GC fingerprint spectrum detection method of traditional Chinese medicine

Abstract

The invention discloses a GC fingerprint spectrum detection method for traditional Chinese medicine, and belongs to the field of traditional Chinese medicine detection. The traditional Chinese medicine is prepared from nine medicinal materials including pericarpium citri reticulatae, thunberg fritillary bulb, selfheal, oroxylum indicum, platycodon grandiflorum, radix scrophulariae, semen coicis, biond magnolia flower and divaricate saposhnikovia root, and is used for treating the child adenoid hypertrophy, and the method comprises the following steps: preparing a test solution, adopting an HP-5 capillary column, performing analysis under the chromatographic conditions that the split ratio is 30: 1, temperature programming is performed, the temperature of a sample inlet is 290 DEG C, and the temperature of a detector is 300 DEG C, and recording a chromatogram for 075 min. A standard fingerprint spectrum containing 19 common peaks is established by measuring a plurality of batches of samples, the attribution of each peak is defined, and five characteristic components such as eucalyptol and beta-bisabolol are identified. The method is good in stability and high in reproducibility, the constructed fingerprint spectrum is high in characteristic, the method can be used for quality consistency evaluation and production process control of traditional Chinese medicine, and safety and effectiveness of clinical medication are guaranteed.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine detection and relates to a gas chromatography (GC) fingerprint detection method for traditional Chinese medicine used to treat adenoid hypertrophy in children. Background Technology

[0002] Xiaoxiaoxian Granules are a Class I new traditional Chinese medicine under development by Jianmin Pharmaceutical Group Co., Ltd. It is an empirical formula developed by Dr. Chen Yonghui of the Children's Hospital Affiliated to the Capital Institute of Pediatrics based on extensive clinical practice. Composed of nine medicinal herbs including tangerine peel, fritillaria thunbergii, prunella vulgaris, oroxylum indicum, and platycodon grandiflorus, it has the effects of resolving phlegm and dissipating nodules, relieving sore throat and clearing nasal passages. It is mainly used for adenoid hypertrophy in children. Early clinical results show that the overall effective rate of this product for adenoid hypertrophy in children with phlegm accumulation is 90.15%, and it can significantly improve the main symptoms such as nasal congestion and snoring.

[0003] Traditional Chinese medicine compound preparations are characterized by synergistic effects of multiple components and targets, making quality control and efficacy assurance a persistent challenge in the industry. Traditional quality control methods typically perform qualitative or quantitative analysis on only one or a few indicator components, which is insufficient to comprehensively reflect the overall chemical composition of the preparation and cannot effectively monitor the consistency and stability of the finished drug quality, potentially affecting the clinical efficacy and safety of the drug.

[0004] Fingerprinting technology is a quality control method capable of comprehensively and macroscopically analyzing the chemical composition characteristics of complex material systems. It acquires overall information about multiple chemical components in a sample, characterizing its intrinsic quality in the form of a chromatogram. It is currently an internationally recognized effective means for quality evaluation of traditional Chinese medicine and natural drugs. Among these methods, gas chromatography (GC) is particularly suitable for the analysis of volatile components in traditional Chinese medicine due to its high separation efficiency, fast analysis speed, high sensitivity, and good reproducibility, and thus occupies an important position in the fingerprinting research of volatile components in traditional Chinese medicine.

[0005] Currently, there are no publicly reported methods for comprehensive quality control of Xiaoxiaosan Granules, especially GC fingerprinting methods that can reflect the overall characteristics of its volatile components. Therefore, establishing a specific and reproducible GC fingerprinting method for Xiaoxiaosan Granules is of great significance for scientifically evaluating its intrinsic quality and ensuring its clinical efficacy. Summary of the Invention

[0006] The purpose of this invention is to overcome the shortcomings of the prior art and provide a GC fingerprint detection method that is highly specific, has good repeatability, and can be used to comprehensively control the quality of Xiaoxian Granules.

