Traditional Chinese medicine identification technical scheme based on multi-dimensional mass spectrum array identity card

Abstract

The invention provides a traditional Chinese medicine identification technical scheme based on a multi-dimensional mass spectrum array identity card, and relates to the technical field of traditional Chinese medicine identification and analysis. According to the technical scheme, the method comprises the following steps: S101, traditional Chinese medicine mass spectrometric detection S102, traditional Chinese medicine mass spectrum digital quantification storage; s103, extracting a multi-dimensional mass spectrum array identity card; s104, carrying out mass spectrometric detection on an unknown traditional Chinese medicine sample; s105, quantitative treatment of unknown traditional Chinese medicine samples; s106, performing ordered comparison on parameter regulation and control; and S107, comparing and feeding back matching results. The core feature of the method is an ordered matching algorithm and a matching degree R feedback algorithm. The method accords with the holistic concept of traditional Chinese medicine, retention time-mass-to-charge ratio-ionic strength reflecting chemical component information is taken as an element individual, array representation formed by multi-dimensional element individuals is taken as a traditional Chinese medicine multi-dimensional mass spectrum array identity card through parameter regulation and control, the attribute information of unknown components is effectively utilized, the specificity is higher, and the method is suitable for popularization and application. Through non-targeted identification, the identification efficiency is remarkably improved.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicine identification and analysis, and particularly to a technical solution for traditional Chinese medicine identification based on a multi-dimensional mass spectrometry array identity card. Technical Background

[0002] Traditional Chinese medicine identification and analysis play an important role in the quality control and evaluation research of traditional Chinese medicine, and are of great significance for distinguishing the authenticity of traditional Chinese medicine, combating crimes of counterfeit and inferior drugs, and strengthening the quality control and supervision of traditional Chinese medicine. At present, the technical methods of traditional Chinese medicine identification have achieved remarkable results relying on liquid chromatography-mass spectrometry analysis. However, more still rely on inspection according to the pharmacopoeia standards, that is, the known targeted identification of samples - based on a single or a few chemical components, formulating corresponding limit indexes to achieve the identification and analysis at the individual level of traditional Chinese medicine. This method breaks the overall concept of traditional Chinese medicine, overgeneralizes, only characterizes traditional Chinese medicine with a single or a few components, lacks exclusive representativeness, and has significantly lower identification efficiency. It fails to make full use of the attribute information of unknown components, and the results are hardly convincing.

[0003] Based on this, to solve the above problems, the present invention discloses a technical method for traditional Chinese medicine identification based on a multi-dimensional mass spectrometry array identity card, that is, starting from the unknown non-targeted identification of samples, through high-resolution mass spectrometry detection, using the multi-dimensional mass spectrometry array information as the quantitative characterization of traditional Chinese medicine, obtaining the "digital identity card" of each traditional Chinese medicine, and based on this, through the orderly comparison with the mass spectrometry information of unknown samples, quickly and accurately realizing the identification and analysis of traditional Chinese medicine. As Figure 1 shown, the technical method specifically includes the following steps:

[0004] S101: Traditional Chinese medicine mass spectrometry detection and analysis

[0004] S102: Digital quantization storage of traditional Chinese medicine mass spectrometry;

[0005] S103: Extraction of multi-dimensional mass spectrometry array identity card;

[0006] S104: Mass spectrometry detection of unknown traditional Chinese medicine samples;

[0007] S105: Quantization processing of unknown traditional Chinese medicine samples;

[0008] S106: Parameter regulation for orderly comparison;

[0009] S107: Feedback of comparison and matching results.

[0010] In the above steps S101 and S102, the traditional Chinese medicine is different traditional Chinese medicines of different species, different effects, different batches or origins;

[0011] In step S101, the traditional Chinese medicine mass spectrometry detection and analysis includes sample pretreatment and detection analysis conditions;

[0012] For the sample pretreatment, the following specific scheme is adopted:

[0013] Precisely weigh 1 - 2 g of the traditional Chinese medicine powder to be tested, add 25 - 100 ml of extraction solvent, extract in the same manner, and perform centrifugation or membrane filtration to obtain the solution of the traditional Chinese medicine sample to be tested;

[0014] Preferably, the extraction solvent is 25 ml of mass spectrometry methanol, the centrifugation parameters are (14000 r / min, 10 min), and it is filtered through a 0.22 μm organic filter membrane;

[0015] For the detection and analysis conditions, the specific scheme is as follows:

[0016] Liquid phase part: Reversed-phase chromatographic column UPLC BEH-C 18 (2.1×100 mm, 1.7 μm); column temperature 30 - 40 °C; the mobile phase uses a 0.01% - 0.1% formic acid in water (C) - acetonitrile (D) solution system, and the gradient elution conditions are: 0 - 23 min, 5% - 95% D; 23 - 26 min, 95% D; 26 - 26.1 min, 95% - 5% D, 26.1 - 30 min, 5% D; injection volume 1 - 5 μl;

[0017] Mass spectrometry part: Capillary: 1.5 - 3.0 kV (ESI+); Sampling Cone: 40 V; Source Offset: 80 V; Desolvation Temperatures: 450 °C; Desolvation Gas: 600 - 900 L / h; Source Temperatures: 110 - 120 °C; MS method: MS E ; Scan range: 100 - 1500; Collision energy: 10 V - 40 V.

