency added by the method of the present invention, an appropriate compound, for example, a library of compounds, can be provided to provide a specific level suitable for the identification and / or confirmation of the sample in the presence of the sample.

[0036] In certain aspects, the at least one compound is 1 to 100 species of compounds, preferably 1 to 80 species of compounds, more preferably 1 to 60 species of compounds, even more preferably 1 to 50 species of compounds, even more preferably 1 to 40 species of compounds, even more preferably 1 to 30 species of compounds, even more preferably 1 to 20 species of compounds, even more preferably 1 to 15 species of compounds, even more preferably 1 to 12 species of compounds, or even more preferably at least 9 species of compounds. The term "at least" means adding more compounds than a specific number to the sample or adding a specific number of types of compounds. types of compounds.

[0037] In a preferred aspect, the at least one compound represents more than 1 species compound. In certain aspects, the at least one compound is specific to a par certain substance / substance group in the sample as for specific ion suppression or ion increasing effect It has. At least one compound added to the sample can be identified in the mass spectrum. than At least one compound can have different chemical or physical properties (e.g., log P, pKa, and molecular weight) and is capable of obtaining orthogonal information from the sample that can be compared with information obtained from various separation mechanisms by conventional chromatography-based methods. In certain embodiments, the present disclosure The compound added by the method provided in can have a high molecular weight (e.g., peptide or protein) or a low molecular weight (e.g., small molecule) determined by a mass spectrometer, different partition coefficients (logP), and / or different pKa. When used in these embodiments, the terms "high" and "low" mean that different masses can be determined by a mass spectrometer, i.e., the levels of more than one compound can be resolved by a mass spectrometer. At least one compound can have a sample concentration that is at least 5 times the limit of quantification after addition to the sample. In particular, at least one compound suppresses and / or increase has a sample concentration after addition to the sample such that the level of at least one compound is reduced by about 20% to about 80% compared to the level of the same compound determined in the absence of the sample.

[0038] In certain embodiments, at least one compound is not contained in the sample prior to addition to the sample. the present disclosure When used in, the term "not contained in the sample" means that the level of at least one compound used for using the method prior to addition of at least one compound to the sample cannot be determined by a mass spectrometer. Thus, the level of at least one compound is below the detection limit in such embodiments. Preferably, at least one compound is not present in the sample.

[0039] As shown below the present disclosure if at least one compound is already contained in the sample prior to adding at least one compound to the sample, the sample is identified and / or confirmation It can be done. In such an embodiment, at least one compound is added to the sample at a concentration such that it has an effect on a level that increases and can be determined by mass spectrometry for the sample. Alternatively, at least one compound may be added to two or more aliquots of the sample at, for example, two different concentrations, and the level difference obtained from the sample is compared with the level difference of the signals in the calibration series.

[0040] Added by the method of the present invention, the present disclosure at least one compound provided (or the present disclosure the kit provided and used) is preferably selected from the group consisting of sulfaguanidine, sodium naproxen, sulfadimethoxine, ciprofloxacin, tetracycline hydrochloride, verapamil hydrochloride, terfenadine, leucine enkephalin acetate hydrate, and reserpine.

[0041] Added by the method of the present invention, the present disclosure at least one compound provided (or the present disclosure the kit provided and used) is preferably selected from the group consisting of leucine enkephalin acetate hydrate, terfenadine, verapamil hydrochloride, tetracycline hydrochloride, diltiazem hydrochloride, lincomycin hydrochloride, buspirone hydrochloride, sarafloxacin hydrochloride hydrate, haloperidol, trazodone hydrochloride, ciprofloxacin, quinine, ranitidine hydrochloride, triclocarban, sulfadimethoxine, trimethoprim, amitriptyline hydrochloride, atenolol, propranolol hydrochloride, sulfathiazole, sulfamethoxazole, cimetidine, salbutamol, melatonin, sodium naproxen, atrazine, sulfaguanidine, metformin hydrochloride, and reserpine. the present disclosure The preferred compounds provided at may be used as any of their other salts.

[0042] In particular, at least one compound is sulfaguanidine, sodium naproxen, sulfadimethoxine, ciprofloxacin, tetracycline hydrochloride, verapamil hydrochloride, terfenadine, leucine enkephalin acetate hydrate, and reserpine. Such compounds are hereinafter preferred compounds used in the examples. The CAS numbers are disclosed below the present disclosure . As long as the compound can be determined at the compound level by a mass spectrometer variable ionization efficiency , any other compound may be used in the manner provided the present disclosure . Thus, in certain embodiments, at least one compound level is adjusted to a particular sample so as to enable the identification and / or confirmation of the sample. Suitable compounds may be identified by screening a compound library by mass spectrometry. The screening method may select compounds that exhibit than changed ionization degree in the sample. In other words, at least one compound may preferably be at least one compound having variable ionization degree in a mass spectrometer such that the sample has an effect on the at least one compound level. For example, the screening method may preferably select compounds that are commercially available, stable, pure, soluble in the sample, and / or contained in the sample, where the sample has a matrix effect on the compound.

[0043] Further, the present disclosure in the method provided, a quencher may additionally be added to the sample, particularly at one or more concentrations suitable for calibrating and quality controlling the method. Such a quencher may be included in a composition or kit provided the present disclosure that contains at least one compound. The quencher may represent a standard. Alternatively, the quencher may enhance the quenching effect.

[0044] In a further embodiment of the present invention, the present disclosure Provide a sample that can be used as a standard for calibrating the methods provided herein. Such calibration samples can be, for example, known compounds, in particular, known variable ionization degree and / or the ion suppression effect on at least one compound, the ion increase effects, etc., of known ionization degree effects, or compounds that are known to have substantially no change in at least one compound level. Thus, using such a sample, the present disclosure the methods provided herein and the present disclosure the apparatus used in the methods provided herein may be calibrated.

[0045] the present disclosure When used in, "mass spectrum" represents a plot of the relative abundance of ions that generate a beam or other collection as a function of its m / z value. the present disclosure In the method provided herein, at least obtain the "mass spectrum" of at least one compound. The determination of the level and acquisition of the mass spectrum may be obtained in positive mode or negative mode. the present disclosure In the method provided herein, the mass spectrum of at least one compound can be obtained, and if necessary, additional mass spectra can also be obtained for the substances of the sample. In a particular aspect of the present invention, more than one compound may be added to the sample, so the mass spectrum of each of at least one compound may be obtained. In a particular aspect, only the level of at least one compound is determined by the method of the present invention. In particular, as an aspect, the level of the substances contained in the sample is not determined. Thus, in a particular aspect, only the level of at least one compound is determined, and the level of the substances contained before the addition of at least one compound is not determined. As described below and above the present disclosure As described in, identify and / or confirmation the sample based on the effect of the sample on at least one compound level. Thus, at least one compound level is for the identification and / or confirmation It may be sufficient to do so. In some aspects of the present invention, the level of the substance contained in the sample may be determined. Such a level may provide an internal standard to the mass spectrum, for example.

[0046] In certain aspects, a mass spectrum is obtained by mass spectrometry using an ionization source and an interface. Preferably, a mass spectrum is obtained by direct injection mass spectrometry. A mass spectrum is obtained by a mass spectrometer connected to an ion source, particularly an electrospray ionization source.