[0007] To achieve the above objectives, the present invention adopts the following technical solution: A GC fingerprinting method for detecting a traditional Chinese medicine, wherein the traditional Chinese medicine is made from nine herbs: tangerine peel, fritillaria thunbergii, prunella vulgaris, oroxylum indicum, platycodon grandiflorus, scrophularia ningpoensis, coix seed, magnolia flower bud, and saposhnikovia divaricata, and is used to treat adenoid hypertrophy in children. The detection method includes the following steps: (1) After grinding the sample to be tested, extract it with methanol first, then extract the extract with ethyl acetate, evaporate the extract to dryness and reconstitute it to prepare the test solution; (2) The test solution is injected into a gas chromatograph for detection. The gas chromatograph contains a flame ionization detector. The chromatographic column is a quartz capillary column with 5% phenyl dimethyl polysiloxane as the stationary phase. The carrier gas is nitrogen. The injection port temperature is 250-290℃. The split injection method is adopted with a split ratio of 30:1. The temperature is programmed as follows: the initial column temperature is 75℃, held for 6 min, then increased to 145℃ at a rate of 5℃ per minute, held for 10 min; then increased to 215℃ at a rate of 7℃ per minute; finally increased to 280℃ at a rate of 9℃ per minute, held for 25 min. The detector temperature is 280-300℃. (3) Record the chromatogram of the test solution from 0 to 75 min. Using the Chinese Pharmacopoeia Commission's Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System, the chromatogram of the test solution is processed by data import, multi-point correction and data matching to obtain the fingerprint spectrum and perform similarity analysis.

[0008] Preferably, the quartz capillary column is of model HP-5.

[0009] More preferably, the quartz capillary column has a length of 30m, an inner diameter of 0.32mm, and a film thickness of 0.25μm.

[0010] Preferably, the injection volume of the test solution is 0.5-2 μl.

[0011] Preferably, the flow rate of the carrier gas is 0.5-1 ml / min.

[0012] Preferably, the fingerprint spectrum contains 19 common characteristic peaks, belonging to seven Chinese medicinal herbs: tangerine peel, fritillaria thunbergii, oroxylum indicum, platycodon grandiflorus, scrophularia ningpoensis, magnolia biondii, and saposhnikovia divaricata. Taking peak 6 as the reference peak, the relative retention times of the 19 common peaks are as follows: 0.7710, 0.8588, 0.8708, 0.8895, 0.9899, ​​1.0000, 1.0590, 1.1514, 1.2737, 1.3426, 1.3708, 2.0228, 2.0594, 2.1091, 2.2980, 2.7151, 2.7353, 2.8329, and 4.8254.

[0013] Preferably, the 19 common peaks belong to the following Chinese medicinal materials: peaks 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 18, and 19 belong to Magnolia biondii; peaks 4, 5, and 7 belong to Citrus reticulata peel; peaks 3, 4, 5, and 15 belong to Saposhnikovia divaricata; peaks 12 and 17 belong to Fritillaria thunbergii; peak 14 belongs to Oroxylum indicum; peak 15 belongs to Platycodon grandiflorus; and peaks 13 and 16 belong to Scrophularia ningpoensis.

[0014] More preferably, among the 19 common peaks, the following characteristic components were identified by comparison with reference standards: peak 3 is β-pinene, peak 5 is limonene, peak 6 is eucalyptol, peak 7 is γ-terpinene, and peak 15 is β-bisabolene.

[0015] The beneficial effects of this invention are: The GC detection method established in this invention enables efficient separation of complex volatile components in Xiaoxiaoxian granules. The chromatographic peaks are sharp and symmetrical, with good resolution and a reasonable analysis time (completed within 75 minutes). The method's stability, repeatability, and precision all meet the requirements of fingerprint spectroscopy technology. The established fingerprint spectrum contains 19 common peaks, comprehensively reflecting the overall characteristics of volatile components in the preparation. Through peak assignment and identification, the chromatographic features are directly correlated with the seven medicinal herbs in the prescription (Magnolia biondii, Citrus reticulata peel, Saposhnikovia divaricata, Fritillaria thunbergii, Oroxylum indicum, Platycodon grandiflorus, and Scrophularia ningpoensis) and specific chemical components (such as eucalyptol and β-bisabolene), demonstrating strong specificity.

[0016] The fingerprint spectrum constructed in this invention can be used for batch-to-batch quality consistency evaluation of Xiaoxian Granules for Children. By calculating the similarity between the fingerprint spectrum of the test sample and the standard spectrum, the stability of product quality can be quickly determined. Simultaneously, this method can be used for the identification and monitoring of raw materials, ensuring product quality from the source and guaranteeing the safety, efficacy, and reliability of clinical medication. This provides a scientific basis and technical support for the formulation of quality standards for Xiaoxian Granules for Children, and has promising prospects for industrial application. Attached Figure Description

[0017] Figure 1 Overlay spectrum of 15 batches of samples of Xiaoxiaoxian Granules (only up to 47 min).