[0018] In step S102, for the digital quantization storage of the traditional Chinese medicine mass spectrometry, the original file format is one of raw,.ddirectories, WIFF, T2D, FID, mzML, UNIFIData, mzXML, MS2 / CMS2 / BMS2, MGF, LCD, etc.;

[0019] In step S102, for the digital quantization storage of the traditional Chinese medicine mass spectrometry, the quantization software involved is one or several of Progenesis QI, UNIFI, MSConvert, MassHunter, LabSolutions, etc.;

[0020] In step S102, the digital quantization storage of traditional Chinese medicine mass spectrometry is performed in one or several of the following storage methods: static files, MongDB databases, MySQL databases, software self-built databases, and cloud databases;

[0021] In step S102, the digital quantization storage of traditional Chinese medicine mass spectrometry has a storage file format of one of the following formats: Excel, txt, csv, JSON, XML, TOML, CSON, YAML, etc.;

[0022] In step S102, the digital quantization storage of traditional Chinese medicine mass spectrometry involves multiple pieces of information in traditional Chinese medicine mass spectrometry, including sample information, processor information, mass-to-charge ratio (m / z), retention time, ion abundance, peak width, full width at half maximum, and neutral loss molecules, etc.

[0023] Preferably, the digital quantization content of traditional Chinese medicine mass spectrometry is mass-to-charge ratio (m / z), retention time, and ion abundance.

[0024] In step S102, the digital quantization storage of traditional Chinese medicine mass spectrometry is characterized by having no less than 200 different traditional Chinese medicines, with no less than 2 batches for each traditional Chinese medicine, and the traditional Chinese medicine varieties can be called and expanded at any time.

[0025] In step S103, the extraction of the multi-dimensional mass spectrometry array identification card is characterized in that the selected parameter thresholds are extracted in the following manner: TCMs-DM (traditional Chinese medicine name) = (I 1 , I 2 , I 3 , I 4 , …… I n-3 , I n-2 , I n-1 , I n ) T (n≥1) Or

[0026] The parameter thresholds are one or more of the retention time, mass-to-charge ratio, and ion intensity thresholds.

[0027] In step S104, the mass spectrometry detection of the unknown traditional Chinese medicine sample is characterized in that the sample pretreatment and detection analysis conditions are the same as those in step S101.

[0028] In step S105, the quantization processing of the unknown traditional Chinese medicine sample is characterized in that the quantization software, storage method, storage file format, and mass spectrometry information are the same as those in step S102.

[0029] In step S106, the parameter regulation performs an ordered comparison, characterized in that the "retention time - mass-to-charge ratio - ion intensity" is regarded as the individual elements in the multi-dimensional mass spectrometry array ID card of traditional Chinese medicine, and the specific parameters are one or several of the parameters such as the retention time deviation threshold, the retention time range, the mass-to-charge ratio deviation threshold, the mass-to-charge ratio range, the ion intensity range, and the output quantity after sorting the ion intensity, etc.;

[0030] Preferably, the retention time deviation threshold is 0.01 - 0.50, the mass-to-charge ratio deviation threshold is 0.005 - 0.05, and the output quantity is 1 - 1000;

[0031] In step S106, the parameter regulation performs an ordered comparison, characterized by an ordered matching algorithm. Based on the multi-dimensional mass spectrometry array ID card of each traditional Chinese medicine, it is compared and matched with the mass spectrometry element data of the unknown traditional Chinese medicine sample in the order of polarity size. When and only when all the selected parameter threshold conditions are met, it is regarded as a successful match. If at least one does not meet the selected parameter threshold, it is regarded as a failed match.

[0032] In step S107, the comparison and matching result feedback is characterized by a matching degree R feedback algorithm;

[0033] The matching degree R feedback algorithm is characterized in that: (1) the "retention time - mass-to-charge ratio - ion intensity" is regarded as the individual elements in the multi-dimensional mass spectrometry array ID card of traditional Chinese medicine. The number of elements successfully matched between the unknown traditional Chinese medicine sample and the multi-dimensional mass spectrometry array ID card of the stored traditional Chinese medicine is denoted as x, and the number of individual elements in the multi-dimensional mass spectrometry array ID card of traditional Chinese medicine under parameter regulation is denoted as y. Then the matching degree R = x / y * 100%; (2) through the ordered matching of the unknown sample with n traditional Chinese medicines in the storage, the corresponding [R 1 , R 2 , R 3 ,......, R n-2 , R n-1 , R n] array is obtained, and based on the TopN rule, the traditional Chinese medicine information with relatively high matching degree is fed back to achieve rapid and non-targeted identification.