[0047] A mass spectrum may be obtained by various MS techniques as described above. Sample preparation methods include techniques related to dissolution, fractionation, digestion of the sample into peptides, depletion, concentration, dialysis, desalting, alkylation and / or peptide reduction. However, the present disclosure One advantage of the method provided in is that these steps are optional because such a method does not require pretreatment as shown below. Selective detection of analyte ions may be performed using tandem mass spectrometry (MS / MS). Tandem mass spectrometry features a mass selection step ( the present disclosure When used, the term "mass selection" refers to the isolation of ions having a specified m / z or narrow m / z range), followed by fragmentation of the selected ions and mass analysis of the resulting product (fragment) ions.

[0048] In certain aspects of the present invention, the sample may be directly injected into the ion source of the mass spectrometer. the present disclosure The accompanying examples of show methods that did not use a separation step. In such aspects, the sample used in step (i) did not perform a separation step. This means, for example, that the sample was not concentrated for a particular substance before adding and determining at least one compound level. For example, the present disclosure the level in step (ii) of the method provided in is determined without using chromatographic separation, such as liquid chromatography.

[0049] In certain embodiments, a mass spectrum may be obtained by matrix-assisted laser desorption ionization (MALDI). As long as the levels of the sample, reference sample, and at least one compound are determined by the same ionization and mass spectrometry method, e.g., MALDI-MS, the effects of the ionization technique and / or mass spectrometry method are the same for the sample level and the reference level. the present disclosure The sample can be identified and / or confirmation done by the method provided in. Thus, the effect of the sample on at least one compound level can also be determined by MALDI-MS. The matrix-assisted laser desorption ionization (MALDI) method can be used the present disclosure by the method provided in.

[0050] Furthermore, as described in Journal of Chromatography A, 1493 (2017) 57 - 63 and Food Chemistry 244 (2018) 128 - 135, online sample preparation for removing specific substance classifications such as salts that may have an impact on the MS system, and post-column addition of specific agents to improve the detection and signal of the target compound are known. Thus, for example, initial sample preparation in the form of a chromatography step is performed to remove potential interfering substance classifications and add at least one compound, similar to the post-column derivatization method. Thus, a mass spectrum may be obtained by applying chromatographic separation and subsequent addition of at least one compound.

[0051] the present disclosure When used in, "sample" refers to a sample containing a substance to which at least one compound is added for identification and / or confirmation adding for the purpose of doing so. the present disclosure The sample used may have a complex composition containing substances such as a number of metabolites, chemicals, lipids, peptides, nucleic acids, and / or proteins. The term "sample" may represent a matrix. Since the provided method is a mass spectrometry method, the sample is a sample suitable for analysis by a mass spectrometer. Thus, the present disclosure the method provided in confirmation is a mass spectrometry method for identifying and / or the present disclosure processing a sample. The sample may be a liquid sample. However, the sample may also be a solid sample suitable for analysis by mass spectrometry or made suitable for analysis by mass spectrometry. Those skilled in the art recognize how to prepare a sample to be suitable for analysis by mass spectrometry. For example, to prepare a (liquid) sample that can be analyzed by a mass spectrometer, the substances contained in the (solid) sample can be extracted or dissolved. For example, as described in Journal of Chromatography A 1617 (2020) 460830, a solid sample can be analyzed by LC-MS after a sample preparation method known to those skilled in the art. For example, a solid sample can be cryogenically pulverized with liquid nitrogen in a cutting mill to avoid heat loss of the target substance, and then an extraction step can be performed, such as solvent extraction or any other extraction / sample preparation method suitable for the target substance in the solid sample. At least one compound can be added to the extract, the present disclosure and the sample can be processed by the method described in the present disclosure . Thus, as long as the sample can be analyzed by mass spectrometry or as long as the mass spectrum of at least one or more compounds added to the sample can be obtained, the present disclosure the sample of the method provided in

[0052] is not particularly limited. the present disclosure As shown in the present disclosure The method provided herein can also identify a water sample. The water sample contains a substance having an effect on at least one compound level. A possible result is whether the sample contains a substance having an effect on at least one compound level. This will apply to the fractions of the sample. Thus, as long as the sample (or the substance contained in the sample) has an effect on at least one compound level as described above, any sample is the present disclosure suitable for use in the method provided herein. One skilled in the art will recognize how to determine whether a sample has an effect on at least one compound level as described above. For example, at least one compound level can be determined without adding at least one compound to the sample. In a further step, the sample can be added to at least one compound. Thus, at least one compound level can be determined in the presence and absence of the sample, and these levels can be compared to each other. The comparison of the levels reveals whether the sample has an effect on at least one compound. Thus, any sample having an effect on at least one compound the present disclosure can be used in the method provided herein. In other words, any sample can be used in the method provided herein that provides a matrix effect on at least one compound the present disclosure can be used in the method provided herein.

[0053] In particular, the sample is selected from the group consisting of wine, alcoholic beverages, foodstuffs, processed foods, tea, coffee, herbal extracts, natural products, natural product extracts, beer, fruit juices (e.g., orange and apple), pharmaceutical compositions, pharmaceutical formulations, body fluids, tissue extracts, blood, plasma, serum, and urine. In particular, this is a foodstuff. In a preferred embodiment, the sample is wine. In a more preferred embodiment, having an effect on at least one compound added to the wine sample the present disclosure The sample that is the matrix referred to herein is wine. The sample may comprise a single wine or may be a mixture of wines. For example, the sample may be a pool sample suitable for use in quality control.

[0054] the present disclosure When used, the term "reference sample" refers to a known sample, e.g., a reference sample, e.g., a standard sample. In certain embodiments, the sample and the reference sample are from the same species. For example, if the sample is a wine sample, the reference sample is also a wine sample, particularly a wine sample of known origin, vintage, etc. In a preferred embodiment, the reference sample is a reference standard sample. In the most preferred embodiment and as shown in the appended examples, the reference sample is a wine sample of a particular grape or a particular vintage of a particular origin. the present disclosure As described in, the sample may be a pharmaceutical composition or a formulation of a pharmaceutical product. In such a case, the present disclosure the method provided in may be used to determine whether the pharmaceutical composition or formulation of the pharmaceutical product to be identified is authentic or whether the sample is a counterfeit or diluted sample. The sample may be a body fluid, blood, plasma, serum, and urine. In such a case, the present disclosure the method provided in may be used in diagnosis, prognosis, risk assessment, risk stratification, monitoring, treatment guidance and / or treatment control of a subject. For example, for example, if a subject is suffering from a particular condition (e.g., a disease or disorder), when a particular condition occurs, a substance may be contained in the sample. Such a substance may affect the ionization degree of at least one compound after addition to the sample. Thus, whether or not a substance is contained in the sample, such a substance may affect the ionization degree It can have an impact. Therefore, the method of the present invention may be used in diagnosis, prognosis, risk assessment, risk stratification, monitoring, treatment guidance, and / or treatment control of a subject. In certain embodiments, a reference sample or reference level is determined by the same mass spectrometry technique as at least one compound level. Thus, the same mass spectrometry technique, e.g., ESI or MALDI and / or direct injection into the ion source of a mass spectrometer, is used for the reference level / sample and at least one compound level.

