A method for determining GC fingerprint of Ganlu Xiaodu Pills and its application

Abstract

The present invention provides a method for determining the GC fingerprint of Ganlu Disinfection Pills. The present invention also provides the use of the above method for determining the GC fingerprint of Ganlu Disinfection Pills in the quality detection of the components in Ganlu Disinfection Pills and its quality detection method. The present invention further provides a method for determining the contents of the five components in the above Ganlu Disinfection Pills. The present invention further provides a method for screening the gas phase fingerprint of multiple medicinal materials in Ganlu Disinfection Pills. The present invention provides a method for determining the GC fingerprint of Ganlu Disinfection Pills and its application, establishes the gas phase fingerprint of Ganlu Disinfection Pills, can quantitatively analyze the five chemical components in Ganlu Disinfection Pills, and confirm the attribution of the chromatographic peaks of each single medicinal material in Ganlu Disinfection Pills, can effectively monitor from the source of the raw materials, strictly control the quality of the original medicinal materials, and can be used for quality control of the production process, thereby ensuring the safety and effectiveness of clinical medication.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicine component detection, and relates to a method for determining a GC fingerprint spectrum of Ganlu Xiaodu Pills and an application thereof. Background Art

[0002] Ganlu Xiaodu Pills are composed of 11 herbs, namely Patchouli, Forsythia, Fritillaria, Artemisia capillaris, Belamcanda, Cardamom, Akebia, Scutellaria, Talcum, Acorus calamus and Mentha. They are aromatic and dehumidifying, heat-clearing and detoxifying medicines, and are included in the 2020 edition of the Pharmacopoeia of the People's Republic of China. They are clinically used for summer heat and dampness accumulation, body heat and limb soreness, chest tightness and abdominal distension, and red urine and jaundice. Clinical studies have found that Ganlu Xiaodu Pills have a good therapeutic effect on infectious diseases such as influenza, mumps, atypical pneumonia, respiratory viral infections, and hand, foot and mouth disease. Experiments have also proved that it has a broad-spectrum antiviral effect and has a certain effect on influenza virus, Coxsackie virus, varicella-zoster virus, etc. It has also been found to be safe and effective for COVID-19.

[0003] Since Ganlu Xiaodu Pills contain many medicinal materials and complex chemical components, it is difficult to fully reflect the material basis and the integrity and complexity of the chemical component group of Ganlu Xiaodu Pills by identifying or testing a single compound, and it is difficult to fully control the intrinsic quality of Ganlu Xiaodu Pills. Traditional Chinese medicine fingerprints have the characteristics of overall, macroscopic, and fuzzy analysis. They can achieve the purpose of overall quality control by describing the overall characteristics of traditional Chinese medicine and using appropriate fuzzy processing methods. Therefore, they have become an effective means of quality control of traditional Chinese medicine. Among them, chromatographic fingerprint analysis can visualize the overall characteristics of various chemical components contained in traditional Chinese medicine, thereby revealing quality problems that are difficult to find in conventional inspections. Fingerprints are an important modern quality control method based on the overall comprehensive effects of multiple components and multiple targets of traditional Chinese medicine, starting from the perspective of "all ingredients", and are more comprehensive for the quality control of non-single ingredient drugs.

[0004] Ganlu Disinfection Pills contain a large amount of volatile components, and there is currently a lack of systematic qualitative research. Therefore, it is necessary to establish the GC fingerprint of Ganlu Disinfection Pills to complete the qualitative analysis of the volatile components in Ganlu Disinfection Pills in order to effectively and comprehensively control the quality of Ganlu Disinfection Pills. Summary of the invention

[0005] In view of the shortcomings of the prior art mentioned above, the purpose of the present invention is to provide a method for determining the GC fingerprint of Ganlu Disinfection Pills and its application. The gas phase fingerprint of Ganlu Disinfection Pills is established by using the pretreatment and gas chromatography method under optimized conditions. The systematic contribution of different types of chemical components to the gas phase fingerprint of Ganlu Disinfection Pills is highlighted from different aspects, the current status of each component in Ganlu Disinfection Pills is reflected more comprehensively, and a variety of specific indicator components in Ganlu Disinfection Pills are detected, providing a reference for the overall control and evaluation of the quality of Ganlu Disinfection Pills.

[0006] To achieve the above object and other related objects, the present invention provides a method for determining the GC fingerprint of Ganlu Xiaodu Pills in a first aspect, comprising the following steps:

[0007] 1) Preparation of test sample: headspace injection of Ganlu Xiaodu Pills sample was performed to obtain the test sample;

[0008] 2) Preparation of reference solution: adding at least one of α-pinene, eucalyptol, linalool, menthol and euphorbia alcohol reference substances to a solvent, dissolving the solution and making up to volume to prepare a reference solution;

[0009] 3) Determination: The test sample and reference solution were determined separately by gas chromatography (GC) under the same chromatographic conditions, the obtained gas phase fingerprint of the test sample was compared with the gas phase fingerprint of the reference solution, the index components in the gas phase fingerprint of the test sample were attributed and located, so as to obtain the gas phase fingerprint of Ganlu Disinfection Pills.

[0010] Preferably, in step 1), the Ganlu Xiaodu Pills sample is the Ganlu Xiaodu Pills powder obtained by crushing the Ganlu Xiaodu Pills raw drug and then sieving it.

[0011] Preferably, in step 1), in the headspace injection, the injection volume is 0.01-0.5 g, preferably 0.05-0.2 g, and more preferably 0.1 g.

[0012] Preferably, in step 1), during the headspace injection, the temperature of the heating box is 100-150°C, preferably 120-140°C, and more preferably 130°C.

[0013] Preferably, in step 1), in the headspace injection, the temperature of the quantitative loop is 120-150°C, preferably 130-140°C, and more preferably 135°C.

[0014] Preferably, in step 1), in the head space injection, the temperature of the transmission line is 130-150°C, preferably 135-145°C, more preferably 140°C.

[0015] Preferably, in step 1), in the head space injection, the head space equilibration time is 10-20 min, preferably 12-18 min, more preferably 15 min.

[0016] Preferably, in step 2), the CAS number of the α-pinene is 80-56-8, the CAS number of the eucalyptol is 470-82-6, the CAS number of the linalool is 78-70-6, the CAS number of the menthol is 89-78-1, and the CAS number of the linalool is 5986-55-0.

[0017] Preferably, in step 2), the solvent is anhydrous ethanol.

[0018] Preferably, in step 3), the gas phase fingerprint spectrum of the test sample is compared with the gas phase fingerprint spectrum of the reference solution, and the corresponding characteristic peaks in the gas phase fingerprint spectrum of the test sample are identified by relative retention time based on the known characteristic peaks in the gas phase fingerprint spectrum of the reference solution, thereby attributing and locating the indicator components in the gas phase fingerprint spectrum of the test sample.

[0019] Preferably, in step 3), the chromatographic column used in the gas chromatography is an Agilent HP-5 chromatographic column (30m×0.25mm, 0.25μm).

[0020] Preferably, in step 3), the detector used in the gas chromatography is a flame ionization detector (FID).

