CN115754031A - GC-MS-based centroid distinguishing method for different medicinal parts of elsholtzia rugulosa - Google Patents
GC-MS-based centroid distinguishing method for different medicinal parts of elsholtzia rugulosa Download PDFInfo
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- 238000000034 method Methods 0.000 title claims abstract description 18
- 241000104430 Elsholtzia rugulosa Species 0.000 title claims abstract description 16
- 238000002290 gas chromatography-mass spectrometry Methods 0.000 title claims abstract description 14
- 241000594402 Damnacanthus Species 0.000 claims abstract description 15
- 241000131458 Elsholtzia Species 0.000 claims description 6
- NVEQFIOZRFFVFW-RGCMKSIDSA-N caryophyllene oxide Chemical compound C=C1CC[C@H]2O[C@]2(C)CC[C@H]2C(C)(C)C[C@@H]21 NVEQFIOZRFFVFW-RGCMKSIDSA-N 0.000 claims description 6
- DCAYPVUWAIABOU-UHFFFAOYSA-N hexadecane Chemical compound CCCCCCCCCCCCCCCC DCAYPVUWAIABOU-UHFFFAOYSA-N 0.000 claims description 6
- 238000002347 injection Methods 0.000 claims description 6
- 239000007924 injection Substances 0.000 claims description 6
- 238000004458 analytical method Methods 0.000 claims description 4
- PJPMEFIJBKCSBT-UHFFFAOYSA-N 2-methoxyethenylcyclopropane Chemical compound COC=CC1CC1 PJPMEFIJBKCSBT-UHFFFAOYSA-N 0.000 claims description 3
- NVEQFIOZRFFVFW-UHFFFAOYSA-N 9-epi-beta-caryophyllene oxide Natural products C=C1CCC2OC2(C)CCC2C(C)(C)CC21 NVEQFIOZRFFVFW-UHFFFAOYSA-N 0.000 claims description 3
- NIDGCIPAMWNKOA-WOJBJXKFSA-N Neophytadiene Natural products [C@H](CCC[C@@H](CCCC(C)C)C)(CCCC(C=C)=C)C NIDGCIPAMWNKOA-WOJBJXKFSA-N 0.000 claims description 3
- RSYBQKUNBFFNDO-UHFFFAOYSA-N caryophyllene oxide Natural products CC1(C)CC2C(=C)CCC3OC3(C)CCC12C RSYBQKUNBFFNDO-UHFFFAOYSA-N 0.000 claims description 3
- 150000002596 lactones Chemical class 0.000 claims description 3
- NIDGCIPAMWNKOA-UHFFFAOYSA-N neophytadiene Chemical compound CC(C)CCCC(C)CCCC(C)CCCC(=C)C=C NIDGCIPAMWNKOA-UHFFFAOYSA-N 0.000 claims description 3
- FATBGEAMYMYZAF-KTKRTIGZSA-N oleamide Chemical compound CCCCCCCC\C=C/CCCCCCCC(N)=O FATBGEAMYMYZAF-KTKRTIGZSA-N 0.000 claims description 3
- FATBGEAMYMYZAF-UHFFFAOYSA-N oleicacidamide-heptaglycolether Natural products CCCCCCCCC=CCCCCCCCC(N)=O FATBGEAMYMYZAF-UHFFFAOYSA-N 0.000 claims description 3
- 230000005540 biological transmission Effects 0.000 claims description 2
- 239000012159 carrier gas Substances 0.000 claims description 2
- 239000001307 helium Substances 0.000 claims description 2
- 229910052734 helium Inorganic materials 0.000 claims description 2
- SWQJXJOGLNCZEY-UHFFFAOYSA-N helium atom Chemical compound [He] SWQJXJOGLNCZEY-UHFFFAOYSA-N 0.000 claims description 2
- 238000001819 mass spectrum Methods 0.000 claims description 2
- 239000002904 solvent Substances 0.000 claims description 2
- 239000000523 sample Substances 0.000 description 17
- 239000000126 substance Substances 0.000 description 5
- 235000019504 cigarettes Nutrition 0.000 description 4
- 238000001514 detection method Methods 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- 239000000796 flavoring agent Substances 0.000 description 3
- 235000019634 flavors Nutrition 0.000 description 3
- 238000005070 sampling Methods 0.000 description 3
- 241001517299 Bulbophyllum Species 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- 244000062748 Eupatorium adenophorum Species 0.000 description 2
- 241000221089 Jatropha Species 0.000 description 2
- 241001048891 Jatropha curcas Species 0.000 description 2
- 230000000975 bioactive effect Effects 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 239000000284 extract Substances 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 208000024827 Alzheimer disease Diseases 0.000 description 1
- 206010012735 Diarrhoea Diseases 0.000 description 1
- 241000131457 Elsholtzia stauntonii Species 0.000 description 1
- 241000735527 Eupatorium Species 0.000 description 1
- 244000062327 Eupatorium odoratum Species 0.000 description 1
- 206010037660 Pyrexia Diseases 0.000 description 1
- 230000001093 anti-cancer Effects 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 230000000840 anti-viral effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 238000012850 discrimination method Methods 0.000 description 1
- 230000000857 drug effect Effects 0.000 description 1
- 235000018927 edible plant Nutrition 0.000 description 1
- OAMZXMDZZWGPMH-UHFFFAOYSA-N ethyl acetate;toluene Chemical class CCOC(C)=O.CC1=CC=CC=C1 OAMZXMDZZWGPMH-UHFFFAOYSA-N 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000003205 fragrance Substances 0.000 description 1