[0055] the present disclosure In the method provided by, the level of at least one compound after addition to the sample is determined. the present disclosure When used in, the term "determining the level of at least one compound after addition to the sample" or grammatical variations thereof may mean determining the level of ions of at least one compound or a fragment thereof. the present disclosure When used in the present disclosure The at least one compound level determined or compared in may represent the level of ions of at least one compound or a fragment thereof. This applies to the reference level. Therefore, the method is: (i) A step of adding at least one compound to a sample, in particular, the at least one compound has variable ionization degree a step with; (ii) Obtaining a mass spectrum of at least one compound and determining the level of ions of at least one compound or a fragment thereof after addition of the sample; (iii) A step of comparing the level of ions of at least one compound or a fragment thereof determined in (ii) with a reference level, where the reference level is the level of ions of the same compound or a fragment thereof determined after addition to a reference sample; (iv) A step of identifying the sample based on the comparison in step (iii) and the effect of the sample on the level of at least one compound or a fragment thereof, and may represent a method comprising.

[0056] the present disclosure When used herein, the term "level" refers to the peak intensity or integral of a peak determined by a mass spectrometer. In particular, the term "level" refers to the relative abundance of ions generated by a mass spectrometer. the present disclosure When used herein, the term "level" refers to one or more levels of at least one compound. One of ordinary skill in the art will appreciate that the level determined by a mass spectrometer for a particular compound may include the level of ions of the particular compound or a fragment thereof. Thus, the level of a (at least one) compound may represent the abundance of ions generated from the (at least one) compound in a mass spectrometer. Thus, one level may be obtained for a particular compound, or more than one level may be obtained for a particular compound in an embodiment of the invention. In certain embodiments, more than one compound is added to the sample. In such embodiments, the level of the compound after addition to the sample is determined in step (ii), and in step (iii) the level of the compound is compared to a corresponding reference level. In particular, the level of each compound after addition to the sample is determined in step (ii), and in step (iii) the level of each compound is compared to each corresponding reference level. In certain alternative embodiments, the term "level" may also represent a mass spectrum. In such embodiments, the invention comprises: (i) adding at least one compound to a sample, wherein in particular the at least one compound variable ionization degree has, the step of; (ii) determining the mass spectrum of the at least one compound after addition to the sample; (iii) comparing the mass spectrum of the at least one compound determined in (ii) to a reference mass spectrum, wherein the reference mass spectrum is the mass spectrum of the same compound determined after addition to a reference sample; (iv) identifying the sample based on the comparison in step (iii) and the effect of the sample on the mass spectrum of the at least one compound, and may represent a method comprising.

[0057] the present disclosure In the method provided, the level of at least one compound determined in (ii) is compared with a reference level. One of ordinary skill in the art will understand that this term can mean comparing the level of ions of at least one compound determined in (ii) with a reference level. In certain embodiments, the level of a particular compound is compared with a reference level or a corresponding reference level. Thus, the level of at least one compound may be compared with a reference level or a corresponding reference level. the present disclosure When used, the term "reference level" is the level of the same compound determined after addition to a reference sample. Thus, the term "reference level" means a level indicative of a known and particular sample. The reference level may be determined by the method of the present invention or may be known. Thus, the reference level can be included in a kit and the determined level of a sample can be compared to such reference level.

[0058] Purely as an example of the method of the present invention and without any limiting features, Compounds X and Y are added to Sample A in step (i) of the method of the present invention. Mass spectra of each of Compounds X and Y are obtained and the levels of Compounds X and Y after addition to the sample (e.g., several levels of each X and Y) are determined in step (ii) of the method of the present invention. In step (iii) of the method of the present invention, the level of Compound X determined in step (ii) is compared with the reference level of Compound X determined after addition to a reference sample. Additionally, the level of Compound Y determined in step (ii) is compared with the reference level of Compound Y determined after addition to a reference sample. Thus, the corresponding levels are compared. Thus, "the reference level is the level of the same compound" means comparing the determined level of at least one compound with the corresponding reference level, e.g., comparing the level of Compound X with the reference level of Compound X.

[0059] In step (iv) of the method of the present invention, the sample is identified based on the comparison in step (iii) and the effect of the sample on at least one compound level. As described above and below the present disclosure as described in However, (1) less whether at least one compound ionization degree can be substantially unchanged, (2) whether it can have no ion suppression effect on at least one compound, or (3) the sample (or a substance contained in the sample) can have a matrix effect on at least one compound such that it can have an ion increase effect. Such an effect affects at least one compound level such that at least one compound level either increases or decreases or is substantially unchanged. The sample is identified by comparing at least one compound level to a reference level. For example and in certain embodiments, a similar or same level of at least one compound compared to the reference level indicates that the sample matches the reference level; or a different level of at least one compound when compared to the reference level indicates that the sample does not match the reference level. the present disclosure When used, "different" means that the level of at least one compound can increase or decrease compared to the reference level. For example, the increase may be due to an ion increase effect or the decrease may be due to an ion suppression effect. As shown, when determining more than one level for a particular compound of at least one compound, the level of the particular compound can be compared to the corresponding reference level. Thus, the method of the present invention can also relate to a method wherein a similar or same level of at least one compound compared to the reference level indicates that the sample matches the reference level; or a difference in level of at least one compound when compared to the reference level indicates that the sample does not match the reference level. As shown above, more than one level can be determined for each compound and more than one compound can be the present disclosure It may also be used in the methods provided. In such embodiments, a similar level or the same level of the compound compared to the reference level indicates that the sample matches the reference level; or a level difference of at least one compound when compared to the reference level indicates that the sample does not match the reference level. One of ordinary skill in the art recognizes which levels are appropriate for comparison with each other and which levels to use to identify a sample since the corresponding reference levels of the reference samples are known. Thus, the most appropriate level (e.g., the most abundant level) can be determined and compared to the corresponding reference level, and based on this, the sample can be identified. In other words, the levels of the compounds may be weighted in the identification of the sample.

[0060] As shown above and below the present disclosure As demonstrated in, the ionization efficiency of at least one compound determines how many ions are generated and whether it can be determined by a mass spectrometer. The sample (or the substance contained in the sample) determines the ionization efficiency of at least one compound. Thus, the method of the present invention is a method that determines the ion suppression efficiency or the increase efficiency of at least one compound, compares the ion suppression efficiency or the increase effect with the ion suppression efficiency or ion increase effect of a reference sample, and based on this, identifies the sample.

[0061] In certain embodiments, the method of the present invention is a method that, in step (iv), identifies a sample based on the ion suppression or ion increase of at least one compound in a specific pattern.