[0018] Figure 2 GC standard fingerprint spectrum of Xiaoxiaoxian Granules for Children. Detailed Implementation

[0019] The present invention will be described in detail below using Xiaoxia Xiaogan Granules as an example through specific embodiments.

[0020] Instruments: Agilent 8860 gas chromatograph (Agilent Technologies, USA), automated liquid sampler, electronic gas path control (EPC) module, flame ionization detector (FID), programmed temperature column oven. Column: HP-5 capillary column (5% phenyl-95% dimethyl polysiloxane) (30m length, 0.32mm inner diameter, 0.25μm film thickness).

[0021] Test reagents: 15 batches of Xiaoxiaqian Granules for Children (250701-705, 250801-804, 250901-904, 251001-1002); ethyl acetate was analytical grade, and water was ultrapure water; γ-terpinene reference standard (Jiangxi Baicaoyuan Biotechnology Co., Ltd., batch number: 004224-202502, purity ≥95%); eucalyptol reference standard (Jiangxi Baicaoyuan Biotechnology Co., Ltd., batch number: 004224-202502, purity ≥95%). 1176-202305, purity ≥98%; β-bisabolene reference standard (Jiangxi Baicaoyuan Biotechnology Co., Ltd., batch number: 005567-202505, purity ≥95%); β-pinene reference standard (China National Institutes for Food and Drug Control, batch number: 111827-201202); limonene reference standard (China National Institutes for Food and Drug Control, batch number: 100470-202304, content 93.7%) The drug is provided by Jianmin Pharmaceutical Group Co., Ltd., and its prescription and process have been filed for another Chinese invention patent on December 15, 2025, patent application number 2025118885421. The ingredients are: 50g dried tangerine peel, 50g magnolia flower bud, 30g saposhnikovia root, 80g fritillaria bulb, 80g prunella vulgaris, 50g oroxylum indicum, 50g platycodon root, 80g scrophularia root, and 100g stir-fried coix seed. The volatile oils of dried tangerine peel, magnolia flower bud, and saposhnikovia root are extracted and set aside. The residue after extracting the volatile oils is mixed with the remaining medicinal materials, decocted with water, filtered, and the filtrate is concentrated into an extract. Sucrose, dextrin, and the volatile oils are then added to make granules.

[0022] Example 1: Establishment of GC fingerprint detection method 1. Detection Method (1) Preparation of test solution: Take 2.0g of test sample, accurately weigh it, add 50ml of methanol, sonicate for 30min, filter, extract the filtrate with ethyl acetate twice, 15ml each time, combine the ethyl acetate solutions, evaporate to dryness, add 2ml of ethyl acetate to the residue to dissolve it, shake well, filter with a microporous membrane, and take the filtrate as the test solution.

[0023] (2) Preparation of reference solutions: Accurately weigh appropriate amounts of β-bisabolene, β-pinene, limonene, γ-terpinene, and eucalyptol reference standards, place them in 2 ml volumetric flasks respectively, add ethyl acetate to dissolve them, and prepare 1 ml reference solutions containing 4.8825 mg / ml, 5.264 mg / ml, 5.0095 mg / ml, 5.697 mg / ml, and 5.0425 mg / ml of β-bisabolene, β-pinene, limonene, γ-terpinene, and eucalyptol respectively.

[0024] (3) Chromatographic conditions: Chromatographic column: HP-5 capillary column (stationary phase: 5% phenyl-95% dimethyl polysiloxane), with dimensions of 30 m × 0.32 mm × 0.25 μm.

[0025] Detector: Flame Ionization Detector (FID), temperature set at 300°C.

[0026] Inlet temperature: 290℃.

[0027] Carrier gas: high-purity nitrogen, flow rate 0.5 ml / min.

[0028] Split mode: Split injection, split ratio set to 30:1.