[0021] Preferably, in step 3), the temperature program used in the gas chromatography method is: the initial temperature is 58-62°C, and the temperature is increased to 68-72°C at a rate of 0.8-1.2°C / min; the temperature is increased to 93-97°C at a rate of 9-11°C / min; the temperature is increased to 99-101°C at a rate of 0.8-1.2°C / min; the temperature is increased to 109-111°C at a rate of 1.8-2.2°C / min, and maintained for 2-4 min; heat to 134-136°C at a rate of 18-22°C / min; heat to 139-141°C at a rate of 0.8-1.2°C / min, hold for 24-26 min; heat to 169-171°C at a rate of 2.8-3.2°C / min, hold for 1.8-2.2 min; heat to 239-241°C at a rate of 24-26°C / min, hold for 4.8-5.2 min.

[0022] More preferably, in the gas chromatography method, the temperature rising program adopted is: the initial temperature is 60°C, and the temperature is raised to 70°C at a rate of 1°C / min; the temperature is raised to 95°C at a rate of 10°C / min; the temperature is raised to 100°C at a rate of 1°C / min; the temperature is raised to 110°C at a rate of 2°C / min, and maintained for 3min; the temperature is raised to 135°C at a rate of 20°C / min; the temperature is raised to 140°C at a rate of 1°C / min, and maintained for 25min; the temperature is raised to 170°C at a rate of 3°C / min, and maintained for 2min; the temperature is raised to 240°C at a rate of 25°C / min, and maintained for 5min.

[0023] Preferably, in step 3), the gas chromatography method further comprises the following detection conditions:

[0024] The sample port temperature is 230-250°C; the injection volume is 0.5-2μL; the carrier gas is high-purity helium, and the carrier gas purity is ≥99.999%; the carrier gas flow rate is 0.1-1.0mL / min; the injection method is split injection, and the split ratio is 5-15:1.

[0025] More preferably, the gas chromatography method further comprises the following detection conditions:

[0026] The sample port temperature was 240°C; the injection volume was 1 μL; the carrier gas was high-purity helium with a carrier gas purity of ≥99.999%; the carrier gas flow rate was 0.4 mL / min; the injection method was split injection with a split ratio of 10:1.

[0027] The second aspect of the present invention provides an application of a method for determining the GC fingerprint of Ganlu Xiaodu Pills in quality detection of components in Ganlu Xiaodu Pills.

[0028] The third aspect of the present invention provides a quality detection method for Ganlu Disinfection Pills, comprising using the aforementioned method for determining the GC fingerprint of Ganlu Disinfection Pills to obtain a gas phase fingerprint of Ganlu Disinfection Pills, and comparing the obtained gas phase fingerprint of Ganlu Disinfection Pills with a standard gas phase fingerprint of Ganlu Disinfection Pills obtained under the same fingerprint detection conditions for similarity.

[0029] Preferably, when the gas phase fingerprint spectrum of Ganlu Xiaodu Pills is compared with the standard gas phase fingerprint spectrum of Ganlu Xiaodu Pills for similarity, the software of "Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System" 2012 version issued by the State Pharmacopoeia Committee is used for comparison. More preferably, the similarity between the gas phase fingerprint spectrum of Ganlu Xiaodu Pills measured by the present invention and the standard gas phase fingerprint spectrum of Ganlu Xiaodu Pills is ≥ 0.98.

[0030] More preferably, when the gas phase fingerprint spectrum of the Ganlu Disinfection Pills is matched with the standard gas phase fingerprint spectrum of the Ganlu Disinfection Pills for fingerprint common peaks, automatic matching is performed after multi-point correction when the time window width is 0.05-0.15min, preferably 0.1min, and the gas phase fingerprint spectrum and the standard gas phase fingerprint spectrum are generated by the average method.

[0031] Preferably, the standard gas phase fingerprint of Ganlu Xiaodu Pills is obtained under the same conditions as the aforementioned method for determining the GC fingerprint of Ganlu Xiaodu Pills, and the standard gas phase fingerprint of Ganlu Xiaodu Pills includes 20 common fingerprint peaks, with peak 18 as the reference peak (S peak, relative retention time of 1.000, relative peak area of ​​1.000), and the relative retention times and relative peak areas of the other 19 common fingerprint peaks are peak 1 (relative retention time of 0.242±0.002, relative peak area of ​​0.106±0.003), peak 2 (relative retention time of 0.274±0.003, relative peak area of ​​0.336±0.010), peak 3 (relative retention time of 0.3 14±0.003, relative peak area 0.148±0.004), peak 4 (relative retention time 0.318±0.003, relative peak area 2.147±0.064), peak 5 (relative retention time 0.348±0.003, relative peak area 0.180±0.005), peak 6 (relative retention time 0.375±0.004, relative peak area 0.174±0.005), peak 7 (relative retention time 0.377±0.004, relative peak area 0.119±0.003), peak 8 (relative retention time 0.446±0.004, relative peak area 0.195±0.006) , peak 9 (relative retention time: 0.450±0.004, relative peak area: 0.201±0.006), peak 10 (relative retention time: 0.453±0.005, relative peak area: 0.198±0.006), peak 11 (relative retention time: 0.462±0.005, relative peak area: 0.978±0.029), peak 12 (relative retention time: 0.467±0.005, relative peak area: 0.900±0.027), peak 13 (relative retention time: 0.706±0.007, relative peak area: 0.324±0.010), peak 14 (relative retention time: 0.789±0.0 The relative peak areas were as follows: peak 15 (relative retention time: 0.803±0.008, relative peak area: 0.566±0.017), peak 16 (relative retention time: 0.818±0.008, relative peak area: 0.191±0.006), peak 17 (relative retention time: 0.832±0.008, relative peak area: 0.258±0.008), peak 19 (relative retention time: 1.031±0.010, relative peak area: 3.396±0.102), peak 20 (relative retention time: 1.088±0.011, relative peak area: 0.752±0.023).

[0032] The specific data of the standard gas phase fingerprint of Ganlu Xiaodao Pills can be found inFigure 2 .

[0033] More preferably, the standard gas phase fingerprint of the Ganlu Disinfection Pills is compared with the gas phase fingerprint of the reference solution, and peak No. 1 is located and determined to be the fingerprint peak of α-pinene, peak No. 4 is the fingerprint peak of eucalyptol, peak No. 7 is the fingerprint peak of linalool, peak No. 9 is the fingerprint peak of menthol, and peak No. 20 is the fingerprint peak of linalool.

[0034] The fourth aspect of the present invention provides a method for determining the contents of five components in Ganlu Xiaodu Pills, comprising the following steps:

[0035] a) Preparation of test sample: the same as step 1) of the method for determining the GC fingerprint of Ganlu Xiaodu Pills;

[0036] b) Preparation of reference solution: the same as step 2) of the method for determining the GC fingerprint of Ganlu Xiaodu Pills;

[0037] c) Determination: The test sample of step a) and the reference solution of step b) were determined respectively by gas chromatography under the same chromatographic conditions as those in the method for determining the fingerprint of Ganlu Xiaodu Pills C, and the contents of the five components in the test sample were calculated by the external standard method.

[0038] Preferably, in step b), the reference solution is prepared by stepwise dilution.

[0039] Preferably, in step b), in the reference solution, the content of α-pinene is in the range of 100 to 1000 μg / mL, the content of eucalyptol is in the range of 100 to 1000 μg / mL, the content of linalool is in the range of 100 to 1000 μg / mL, the content of menthol is in the range of 50 to 500 μg / mL, and the content of spatholobiol is in the range of 50 to 500 μg / mL.