[0062] In certain embodiments of the method of the present invention, one or more dose-response curves are determined. Dilution of the sample and / or at least one compound enables the creation of multiple dose-response curves. the present disclosure When used, the dose-response curve represents a function of the level to be determined. The dose-response curve can be obtained based on the levels determined by the method of the present invention. In certain embodiments, the dose-response curve is based on the concentration of at least one compound; or the dose-response curve is based on the level of at least one compound after addition of the sample to different concentrations thereof. For such embodiments, in step (i) of the method of the present invention, different concentrations of the at least one compound are added to the sample, and in step (ii), the levels of the different concentrations of the at least one compound are determined after addition to the sample. Alternatively, in step (i) of the method of the present invention, at least one compound is added to different concentrations of a sample (e.g., different dilutions of the sample), and in step (ii), the level of at least one compound is determined after addition to the sample. Such levels can be used to create a dose-response as described above. As described above, at least one compound can be added to the sample at different concentrations. Suitably, the at least one compound has a sample concentration that is at least 5 times the limit of quantification after addition to the sample. Thus, starting from such a concentration, at least one compound can be added at higher concentrations. In certain embodiments, the at least one compound suppresses and / or increase has a sample concentration after addition to the sample, as appropriate.

[0063] A dose-response curve can be obtained based on the level of at least one compound after addition to samples having different concentrations, and the dose-response curve is compared to the dose-response curve of a reference sample, the dose-response curve of the reference sample being based on the reference level of at least one compound after addition to reference samples having different concentrations.

[0064] the present disclosure In the method provided, at least one compound provides an optimal ion suppression effect and ions for the sample increase It may be adjusted for the effect. Preferably, the concentration of at least one compound is selected in such a way that the level of at least one compound is at least 5 times the limit of quantification, and the sample causes an effect of signal enhancement of about 20% or less or signal reduction of about 80% or less.

[0065] In certain embodiments, the identification of the sample is the identification of a dilution of the sample. Such a method is particularly suitable for revealing whether the sample is the same as the original reference sample or whether the sample is diluted compared to the reference sample. Dilution of the sample may be important if the sample is, for example, an expensive wine or if the sample is a pharmaceutical composition, and the sample concentration may determine medical effectiveness. In such embodiments, the dilution of the sample may be identified based on a comparison of the dose-response curve of the sample to the dose-response curve of the reference sample. In certain embodiments, the dose-response curve may be based on the levels of different concentrations of at least one compound determined in the sample and determined in the reference sample. Alternatively, the dose-response curve may be based on the levels of at least one compound determined at different concentrations of the sample and determined at different concentrations of the reference sample.

[0066] The present invention further relates to a method for identifying a dilution of a sample by comparing the dose-response curve of a reference sample to a constant or various concentrations of at least one compound.

[0067] As shown in the attached examples, the present disclosure the method provided in may preferably be carried out in the order of steps (i), (ii), (iii) and (iv) described above the present disclosure .

[0068] The present invention further relates to the use of the method and the present disclosure the use of at least one compound in the method provided in. The present invention further relates to the kit and the present disclosure the use of the kit in the method provided in. The above and below the present disclosure All the explanations, definitions, and information provided herein also apply to aspects of the present invention. Further specific embodiments of these aspects are described below.

[0069] The present invention relates to the present disclosure the use of at least one compound described in. In particular, the present invention relates to variable ionization efficiency the use of at least one compound having. The present invention relates to 1 to 100 species of compounds, preferably 1 to 80 species of compounds, more preferably 1 to 60 species of compounds, even more preferably 1 to 50 species of compounds, even more preferably 1 to 40 species of compounds, even more preferably 1 to 30 species of compounds, even more preferably 1 to 20 species of compounds, even more preferably 1 to 15 species of compounds, even more preferably 1 to 12 species of compounds, or even more preferably at least 9 species of compounds, or 29 species of compounds, and further relates to a composition containing. The composition of the present invention may contain compounds of different classifications of compounds. In the attached examples, a composition containing 9 species of compounds was used. Further, in the attached examples, a composition containing 29 species of compounds or 1 species compounds was used. Therefore, such a composition is preferred. A composition containing at least one compound and the present disclosure its use in the method provided in are preferred. In particular, the present disclosure The composition provided thereby contains sulfaguanidine, sodium naproxen, sulfadimethoxine, ciprofloxacin, tetracycline hydrochloride, verapamil hydrochloride, terfenadine, leucine enkephalin acetate hydrate, and reserpine. Further, the composition may contain leucine enkephalin acetate hydrate, terfenadine, verapamil hydrochloride, tetracycline hydrochloride, diltiazem hydrochloride, lincomycin hydrochloride, buspirone hydrochloride, sarafloxacin hydrochloride hydrate, haloperidol, trazodone hydrochloride, ciprofloxacin, quinine, ranitidine hydrochloride, triclocarban, sulfadimethoxine, trimethoprim, amitriptyline hydrochloride, atenolol, propranolol hydrochloride, sulfathiazole, sulfamethoxazole, cimetidine, salbutamol, melatonin, sodium naproxen, atrazine, sulfaguanidine, metformin hydrochloride, and reserpine or any other salts thereof.

[0070] This composition may preferably be provided in the kit of the present invention. At least one compound may be included in the composition or added separately to the sample.

[0071] The present invention further relates to a kit and the present disclosure the use of the kit in the method provided thereby. The kit includes at least one compound described in any one of the methods in the previous section, and optionally the present disclosure includes. Thus, the reference level may be provided in the kit, for example, in the form of an instruction manual. In a further aspect, in particular, reference sample of the reference level to calibrate the device used in the method provided thereby, the present disclosure a kit including a standard sample provided thereby is provided. Thus, such a kit may include a standard sample described in the present disclosure including a known compound, and in addition, an instruction manual for calibrating a device suitable for use in the provided method. the present disclosure

[0072] the present disclosure When used, the terms "comprising" and "including" or grammatical variations thereof are considered to define at least the specified features, integers, steps or components, but not to exclude the addition of one or more additional features, integers, steps, components or groups thereof. This term encompasses the terms "consisting of" and "consisting essentially of", which are understood to define only the specified features, integers, steps or components that exclude any additional features.

[0073] Accordingly, the terms "comprising" / "including" / "having" mean that additional components (or similarly features, integers, steps, etc.) can / may be present.

[0074] The term "consisting of" means that no additional components (or similarly features, integers, steps, etc.) are present.

[0075] The term "method" represents a method, means, technique and procedure for achieving a given task, including, but not limited to, methods, means, techniques and procedures developed from, or readily developed from, methods, means, techniques and procedures known to practitioners of chemical, biological and biophysical technologies.

[0076] The term "about" preferably represents ±10% of the specified numerical value, more preferably ±5% of the specified numerical value, and in particular the exact numerical value shown.

[0077] the present disclosure When used, the term "about" represents ±10% of the indicated numerical value, particularly ±5% of the indicated numerical value. When the term "about" is used, a specific indication of the exact numerical value shown is also included. When the term "about" is used in connection with a parameter that is quantified as an integer, such as the number of nucleotides in a given spread, the number corresponding to ±10% or ±5% of the specified numerical value shall be rounded to the nearest integer. For example, the expression "about 25 amino acids" represents a range of 23 to 28 amino acids, particularly a range of 24 to 26 amino acids, and preferably represents the specific value of 25 amino acids.