[0029] Temperature program: Initial column temperature 75℃, hold for 6 minutes; increase to 145℃ at a rate of 5℃ / min, hold for 10 minutes; then increase to 215℃ at a rate of 7℃ / min; finally increase to 280℃ at a rate of 9℃ / min, hold for 25 minutes.

[0030] Theoretical plate number: calculated based on the reference peak (eucalyptol peak), should be no less than 100,000.

[0031] (4) Sample determination: Take the sample of Xiaoxiaoxin Granules to be tested, prepare the test solution, accurately pipette 1 μl of the reference solution and the sample solution, and determine it under the above chromatographic conditions. Record the chromatogram from 0 to 75 min. Calculate the relative retention time and relative peak area in the chromatogram with peak 6 as the reference peak.

[0032] 2. Parameter selection and optimization (1) Determination of the split ratio The chromatograms obtained at three split ratios of 20:1, 30:1 and 50:1 were compared and analyzed. The elution time, resolution and peak area of ​​the main peaks are shown in Table 1.

[0033] Table 1 Comparison of fingerprint peak retention time, separation, and peak area for different split ratios

[0034] As shown in the table above, when the split ratio is 30:1, the separation of each chromatographic peak is better and the number of fingerprint characteristic spectra is greater. Therefore, the split ratio is determined to be 30:1.

[0035] (2) Optimization of programmed heating conditions Three different programmed heating conditions were investigated, and their specific conditions and corresponding separation effects are shown in Table 2.

[0036] Program 1: Initial column temperature 75℃, hold for 6 min; increase temperature to 145℃ at a rate of 5℃ / min and hold for 10 min; then increase temperature to 215℃ at a rate of 7℃ / min; finally increase temperature to 280℃ at a rate of 9℃ / min and hold for 25 min.

[0037] Procedure 2: Initial column temperature 75℃, hold for 10 min; increase temperature to 115℃ at a rate of 5℃ / min and hold for 15 min; then increase temperature to 170℃ at a rate of 8℃ / min; finally increase temperature to 250℃ at a rate of 15℃ / min, then increase temperature to 280℃ at a rate of 10℃ / min and hold for 15 min.

[0038] Procedure 3: Initial column temperature 75℃, hold for 10 min; increase temperature to 130℃ at a rate of 5℃ / min and hold for 9 min; then increase temperature to 260℃ at a rate of 8℃ / min; finally increase temperature to 280℃ at a rate of 5℃ / min and hold for 15 min.

[0039] Table 2 Comparison of fingerprint peak retention time, resolution, and peak area under different temperature ramping programs

[0040] Comparing the chromatograms obtained under the three programmed temperature conditions, Method 1 was selected as the final programmed temperature condition, with a total recording time of 0–75 min. The volatile components in the formulation were detected more comprehensively, the chromatographic peaks were better separated, and the fingerprint characteristics were clearer.

[0041] (3) Determination of the optimal detector temperature The flame ionization detector (FID) temperature is one of the important parameters affecting the gas chromatography response and baseline stability. During the optimization of gas chromatography conditions, the chromatograms obtained under FID temperatures of 280℃ and 300℃ were compared and analyzed. The specific conditions and corresponding separation effects are shown in Table 3.

[0042] Table 3 Comparison of fingerprint peak retention time, resolution, and peak area at different detector temperatures

[0043] The results showed that at 300℃, the chromatographic peaks of each component were sharp and symmetrical, with good separation, large peak areas, and the ability to accommodate the response intensity of both low-boiling and high-boiling components. Considering the number of chromatographic peaks, peak shape, separation, and signal response, 300℃ was ultimately determined to be the optimal detection temperature for fingerprint analysis using the FID (Finger ID).

[0044] Example 2, Methodological Investigation Precision, stability, and repeatability tests were conducted according to the requirements of fingerprint spectroscopy.

[0045] 1. Stability Test: The test solution was measured at 0, 4, 8, 12, 18, and 24 hours according to the chromatographic conditions described in Example 1. The relative retention time and relative peak area RSD of each common peak were examined. The results are shown in Tables 4 and 5. The RSD of the relative retention time of each chromatographic peak was less than 1.0, and the RSD of the relative peak area of ​​each chromatographic peak was less than 4.0, which meets the requirements of fingerprint chromatographic technology.

