CN115436517A - GC-MS-based linear discrimination method for origin tracing of cloud-produced wild dams - Google Patents

GC-MS-based linear discrimination method for origin tracing of cloud-produced wild dams Download PDF

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CN115436517A
CN115436517A CN202211080571.1A CN202211080571A CN115436517A CN 115436517 A CN115436517 A CN 115436517A CN 202211080571 A CN202211080571 A CN 202211080571A CN 115436517 A CN115436517 A CN 115436517A
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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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Abstract

The invention analyzes the samples of the cloud production elsholtzia rugulosa in different production areas by GC-MS, and establishes 4 linear discriminant functions for identifying the elsholtzia rugulosa in different production areas, if the four function values are compared, if Y is Principle of the major theory If the numerical value is larger, the Elsholtzia rugulosa in the sample is the Geranium of the city of the university, if Y Rich people If the value is larger, the wild dam in the sample is Fumin county wild dam, and if Y is greater Yongsheng If the value is larger, the weiba in the sample is Yongsheng county weiba, and if Y is greater Lijiang river If the numerical value is larger, the abutment in the sample is Rijiang county abutment.

Description

GC-MS-based linear discrimination method for origin tracing of cloud-produced wild dams
Technical Field
The invention relates to an analytical method for tracing the origin of a wild dam, in particular to a GC-MS-based linear discrimination method for tracing the origin of a cloud-produced wild dam.
Background
Elsholtzia rugulosa belongs to a medicine and food homologous plant of Labiatae, and has a special fragrance. It is mainly distributed in the southwest area of China, especially Yunnan province. The elsholtzia rugulosa is widely used for herbal tea, medicinal materials and honey plants in Yunnan province. In Yunnan province, minority nationalities use it to treat cold, fever, flu and diarrhea. In addition to the above medicinal functions, recent studies have shown that many compounds isolated from the elsholtzia have anticancer effects and can even alleviate the symptoms of alzheimer's disease. And identifying various non-volatile components such as terpenes, polyphenols, flavonoids and the like by a phytochemical method.
In addition, the elsholtzia rugulosa is used as a characteristic natural plant with stronger regional property, and the extract of the elsholtzia rugulosa has the application effects of increasing aroma, improving quality, correcting taste and the like in cigarettes, has the characteristics of excellent natural cigarette flavor, and is a potential resource of natural flavor added to Chinese-style cigarettes. At present, the major sources of the cloud-produced wild dams in the market are the geographic city, the Yongsheng county, the Lijiang city and the Fumin county. Since elsholtzia rugulosa is a herbaceous plant, its safety, authenticity and medicinal effect are all affected by geographical origin. Therefore, it is necessary to trace the source of 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 the herbal medicine, and further the source tracing can be realized by combining a chemometrics method.
The present invention has been made to solve the above problems.
Disclosure of Invention
The purpose of the invention is realized by the following technical scheme.
The invention provides a GC-MS-based linear discrimination method for tracing the origin of a cloud-produced wild dam, which comprises the following steps:
step (1): analyzing samples of the Yunyuan wild dams in different producing areas by GC-MS;
step (2): identifying the wild dams in the geographic city, the Yongsheng county, the Lijiang county and the Fumin county based on the following discriminant functions;
Y theory of the major organization ＝-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 1 Is the content of 2-methylpropyl 3-methylbutyrate mu g/g, X 2 The content of artemisone is mu g/g, X 3 The content of methyl 5-hydroxyiminovalerate is mu g/g,X 4 is 2- [2-H3]Content of methylpyrazine [ mu ] g/g, X 5 The content of humulol-II is mu g/g, X 6 Is cumin phenol content mu g/g, X 7 Is 6,10,14-trimethyl-2-pentadecanone in the content of mu g/g, X 8 Is the content of eicosyl vinyl carbonate (mu g/g), X 9 Is 2-methyl-eicosane content mu g/g, X 10 The content of the 2-methyl-octacosane is mu g/g;
comparing the four, if Y Theory of the major organization If the numerical value is larger, the wild dam in the sample is the wild dam in the physical city, and if Y is larger Rich people If the value is larger, the wild dam in the sample is Fumin county wild dam, and if Y is greater Yongsheng If the value is larger, the weiba in the sample is Yongsheng county weiba, and if Y is greater Lijiang river If the numerical value is larger, the wild dam in the sample is the wild dam 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 into a plurality of parts, sampling each part according to a certain weight, and bagging to be detected;
(12) Sample pretreatment
0.1g of a sample was weighed into a 10ml centrifuge tube, and 1.5ml of a 1. Mu.g/ml deuterated toluene-ethyl acetate solution was added thereto. The mixture was sonicated for 10 minutes, centrifuged at 4000rpm for 8 minutes, filtered, and 500. Mu.L of the supernatant was transferred to a chromatography vial in preparation for GC-MS analysis.