[0078] In certain embodiments, the present invention relates to the following: 1. A method for identifying and / or confirmation performing, said method comprising: (i) adding at least one compound to a sample, wherein in particular, the at least one compound variable ionization degree has, the step; (ii) obtaining a mass spectrum of the at least one compound and determining the level of the at least one compound after addition of the sample; (iii) comparing the level of the at least one compound determined in (ii) with a reference level, wherein the reference level is the level of the same compound determined after addition to a reference sample, the step; (iv) identifying the sample based on the comparison in step (iii) and the effect of the sample on the level of the at least one compound, and, a method. 2. The method according to item 1, wherein the level is a mass spectrometry signal level, and in particular, the level is the abundance determined in the mass spectrum. 3. The method according to item 1 or 2, wherein the sample has ion suppression for the at least one compound, or has an ion increase effect, or the sample does not substantially alter the ionization degree of the at least one compound. 4. Determine the ion suppression efficiency or the ion increase efficiency of the sample with respect to the at least one compound, and compare the ion suppression efficiency or the ion increase effect with the ion suppression efficiency or the ion increase effect of the reference sample with respect to the same at least one compound, and identify the sample based thereon. The method according to any one of items 1 to 3. 5. When compared with the reference level, a similar level or the same level of the at least one compound indicates that the sample matches the reference sample; or A different level of the at least one compound compared with the reference level indicates that the sample does not match the reference sample. The method according to any one of items 1 to 4. 6. In step (i), add 1 to 15 compounds. The method according to any one of items 1 to 5. 7. The compound is for ion suppression and / or ion increase effect is sensitive to of a specific substance contained in the sample. The method according to any one of items 1 to 6. 8. In step (ii), determine the level of each of the at least one compound, and in step (iii), compare the level of each of the at least one compound with each reference level. The method according to any one of items 1 to 7. 9. The sample is a pooled sample suitable for quality control purposes. The method according to any one of items 1 to 8. 10. The sample is selected from the group consisting of wine, alcoholic beverages, food products, processed foods, tea, coffee, herbal extracts, natural products, natural product extracts, beer, fruit juices (e.g., orange and apple), pharmaceutical compositions, pharmaceutical preparations, body fluids, tissue extracts, blood, plasma, serum, and urine. In particular, the sample is wine. The method according to any one of items 1 to 9. 11. The reference sample enables the identification and / or confirmation of the sample. The method according to any one of items 1 to 10. 12. The method according to any one of items 1 to 11, wherein the reference sample is a reference sample or a sample having a known composition. 13. The method according to any one of items 1 to 12, wherein the reference sample is a wine sample of a specific vintage, a wine sample of a specific grape variety, a wine sample of a specific region, or a wine sample of a specific producer. 14. In the method according to any one of items 1 to 13 variable ionization degree Use of said at least one compound having 15. In the method according to any one of items 1 to 14 variable ionization degree Use of a kit comprising and optionally at least one compound having a reference level of said reference sample. 16. The sample is a known variable ionization degree and / or a known ion suppression effect on said at least one compound, a known ion increase effect on said at least one compound, etc. ionization degree effect, or a known compound known not to substantially change the level of said at least one compound, and calibrating the method and / or the apparatus used in any one of items 1 to 13 using said sample. The method according to any one of items 1 to 13.

Brief Description of the Drawings

[0079] The present invention will be further described by referring to the following non-limiting drawings and examples. [Figure 1] Score plot of the first two preferred compounds from Example 1. [Figure 2] Score plot of the first two preferred compounds from Example 1. [Figure 3] Results of hierarchical cluster analysis (similarity is represented by the x-axis). [Figure 4] Score plot of principal component analysis (PC1 vs. PC2). The 95% confidence limits of the samples from each source do not overlap with the representation of 100% separation of the sources. [Figure 5] Score plot of principal component analysis (PC1 vs. PC3). The 95% confidence limits of the samples from each source do not overlap with the representation of 100% separation of the sources. [Figure 6] Score plot of principal component analysis (PC1 vs. PC3 vs. PC2). All clusters are separated in the three-dimensional projection. [Figure 7] Score plot of principal component analysis (PC1 vs. PC2) for a set of whisky and gin samples. Gin samples (lower left cluster) and whisky samples (upper right, two clusters) are clearly separated. [Figure 8] Effect of atrazine addition to wine samples - signal change of the atrazine peak.

Example

[0080] Example 1 The following compounds were used in the preferred method: - Sulfaguanidine - CAS No. 57 - 67 - 0 - Naproxen sodium - CAS No. 26159 - 34 - 2 - Sulfadimethoxine - CAS No. 122 - 11 - 2 - Ciprofloxacin - CAS No. 85721 - 33 - 1 - Tetracycline hydrochloride - CAS No. 64 - 75 - 5 - Verapamil hydrochloride - CAS No. 152 - 11 - 4 - Terfenadine - CAS No. 50679 - 08 - 8 - Leucine enkephalin acetate hydrate - MDL No. MFCD11045938 - Reserpine - CAS No. 50 - 55 - 5 Additional chemicals used: Domestic water supply system (Sartorius Arium Lab Water System, 18.2 MΩ) Ethanol - CAS No. 64 - 17 - 5 (Merck, LiChrosolv) Acetonitrile - CAS No. 75 - 05 - 8 (Fisher Chemicals, LC - MS grade) Formic acid - CAS No. 64 - 18 - 6 (Fluka, MS grade) Liquid handling system and MS: Binary pump model G1312B (Agilent Technologies) Autosampler model G1329A (Agilent Technologies) Triple quadrupole mass spectrometer model 6460A (Agilent Technologies) Electrospray source model G1958B (Agilent Technologies)

[0081] Sample preparation Chemistry purpose Compositions containing compounds were prepared by adding 10 μg of each compound to 1 mL of acetonitrile / water (50 / 50 (v / v)).

[0082] From a set of 3 different wines, each represented by 5 bottles, 1 mL samples were taken from each bottle using a Coravin (trademark) model 2 wine system and placed into 1.5 mL Eppendorf PCR tubes. 15 tubes were centrifuged at 12,500 g for 10 minutes. A solution of water / ethanol 85 / 15 (v / v) was prepared and degassed with nitrogen for 5 minutes. The wine samples were placed into 15 different HPLC vials and diluted with water / ethanol 85 / 15 (v / v) by adding 16 volumes of water / ethanol per volume of wine. After this step, 10 μL volumes of the chemistry purpose Compositions containing compounds were added to the diluted wine and mixed 20 times using the push - pull function of a pipette. For example: 31 μL of wine, 10 μL of the composition containing the compound, and 459 μL of water / ethanol 85 / 15 (v / v) were mixed. The samples were randomly placed into the autosampler and then measured by an LC - MS system.