[0046] Table 4. Relative retention times for stability studies

[0047] Table 5 Relative peak area for stability assessment

[0048] 2. Repeatability Test: Six samples from the same batch were taken, and six test solutions were prepared according to the test solution preparation method in Example 1. The relative retention time and relative peak area RSD of each common peak were examined. The results showed that the RSD of the relative retention time of each chromatographic peak was less than 1.0, and the RSD of the relative peak area of ​​each chromatographic peak was less than 3.0, which met the requirements of fingerprint chromatographic technology.

[0049] 3. Precision Test: The test solution was injected six times consecutively under the chromatographic conditions described above, and the relative retention time and relative peak area of ​​each common peak were examined. The results showed that the RSD of the relative retention time of each chromatographic peak was less than 1.0, and the RSD of the relative peak area of ​​each chromatographic peak was less than 4.0, which meets the requirements of fingerprint chromatographic technology.

[0050] Example 3: Establishment of fingerprint chromatogram and assignment of chromatographic peaks 1. Establishment of fingerprint spectrum Fifteen batches of Xiaoxiaoxian Granules samples were collected, and test solutions were prepared. Chromatograms were measured and recorded at 75 min. All chromatographic peaks were concentrated within 75 min. Comparison of the chromatograms from the 15 batches identified 19 common peaks. Peak 6, with a retention time of 13.261 min, was selected as the reference peak. The chromatograms of the 15 batches of Xiaoxiaoxian Granules were imported into the "Traditional Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System (Version 2012A)" issued by the National Pharmacopoeia Commission. A time window width of 0.1 min was selected. After multi-point correction and data matching, a common pattern for chromatographic fingerprints was generated. The similarity calculation results of the fingerprints of the 15 batches are shown in Table 6. The superimposed fingerprints are shown in... Figure 1 .

[0051] Table 6. Similarity of 15 batches of samples

[0052] Establish the peak retention times and peak areas in gas chromatography, collect all detection data, calculate the average relative retention time and average relative peak area of ​​each corresponding peak, as well as the standard deviation, etc., and divide these values ​​by the average retention time and average peak area of ​​the reference peak to obtain the relative peak retention time and relative peak area values ​​for the standard fingerprint chromatogram. Derive the calculation results, formulate the standard fingerprint chromatogram data, and plot the standard fingerprint chromatogram. (See [link to standard fingerprint chromatogram]). Figure 2 The results are shown in Table 7.

[0053] Table 7 Standard fingerprint data

[0054] 2. Correlation between medicinal materials and fingerprint spectra Each herb in the prescription was granulated using the same process as Xiaoxia Xiaogan Granules. In addition, samples lacking any of the individual herbs were prepared. The chromatographic peaks were analyzed and assigned according to standard fingerprint chromatographic conditions. The results are shown in Table 8. The established fingerprint chromatographic peaks were assigned to the seven herbs in the prescription. Except for Prunella vulgaris and Coix lacryma-jobi, all other herbs had corresponding fingerprint peaks. Among them, Magnolia biondii, Citrus reticulata peel, and Saposhnikovia divaricata, which contain a large amount of volatile oil components, had a relatively large number of fingerprint peaks. The other four herbs, Fritillaria thunbergii, Oroxylum indicum, Platycodon grandiflorus, and Scrophularia ningpoensis, also had corresponding volatile component fingerprint peaks.

[0055] Table 8. Correlation analysis between pediatric anti-inflammatory granules and medicinal materials.

[0056] Furthermore, to verify the accuracy of the peak assignment results, a reference standard containing known volatile components in the formulation was introduced for comparative analysis. By comparing the retention time and peak shape characteristics of the sample chromatogram and the reference standard under the same chromatographic conditions, the assignment relationship of each peak was determined. The results showed that: peak 3 (11.665 min) was consistent with the retention time of β-pinene reference standards in Magnolia biondii and Saposhnikovia divaricata, and was a common peak of Magnolia biondii and Saposhnikovia divaricata; peak 5 (13.261 min) was consistent with the retention time of limonene reference standards in Magnolia biondii, Citrus reticulata peel and Saposhnikovia divaricata, and was a common peak of all three; peak 6 (13.396 min) was consistent with the peak of eucalyptol reference standard in Magnolia biondii, and was a characteristic peak of Magnolia biondii; peak 7 (14.186 min) was consistent with the peak of γ-terpinene reference standards in Magnolia biondii and Citrus reticulata peel, and was a common peak of Magnolia biondii and Citrus reticulata peel; peak 15 (30.784 min) was consistent with the peak of β-bisabolene reference standards in Saposhnikovia divaricata and Platycodon grandiflorus, and was a characteristic peak of Saposhnikovia divaricata and Platycodon grandiflorus.