[0040] Preferably, in step c), the external standard method refers to: respectively taking a series of different volumes of the reference solution of step b), preparing a series of solutions of different concentrations, using gas chromatograph for sample analysis, obtaining the linear relationship between the content and peak area of ​​the five components in the reference solution, using the chromatographic peak area of ​​each component to correspond to its corresponding content, drawing the corresponding standard working curve, and calculating the regression equation of each standard working curve. Then the test sample is detected by gas chromatograph, and the chromatographic peak areas of the five components in the test sample are respectively substituted into the regression equation of each standard working curve to obtain the content of the corresponding component.

[0041] More preferably, in the standard working curve, the peak area is used as the ordinate and the content of each reference solution is used as the abscissa.

[0042] The fifth aspect of the present invention provides a method for screening gas phase fingerprints of multiple medicinal materials in Ganlu Xiaodu Pills, comprising the following steps:

[0043] A) Preparation of single medicinal material samples: prepare any one or more of the 11 medicinal material samples of Agastache rugosa, Forsythia suspensa, Fritillaria cirrhosa, Artemisia capillaris, Belamcanda chinensis, Cardamom, Akebia caulis, Scutellaria baicalensis, Talcum, Acorus calamus, and Mentha arvense according to step 1) of the method for determining the GC fingerprint of Ganlu Xiaodu Pills, and obtain at least one single medicinal material sample respectively;

[0044] B) Preparation of missing negative samples: remove any one of the 11 medicinal material samples of Patchouli, Forsythia, Fritillaria, Artemisia capillaris, Belamcanda, Cardamom, Akebia, Scutellaria, Talcum, Acorus calamus, and Mentha, and prepare according to step 1) of the method for determining the GC fingerprint of Ganlu Xiaodu Pills, and obtain 11 kinds of missing negative samples respectively;

[0045] C) Determination: The single medicinal material sample and 11 kinds of negative samples are respectively determined by gas chromatography (GC) under the same chromatographic conditions as in step 3) of the method for determining the GC fingerprint of Ganlu Xiaodu Pills, and the gas phase fingerprints of the single medicinal material sample and the 11 kinds of negative samples are respectively obtained;

[0046] D) Obtaining the standard gas phase fingerprint: using the same steps as the method for determining the GC fingerprint of Ganlu Xiaodu Pills, the standard gas phase fingerprint of Ganlu Xiaodu Pills is obtained;

[0047] E) Quality inspection: The gas phase fingerprint of the single medicinal material sample and the gas phase fingerprint of 11 types of negative samples were compared with the standard gas phase fingerprint of Ganlu Disinfection Pills, and the corresponding characteristic peaks of the single medicinal material sample in the standard gas phase fingerprint of Ganlu Disinfection Pills were identified by relative retention time, thereby attributing and locating the characteristic peaks in the gas phase fingerprint of the single medicinal material sample.

[0048] Preferably, in step A), the patchouli is the dried aerial part of Pogostemon cablin (Blanco) Benth. or Agastache rugosa (Fisch. Et Mey.) O.Ktze. of the Lamiaceae family.

[0049] Preferably, in step A), the forsythia is the dried fruit of Forsythia suspensa (Thunb.) Vahl of the Oleaceae family.

[0050] Preferably, in step A), the Fritillaria cirrhosa is the dried bulb of Fritillaria cirrhosa D.Don, Fritillaria unibracteata Hsiao et KCHsia, Fritillaria przewalskii Maxim. or Fritillaria delavayi Franch. of the Liliaceae family.

[0051] Preferably, in step A), the Artemisia scoparia is the dried aerial part of Artemisia scoparia Waldst.etKit. or Artemisia capillaris Thunb. of the Asteraceae family.

[0052] Preferably, in step A), the Belamcanda chinensis is the dried rhizome of Belamcanda chinensis (L.) DC. of the Iridaceae family.

[0053] Preferably, in step A), the cardamom is the dried mature fruit of Amomum kravanh Pierre ex Gagnep. or Amomum compactum Soland ex Maton, a plant of the Zingiberaceae family.

[0054] Preferably, in step A), the akebia is Akebia quinata (Thunb.) Decne., Akebia trifoliata (Thunb.) Koidz. or Akebia trfioliata of the Akebiaceae family.

[0055] Dried vines of (Thunb.) Koidz. var. australis (Diels) Rehd.

[0056] Preferably, in step A), the scutellaria baicalensis is the dried root of Scutellaria baicalensis Georgi, a plant of the Lamiaceae family.

[0057] Preferably, in step A), the talc is a silicate mineral talc, mainly containing a mineral containing hydrous magnesium silicate. The hydrous magnesium silicate is [Mg3(Si4O 10 )(OH)2】, which is included in the "Chinese Pharmacopoeia (2020 Edition) Volume 1".

[0058] Preferably, in step A), the Acorus tatarinox is the dried rhizome of Acorus tatarinoxjuilSchott, a plant of the Araceae family.

[0059] Preferably, in step A), the mint is the dried aerial part of Mentha haplocalyx Briq., a plant of the genus Mentha in the family Lamiaceae.

[0060] Preferably, in step E), the present invention locates the attribution of characteristic peaks of the measured gas fingerprint of the single medicinal material sample, the gas fingerprint of 11 kinds of missing negative samples and the standard gas fingerprint of Ganlu Xiaodu Pills, uses the "Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System" 2012 version software issued by the State Pharmacopoeia Committee for analysis and processing, and confirms the attribution of the characteristic peaks of each single medicinal material of Ganlu Xiaodu Pills by the relative retention time of each characteristic peak on the standard gas fingerprint of Ganlu Xiaodu Pills.

[0061] Preferably, in step E), among the single medicinal material samples, the fingerprint of the Patchouli sample includes two common fingerprint peaks, and the two common fingerprint peaks are peak 4 and peak 20. The specific results are shown in Table 1.

[0062] Preferably, in step E), among the single medicinal material samples, the fingerprint spectrum of the Forsythia suspensa sample includes 14 common fingerprint peaks, and the 14 common fingerprint peaks are peaks 1 to 12, peak 19, and peak 20. The specific results are shown in Table 1.

[0063] Preferably, in step E), among the single medicinal material samples, the fingerprint of the Artemisia capillaris sample includes 10 common fingerprint peaks, and the 10 common fingerprint peaks are peaks 4 to 12 and peak 19. The specific results are shown in Table 1.

[0064] Preferably, in step E), among the single medicinal material samples, the fingerprint spectrum of the Belamcanda chinensis sample includes 9 common fingerprint peaks, and the 9 common fingerprint peaks are peak 4, peaks 6-12, and peak 19. The specific results are shown in Table 1.

[0065] Preferably, in step E), among the single medicinal material samples, the fingerprint of the cardamom sample includes 5 common fingerprint peaks, and the 5 common fingerprint peaks are peak 3, peak 4, peak 11, peak 19, and peak 20. The specific results are shown in Table 1.

[0066] Preferably, in step E), among the single medicinal material samples, the fingerprint of the Akebia sample includes 4 common fingerprint peaks, and the 4 common fingerprint peaks are peak 11, peak 18, peak 19, and peak 20. The specific results are shown in Table 1.