- 235000015092 herbal tea Nutrition 0.000 description 1
- 235000008216 herbs Nutrition 0.000 description 1
- 206010022000 influenza Diseases 0.000 description 1
- 238000010813 internal standard method Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 201000009240 nasopharyngitis Diseases 0.000 description 1
- 238000010606 normalization Methods 0.000 description 1
- 238000003672 processing method Methods 0.000 description 1
- 238000013139 quantization Methods 0.000 description 1
- 239000012488 sample solution Substances 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
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Abstract
The method comprises the steps of analyzing a sample containing a medicinal part of the wild dam by using GC-MS, and identifying the wild dam flowers, the wild dam cotyledons or the wild dam cotyledons by establishing a discrimination function and a centroid equation; respectively calculating discriminant functions (X, Y) of the wild damnacanthus flower, the wild damnacanthus leaf or the wild damnacanthus stem to the mass centers (X) of different wild damnacanthus parts i ,Y i ) The part corresponding to the smallest value is the medicinal part of the elsholtzia rugulosa in the sample.
Description
Technical Field
The invention relates to a method for detecting different medicinal parts of an elsholtzia rugulosa, in particular to a GC-MS-based method for distinguishing the mass centers of the different medicinal parts of the elsholtzia rugulosa.
Background
The elsholtzia rugulosa is a medicinal and edible plant with special fragrance. It is mainly distributed in the southwest area of China, especially Yunnan province. In Yunnan province, it is used with herbal tea, herbs and honey-sourced plants. It is used by local Yi nationality and other minority nationalities to treat common cold, fever, influenza and diarrhea. Many compounds isolated from Elsholtzia rugulosa have been reported to have antiviral, anticancer, anti-inflammatory activities and even alleviate the symptoms of Alzheimer's disease. In addition, the elsholtzia rugulosa is used as a characteristic natural plant with stronger regionality, the extract of the elsholtzia rugulosa has the application effects of increasing aroma, improving quality, correcting flavor and the like in cigarettes, has the characteristics of excellent natural cigarette flavor deeply, and is a potential resource of natural flavor added by Chinese cigarettes.
The different parts of the jatropha have different drug effect results, wherein the relative content of bioactive substances in the flowers of the jatropha is the highest, the relative content of the bioactive substances in the leaves is the lowest, and the relative content of the stems is the lowest. Meanwhile, the yield of the extracts of different parts and the adding effect in cigarettes are also greatly different. Driven by commercial interest, many illicit individuals impersonate or adulterate the jatropha curcas flower products with the jatropha curcas stem and leaf products to obtain illicit profits. Therefore, it is necessary to distinguish different parts such as the stems and leaves of the wild damnacanthus.
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 different medicinal parts of an elsholtzia rugulosa, which comprises the following steps of:
step (1): analyzing a sample containing the medicinal part of the elsholtzia by GC-MS;
step (2): identifying the wild damnacanthus, wild damnacanthus leaves or wild damnacanthus stems based on the following discriminant functions and a centroid equation;
a discriminant function:
X＝-27.379C 1 -46.145C 2 +14.698C 3 +56.639C 4 +0.229C 5 +9.015C 6 -153.053；
Y＝0.603C 1 -2.949C 2 +0.330C 3 +13.730C 4 +0.052C 5 -0.073C 6 -6.772；
wherein: c 1 Is the content of the caryophyllene oxide mu g/g, C 2 Is 18-octadecan-9-ene lactone microgram/gram, C 3 Is the content of 1- (2-methoxyvinyl) cyclopropane mu g/g, C 4 Is the content of neophytadiene mu g/g, C 5 The content of hexadecane is mu g/g, and the content of C6 is mu g/g of oleamide;
the corresponding mass center equation of different medicinal parts of the elsholtzia rugulosa is as follows:
according to the formula:
calculating (X, Y) to different parts of the dam (X) i ,Y i ) The part corresponding to the smallest value is the medicinal part of the elsholtzia rugulosa in the sample.
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 flask 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) in a direct sample injection manner.