(13) GC-MS detection
And (4) 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:
Figure BDA0003832966640000021
wherein: c i Represents the measured content of the volatile component; a. The i Is the peak area of the corresponding compound; a. The 0 Peak areas of the internal standard are shown; m is a group of 0 Represents the mass of the internal standard; m is the mass of the sample taken. The data were then used for statistical analysis.
The content value of the volatile components is taken as the arithmetic mean value of a plurality of 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 according to the detection principle.
Preferably, the samples taken in step (1) are divided into no less than 6 parts on average, and each part is not less than 0.1 g.
Preferably, in step (1), the instrumental analysis parameters are as follows:
chromatographic conditions are as follows:
mass spectrum conditions: the column was a DB-35GC column (30 m.times.0.25 mm.times.0.25 μm) (Agilent, USA). The injection port temperature was 280 ℃. The carrier gas was helium and the flow rate was 1ml/min. The sample volume is 1.0 μ L, and the split injection is performed, wherein the split ratio is 10. The initial temperature was 80 deg.C, heated at a rate of 60 deg.C/min to 275 deg.C, then raised at a rate of 1 deg.C/min to 295 deg.C, and held for 1 minute.
The transmission line and ion source temperatures are 300 ℃ and 280 ℃ respectively; scanning mode: a full scan mode; ionization mode is 70ev electron impact; the solvent delay time was 2min. Of course, other instrumental analysis parameters may be used in step (1) as long as the content of the characteristic substance of interest of the present invention can be determined.
Compared with the prior art, the invention has the following beneficial effects:
1. the method realizes accurate identification of medicinal parts and products of different wild dams through GC-MS analysis for the first time, and can carry out primary analysis on volatile medicinal components of the products.
2. The invention firstly traces the source of the different producing areas of the elsholtzia by establishing a discriminant function relationship, the method is simple and convenient, and only needs to carry out the tracing on the 2-methylpropyl 3-methylbutyrate, the artemisone, the 5-hydroxyiminomethyl valerate and the 2- [2-H3 ]]Methyl pyrazine, humenol-II, cumin phenol, 6,10, 14-trimethyl-2-pentadecanone, eicosyl vinyl carbonateSubstituting the contents of the ester, the 2-methyl-eicosane and the 2-methyl-octacosane into the four discriminant equations in the step (2) to respectively obtain Y Theory of the major organization 、Y Rich people 、Y Yongsheng And Y Lijiang river The specific production place of the cloud wild dam can be judged to be the big reason city, the Yongsheng county, the Lijiang county or the Fumin county by comparing the four function values, the result is reliable, and the cloud wild dam can be widely applied in actual life.
Detailed Description
The present invention will be described below with reference to specific examples, but the embodiments of the present invention are not limited thereto. The experimental methods not specified in the examples are generally commercially available according to the conventional conditions and the conditions described in the manual, or according to the general-purpose equipment, materials, reagents and the like used under the conditions recommended by the manufacturer, unless otherwise specified.
Example 1
According to the step (1), 6 samples of Dairy city abutment, lijiang county abutment, fumin abutment 1, yongsheng abutment and Fumin abutment 2 are respectively taken, and are analyzed and processed according to the steps to obtain corresponding GC-MS data shown in a table 1:
TABLE 1 GC-MS data for Pachyrhizus samples from different origins
Figure BDA0003832966640000041
Figure BDA0003832966640000051
Figure BDA0003832966640000061
The corresponding component content values are respectively substituted into the discrimination equation in the step (2), and the obtained results are shown in table 2:
TABLE 2 sample discrimination equation values and discrimination results
Figure BDA0003832966640000062
Figure BDA0003832966640000071
The experimental result shows that the judgment result of the judgment 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 triband of the city of the big reason, 4 triband of the county of Lijiang county, 4 Yongsheng triband 4 samples of the wild dam of the county of Fumin are respectively selected, the samples are not visually distinguished after sampling and sample preparation, and then under the condition that the sources of the specific samples are not informed to detection personnel, the detection personnel are asked to judge the positions of the samples purely based on the method of the invention, and the judgment results are shown in the following table 3:
table 3 verification results of the method
Figure BDA0003832966640000072
As can be seen from Table 3, the prediction accuracy of the Hongkui of Yusheng Dairy City, the Yongsheng Fengkui of Lijiang county and the Fumin Fengkui of the method of the present invention is 100%, and the method can be used for the accurate prediction of the Yusheng Fengkui production area.