[0067] Preferably, in step E), among the single medicinal material samples, the fingerprint spectrum of the Radix Scutellariae Root sample includes 15 common fingerprint peaks, and the 15 common fingerprint peaks are peak 4, peak 6, and peaks 8 to 20. The specific results are shown in Table 1.

[0068] Preferably, in step E), among the single medicinal material samples, the fingerprint of the Acorus tatarinowii sample includes 14 common fingerprint peaks, and the 14 common fingerprint peaks are peak 4, peak 6, and peaks 8 to 19. The specific results are shown in Table 1.

[0069] Preferably, in step E), among the single medicinal material samples, the fingerprint of the mint sample includes 4 common fingerprint peaks, and the 4 common fingerprint peaks are peak 4, peak 9, peak 11, and peak 12. The specific results are shown in Table 1.

[0070] Preferably, in step E), among the single medicinal material samples, the fingerprints of the Fritillaria cirrhosa sample and the talc sample do not include a common fingerprint peak. Specific results are shown in Table 1.

[0071] Table 1

[0072] Medicinal Materials Number of peaks Total fingerprint peak number Patchouli 2 4、20 Forsythia 14 1-12、19、20 Fritillaria cirrhosa 0 — Artemisia capillaris 10 4-12、19 cumshot 9 4、6-12、19 Cardamom 5 3、4、11、19、20 Akebia 4 11、18、19、20 Scutellaria baicalensis 15 4、6、8-20 talc 0 — Acorus 14 4、6、8-19 Mint 4 4、9、11、12

[0073] Among the above-mentioned common fingerprint peaks, peak 1 is determined to be the fingerprint peak of α-pinene, peak 4 is the fingerprint peak of eucalyptol, peak 7 is the fingerprint peak of linalool, peak 9 is the fingerprint peak of menthol, and peak 20 is the fingerprint peak of eucalyptol.

[0074] The water used in the present invention is all purified water.

[0075] As described above, the present invention provides a method for determining the GC fingerprint of Ganlu Xiaodu Pills and its application, and adopts headspace powder sampling and gas chromatography with optimized reaction conditions to establish a GC fingerprint method for Ganlu Xiaodu Pills. The present invention first established a GC fingerprint detection method for Ganlu Xiaodu Pills, which has good precision and repeatability, and determined 20 common peaks. The RSD of the relative retention time of each common peak is less than 0.04%. The similarity evaluation results of multiple batches of Zhengqi Tablets samples are all greater than 0.98, and the stability of the test solution can be maintained within 24 hours.

[0076] The method provided by the present invention can also quantitatively analyze five chemical components (α-pinene, eucalyptol, linalool, menthol, and linalool) in Ganlu Xiaodu Pills, thereby achieving one-time sample analysis and completing qualitative and quantitative analysis simultaneously.

[0077] The method provided by the present invention can also confirm the attribution of multiple single medicinal material chromatographic peaks in Ganlu Xiaodu Pills, and the gas phase fingerprint spectrum shows the characteristic peaks of Patchouli, Forsythia, Fritillaria, Artemisia Capillaris, Belamcanda, Cardamom, Akebia, Scutellaria, Acorus calamus, and Mentha. The method can realize effective monitoring from the source of raw materials, strictly control the quality of raw medicinal materials, and can be used for quality control of the production process, thereby ensuring the safety and effectiveness of clinical medication. BRIEF DESCRIPTION OF THE DRAWINGS

[0078] Figure 1 It shows the superimposed GC fingerprints of 15 batches of Ganlu Disinfection Pills in the present invention, wherein S1 to S15 are samples of 15 batches of Ganlu Disinfection Pills.

[0079] Figure 2 The standard GC fingerprint of the Ganlu Disinfection Pills of the present invention is shown, wherein 1: α-pinene, 4: eucalyptol, 7: linalool, 9: menthol, and 20: linalool.

[0080] Figure 3 The GC fingerprint of the reference substance of Ganlu Xiaodu Pills in the present invention is shown, wherein 1: α-pinene, 4: eucalyptol, 7: linalool, 9: menthol, and 20: linalool.

[0081] Figure 4A , 4B The GC fingerprint of Ganlu Xiaodu Pills in the present invention and the GC fingerprint of each single herbal medicine are shown as a comparative superimposed chromatogram, wherein: Figure 4A To compare the overall peaks of the superimposed chromatograms 1-20, Figure 4B For comparison, the local map of the common peaks of superimposed chromatograms 1-12; S1: Patchouli; S2: Forsythia; S3: Fritillaria; S4: Artemisia capillaris; S5: Belamcanda chinensis; S6: Cardamom; S7: Akebia; S8: Scutellaria baicalensis; S9: Talc; S10: Acorus calamus; S11: Mentha; S12: standard fingerprint of Ganlu Xiaodu Pills. DETAILED DESCRIPTION

[0082] The present invention is further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and are not used to limit the scope of protection of the present invention.

[0083] The following describes the embodiments of the present invention through specific examples, and those skilled in the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and the details in this specification can also be modified or changed in various ways based on different viewpoints and applications without departing from the spirit of the present invention.

[0084] The reagents and instruments used in the following examples are as follows:

[0085] 1. Reagents

[0086] Samples: 15 batches of Ganlu Disinfection Pills (batch numbers S1-20220201, S2-20220202, S3-20220203, S4-20220204, S5-20220205, S6-20220301, S7-20220311, S8-20220315, S9-20220324, S10-20220325, S11-20220506, S12-20220507, S13-20220509, S14-20220511, S15-20220512) were provided by Shanghai Hutchison Pharmaceuticals Co., Ltd.

[0087] Patchouli, Forsythia suspensa, Fritillaria cirrhosa, Artemisia capillaris, Belamcanda chinensis, Cardamom, Akebia trifoliata, Scutellaria baicalensis, Talc, Acorus calamus and Menthus sinensis were provided by Shanghai Hutchison Pharmaceuticals Co., Ltd. They are all commonly used Chinese medicinal materials and can be purchased from the market.

[0088] Reference substances: α-pinene (mass fraction 90.8%), eucalyptol (mass fraction 100%), linalool (mass fraction 99.7%), menthol (mass fraction 99.8%), and linalool (mass fraction 99.1%), all purchased from Shanghai Shidande Standard Technology Service Co., Ltd.

[0089] Reagents: Ultrapure water (prepared by Milli-Q ultrapure water treatment system).

[0090] 2. Instruments

[0091] 7890A gas chromatograph (USA Agilent Co. Ltd); AL204 and XS205 analytical electronic balances (METTLER TOLEDO Instrument Shanghai Co. Ltd); Milli-Q water purifier (USA Millipore Co. Ltd).

[0092] Example 1

[0093] 1. Sample pretreatment

[0094] Preparation of the test sample: Take 0.1g of Ganlu Xiaodu Pill powder sample, weigh it accurately, and place it in a 20mL headspace injection bottle for headspace injection to obtain test sample 1#. During the headspace injection, the heating box temperature is 130℃, the quantitative loop temperature is 135℃, the transmission line temperature is 140℃, and the headspace equilibrium time is 15min.