[0083] Mass spectrometry method The modular sample introduction system consists of a degassing device, a binary pump, and an autosampler. Due to a constant backpressure, a restriction capillary (0.12 mm ID, 2000 mm) was placed between the pump and the autosampler. The autosampler valve was directly connected to the electrospray source (ESI) using a 0.12 mm ID capillary without using a column for compound separation. Mobile phase A consisted of water (channel A), and mobile phase B consisted of acetonitrile, both containing 0.1% formic acid. Isocratic mixing was performed at a constant flow rate of 600 μL / min using 25% B. Measurements were carried out using 1 μL in the flow injection analysis mode, and injection was performed after 0.2 minutes. The stop time was set to 0.7 minutes. The ESI source was operated in the positive mode using the following parameter settings: nebulizer pressure 45 psig (approx. 3.1×10 5 Pa[G]), nozzle voltage 0 V, sheath gas flow 12 L / min, sheath gas temperature 375 °C, drying gas flow 8 L / min, drying gas temperature 350 °C, and capillary voltage 3000 V. The mass spectrometer was operated in the multiple reaction monitoring mode with a dwell time of 40 milliseconds per transition. The resolution of the first and second quadrupoles was set to a unit resolution of 0.7 m / z FWHM (full width at half maximum). The cell acceleration voltage was fixed at 4 V for all compounds, and the delta electro multiplier voltage was set to 0. The system was controlled using MassHunter Acquisition version 10, and data analysis was performed using MassHunter Quantitative Analysis version 10 and Microsoft Excel 2016.

[0084] The peak height of the compound was used for subsequent multivariate data analysis. The peak heights of two subsequent measurements of the same sample were integrated, and a data matrix was created using the peak height as a variable (column) and the integrated measurement values as rows. The variables were then set to the first scale (subtracting the mean of each variable and then dividing by the standard deviation of each variable). Principal component analysis (PCA) was performed using the software Metaboanalyst 4.0. The scores were plotted against the first two preferred compounds shown in Figures 1 and 2.

[0085] Mass spectrometry parameters

Table 1

[0086] Example 2: Identification of water samples Water samples were analyzed from six different sources. 1. Aquina bottled mineral water 2. Arkina bottled mineral water 3. Tap water from Muttenz (Switzerland) called 「Normale」 4. Tap water from Effingen-Kirchen (Germany) 5. Deionized water 6. Nanopur water (purified using a Nanopur lab water system)

[0087] Three samples from each source were analyzed by adding the following 29 compounds (integration of 3 injections per sample).

[0088] The following compounds were used in the preferred method: - Leucine enkephalin acetate hydrate MDL N MFCD11045938 - Terfenadine - CAS number 50679-08-8 - Verapamil hydrochloride - CAS number 152-11-4 - Tetracycline hydrochloride - CAS number 64-75-5 - Diltiazem hydrochloride - CAS number 33286-22-5 - Lincomycin hydrochloride - CAS number 859-18-7 - Buspirone hydrochloride - CAS number 33386-08-2 - Sarafloxacin hydrochloride hydrate - CAS number 91296-87-6 - Haloperidol - CAS number 52-86-8 - Trazodone hydrochloride - CAS number 25332-39-2 - Ciprofloxacin - CAS No. 85721-33-1 - Kinin - CAS No. 130-95-0 - Ranitidine hydrochloride - CAS No. 66357-59-3 - Trichlocarban - CAS No. 101-20-2 - Sulfadimethoxine - CAS No. 122-11-2 - Trimethoprim - CAS No. 738-70-5 - Amitriptyline hydrochloride - CAS No. 549-18-8 - Atenolol - CAS No. 29122-68-7 - Propranolol hydrochloride - CAS No. 318-98-9 - Sulfathiazole - CAS No. 72-14-0 - Sulfamethoxazole - CAS No. 723-46-6 - Cimetidine - CAS No. 51481-61-9 - Salbutamol - CAS No. 18559-94-9 - Melatonin - CAS No. 73-31-4 - Naproxen sodium - CAS No. 26159-34-2 - Atrazine - CAS No. 1912-24-9 - Sulfaguanidine - CAS No. 57-67-0 - Metformin hydrochloride - CAS No. 1115-70-4 - Reserpine - CAS No. 50-55-5 Additional chemicals used: Domestic water supply system (Sartorius Arium Lab Water System, 18.2 MΩ) Ethanol - CAS No. 64-17-5 (Merck, LiChrosolv) Acetonitrile - CAS No. 75-05-8 (Fisher Chemicals, LC-MS grade) Formic acid - CAS No. 64-18-6 (Fluka, MS grade) Liquid handling system and MS: Binary pump model G1312B (Agilent Technologies) Autosampler model G1329A (Agilent Technologies) Triple quadrupole mass spectrometer model 6460A (Agilent Technologies) Electrospray source model G1958B (Agilent Technologies)

[0089] Sample preparation Six different water samples were collected. From each sample, 800 μL of three aliquots were mixed with 200 μL of a compound mixture (a dilution mixture of 29 compounds with adjusted concentrations in an 85 / 15 (v / v) water / ethanol solution). Samples were measured by the LC-MS system in a completely random order.

[0090] Liquid handling, mass spectrometry and data analysis methods The modular sample introduction system consists of a degasser, a binary pump and an autosampler. Due to a constant backpressure, a restriction capillary (0.12 mm ID, 2000 mm) was placed between the pump and the autosampler. The autosampler valve was directly connected to the electrospray source (ESI) using a 0.12 mm ID capillary without using a column for compound separation. Mobile phase A consisted of water (channel A), and mobile phase B consisted of acetonitrile, both containing 0.1% formic acid. Isocratic mixing was performed at a constant flow rate of 600 μL / min using 25% B. Measurements were performed using 1 μL in flow injection analysis mode, and injection was performed after 0.2 min. The stop time was set to 0.7 min. The ESI source was operated in positive mode using the following parameter settings: nebulizer pressure 45 psig (approx. 3.1×10 5Pa[G]), nozzle voltage 0 V, gas flow 12 L / min, gas temperature 350 °C, capillary voltage 3000 V. The mass spectrometer was operated in multiple reaction monitoring mode at a dwell time of 20 milliseconds per transition. The resolution of the first and second quadrupoles was set to a unit resolution of 0.7 m / z FWHM (full width at half maximum). The cell acceleration voltage was fixed at 4 V for all compounds, and the delta electro multiplier tube voltage was set to 0. The system was controlled using MassHunter Acquisition version 10, and data analysis was performed using MassHunter Quantitative Analysis version 10 and Microsoft Excel 2016.

[0091] The peak height of the compound was used for subsequent multivariate data analysis. The peak heights of two subsequent measurements of the same sample were integrated, and a data matrix was created using the peak height as a variable (column) and the integrated measurement value as a row. Next, the variable was set to the first scale (subtracting the mean of each variable and then dividing by the standard deviation of each variable).

[0092] Subsequently, multivariate data analysis (hierarchical cluster analysis HCA, principal component analysis PCA, and linear discriminant analysis) was performed using the measured peak heights of all 29 compounds.

[0093] Qualitative analysis (HCA and PCA, see Figures 3 - 5) showed the separation of all 6 samples of water. Furthermore, similar sources (both tap water source on one side and Nanopur / deionized water on the other) were more comparable to other sources of water. In particular, the analyzed tap water samples (Muttenz or Effingen-Kirchen) were similar. One sample of Muttenz in three was clustered in the Effingen-Kirchen cluster, which may be due to the fact that the similarity and cluster analysis of tap water at both locations are unoptimized unsupervised similarity searches for discriminating different classifications. Nevertheless, the supervised pattern recognition analysis using LDA of the samples showed 100% classification of 10-fold cross-validation prediction of the samples, so the distinction and thus the identification of all water samples were observed.

[0094] Briefly, all sources of water were successfully discriminated using the method described in the present disclosure .

