CN115436517B - GC-MS (gas chromatography-mass spectrometry) -based linear discrimination method for origin tracing of cloud-produced dam - Google Patents

GC-MS (gas chromatography-mass spectrometry) -based linear discrimination method for origin tracing of cloud-produced dam Download PDF

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CN115436517B
CN115436517B CN202211080571.1A CN202211080571A CN115436517B CN 115436517 B CN115436517 B CN 115436517B CN 202211080571 A CN202211080571 A CN 202211080571A CN 115436517 B CN115436517 B CN 115436517B
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dam
seeds
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content
county
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CN115436517A (en
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熊文
朱保昆
吴亿勤
王晋
蔡炳彪
李勇
赵辉
武凯
王颖琦
杨国荣
于春霞
倪朝敏
张涛
赵颖
孔留艳
朱涔铭
王明敬
卢婷
蔡昊城
倪旭东
张超
李娟�
李韶阳
许健
张承明
杨光宇
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China Tobacco Yunnan Industrial Co Ltd
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    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/04—Preparation or injection of sample to be analysed
    • G01N30/06—Preparation
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/62—Detectors specially adapted therefor
    • G01N30/72—Mass spectrometers
    • G01N30/7206—Mass spectrometers interfaced to gas chromatograph
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    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/86—Signal analysis
    • G01N30/8675—Evaluation, i.e. decoding of the signal into analytical information
    • G01N30/8679—Target compound analysis, i.e. whereby a limited number of peaks is analysed
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/86—Signal analysis
    • G01N30/8696—Details of Software
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    • G06—COMPUTING; CALCULATING OR COUNTING
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    • G06F17/00—Digital computing or data processing equipment or methods, specially adapted for specific functions
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    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/04—Preparation or injection of sample to be analysed
    • G01N2030/042—Standards
    • G01N2030/045—Standards internal
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/04—Preparation or injection of sample to be analysed
    • G01N30/06—Preparation
    • G01N2030/062—Preparation extracting sample from raw material
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/04—Preparation or injection of sample to be analysed
    • G01N30/06—Preparation
    • G01N2030/065—Preparation using different phases to separate parts of sample
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

According to the invention, samples of the cloudy dam seeds of different producing areas are analyzed by GC-MS, 4 linear discriminant functions for identifying the cloudy dam seeds of different producing areas are established, and compared with four function values, if Y Marble (L.) Gaertn If the value is larger, the dam seeds in the sample are the dam seeds in Mars, if Y Rich people If the value is larger, the wild dam seeds in the sample are those in Fu Min county, if Y Yongsheng If the value is larger, the wild dam seeds in the sample are Yongsheng county wild dam seeds, if Y Lijiang river If the value is larger, the dam seeds in the sample are those in Lijiang county.