[0095] Preparation of reference solution: Take reference substances of α-pinene, eucalyptol, linalool, menthol and scholorhexyl alcohol, accurately weigh them, add anhydrous ethanol and dissolve them in a 100mL volumetric flask to make up the volume, so that each 1ml contains 800μg of α-pinene, eucalyptol, linalool, and 200μg of menthol and scholorhexyl alcohol.

[0096] 2. Chromatographic conditions

[0097] The chromatographic conditions of the gas chromatography method are: Agilent HP-5 chromatographic column (column length 30m×inner diameter 0.25mm, filler particle size 0.25μm); the detector is a flame ionization detector (FID); the sample port temperature is 240°C; the injection volume is 1μL; the carrier gas is high-purity helium, and the carrier gas purity is ≥99.999%; the carrier gas flow rate is 0.4mL / min; the injection method is split injection, and the split ratio is 10:1.

[0098] The temperature rising program adopted is as follows: the initial temperature is 60℃, and the temperature is raised to 70℃ at a rate of 1℃ / min; the temperature is raised to 95℃ at a rate of 10℃ / min; the temperature is raised to 100℃ at a rate of 1℃ / min; the temperature is raised to 110℃ at a rate of 2℃ / min, and kept for 3min; the temperature is raised to 135℃ at a rate of 20℃ / min; the temperature is raised to 140℃ at a rate of 1℃ / min, and kept for 25min; the temperature is raised to 170℃ at a rate of 3℃ / min, and kept for 2min; the temperature is raised to 240℃ at a rate of 25℃ / min, and kept for 5min.

[0099] 3. Determination

[0100] The gas chromatography method with the chromatographic conditions in the above step 2 is used to measure the test sample and the reference solution in the above step 1 respectively, and the gas phase fingerprint of the test sample and the gas phase fingerprint of the reference solution are obtained. The gas phase fingerprint of the test sample is compared with the gas phase fingerprint of the reference solution, and the corresponding characteristic peaks in the gas phase fingerprint of the test sample are identified by relative retention time according to the known characteristic peaks in the gas phase fingerprint of the reference solution, so as to attribute and locate the index components in the gas phase fingerprint of the test sample, and obtain the gas phase fingerprint of Ganlu Xiaodu Pills GC.

[0101] Example 2

[0102] 1. Sample pretreatment

[0103] Preparation of the test sample: Take 0.1g of Ganlu Xiaodu Pill powder sample, weigh it accurately, and place it in a 20mL headspace injection bottle for headspace injection to obtain test sample 2#. During the headspace injection, the heating box temperature is 135℃, the quantitative loop temperature is 138℃, the transfer line temperature is 142℃, and the headspace equilibrium time is 18min.

[0104] Preparation of reference solution: Take appropriate amount of α-pinene, eucalyptol, linalool, menthol, and scholorhexyl alcohol, weigh accurately, add anhydrous ethanol and dissolve in a 100mL volumetric flask to make up to volume, so that each 1ml contains 800μg of α-pinene, eucalyptol, linalool, 200μg of menthol and scholorhexyl alcohol.

[0105] 2. Chromatographic conditions

[0106] The chromatographic conditions of the gas chromatography method are: Agilent HP-5 chromatographic column (column length 30m×inner diameter 0.25mm, filler particle size 0.25μm); the detector is a flame ionization detector (FID); the sample port temperature is 245°C; the injection volume is 2μL; the carrier gas is high-purity helium, and the carrier gas purity is ≥99.999%; the carrier gas flow rate is 0.6mL / min; the injection method is split injection, and the split ratio is 12:1.

[0107] The temperature rising program adopted was as follows: the initial temperature was 62°C, and the temperature was raised to 72°C at a rate of 0.8°C / min; the temperature was raised to 97°C at a rate of 11°C / min; the temperature was raised to 101°C at a rate of 1.2°C / min; the temperature was raised to 111°C at a rate of 2.2°C / min, and kept for 2 min; the temperature was raised to 136°C at a rate of 18°C / min; the temperature was raised to 141°C at a rate of 0.8°C / min, and kept for 26 min; the temperature was raised to 171°C at a rate of 3.2°C / min, and kept for 2.2 min; the temperature was raised to 241°C at a rate of 24°C / min, and kept for 5.2 min.

[0108] 3. Determination

[0109] The specific determination process is the same as step 3 in Example 1.

[0110] Example 3

[0111] The method for determining the UC fingerprint of Ganlu Xiaodu Pills established in the above Example 1 was used to detect 15 batches of Ganlu Xiaodu Pills (S1-S15), and the chromatograms were recorded. The fingerprint data of the test sample was imported into the 2012 version of the software "Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System" issued by the State Pharmacopoeia Committee, and automatic matching was performed after multi-point correction (time window of 0.1 min). The gas phase fingerprint of the test sample and the standard gas phase fingerprint of Ganlu Xiaodu Pills were generated by the average method. The specific spectra are shown in Figure 1 , 2 The gas phase fingerprint of the test sample was compared with the standard gas phase fingerprint of Ganlu Disinfection Pills obtained under the same fingerprint detection conditions, and the similarity was calculated. The specific calculation results are shown in Table 2. As shown in Table 2, the similarity of the 15 batches of samples is above 0.98, which meets the requirements.

[0112] Table 2 Similarity results

[0113]

[0114] Depend on Figure 2The standard gas phase fingerprint of Ganlu Xiaodu Pills can be seen. According to the relative retention time of each chromatographic peak in the chromatogram, the common peaks are determined, and 20 common peaks are selected as characteristic fingerprint peaks. Peak 18 is used as the reference peak (S), and the relative retention time and relative peak area of ​​each chromatographic peak are calculated. The calculation results are shown in Table 3. The results show that the standard gas phase fingerprint of Ganlu Xiaodu Pills includes 20 common fingerprint peaks, with peak 18 as the reference peak (S peak, relative retention time of 1.000, relative peak area of ​​1.000), and the relative retention time and relative peak area of ​​the other 19 common fingerprint peaks are peak 1 (relative retention time of 0.242, relative peak area of ​​0.106), peak 2 (relative retention time of 0.274, relative peak area of ​​0.336), peak 3 (relative retention time of 0. 314, relative peak area 0.148), peak 4 (relative retention time 0.318, relative peak area 2.147), peak 5 (relative retention time 0.348, relative peak area 0.180), peak 6 (relative retention time 0.375, relative peak area 0.174), peak 7 (relative retention time 0.377, relative peak area 0.119), peak 8 (relative retention time 0.446, relative peak area 0.195), peak 9 (relative retention time 0.349, relative peak area 0.180), peak 10 (relative retention time 0.344, relative peak area 0.181), peak 11 (relative retention time 0.342, relative peak area 0.180), peak 12 (relative retention time 0.340, relative peak area 0.181), peak 13 (relative retention time 0.344, relative peak area 0.181), peak 14 (relative retention time 0.340, relative peak area 0.181), peak 15 (relative retention time 0.340, relative peak area 0.181), peak 16 (relative retention time 0.340, relative peak area 0.181), peak 17 (relative retention time 0.340, relative peak area 0.181), peak 18 (relative retention time 0.340, relative peak area Peak 10 (relative retention time 0.453, relative peak area 0.19), peak 11 (relative retention time 0.462, relative peak area 0.978), peak 12 (relative retention time 0.467, relative peak area 0.900), peak 13 (relative retention time 0.706, relative peak area 0.324), peak 14 (relative retention time 0.789, relative peak area 0.944), peak 15 (relative retention time 0.890, relative peak area 0.822), peak 16 (relative retention time 0.842, relative peak area 0.830), peak 17 (relative retention time 0.864, relative peak area 0.871), peak 18 (relative retention time 0.844, relative peak area 0.863), peak 19 (relative retention time 0.822, relative peak area 0.831), peak 20 (relative retention time 0.844, relative peak area 0.863), peak 21 (relative retention time 0.844, relative peak area 0.863), peak 22 (relative retention time 0.844, relative peak area 0.863), peak 23 (relative retention time 0.844, relative peak area , relative peak area is 0.257), peak 15 (relative retention time is 0.803, relative peak area is 0.566), peak 16 (relative retention time is 0.818, relative peak area is 0.191), peak 17 (relative retention time is 0.832, relative peak area is 0.258), peak 19 (relative retention time is 1.031, relative peak area is 3.396), peak 20 (relative retention time is 1.088, relative peak area is 0.752).

