CN109668969B - Gas chromatography detection method of traditional Chinese medicine composition - Google Patents
Gas chromatography detection method of traditional Chinese medicine composition Download PDFInfo
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Abstract
The invention provides a detection method of a traditional Chinese medicine composition, which is used for quantitatively determining main representative chemical components in a preparation by using a Gas Chromatography (GC) aiming at volatile components in the traditional Chinese medicine composition. The detection method adopts a gas chromatography, and the temperature rise program comprises the following steps: the initial column temperature is 150-180 ℃, the temperature is kept for 3-10 min, the temperature is increased to 200-210 ℃ at the speed of 5-15 ℃/min, and the temperature is kept for 3-10 min.
Description
Technical Field
The invention relates to a detection method of a traditional Chinese medicine composition, and belongs to the field of traditional Chinese medicines.
Background
The quality control means of the Chinese medicinal compound preparation mainly carries out qualitative and quantitative analysis on microscopic identification, inspection, content measurement and the like of individual raw medicinal materials in the preparation. However, the traditional Chinese medicine compound preparation usually comprises a plurality of medicinal materials, and some preparations even comprise animal medicines or mineral medicines, and the complexity of chemical components contained in the traditional Chinese medicine compound preparation is far higher than that of a single medicinal material. In fact, more and more pharmaceutical researchers recognize that the quality consistency and effectiveness of the traditional Chinese medicine compound preparation can not be comprehensively and effectively reflected by utilizing microscopic identification, thin-layer chromatography identification and quantitative analysis of single index components. From the section of Chinese pharmacopoeia 2015, the quality control standard of the Chinese herbal compound preparation is improved to a certain extent. For example, quality control standards are consolidated for the same formulation with different quality control standards; thin-layer identification indexes are added for some traditional Chinese medicine compound preparations without thin-layer identification items (or the thin-layer identification items are less and the thin-layer identification items are not specific to monarch drugs); the content measurement of a plurality of index components with related drug effects is increased from the original single index component content measurement; the variety of the preparation for content determination by a thin-layer scanning method is reduced; enhances the application of the gas chromatography to Chinese medicinal compound preparations containing volatile chemical components, and the like; the improvement of the quality control methods undoubtedly reflects the continuous improvement of the quality standard and the quality control force of the Chinese herbal compound preparation. However, the quality evaluation and control problem of the compound Chinese medicine preparation is still a bottleneck which troubles the development and internationalization of the Chinese medicine, and the development of new quality evaluation and control modes and method researches of the compound Chinese medicine preparation is urgent.
Disclosure of Invention
Aiming at the problems in the prior art, the invention provides a detection method of a traditional Chinese medicine composition, which is characterized in that the main representative chemical components in the preparation are quantitatively determined by adopting a Gas Chromatography (GC) aiming at the volatile components, and the quality of the preparation is evaluated by multi-component chemical rapid analysis.
The purpose of the invention is realized by the following technical scheme:
a detection method of a traditional Chinese medicine composition comprises the following steps:
step a, preparing a test solution: adding organic solvent into the traditional Chinese medicine composition for extraction to obtain a test solution;
step b, preparation of a reference substance solution: dissolving control products of patchouli alcohol and menthol in organic solvent to obtain control solution;
step c, detection: injecting the reference solution and the sample solution into a gas chromatograph respectively, and measuring according to the gas chromatography, wherein the chromatographic conditions are as follows: the initial column temperature is 150-180 ℃, the temperature is kept for 3-10 min, the temperature is increased to 200-210 ℃ at the speed of 5-15 ℃/min, and the temperature is kept for 3-10 min.
Preferably, the temperature raising procedure is: the initial column temperature is 170 ℃, the temperature is kept for 5min, the temperature is increased to 180 ℃ at the speed of 10 ℃/min, and the temperature is kept for 5 min; or, the initial column temperature is 170 ℃, the temperature is kept for 5min, the temperature is raised to 200 ℃ at the speed of 10 ℃/min, and the temperature is kept for 5 min.
In a specific embodiment, the organic solvent in step a is any one or more of methanol, ethanol, acetone, dichloromethane and chloroform, preferably a chloroform solution; the extraction method comprises ultrasonic extraction or reflux extraction; the concentration of the test solution is 10-20 mg/mL, preferably 12-14 mg/mL.