[0034] The present invention has the following beneficial effects:

[0035] The present invention provides a technical solution for the identification of traditional Chinese medicine based on the multi-dimensional mass spectrometry array ID card. This solution conforms to the overall concept of traditional Chinese medicine. Taking the "retention time - mass-to-charge ratio - ion intensity" that reflects the chemical composition information as the individual elements, and through parameter regulation, the array representation formed by multi-dimensional individual elements is used as the multi-dimensional mass spectrometry array ID card of traditional Chinese medicine, realizing the effective utilization of the unknown component attribute information, with stronger specificity. Through non-targeted identification, the identification efficiency is significantly improved. Description of the Drawings

[0036] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the accompanying drawings required in the description of the embodiments or the prior art. Obviously, the accompanying drawings in the following description are only some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained based on these drawings.

[0037] Figure 1 Schematic diagram of the technical method steps of the present invention;

[0038] Figure 2 Matching identification results of unknown traditional Chinese medicines in the embodiment;

[0039] The following will clearly and completely describe the technical solutions in the embodiments of the present invention with reference to the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, rather than all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts belong to the scope of protection of the present invention. Embodiment

[0041] This embodiment provides a technical solution for the identification of Rhizoma Menispermi based on a multi-dimensional mass spectrometry array identity card, which specifically includes the following steps:

[0042] S101: Detection and analysis of traditional Chinese medicine mass spectrometry;

[0043] Take about 1.0 g of powder of 256 traditional Chinese medicines such as Rhizoma Menispermi, Radix Ginseng, Pericarpium Papaveris, Radix Saposhnikoviae, Herba Senecionis Scandentis, Cortex Phellodendri, Semen Sojae Nigrum, Pericarpium Arecae, Herba Angelicae Pubescentis, Bulbus Fritillariae Thunbergii, Rhizoma Coptidis Deltoideae, Radix Scutellariae, etc. (at least 2 batches of different batches for each traditional Chinese medicine), add 25 ml of mass spectrometry methanol respectively, ultrasonically extract for 30 min at 500 W and 40 KHz, filter through a 0.22 μm organic filter membrane to obtain the sample solution to be detected, and perform detection and analysis according to the following liquid chromatography-mass spectrometry conditions: reverse-phase chromatographic column UPLC BEH-C 18(2.1×100 mm, 1.7 μm); Column temperature: 35 °C; Mobile phase: 0.01% formic acid in water (C) - acetonitrile (D) solution system, gradient elution conditions: 0 - 23 min, 5% - 95% D; 23 - 26 min, 95% D; 26 - 26.1 min, 95% - 5% D, 26.1 - 30 min, 5% D; Injection volume: 2 μl; Capillary: 3.0 kV (ESI+); Sampling Cone: 40 V; Source Offset: 80 V; Desolvation Temperatures: 450 °C; Desolvation Gas: 900 L / h; Source Temperatures: 115 °C; MS method: MS E ; Scan range: 100 - 1500; Collision energy: 10 V - 40 V.

[0044] S102: Digital quantization storage of traditional Chinese medicine mass spectrometry;

[0045] Using Progenesis QI and UNIFI software, convert the mass spectrometry spectra of the obtained 256 traditional Chinese medicines into CSV files containing mass spectrometry data such as mass-to-charge ratio, ion intensity, retention time, etc., and store them as static files.

[0046] S103: Extraction of multi-dimensional mass spectrometry array identity cards;

[0047] For the same traditional Chinese medicine in different batches, select the retention time threshold as 0.05 and the mass-to-charge ratio threshold as 0.05, and extract the multi-dimensional mass spectrometry array identity cards in the following way:

[0048] S104: Mass spectrometry detection of unknown traditional Chinese medicine samples;

[0049] Take about 1.0 g of traditional Chinese medicine powder of unknown variety, add 25 ml of mass spectrometry methanol, extract ultrasonically at 500 W and 40 KHz for 30 min, filter through a 0.22 μm organic filter membrane to obtain the sample solution to be measured, and perform detection and analysis under the following conditions: Reverse-phase chromatographic column UPLC BEH-C 18(2.1×100 mm, 1.7 μm); Column temperature: 35 °C; The mobile phase used was a 0.01% formic acid in water (C) - acetonitrile (D) solution system, and the gradient elution conditions were: 0 - 23 min, 5% - 95% D; 23 - 26 min, 95% D; 26 - 26.1 min, 95% - 5% D, 26.1 - 30 min, 5% D; Injection volume: 2 μl; Capillary: 3.0 kV (ESI+); Sampling Cone: 40 V; Source Offset: 80 V; Desolvation Temperatures: 450 °C; Desolvation Gas: 900 L / h; Source Temperatures: 115 °C; MS method: MS E ; Scan range: 100 - 1500; Collision energy: 10 V - 40 V.