[0115] As shown in Table 3, the RSD of the relative retention time of different samples is 0.00%; the RSD of the relative peak area is between 0.31% and 28.12%. It can be seen that the peak time of the common substances in each batch of samples is basically the same, the composition is relatively stable, but the amount varies greatly.

[0116] Table 3 Relative retention time and relative peak area of ​​common peaks

[0117]

[0118] At the same time, through Figure 3 By comparing the chromatogram of the reference substance shown in the figure, it was determined that in the standard gas phase fingerprint of Ganlu Disinfection Pills, peak 1 is α-pinene, peak 4 is eucalyptol, peak 7 is linalool, peak 9 is menthol, and peak 20 is linalool.

[0119] Example 4

[0120] The method for determining the GC fingerprint of Ganlu Xiaodu Pills in the present invention was methodologically verified, and the performance index results are as follows.

[0121] 1. Precision

[0122] Take 1 sample of Ganlu Xiaodu Pills (batch number 20220201), prepare the test sample according to step 1 in the above embodiment 1, detect according to the chromatographic conditions of step 2 in the above embodiment 1, inject 6 times continuously, record the chromatogram, take peak 18 as the reference peak, examine the relative retention time of the common peaks specified in the fingerprint, and the results show that the RSD of the relative retention time of the 20 common peaks is less than 0.04%, and the RSD of the relative retention area is less than 3%. The results show that the instrument has good precision.

[0123] 2. Repeatability

[0124] Take 6 samples of Ganlu Xiaodu Pills (batch number 20220201), prepare the test sample according to step 1 in the above embodiment 1, detect according to the chromatographic conditions of step 2 in the above embodiment 1, take peak 18 as the reference peak, calculate the relative retention time of each common peak and the RSD of the relative peak area, and the results show that the relative retention time RSD of the 20 common peaks is less than 0.04%, and the relative peak area RSD is less than 3%, indicating that the method of the present invention has good repeatability and high accuracy.

[0125] 3. Stability

[0126] Take a sample of Ganlu Xiaodu Pills (batch number 20220201), prepare the test sample according to step 1 in the above Example 1, and place it for 0h, 3h, 6h, 9h, 12h, and 24h respectively, and then detect it according to the chromatographic conditions of step 2 in the above Example 1, with peak 18 as the reference peak. As a result, the relative retention time RSD of the 20 common peaks is less than 0.04%, and the relative peak area RSD is less than 5%, indicating that the test sample has good stability within 24h.

[0127] Example 5

[0128] 1. Sample pretreatment

[0129] Preparation of the test sample: The preparation process of the test sample is the same as that in step 1 of Example 1.

[0130] Preparation of reference solution: The preparation process of reference solution in step 1 of Example 1 is the same. The reference solution is prepared by stepwise dilution. In the reference solution, the content of α-pinene is in the range of 100 to 1000 μg / mL, the content of eucalyptol is in the range of 100 to 1000 μg / mL, the content of linalool is in the range of 100 to 1000 μg / mL, the content of menthol is in the range of 50 to 500 μg / mL, and the content of eucalyptol is in the range of 50 to 500 μg / mL.

[0131] 2. Chromatographic conditions

[0132] The chromatographic conditions of the gas chromatography are the same as those of the gas chromatography in step 2 in Example 1.

[0133] 3. Determination

[0134] Using the external standard method, a series of different volumes of reference solution were pipetted to prepare a series of solutions with different concentrations, and the samples were analyzed by gas chromatography to draw a standard working curve. Then, the obtained test sample was analyzed by gas chromatography after headspace injection, and the analysis results were substituted into the standard working curve to obtain the contents of the five components in the test sample.

[0135] Specifically, a series of different volumes of reference solution were pipetted to prepare a series of solutions with different concentrations, and gas chromatography was used for sample analysis to obtain the linear relationship between the content and peak area of ​​the five components in the reference solution. The chromatographic peak area of ​​each component corresponded to its corresponding content, and the corresponding standard working curve was drawn to calculate the regression equation of each standard working curve. The headspace of the test sample was then sampled and tested by gas chromatography, and the chromatographic peak areas of the five components in the test sample were respectively substituted into the regression equation of each standard working curve to obtain the content of the corresponding component.

[0136] Example 6

[0137] The test sample was prepared by using step 1 in the above embodiment 1.

[0138] Take Patchouli, Forsythia suspensa, Fritillaria cirrhosa, Artemisia capillaris, Belamcanda chinensis, Cardamom, Akebia trifoliata, Scutellaria baicalensis, Talcum, Acorus calamus, and Mentha arvense respectively, and adopt step 1 in the above embodiment 1 to prepare 11 single medicinal material samples.

[0139] Prepare the missing negative samples, which are obtained by removing one medicinal material from the 11 medicinal material samples, and adopt step 1 in the above embodiment 1 to prepare 11 kinds of missing negative samples.

[0140] According to step 2 in Example 1, the test sample, 11 single medicinal material samples, and 11 missing negative samples were measured respectively, and the gas phase fingerprints of the test sample, 11 single medicinal material samples, and 11 missing negative samples were obtained respectively. The obtained gas phase fingerprints of the test sample, 11 single medicinal material samples, and 11 missing negative samples were introduced into the "Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System" 2012 version software issued by the State Pharmacopoeia Committee for analysis and processing, and at the same time, according to step 3 in Example 1, the standard gas phase fingerprint of Ganlu Xiaodu Pills was obtained.

[0141] The gas phase fingerprints of the test sample, 11 single medicinal material samples, and 11 missing negative samples were compared with the standard gas phase fingerprint of Ganlu Xiaodu Wan, and the corresponding characteristic peaks of the 11 single medicinal material samples in the standard gas phase fingerprint of Ganlu Xiaodu Wan were identified by relative retention time, so as to attribute and locate the characteristic peaks in the gas phase fingerprint of the 11 single medicinal material samples. The specific results are shown in Figure 4A , Figure 4B , Table 1. The gas phase fingerprints of the 11 negative samples can be used for comparison to confirm the source of the characteristic peaks.