[0057] The above results show that the present invention has successfully constructed the GC fingerprint spectrum of Xiaoxiaoxian Granules, which can be used for qualitative identification of the seven medicinal materials in the prescription, thereby reflecting the quality of traditional Chinese medicine in a more comprehensive way. The attribution of each common peak and characteristic peak is clear, and it can better reflect the composition characteristics of each medicinal material in the finished product.

Claims

1. A GC fingerprinting method for detecting a traditional Chinese medicine, wherein the traditional Chinese medicine is made from nine medicinal materials: tangerine peel, fritillaria thunbergii, prunella vulgaris, oroxylum indicum, platycodon grandiflorus, scrophularia ningpoensis, coix seed, magnolia flower bud, and saposhnikovia divaricata, and is used to treat adenoid hypertrophy in children, characterized in that... The detection method includes the following steps: (1) After grinding the sample to be tested, extract it with methanol first, then extract the extract with ethyl acetate, evaporate the extract to dryness and reconstitute it to prepare the test solution; (2) The test solution is injected into a gas chromatograph for detection. The gas chromatograph contains a flame ionization detector. The chromatographic column is a quartz capillary column with 5% phenyl dimethyl polysiloxane as the stationary phase. The carrier gas is nitrogen. The injection port temperature is 250-290℃. The split injection method is adopted with a split ratio of 30:

1. The temperature is programmed as follows: the initial column temperature is 75℃, held for 6 min, then increased to 145℃ at a rate of 5℃ per minute, held for 10 min; then increased to 215℃ at a rate of 7℃ per minute; finally increased to 280℃ at a rate of 9℃ per minute, held for 25 min. The detector temperature is 280-300℃. (3) Record the chromatogram of the test solution from 0 to 75 min. Using the Chinese Pharmacopoeia Commission's Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System, the chromatogram of the test solution is processed by data import, multi-point correction and data matching to obtain the fingerprint spectrum and perform similarity analysis.

2. The detection method as described in claim 1, characterized in that: The quartz capillary column is model HP-5.

3. The detection method as described in claim 2, characterized in that: The quartz capillary column has a length of 30m, an inner diameter of 0.32mm, and a film thickness of 0.25μm.

4. The detection method as described in claim 1, characterized in that: The injection volume of the test solution is 0.5-2 μl.

5. The detection method as described in claim 1, characterized in that: The flow rate of the carrier gas is 0.5-1 ml / min.

6. The detection method as described in claim 1, characterized in that: The fingerprint spectrum contains 19 common characteristic peaks, belonging to seven Chinese medicinal herbs: tangerine peel, fritillaria thunbergii, oroxylum indicum, platycodon grandiflorus, scrophularia ningpoensis, magnolia biondii, and saposhnikovia divaricata. Taking peak 6 as the reference peak, the relative retention times of the 19 common peaks are as follows: 0.7710, 0.8588, 0.8708, 0.8895, 0.9899, ​​1.0000, 1.0590, 1.1514, 1.2737, 1.3426, 1.3708, 2.0228, 2.0594, 2.1091, 2.2980, 2.7151, 2.7353, 2.8329, and 4.8254.

7. The detection method as described in claim 6, characterized in that, The 19 common peaks belong to the following Chinese medicinal materials: peaks 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 18, and 19 belong to Magnolia biondii; peaks 4, 5, and 7 belong to Citrus reticulata peel; peaks 3, 4, 5, and 15 belong to Saposhnikovia divaricata; peaks 12 and 17 belong to Fritillaria thunbergii; peak 14 belongs to Oroxylum indicum; peak 15 belongs to Platycodon grandiflorus; and peaks 13 and 16 belong to Scrophularia ningpoensis.

8. The detection method as described in claim 6, characterized in that, Among the 19 common peaks, the following characteristic components were identified by comparison with reference standards: peak 3 is β-pinene, peak 5 is limonene, peak 6 is eucalyptol, peak 7 is γ-terpinene, and peak 15 is β-bisabolene.

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