Description

GC-MS (gas chromatography-mass spectrometry) -based linear discrimination method for origin tracing of cloud-produced dam
Technical Field
The invention relates to an analysis method for source tracing of a dam production area, in particular to a linear discrimination method for source tracing of a cloud dam production area based on GC-MS.
Background
The wild dam belongs to a medicinal and edible plant of Labiatae, and has a special fragrance. It is mainly distributed in southwest areas of China, especially in Yunnan province. The dam seed in Yunnan province is widely used for herbal tea, medicinal materials and honey plants. In Yunnan, it is used by minority nations to treat cold, fever, influenza and diarrhea. In addition to the above medicinal functions, recent studies have shown that many compounds isolated from the stageflower dam have anticancer effects and even can alleviate the symptoms of Alzheimer's disease. The phytochemistry method is used for identifying various non-volatile components such as terpenes, polyphenols, flavonoids, etc.
In addition, the wild dam is taken as a special natural plant with stronger territory, the extract of the wild dam has the application effects of enhancing aroma, improving quality, correcting taste and the like in cigarettes, has the characteristics of excellent natural tobacco flavor, and is a natural flavor potential resource added in Chinese cigarettes. The main sources of the cloud-produced dam in the current market are marble city, yongsheng county, lijiang city and Fumin county. Because the wild dam is used as a herbaceous plant, the safety, the authenticity and the medicinal effect of the wild dam are all affected by geographical sources. Therefore, it is necessary to trace the origin.
Gas chromatography-mass spectrometry (GC-MS) has good reproducibility, high sensitivity, and excellent separation and identification of volatile components. GC-MS measurement of volatile components can be used for preliminary evaluation of the efficacy of herbal medicines, and further combining chemometric methods to realize source tracing.
The present invention has been made to solve the above problems.
Disclosure of Invention
The aim of the invention is achieved by the following technical scheme.
The invention provides a GC-MS (gas chromatography-mass spectrometry) -based linear discrimination method for origin tracing of cloud-produced elsholtzia stauntoni seeds, which comprises the following steps:
step (1): analyzing samples of the cloudy elsholtzia rugulosa of different production areas by using GC-MS;
step (2): identifying dam seeds of the marble city, yongsheng county, lijiang county and Fumin county based on the following discriminant function;
Y marble (L.) Gaertn ＝-11687.685X 1 +75.673X 2 +2229.3X 3 +373.426X 4 -17.858X 5 +-5.691X 6 -884.701X 7 +39.451X 8 -2831.515X 9 +106.726X 10 -207.288；
Y Rich people ＝767152.88X 1 -8061.008X 2 -146394.36X 3 -6393.556X 4 +3925.007X 5 +3399.196X 6 +34040.447X7+5303.425X8+184467.643X9-7677.192X10-492593.945；
Y Lijiang river ＝87783.573X 1 -495.56X 2 -17738.558X 3 -3029.5X 4 -30.16X 5 -59.017X 6 +7333.816X 7 -652.077X 8 +22526.668X 9 -589.39X 10 -9916.409；
Y Yongsheng ＝-25471.496X 1 +368.086X 2 +4508.273X 3 -229.34X 4 -242.539X 5 -202.27X 6 -464.837X 7 -440.28X 8 -6030.808X 9 +346.746X 10 -810.274
Wherein: x is X 1 Is 3-methyl butyric acid 2-methyl propyl ester content mug/g, X 2 The content of arteannuin in [ mu ] g/g, X 3 For 5-hydroxy iminovaleric acid methyl ester content mug/g, X 4 2- [2-H3 ]]Methyl pyrazine content μg/g, X 5 The content of humulone-II is mu g/g and X 6 The content of the fennel phenol is mu g/g, X 7 Is 6,10, 14-trimethyl-2-pentadecanone content mug/g, X 8 At the eicosanyl vinyl carbonate content of mu g/g, X 9 Is 2-methyl-eicosane content mug/g, X 10 Is 2-methyl-octacosane content mug/g;
in comparison with the four, if Y Marble (L.) Gaertn If the value is larger, the dam seeds in the sample are the dam seeds in Mars, if Y Rich people If the value is larger, the wild dam seeds in the sample are those in Fu Min county, if Y Yongsheng If the value is larger, the wild dam seeds in the sample are Yongsheng county wild dam seeds, if Y Lijiang river If the value is larger, the dam seeds in the sample are those in Lijiang county.
Preferably, the step (1) may specifically include the following steps:
(11) Sample collection
Collecting a certain amount of sample to be detected, uniformly dividing the sample into a plurality of parts, sampling each part according to a certain weight, and bagging the sample to be detected;
(12) Sample pretreatment
0.1g of the sample was weighed into a 10ml centrifuge tube and 1.5ml of 1. Mu.g/ml deuterated toluene-ethyl acetate solution was added. The mixture was sonicated for 10 minutes, centrifuged at 4000rpm for 8 minutes, filtered, 500 μl of supernatant was removed and transferred to a chromatographic vial in preparation for GC-MS analysis.
(13) GC-MS detection
And (3) carrying out GC-MS detection on the sample solution obtained in the step (12) by a direct sample injection mode.
(14) Data processing
Normalization: quantization was performed using an internal standard method. The content of each volatile component was calculated as follows:
wherein: c (C) i Representing the measured volatile component content; a is that i Peak area of the corresponding compound; a is that 0 Represents the peak area of the internal standard; m is M 0 Indicating the mass of the internal standard; m is the mass of the sample taken. The data was then used for statistical analysis.
The volatile component content value is taken as the arithmetic average of the multiple detections.
Of course, other sampling conditions, sample pretreatment conditions or data processing methods may be used in step (1), as long as the content of each substance can be detected in accordance with the detection principle.
Preferably, the average fraction of the sample obtained in step (1) is not less than 6 parts, each part being not less than 0.1 g.
Preferably, in step (1), the instrument analysis parameters are as follows:
chromatographic conditions:
mass spectrometry conditions: the column was a DB-35GC column (30 m.times.0.25 mm.times.0.25 μm) (Agilent, USA). The sample inlet temperature was 280 ℃. The carrier gas was helium at a flow rate of 1ml/min. The sample injection volume is 1.0 mu L, the split sample injection is carried out, and the split ratio is 10:1. The initial temperature was 80 ℃, heated to 275 ℃ at a rate of 60 ℃/min, then raised to 295 ℃ at a rate of 1 ℃/min, and held for 1 minute.
The temperature of the transmission line and the ion source are 300 ℃ and 280 ℃ respectively; scanning mode: a full sweep mode; ionization mode is 70ev electron collisions; the solvent delay time was 2min. Of course, other instrumental analysis parameters may be used in step (1) as long as the characteristic substance content of interest according to the present invention can be measured.
Compared with the prior art, the invention has the following beneficial effects:
1. according to the invention, the GC-MS analysis is performed for the first time to realize the accurate identification of the medicinal parts and products of different stagedam seeds, and meanwhile, the volatile medicinal components of the products can be primarily analyzed.
2. The invention firstly traces the source of different producing areas of the dam by establishing a discriminant function relationship, and the method is simple and convenient and only needs to carry out the steps of 2-methylpropyl 3-methylbutyrate, arteannuin, 5-hydroxyiminovalerate and 2- [2-H3]Substituting the contents of methyl pyrazine, humulone-II, cumylphenol, 6,10, 14-trimethyl-2-pentadecanone, eicosanyl vinyl carbonate, 2-methyl-eicosane and 2-methyl-octacosane into the four discrimination equations in the step (2) to obtain Y respectively Marble (L.) Gaertn 、Y Rich people 、Y Yongsheng And Y Lijiang river By comparing the values of the four function values, the specific production areas of the cloud production dam can be judged to be the marble city, yongsheng county, lijiang county or Fumin county, the result is reliable, and the cloud production dam can be widely applied to actual life.
Detailed Description
The present invention will be described with reference to specific examples, but embodiments of the present invention are not limited thereto. Experimental methods, in which specific conditions are not specified in examples, are generally available commercially according to conventional conditions as well as those described in handbooks, or according to general-purpose equipment, materials, reagents, etc. used under conditions suggested by manufacturers, unless otherwise specified.
Example 1
According to the step (1), 6 samples of the Marble's dam, the Lijiang county's dam, the Richmanspipe dam 1, the Yongsheng dam and the Richmanspipe dam 2 are respectively taken, and are analyzed and processed according to the steps, so that corresponding GC-MS data are obtained as shown in the table 1:
TABLE 1 GC-MS data for samples of cloudy dam of different origin
Substituting the content values of the corresponding components into the discrimination equations in the step (2), and obtaining the results shown in table 2:
table 2 sample discrimination equation value and discrimination result
Experimental results show that the judging result of the judging method is consistent with the actual production place of the sample, and the method has better reliability.
In order to further verify the reliability of the method, 4 Marble stagedam seeds, 4 Lijiang county stagedam seeds, 4 Yongsheng stagedam seeds and 4 Fumin stagedam seed samples are respectively selected, the samples are not visually distinguished after sampling and sample preparation, and then under the condition that a detector is not informed of the specific sample sources, the detector is asked to carry out part discrimination on each sample purely based on the method, and the discrimination results are shown in the following table 3:
table 3 results of verification of the method
As can be seen from Table 3, the prediction accuracy of the dam in the cloud produced marble city, the dam in Lijiang county, and the dam in the rich people are 100%, and can be used for accurately predicting the production area of the dam in cloud production.