[0142] Depend on Figure 4A , Figure 4B As shown in Table 1, peaks 1 and 2 are derived from Artemisia capillaris; peak 3 is derived from Forsythia suspensa and cardamom; peak 4 is derived from various medicinal materials (except talc and Fritillaria cirrhosa); peak 5 is derived from Forsythia suspensa and Artemisia capillaris; peak 6 is derived from Forsythia suspensa, Artemisia capillaris, Belamcanda chinensis, Scutellaria baicalensis and Acorus tatarinowii; peak 7 is derived from Forsythia suspensa, Artemisia capillaris, Belamcanda chinensis; peak 8 is derived from Forsythia suspensa, Artemisia capillaris, Belamcanda chinensis, Scutellaria baicalensis and Acorus tatarinowii; peak 9 is derived from Forsythia suspensa, Artemisia capillaris, Belamcanda chinensis, Scutellaria baicalensis, Acorus tatarinowii and Mentha chinensis; peak 10 is derived from Peak No. 11 comes from various medicinal materials (except Patchouli, Fritillaria cirrhosa and talc); Peak No. 12 comes from Forsythia suspensa, Capillaries, Belamcanda radix, Scutellaria baicalensis, Acorus calamus and Mentha arvense; Peak No. 13-17 come from Scutellaria baicalensis and Acorus calamus; Peak No. 18 comes from Akebia trifoliata, Scutellaria baicalensis and Acorus calamus; Peak No. 19 comes from various medicinal materials (except Patchouli trifoliata, Fritillaria cirrhosa, talc and Mentha arvense); Peak No. 20 comes from Patchouli trifoliata, Forsythia suspensa, Cardamom, Akebia trifoliata and Scutellaria baicalensis.

[0143] Specifically, the fingerprint of the Agastache rugosa sample includes 2 common fingerprint peaks, which are peak 4 and peak 20; the fingerprint of the Forsythia suspensa sample includes 14 common fingerprint peaks, which are peaks 1-12, peak 19, and peak 20; the fingerprint of the Artemisia capillaris sample includes 10 common fingerprint peaks, which are peaks 4-12 and peak 19; the fingerprint of the Belamcanda chinensis sample includes 9 common fingerprint peaks, which are peak 4, peaks 6-12, and peak 19; the fingerprint of the Cardamom sample includes 5 common fingerprint peaks, which are peaks 3, 4, 11, 1 The fingerprint of the Akebia sample includes 4 common fingerprint peaks, which are Peak 11, Peak 18, Peak 19 and Peak 20; the fingerprint of the Scutellaria sample includes 15 common fingerprint peaks, which are Peak 4, Peak 6, and Peaks 8-20; the fingerprint of the Acorus calamus sample includes 14 common fingerprint peaks, which are Peak 4, Peak 6, and Peaks 8-19; the fingerprint of the mint sample includes 4 common fingerprint peaks, which are Peak 4, Peak 9, Peak 11 and Peak 12; the fingerprints of the Fritillaria cirrhosa sample and the talc sample do not include common fingerprint peaks.

[0144] Among the above common fingerprint peaks, Figure 3 As shown, after comparison, it is determined that peak 1 is the fingerprint peak of α-pinene, peak 4 is the fingerprint peak of eucalyptol, peak 7 is the fingerprint peak of linalool, peak 9 is the fingerprint peak of menthol, and peak 20 is the fingerprint peak of linalool.

[0145] It can be seen that the chemical characteristic peaks of the Chinese medicinal materials in Ganlu Xiaodu Pills are well reflected in the fingerprint spectrum and their attribution is confirmed.

[0146] In summary, the present invention provides a method for determining the UC fingerprint of Ganlu Xiaodu Pills and its application, establishes the gas phase fingerprint of Ganlu Xiaodu Pills, can quantitatively analyze the five chemical components in Ganlu Xiaodu Pills, and confirm the attribution of the chromatographic peaks of each single medicinal material in Ganlu Xiaodu Pills, can effectively monitor the source of raw materials, strictly control the quality of raw medicinal materials, and can be used for quality control of the production process, thereby ensuring the safety and effectiveness of clinical medication. Therefore, the present invention overcomes various shortcomings in the prior art and has a high industrial utilization value.

[0147] The above embodiments are merely illustrative of the principles and effects of the present invention, and are not intended to limit the present invention. Anyone familiar with the art may modify or alter the above embodiments without departing from the spirit and scope of the present invention. Therefore, all equivalent modifications or alterations made by a person of ordinary skill in the art without departing from the spirit and technical concept disclosed by the present invention shall still be covered by the claims of the present invention.

Claims

1. A method for determining the GC fingerprint of Ganlu Xiaodu Pills, comprising the following steps: 1) Preparation of test sample: headspace injection of Ganlu Xiaodu Pills sample was performed to obtain the test sample; 2) Preparation of reference solution: adding at least one of α-pinene, eucalyptol, linalool, menthol and euphorbia alcohol reference substances to a solvent, dissolving the solution and making up to volume to prepare a reference solution; 3) Determination: The test sample and the reference solution are respectively determined by gas chromatography under the same chromatographic conditions, the obtained gas phase fingerprint of the test sample is compared with the gas phase fingerprint of the reference solution, the index components in the gas phase fingerprint of the test sample are attributed and located, so as to obtain the gas phase fingerprint of Ganlu Xiaodu Pills; In step 1) or 2), the following conditions are included: A1) In step 1), the Ganlu Xiaodu Pills sample is the Ganlu Xiaodu Pills powder obtained by crushing the Ganlu Xiaodu Pills crude drug and then sieving it; A2) In step 1), the headspace injection conditions are: injection volume 0.01-0.5 g; heating box temperature 100-150°C; The temperature of the quantitative loop is 120-150°C; the temperature of the transfer line is 130-150°C; the headspace equilibrium time is 10-20 minutes; A3) In step 2), the solvent is anhydrous ethanol; In step 3), the detection conditions of the gas chromatography method are as follows: the chromatographic column is an Agilent HP-5 chromatographic column, 30m×0.25mm, 0.25μm; the detector is a flame ionization detector; the temperature program used is: the initial temperature is 58-62°C, and the temperature is increased to 68-72°C at a rate of 0.8-1.2°C / min; the temperature is increased to 93-97°C at a rate of 9-11°C / min; the temperature is increased to 99-101°C at a rate of 0.8-1.2°C / min; the temperature is increased to 109-110°C at a rate of 1.8-2 ... 111℃, hold for 2-4min; heat to 134-136℃ at a rate of 18-22℃ / min; heat to 139-141℃ at a rate of 0.8-1.2℃ / min, hold for 24-26min; heat to 169-171℃ at a rate of 2.8-3.2℃ / min, hold for 1.8-2.2min; heat to 239-241℃ at a rate of 24-26℃ / min, hold for 4.8-5.2min.

2. The method for determining the GC fingerprint of Ganlu Xiaodu Pills according to claim 1, characterized in that: In step 3), the gas chromatography method also includes the following detection conditions: the sample port temperature is 230-250°C; the injection volume is 0.5-2μL; the carrier gas is high-purity helium, and the carrier gas purity is ≥99.999%; the carrier gas flow rate is 0.1-1.0mL / min; the injection method is split injection, and the split ratio is 5-15:

1.

3. Use of the method for determining the GC fingerprint of Ganlu Xiaodu Pills according to any one of claims 1-2 in the quality detection of the ingredients in Ganlu Xiaodu Pills.