In a specific embodiment, the organic solvent in step b is any one or more selected from methanol, ethanol, acetone, dichloromethane, chloroform and n-hexane, and n-hexane is preferred. Further, the concentrations of patchouli alcohol and menthol in the reference solution are 0.1-0.2 mg/mL and 3-10 mg/mL respectively; further, the concentrations of patchouli alcohol and menthol in the control solution were 0.1mg/mL and 5mg/mL, respectively.
In a specific embodiment, the temperature of the injection port is 220-230 ℃, the temperature of the detector is 250-260 ℃, the split ratio is (3-10): 1, and the carrier gas is N2。
The traditional Chinese medicine composition comprises the following raw materials: 200-300 parts of fructus forsythiae, 200-300 parts of honeysuckle, 50-100 parts of fried ephedra, 50-100 parts of fried bitter almond, 200-300 parts of gypsum, 200-300 parts of isatis root, 200-300 parts of rhizoma dryopteris crassirhizomae, 200-300 parts of houttuynia cordata, 50-100 parts of pogostemon cablin, 20-70 parts of rheum officinale, 50-100 parts of rhodiola rosea, 5-10 parts of menthol and 50-100 parts of liquorice.
Preferably, the traditional Chinese medicine composition comprises the following raw materials: 255 parts of fructus forsythiae, 255 parts of honeysuckle, 85 parts of roasted ephedra, 85 parts of fried bitter almond, 255 parts of gypsum, 255 parts of isatis root, 255 parts of male fern rhizome, 255 parts of heartleaf houttuynia herb, 85 parts of cablin potchouli herb, 51 parts of rhubarb, 85 parts of rhodiola rosea, 7.5 parts of menthol and 85 parts of liquorice.
The preparation method of the traditional Chinese medicine composition comprises the following steps: distilling herba Agastaches with water to obtain volatile oil, collecting volatile oil, and filtering water extractive solution; extracting fructus forsythiae, herba Ephedrae preparata, herba Houttuyniae, and radix et rhizoma Rhei with 70% ethanol twice (2 hr for the first time and 1.5 hr for the second time), filtering the extractive solutions, mixing, and recovering ethanol; decocting honeysuckle, gypsum, isatis root, male fern rhizome, liquorice and rhodiola rosea in water until boiling, adding fried bitter apricot seed, decocting twice, the first time is 1.5 hours, the second time is 1 hour, filtering decoction, combining filtrates, adding an aqueous solution prepared by extracting oil from pogostemon cablin, concentrating until the relative concentration is 1.10-1.15 (60 ℃), adding ethanol until the ethanol content reaches 70%, refrigerating at 4 ℃ for 24 hours, filtering, recovering ethanol from the filtrate, combining with the ethanol extract prepared by extracting the four ingredients of forsythia and the like, concentrating until the relative density is 1.10-1.15 (60 ℃), spray-drying, mixing with a proper amount of starch, granulating, drying, sieving, screening out a proper amount of fine powder, dissolving menthol and pogostemon cablin volatile oil in ethanol, spraying into the fine powder, mixing, sealing for 30 minutes, and encapsulating to prepare 1000 granules.
The structural formula of the component to be detected is shown in the table 1:
TABLE 1 structural formula of the component to be measured
Drawings
FIG. 1 is a GC chromatogram of a sample solution under chromatographic conditions (I);
FIG. 2 is a GC chromatogram of a sample solution under chromatographic conditions (II);
FIG. 3 is a GC chromatogram of a sample solution according to chromatographic conditions c;
FIG. 4 is a GC chromatogram of a sample solution under chromatographic conditions (IV);
FIG. 5 is a GC chromatogram of a sample solution under a chromatographic condition (v);
FIG. 6 is a GC chromatogram of a sample solution under the condition of the chromatography condition;
FIG. 7 is a GC chromatogram of a sample solution under the chromatographic condition (c);
FIG. 8 is a GC chromatogram of the mixed control solution under the chromatographic conditions (v).