[0095]

Table 2

[0096] Results

Table 3

[0097] Example 3: Identification of Gin and Whiskey Thirty-one different whiskey samples and sixteen different gin samples were investigated. Five aliquots were taken from each sample. The samples were measured using 29 compounds and classified using linear discriminant analysis (10-fold cross-validation using 29 peak heights).

[0098] Ninety-seven percent of the aliquots were assigned to the correct gin or whiskey sample. This demonstrates that the method provided in the present disclosure can also identify / confirmation gin and whiskey.

[0099] The following compounds were used in the preferred method: - Leucine enkephalin acetate hydrate MDL N MFCD11045938 - Terfenadine - CAS number 50679-08-8 - Verapamil hydrochloride - CAS number 152-11-4 - Tetracycline hydrochloride - CAS number 64-75-5 - Diltiazem hydrochloride - CAS number 33286-22-5 - Lincomycin hydrochloride - CAS number 859-18-7 - Buspirone hydrochloride - CAS number 33386-08-2 - Sarafloxacin hydrochloride hydrate - CAS number 91296-87-6 - Haloperidol - CAS No. 52 - 86 - 8 - Trazodone hydrochloride - CAS No. 25332 - 39 - 2 - Ciprofloxacin - CAS No. 85721 - 33 - 1 - Kinin - CAS No. 130 - 95 - 0 - Ranitidine hydrochloride - CAS No. 66357 - 59 - 3 - Triclocarban - CAS No. 101 - 20 - 2 - Sulfadimethoxine - CAS No. 122 - 11 - 2 - Trimethoprim - CAS No. 738 - 70 - 5 - Amitriptyline hydrochloride - CAS No. 549 - 18 - 8 - Atenolol - CAS No. 29122 - 68 - 7 - Propranolol hydrochloride - CAS No. 318 - 98 - 9 - Sulfathiazole - CAS No. 72 - 14 - 0 - Sulfamethoxazole - CAS No. 723 - 46 - 6 - Cimetidine - CAS No. 51481 - 61 - 9 - Salbutamol - CAS No. 18559 - 94 - 9 - Melatonin - CAS No. 73 - 31 - 4 - Naproxen sodium - CAS No. 26159 - 34 - 2 - Atrazine - CAS No. 1912 - 24 - 9 - Sulfaguanidine - CAS No. 57 - 67 - 0 - Metformin hydrochloride - CAS No. 1115 - 70 - 4 - Reserpine - CAS No. 50 - 55 - 5 Additional chemicals used: Domestic water supply system (Sartorius Arium Lab Water System, 18.2 MΩ) Ethanol - CAS No. 64 - 17 - 5 (Merck, LiChrosolv) Acetonitrile - CAS No. 75 - 05 - 8 (Fisher Chemicals, LC - MS grade) Formic acid - CAS No. 64 - 18 - 6 (Fluka, MS grade) Liquid handling system and MS: Binary pump model G1312B (Agilent Technologies) Autosampler model G1329A (Agilent Technologies) Triple quadrupole mass spectrometer model 6460A (Agilent Technologies) Electrospray source model G1958B (Agilent Technologies)

[0100] Sample preparation From each whiskey or gin sample, 100 μL aliquots were taken into 1.5 mL HPLC vials and mixed with 400 μL of a compound mixture. A compound mixture of 29 compounds with adjusted concentrations in an 85 / 15 (v / v) water / ethanol solution was used for each compound (see table below).

[0101] Liquid handling, mass spectrometry and data analysis methods The modular sample introduction system consists of a degasser, a binary pump and an autosampler. For a constant backpressure, a restriction capillary (0.12 mm ID, 2000 mm) was placed between the pump and the autosampler. The autosampler valve was directly connected to the electrospray source (ESI) using a 0.12 mm ID capillary without using a column for compound separation. Mobile phase A consisted of water (channel A) and mobile phase B consisted of acetonitrile, both containing 0.1% formic acid. Isocratic mixing was performed at a constant flow rate of 600 μL / min using 25% B. Measurements were carried out using 1 μL in flow injection analysis mode and injection was performed after 0.2 min. The stop time was set to 0.7 min. The ESI source was operated in positive mode using the following parameter settings: nebulizer pressure 45 psig (approx. 3.1×10 5Pa[G]), nozzle voltage 0 V, sheath gas flow rate 12 L / min, sheath gas temperature 350 °C, capillary voltage 3000 V. The mass spectrometer was operated in multiple reaction monitoring mode with a dwell time of 20 milliseconds per transition. The resolution of the first and second quadrupoles was set to a unit resolution of 0.7 m / z FWHM (full width at half maximum). The cell acceleration voltage was fixed at 4 V for all compounds, and the delta electro multiplier tube voltage was set to 0. The system was controlled using MassHunter Acquisition version 10, and data analysis was performed using MassHunter Quantitative Analysis version 10 and Microsoft Excel 2016.

[0102] The peak height of the compound was used for subsequent multivariate data analysis. The peak heights of two subsequent measurements of the same sample were integrated, and a data matrix was created using the peak height as a variable (column) and the integrated measurement values as rows. Next, the variables were set to the first scale (subtracting the mean of each variable and then dividing by the standard deviation of each variable).

[0103] Subsequently, multivariate data analysis (hierarchical cluster analysis HCA, principal component analysis PCA, and linear discriminant analysis) was performed using the measured peak heights of all 29 compounds.

[0104]

Table 4

[0105] Example 4: Analysis using only one compound Analysis of blended gin vs. whiskey The combined whiskey and gin datasets were reanalyzed, and at this time, the classification "whiskey" vs. "gin" had to be accurately assigned to each aliquot. An LDA model was constructed and the samples were classified using a 10-fold cross-validation procedure.

[0106] Regarding the model using the peak heights of all 29 compounds, a 100% accurate prediction of the classification "whiskey" vs. "gin" was achieved.

[0107] Regarding the LDA model using the peak height of only one compound, a prediction performance of 68.5% - 100% was achieved (average 87.5%, standard deviation 7.1%). Two markers (kinin and sarafloxacin) enabled 100% accurate classification each as they were. This demonstrates that a single compound approach can be used for prediction in a related small number of classifications.

[0108] Example 5: Identification of Robustness by Spike of Atrazine and Kinin To investigate the influence of the same chemical admixture used as a compound in the sample on the compound signal, a spike experiment was conducted using atrazine and kinin (both atrazine and kinin were compounds used in the experiment). Thus, atrazine and kinin were already contained in the sample, and additional atrazine and kinin were added to the sample at different concentrations as representatives of at least one compound. · Atrazine test: Three samples of wine were prepared: one original wine sample, one sample spiked with 15.16 ng / mL atrazine, and one sample containing 30.32 ng / mL atrazine. Thus, atrazine was added to three wine samples at different concentrations as shown. · Kinin test: Two samples of wine were prepared: one original wine sample, one sample spiked with 0.25 mg / mL kinin. Thus, kinin was added to one of the wine samples.

[0109] The samples were measured three times and the peaks were analyzed. In the atrazine test, a spike of 15.16 ng / mL atrazine resulted in a non-significant increase in the atrazine peak height, while a spike of 30.32 ng / mL atrazine resulted in a significant increase in the atrazine peak height (see Figure 8). The peak heights of all other compounds were not affected.