4. A quality detection method for Ganlu Disinfection Pills, comprising obtaining a gas phase fingerprint of Ganlu Disinfection Pills by using the method for determining the GC fingerprint of Ganlu Disinfection Pills as described in any one of claims 1-2, and comparing the obtained gas phase fingerprint of Ganlu Disinfection Pills with a standard gas phase fingerprint of Ganlu Disinfection Pills obtained under the same fingerprint detection conditions for similarity.

5. The quality inspection method of Ganlu disinfection pills according to claim 4, characterized in that: The standard gas phase fingerprint of Ganlu Disinfection Pills is obtained under the same conditions as the method for determining the GC fingerprint of Ganlu Disinfection Pills according to any one of claims 1-2, wherein the standard gas phase fingerprint of Ganlu Disinfection Pills includes 20 common fingerprint peaks, with peak No. 18 as the reference peak S peak, the relative retention time is 1.000, and the relative peak area is 1.000; the relative retention times and relative peak areas of the other 19 common fingerprint peaks are: Peak 1, relative retention time 0.242 ± 0.002, relative peak area 0.106 ± 0.003; Peak 2, relative retention time 0.274 ± 0.003, relative peak area 0.336 ± 0.010; Peak 3, relative retention time 0.314 ± 0.003, relative peak area 0.148 ± 0.004; Peak 4, relative retention time 0.318 ± 0.003, relative peak area 2.147 ± 0.064; Peak 5, relative retention time 0.348 ± 0.003, relative peak area 0.180 ± 0.005; Peak 6, relative retention time 0.375 ± 0.004, relative peak area 0.174 ± 0.005; Peak 7, relative retention time 0.377 ± 0.004, relative peak area 0.119 ± 0.003; Peak 8, relative retention time 0.446 ± 0.004, relative peak area 0.195 ± 0.006; Peak 9, relative retention time 0.450 ± 0.004, relative peak area 0.201 ± 0.006; Peak 10, relative retention time 0.453 ± 0.005, relative peak area 0.198 ± 0.006; Peak 11, relative retention time 0.462 ± 0.005, relative peak area 0.978 ± 0.029; Peak 12, relative retention time 0.467 ± 0.005, relative peak area 0.900 ± 0.027; Peak 13, relative retention time 0.706 ± 0.007, relative peak area 0.324 ± 0.010; Peak 14, relative retention time 0.789 ± 0.008, relative peak area 0.257 ± 0.008; Peak 15, relative retention time 0.803 ± 0.008, relative peak area 0.566 ± 0.017; Peak 16, relative retention time 0.818 ± 0.008, relative peak area 0.191 ± 0.006; Peak 17, relative retention time 0.832 ± 0.008, relative peak area 0.258 ± 0.008; Peak 19, relative retention time 1.031 ± 0.010, relative peak area 3.396 ± 0.102; Peak No. 20, the relative retention time is 1.088±0.011, and the relative peak area is 0.752±0.

023.

6. A method for determining the contents of five components in Ganlu Xiaodu Pills, comprising the following steps: a) Preparation of the test sample: the same as step 1) of the method for determining the GC fingerprint of Ganlu Xiaodu Pills according to any one of claims 1-2; b) Preparation of reference solution: the same as step 2) of the method for determining the GC fingerprint of Ganlu Xiaodu Pills according to any one of claims 1-2; c) Determination: Using gas chromatography under the same chromatographic conditions as in the method for determining the fingerprint of Ganlu Xiaodu Pills C as described in any one of claims 1-2, respectively determine the test sample of step a) and the reference solution of step b), and calculate the contents of the five components in the test sample by the external standard method.

7. A method for screening gas phase fingerprints of multiple medicinal materials in Ganlu Xiaodu Pills, comprising the following steps: A) Preparation of single medicinal material samples: prepare any one or more of the 11 medicinal material samples of Agastache rugosa, Forsythia suspensa, Fritillaria cirrhosa, Artemisia capillaris, Belamcanda chinensis, Cardamom, Akebia caulis, Scutellaria baicalensis, Talcum, Acorus calamus, and Mentha arvense according to step 1) of the method for determining the GC fingerprint of Ganlu Xiaodu Pills according to any one of claims 1 to 2, and obtain at least one single medicinal material sample respectively; B) Preparation of missing negative samples: remove any one of the 11 medicinal material samples of Patchouli, Forsythia, Fritillaria, Artemisia capillaris, Belamcanda, Cardamom, Akebia, Scutellaria, Talcum, Acorus calamus, and Mentha, and prepare according to step 1) of the method for determining the GC fingerprint of Ganlu Xiaodu Pills according to any one of claims 1-2, to obtain 11 missing negative samples respectively; C) Determination: using gas chromatography under the same chromatographic conditions as in step 3) of the method for determining the GC fingerprint of Ganlu Xiaodu Pills according to any one of claims 1-2 to respectively determine the single medicinal material sample and 11 kinds of missing negative samples, and obtain gas phase fingerprints of the single medicinal material sample and 11 kinds of missing negative samples respectively; D) Obtaining the standard gas phase fingerprint: using the same steps as the method for determining the GC fingerprint of Ganlu Xiaodu Pills according to any one of claims 1-2 to obtain the standard gas phase fingerprint of Ganlu Xiaodu Pills; E) Quality inspection: The gas phase fingerprint of the single medicinal material sample and the gas phase fingerprint of 11 types of negative samples were compared with the standard gas phase fingerprint of Ganlu Disinfection Pills, and the corresponding characteristic peaks of the single medicinal material sample in the standard gas phase fingerprint of Ganlu Disinfection Pills were identified by relative retention time, thereby attributing and locating the characteristic peaks in the gas phase fingerprint of the single medicinal material sample.

8. The method for screening gas phase fingerprints of multiple medicinal materials in Ganlu Xiaodu Pills according to claim 7, characterized in that: In step E), any one or more of the following conditions are included: B1) The fingerprint of the Agastache rugosa sample includes 2 common fingerprint peaks, which are peak 4 and peak 20; B2) The fingerprint of the Forsythia suspensa sample includes 14 common fingerprint peaks, which are peaks 1-12, peak 19, and peak 20; B3) The fingerprint of the Artemisia capillaris sample includes 10 common fingerprint peaks, and the 10 common fingerprint peaks are peaks 4-12 and peak 19; B4) The fingerprint of the Belamcanda chinensis sample includes 9 common fingerprint peaks, wherein the 9 common fingerprint peaks are peak 4, peaks 6-12, and peak 19; B5) The fingerprint of the cardamom sample includes 5 common fingerprint peaks, and the 5 common fingerprint peaks are peak 3, peak 4, peak 11, peak 19, and peak 20; B6) The fingerprint of the Akebia sample includes 4 common fingerprint peaks, which are peak 11, peak 18, peak 19, and peak 20; B7) The fingerprint of the Scutellaria baicalensis sample includes 15 common fingerprint peaks, and the 15 common fingerprint peaks are peak 4, peak 6, and peaks 8-20; B8) The fingerprint of the Acorus tatarinowii sample includes 14 common fingerprint peaks, and the 14 common fingerprint peaks are peak 4, peak 6, and peaks 8-19; B9) The fingerprint of the mint sample includes 4 common fingerprint peaks, which are peak 4, peak 9, peak 11 and peak 12; B10) The fingerprints of Fritillaria cirrhosa and talc samples do not include common fingerprint peaks.

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