[0050] S105: Quantitative processing of unknown traditional Chinese medicine samples;

[0051] Using Progenesis Q, convert the mass spectrometry spectrum of the unknown traditional Chinese medicine into a CSV file containing mass spectrometry data such as mass-to-charge ratio, ion intensity, and retention time.

[0052] S106: Parameter regulation for orderly comparison;

[0053] The selected orderly comparison parameters were a retention time threshold of 0.1, a mass-to-charge ratio threshold of 0.02, and the top 400 element individuals in terms of ion intensity ranking. The output number of matching results was 7. The mass spectrometry data of the unknown traditional Chinese medicine was orderly compared with the multi-dimensional mass spectrometry array ID cards of each traditional Chinese medicine stored.

[0054] S107: Feedback of comparison matching results;

[0055] According to the orderly comparison results and the matching degree R feedback algorithm, obtain the matching degree R of the unknown traditional Chinese medicine with each traditional Chinese medicine stored i , and based on the TopN rule, sort to obtain the top 7 and their corresponding traditional Chinese medicines. The results are as Figure 2 shown. It was determined that the unknown variety of traditional Chinese medicine was Menispermi Rhizoma, with a matching degree of 94%, which was 3.2 times that of Papaveris Pericarpium with a matching degree of 29.25% ranking second. It had strong specificity, could quickly and efficiently complete the authenticity identification analysis, and could significantly improve the efficiency.

Claims

1. A technical solution for traditional Chinese medicine identification based on a multi-dimensional mass spectrometry array identity card, characterized in that, it includes the following steps: S101: Traditional Chinese medicine mass spectrometry detection and analysis; S102: Digital quantization storage of traditional Chinese medicine mass spectrometry; S103: Extraction of multi-dimensional mass spectrometry array identity card; S104: Mass spectrometry detection of unknown traditional Chinese medicine samples; S105: Quantization processing of unknown traditional Chinese medicine samples; S106: Parameter regulation for orderly comparison; S107: Feedback of comparison and matching results.

2. The technical solution for traditional Chinese medicine identification based on a multi-dimensional mass spectrometry array identity card according to claim 1, characterized in that, the digital quantization storage of traditional Chinese medicine mass spectrometry in step S102 stores no less than 200 different traditional Chinese medicines, with no less than 2 batches for each traditional Chinese medicine, and the varieties of traditional Chinese medicines can be called and expanded at any time.

3. The technical solution for traditional Chinese medicine identification based on a multi-dimensional mass spectrometry array identity card according to claim 1, characterized in that, in step S106, the parameter regulation for orderly comparison takes "retention time—mass-to-charge ratio—ion intensity" as the element individuals in the multi-dimensional mass spectrometry array identity card of traditional Chinese medicine, and the specific parameters are one or several of the parameters such as retention time deviation threshold, retention time range, mass-to-charge ratio deviation threshold, mass-to-charge ratio range, ion intensity range, and the output quantity after ion intensity sorting.

4. For the parameter regulation for orderly comparison according to claim 3, characterized in that the orderly matching algorithm takes the multi-dimensional mass spectrometry array identity card of each traditional Chinese medicine as the benchmark, and compares and matches the mass spectrometry element data of the unknown traditional Chinese medicine sample in the order of polarity size. When and only when all the selected parameter threshold conditions are met, it is regarded as a successful match. If at least one does not meet the selected parameter threshold, it is regarded as a failed match.

5. The technical solution for traditional Chinese medicine identification based on a multi-dimensional mass spectrometry array identity card according to claim 1, characterized in that, for the feedback of comparison and matching results in step S107, characterized in that it is a matching degree R feedback algorithm. (1) Taking "retention time—mass-to-charge ratio—ion intensity" as the element individuals in the multi-dimensional mass spectrometry array identity card of traditional Chinese medicine, the number of elements successfully matched between the unknown traditional Chinese medicine sample and the multi-dimensional mass spectrometry array identity card of the stored traditional Chinese medicine is denoted as x, and the number of element individuals in the multi-dimensional mass spectrometry array identity card of traditional Chinese medicine under parameter regulation is denoted as y, then the matching degree R = x / y × 100%. (2) By performing an orderly matching of the unknown sample with n traditional Chinese medicines in storage, the corresponding [R 1 , R 2 , R 3 ,......, R n-2 , R n-1 , R n array is obtained. Based on the TopN rule, the traditional Chinese medicine information with relatively high matching degree is feedback, realizing rapid and non-targeted identification.

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