[0110] In the kinin test, the addition of 0.25 mg / ml caused a significant increase in the kinin peak height, but the other peaks were not affected.

[0111] Therefore, the present disclosure spiking with a substance representative of a specific compound of the analytical system proposed in will cause an increase in these compounds. Therefore, if at least one compound is already present in the sample, the sample can also be identified, and the same compound is added to the sample so that the compound level increases. Furthermore, the potential effects of atrazine or kinin contamination in wine or other samples will not affect the overall identification. This may affect the signal of this marker, contribute to the identification of such contaminated sample batches, and thus further improve the sample identification / confirmation can be improved.

Claims

1. A method for identifying and / or verifying a sample, the method comprising: (i) adding at least one compound to the sample, wherein the at least one compound has a variable ionization degree or a property of variably ionizing; (ii) after the addition to the sample in (i), obtaining a mass spectrum of the at least one compound and determining the level of the at least one compound; (iii) comparing the level of the at least one compound determined in (ii) with a reference level, wherein the reference level is the level of the same compound determined after addition to a reference sample; (iv) identifying the sample based on the comparison in (iii) and the effect of the sample on the level of the at least one compound. A method comprising the above.

2. The method according to claim 1, wherein the level is a mass spectrometry signal level or the level is an abundance determined in the mass spectrum.

3. The method according to claim 1 or 2, wherein the sample has an ion suppression effect or an ion enhancement effect on the at least one compound, or the sample does not substantially change the ionization degree of the at least one compound.

4. Determining the ion suppression efficiency or ion enhancement efficiency of the sample with respect to the at least one compound, comparing the ion suppression efficiency or ion enhancement efficiency with the ion suppression efficiency or ion enhancement efficiency of the reference sample with respect to the same at least one compound, and identifying the sample based on the comparison result. The method according to any one of claims 1 to 3.

5. When compared with the reference level, a similar level or the same level of the at least one compound indicates that the sample matches the reference sample; or The method according to any one of claims 1 to 4, wherein the level of the at least one compound being compared to the reference level being different indicates that the sample does not match the reference sample.

6. The method according to any one of claims 1 to 5, wherein in (iv), the sample is identified based on a specific pattern of ion suppression or ion enhancement of the at least one compound.

7. The method according to any one of claims 1 to 6, wherein in (i), the sample is used without separation, and in (ii), the level is determined without performing chromatographic separation.

8. The method according to claim 7, wherein the mass spectrum is obtained by a mass spectrometer connected to an ion source.

9. The method according to claim 8, wherein the ion source is an electrospray ionization source.

10. In (i), different concentrations of the at least one compound are added to the sample, and in (ii), the levels of the different concentrations of the at least one compound are determined after the addition to the sample; or In (i), the at least one compound is added to different concentrations of the sample, and in (ii), the levels of the at least one compound after the addition to the sample are determined. The method according to any one of claims 1 to 9.

11. The method includes creating a dose-response curve, The method according to any one of claims 1 to 10, wherein the dose-response curve is based on levels of different concentrations of the at least one compound.

12. The method includes creating a dose-response curve, The dose-response curve is based on the levels of the at least one compound after addition to different concentrations of the sample. The method according to any one of claims 1 to 10.

13. The method according to any one of claims 1 to 12, wherein in (i), 1 to 100 compounds, or 1 to 15 compounds are added.

14. The method according to any one of claims 1 to 13, wherein the compound is sensitive to the ion suppression and / or ion enhancement effect of a specific substance contained in the sample.

15. The method according to any one of claims 1 to 14, wherein the compound is identifiable based on the mass spectrum.

16. The method according to any one of claims 1 to 15, wherein the compound is two or more compounds having different molecular weights, different partition coefficients (logP), and / or different pKa values.

17. The method according to any one of claims 1 to 16, wherein in (ii), the level of the at least one compound is determined, and in (iii), the level of the at least one compound is compared with each reference level.

18. The method according to any one of claims 1 to 17, wherein the at least one compound is not contained in the sample.

19. The method according to any one of claims 1 to 18, wherein the at least one compound is selected from the group consisting of sulfaguanidine, sodium naproxen, sulfadimethoxine, ciprofloxacin, tetracycline hydrochloride, verapamil hydrochloride, promethazine, leucine enkephalin acetate hydrate, and reserpine.

20. The method according to any one of claims 1 to 19, wherein the at least one compound is sulfaguanidine, sodium naproxen, sulfadimethoxine, ciprofloxacin, tetracycline hydrochloride, verapamil hydrochloride, promethazine, leucine enkephalin acetate hydrate, and reserpine.

21. The method according to any one of claims 1 to 20, wherein the at least one compound has a concentration in the sample that is at least five times the limit of quantification after addition to the sample.

22. The method according to any one of claims 1 to 21, wherein the at least one compound has a concentration in the sample after addition to the sample such that the level of the at least one compound is suppressed and / or increased by about 20% to about 80% compared to the level of the same compound determined in the absence of the sample.

23. The method according to any one of claims 1 to 22, wherein a further level of a standard is determined to calibrate the method according to any one of claims 1 to 22.

24. The method according to any one of claims 1 to 23, wherein the sample is a pooled sample for quality control.

25. The method according to any one of claims 1 to 24, wherein the sample is selected from the group consisting of wine, alcoholic beverages, foodstuffs, processed foods, tea, coffee, herbal extracts, natural products, natural product extracts, beer, fruit juices (e.g., orange and apple), pharmaceutical compositions, pharmaceutical formulations, body fluids, tissue extracts, blood, plasma, serum, and urine, and in particular, the sample is wine.

26. The method according to any one of claims 1 to 25, wherein the reference sample enables the identification and / or confirmation of the sample.

27. The method according to any one of claims 1 to 26, wherein the sample and the reference sample are from the same type.

28. The method according to any one of claims 1 to 27, wherein the reference sample is a reference sample or a sample having a known composition.

29. The method according to any one of claims 1 to 28, wherein the reference sample is a wine sample of a specific vintage, a wine sample of a specific grape variety, a wine sample of a specific region, or a wine sample of a specific producer.

30. The identification of the sample is the identification of a dilution of the sample, and the dilution of the sample is identified based on a comparison of the dose-response curve of the sample and the dose-response curve of the reference sample, the dose-response curve being based on different concentration levels of the at least one compound or a constant concentration level of the at least one compound. The method according to any one of claims 1 to 29.

31. The method according to any one of claims 1 to 30, wherein the method includes calibration, and the sample includes a known compound having a known variable ionization degree and / or a known ionization suppression effect on the at least one compound, a known ionization enhancement effect on the at least one compound, etc., or a known compound that is known not to substantially change the level of the at least one compound, and the method according to any one of claims 1 to 30 is calibrated using the sample.

32. Use of a variable ionization degree or the at least one compound having a variably ionizable property in the method according to any one of claims 1 to 31.

33. Use of a variable ionization degree or the at least one compound having a variably ionizable property, and optionally a reference sample at a reference level, in the kit provided in the method according to any one of claims 1 to 31.

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