Claims (3)

1. A GC-MS-based linear discrimination method for tracing the origin of a cloud-produced wild dam is characterized by comprising the following steps:
step (1): analyzing samples of the cloaca yunnanensis in different producing areas by GC-MS;
step (2): identifying the wild dams in the geographic city, the Yongsheng county, the Lijiang county and the Fumin county based on the following discriminant functions;
Y principle of the major theory ＝-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 1 The content of 3-methyl butyric acid 2-methyl propyl ester is mu g/g, X 2 The content of artemisone is mu g/g and X 3 The content of methyl 5-hydroxyiminovalerate is [ mu ] g/g, X 4 Is 2- [2-H3]Content of methylpyrazine [ mu ] g/g, X 5 The content of humulol-II is mu g/g, X 6 Is cumin phenol content mu g/g, X 7 Is 6,10,14-trimethyl-2-pentadecanone content mu g/g, X 8 Is the content of eicosyl vinyl carbonate (mu g/g), X 9 Is 2-methyl-eicosane content mu g/g, X 10 The content of the 2-methyl-octacosane is mu g/g;
comparing the four, if Y Theory of the major organization If the numerical value is larger, the Elsholtzia rugulosa in the sample is the Geranium of the city of the university, if Y Rich people If the value is larger, the wild dam in the sample is Fumin county wild dam, and if Y is greater Yongsheng If the value is larger, the weiba in the sample is Yongsheng county weiba, and if Y is greater Lijiang river If the numerical value is larger, the wild dam in the sample is the wild dam in Lijiang county.
2. The method of claim 1, wherein the samples taken in step (1) are divided into equal parts of no less than 6 parts, each of no less than 0.1 g.
3. The method of claim 1, wherein the machine analysis parameters in step (1) are as follows:
chromatographic conditions are as follows:
the chromatographic column is a DB-35GC chromatographic column (30 m.times.0.25 mm.times.0.25 μm); the temperature of a 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, and the split flow injection is performed, wherein the split flow ratio is 10; the initial temperature was 80 ℃, heated to 275 ℃ at a rate of 60 ℃/min, then ramped up to 295 ℃ at a rate of 1 ℃/min, and held for 1 minute;
mass spectrum conditions:
the transmission line and the ion source are respectively at 300 ℃ and 280 ℃; scanning mode: a full scan mode; ionization mode is 70ev electron impact; the solvent delay time was 2min.
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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20150315626A1 (en) * 2014-04-30 2015-11-05 The Nemours Foundation Mucopolysaccharidosis iva/vii screening and treatment method
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

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20150315626A1 (en) * 2014-04-30 2015-11-05 The Nemours Foundation Mucopolysaccharidosis iva/vii screening and treatment method
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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