(14) Data processing
Normalization: quantization was performed using an internal standard method. The content of each volatile component was calculated as follows:
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 area of the internal standard substance is shown; m 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 injection 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 to 275 deg.C at a rate of 60 deg.C/min, then raised to 295 deg.C at a rate of 1 deg.C/min, and held for 1 minute.
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. 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 invention realizes the accurate identification of the medicinal parts and products of different elsholtzia rugulosa by GC-MS analysis for the first time, and simultaneously can carry out primary analysis on volatile medicinal components of the products.
2. The method is simple and convenient, and only needs to substitute the content of the caryophyllene oxide, the content of 18-octadecane-9-ene lactone, the content of 1- (2-methoxy vinyl) cyclopropane, the content of neophytadiene, the content of hexadecane and the content of oleamide into the discrimination function in the step (2) to obtain coordinates (X and Y), then the centroid distances from (X and Y) to different medicinal parts of the elsholtzia are calculated according to a distance formula, the centroid distances of different elsholtzia are compared, the part corresponding to the minimum centroid distance is the medicinal part of the elsholtzia in the sample, and the result is reliable, so that the method 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), dividing 4 samples of the eupatorium adenophorum leaves 1, the eupatorium stems 1, the eupatorium floribundum flowers 1, the eupatorium adenophorum leaves 2 and the like into 6 parts respectively, analyzing and processing to obtain corresponding GC-MS data shown in a table 1:
TABLE 1 GC-MS data for samples from different parts of the Elsholtzia rugulosa
ND means that the component is not detected, and the content is calculated as 0. Mu.g/g.
The corresponding discriminant function index components are shown in table 2 below:
TABLE 2 content of discriminant function index of different samples
The results obtained by substituting the corresponding component content values into step (2) are shown in Table 3:
TABLE 3 sample discrimination equation values and discrimination results
| ID | D Leaf of Chinese character | D Stem of a tree | D Flower (A. B. A. B. A | Discrimination knotFruit (A. A. B. D. B |
| Wild dam cotyledon 1 | 6.14 | 68.10 | 411.43 | Leaf of Bulbophyllum |
| Wild dam stem 1 | 61.24 | 7.81 | 476.08 | Caulis Seu folium Euonymi Fortunei |
| Wild damnacanthus 1 | 416.86 | 476.48 | 0.53 | All-grass of Bulbophyllum |
| Wild dam cotyledon 2 | 2.22 | 59.92 | 417.43 | Leaf of Elsholtzia Stauntoni |
The experimental result shows that the judgment result of the judgment method is consistent with the actual part of the sample, and the method has better reliability.
In order to further verify the reliability of the method, 4 wild damnacanthus stems, 4 wild damnacanthus leaves and 4 wild damnacanthus flower samples are respectively selected, the samples cannot be visually distinguished directly after sampling and sample preparation, and then under the condition that a detector is not informed of a specific sample source, the detector is asked to perform part judgment on each sample purely based on the method provided by the invention, and the judgment results are shown in the following table 4:
table 4 verification results of the method
The identification result based on the method of the invention is completely consistent with the sample source, and as can be seen from table 4, the prediction accuracy of the wild dam stem, the wild dam cotyledon and the wild dam flower of the method of the invention is 100%, and the method can be used for accurately predicting the medicinal part of the wild dam.
Claims (3)
1. A GC-MS-based centroid distinguishing method for different medicinal parts of elsholtzia rugulosa is characterized by comprising the following steps:
step (1): analyzing a sample containing the medicinal part of the elsholtzia by GC-MS;
step (2): identifying the wild damnacanthus, wild damnacanthus leaves or wild damnacanthus stems based on the following discriminant functions and a centroid equation;
a discriminant function:
X＝-27.379C 1 -46.145C 2 +14.698C 3 +56.639C 4 +0.229C 5 +9.015C 6 -153.053；
Y＝0.603C 1 -2.949C 2 +0.330C 3 +13.730C 4 +0.052C 5 -0.073C 6 -6.772；
wherein: c 1 Is the content of the caryophyllene oxide mu g/g, C 2 Is 18-octadecan-9-ene lactone microgram/g, C 3 Is the content of 1- (2-methoxyvinyl) cyclopropane mu g/g, C 4 The content of neophytadiene is mu g/g, C 5 The content of hexadecane is mu g/g, and the content of C6 is mu g/g of oleamide;
the centroid equation corresponding to different medicinal parts of elsholtzia is as follows:
according to the formula:
calculating (X, Y) to different parts of the dam (X) i ,Y i ) The part corresponding to the smallest value is the medicinal part of the elsholtzia rugulosa in the sample.
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 part being not less than 0.1 g.
3. The method of claim 1, wherein the instrument 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 sample injection is carried out, wherein the split ratio is 10; 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 spectrum conditions:
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.
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