Claims (2)

1. A GC-MS-based linear discrimination method for origin tracing of Yunnan dam seeds is characterized by comprising the following steps:
step (1): samples of the dam seeds produced in Yunnan province at different producing places were analyzed by GC-MS, and the instrument analysis parameters are as follows:
chromatographic conditions:
the chromatographic column is a DB-35GC chromatographic column with the specification of 30m multiplied by 0.25mm multiplied by 0.25 mu m; the temperature of the sample inlet is 280 ℃; the carrier gas is helium, and the flow rate is 1ml/min; the sample injection volume is 1.0 mu L, the split sample injection is carried out, and the split ratio is 10:1; the initial temperature was 80 ℃, heated to 275 ℃ at a rate of 60 ℃/min, then raised to 295 ℃ at a rate of 1 ℃/min, and held for 1 minute;
mass spectrometry conditions:
the temperature of the transmission line and the ion source are 300 ℃ and 280 ℃ respectively; scanning mode: a full sweep mode; ionization mode is 70ev electron collisions; the solvent delay time is 2min;
step (2): identifying dam seeds of the marble city, yongsheng county, lijiang county and Fumin county based on the following discriminant function;
Y marble (L.) Gaertn = -11687.685X 1 +75.673X 2 +2229.3X 3 +373.426X 4 -17.858X 5 +-5.691X 6
-884.701X 7 +39.451X 8 -2831.515X 9 +106.726X 10 -207.288；
Y Rich people = 767152.88X 1 -8061.008X 2 -146394.36X 3 -6393.556X 4 +3925.007X 5
+3399.196X 6 +34040.447X7+5303.425X8+184467.643X9-7677.192X10-492593.945；
Y Lijiang river = 87783.573X 1 -495.56X 2 -17738.558X 3 -3029.5X 4 -30.16X 5 -59.017X 6
+7333.816X 7 -652.077X 8 +22526.668X 9 -589.39X 10 -9916.409；
Y Yongsheng = -25471.496X 1 +368.086X 2 +4508.273X 3 -229.34X 4 -242.539X 5 -202.27X 6
-464.837X 7 -440.28X 8 -6030.808X 9 +346.746X 10 -810.274；
Wherein: x is X 1 Is 3-methyl butyric acid 2-methyl propyl ester content mug/g, X 2 The content of arteannuin in [ mu ] g/g, X 3 For 5-hydroxy iminovaleric acid methyl ester content mug/g, X 4 2- [2-H3 ]]Methyl pyrazine content μg/g, X 5 The content of humulone-II is mu g/g and X 6 The content of the fennel phenol is mu g/g, X 7 Is 6,10, 14-trimethyl-2-pentadecanone content mug/g, X 8 At the eicosanyl vinyl carbonate content of mu g/g, X 9 Is 2-methyl-eicosane content mug/g, X 10 Is 2-methyl-octacosane content mug/g;
in comparison with the four, if Y Marble (L.) Gaertn If the value is larger, the dam seeds in the sample are the dam seeds in Mars, if Y Rich people If the value is larger, the wild dam seeds in the sample are those in Fu Min county, if Y Yongsheng If the value is larger, the wild dam seeds in the sample are Yongsheng county wild dam seeds, if Y Lijiang river If the value is larger, the dam seeds in the sample are those in Lijiang county.
2. The method according to claim 1, wherein the average fraction of the sample obtained in the step (1) is not less than 6 parts and not less than 0.1g per part.
CN202211080571.1A 2022-09-05 2022-09-05 GC-MS (gas chromatography-mass spectrometry) -based linear discrimination method for origin tracing of cloud-produced dam Active CN115436517B (en)

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Publication number Priority date Publication date Assignee Title
KR20190103934A (en) * 2018-02-28 2019-09-05 (주) 제노텍 Qualitative or quantitative mutant genotyping methods and real-time PCR kits for performing the methods
CN110596078A (en) * 2019-08-14 2019-12-20 暨南大学 Method for tracing and identifying origin of geographical marked mandarin fish
CN111551644A (en) * 2020-04-03 2020-08-18 黄埔海关技术中心 Method for tracing origin of imported fragrant rice based on ion mobility spectrometry technology
CN113776913A (en) * 2021-09-18 2021-12-10 秦皇岛海关技术中心 Method for identifying producing area of black mountain brown shell eggs

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9982288B2 (en) * 2014-04-30 2018-05-29 The Nemours Foundation Mucopolysaccharidosis IVA/VII screening and treatment method

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20190103934A (en) * 2018-02-28 2019-09-05 (주) 제노텍 Qualitative or quantitative mutant genotyping methods and real-time PCR kits for performing the methods
CN110596078A (en) * 2019-08-14 2019-12-20 暨南大学 Method for tracing and identifying origin of geographical marked mandarin fish
CN111551644A (en) * 2020-04-03 2020-08-18 黄埔海关技术中心 Method for tracing origin of imported fragrant rice based on ion mobility spectrometry technology
CN113776913A (en) * 2021-09-18 2021-12-10 秦皇岛海关技术中心 Method for identifying producing area of black mountain brown shell eggs

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