Detailed Description
Example 1
The formula is as follows: 255g of fructus forsythiae, 255g of honeysuckle, 85g of mix-fried ephedra, 85g of stir-fried bitter almond, 255g of gypsum, 255g of isatis root, 255g of male fern rhizome, 255g of heartleaf houttuynia herb, 85g of patchouli, 51g of rhubarb, 85g of rhodiola rosea, 7.5g of menthol and 85g of liquorice;
the preparation method comprises the following steps: distilling herba Agastaches with water to obtain volatile oil, collecting volatile oil, and filtering water extractive solution; extracting fructus forsythiae, herba Ephedrae preparata, herba Houttuyniae, and radix et rhizoma Rhei with 70% ethanol twice (2 hr for the first time and 1.5 hr for the second time), filtering the extractive solutions, mixing, and recovering ethanol; decocting honeysuckle, gypsum, isatis root, male fern rhizome, liquorice and rhodiola rosea in water until boiling, adding fried bitter apricot seed, decocting twice, the first time is 1.5 hours, the second time is 1 hour, filtering decoction, combining filtrates, adding an aqueous solution prepared by extracting oil from pogostemon cablin, concentrating until the relative concentration is 1.10-1.15 (60 ℃), adding ethanol until the ethanol content reaches 70%, refrigerating at 4 ℃ for 24 hours, filtering, recovering ethanol from the filtrate, combining with the ethanol extract prepared by extracting the four ingredients of forsythia and the like, concentrating until the relative density is 1.10-1.15 (60 ℃), spray-drying, mixing with a proper amount of starch, granulating, drying, sieving, screening out a proper amount of fine powder, dissolving menthol and pogostemon cablin volatile oil in ethanol, spraying into the fine powder, mixing, sealing for 30 minutes, and encapsulating to prepare 1000 granules.
The detection method comprises the following steps:
1. experimental Material
1.1 Experimental instruments
7890A GC system (Agilent, USA), 7693Auto Sampler, FID detector; AL-204 electronic balance (Switzerland); ultrasonic instrument (Nanjing Xinchen Biotechnology Ltd., 250W,40 KHz).
1.2 drugs and reagents
40 batches of the Chinese medicinal composition capsules (No. S01-S40) are provided by Shijiazhuang Ling pharmaceutical industry; the capsule contents were placed under high temperature (60 ℃), high humidity (RH 95%), light (4500lx) conditions for 20d samples A1, A2, A3 (Nos. S41-S43), respectively;
patchouli alcohol (batch No. 110772-; ethanol, chloroform, and n-hexane were all analytical chemicals (Guangdong Guanghua science and technology Co., Ltd.). High purity nitrogen and oxygen.
2. Experimental methods
2.1 inspection of chromatographic conditions
① Agilent 19095N-123 INNOWAX capillary column (30m × 0.53.53 mm,1 μm), initial column temperature 110 deg.CKeeping for 15min, with a sample inlet temperature of 220 deg.C, a detector temperature of 250 deg.C, a sample injection amount of 1.0 μ L, a split ratio of 3: 1, and N as carrier gas2Column flow 4mL/min, air flow 350mL/min, H2The flow rate was 35 mL/min.
② Agilent 19095N-123 INNOWAX capillary column (30m × 0.53.53 mm,1 μm), initial column temperature of 120 deg.C, holding time of 15min, sample inlet temperature of 220 deg.C, detector temperature of 250 deg.C, sample amount of 1.0 μ L, split ratio of 3: 1, and carrier gas of N2Column flow 4mL/min, air flow 350mL/min, H2The flow rate was 35 mL/min.
③ Agilent 19095N-123 INNOWAX capillary column (30m × 0.53.53 mm,1 μm), initial column temperature of 130 deg.C, holding for 5min, raising to 160 deg.C at 15 deg.C/min, holding for 10min, sample inlet temperature of 220 deg.C, detector temperature of 250 deg.C, sample injection amount of 1.0 μ L, split ratio of 3: 1, and carrier gas of N2Column flow 4mL/min, air flow 350mL/min, H2The flow rate was 35 mL/min.
④ Agilent 19095N-123 INNOWAX capillary column (30m × 0.53.53 mm,1 μm), initial column temperature of 130 deg.C, holding for 5min, raising to 200 deg.C at 15 deg.C/min, holding for 5min, sample inlet temperature of 220 deg.C, detector temperature of 250 deg.C, sample injection amount of 1.0 μ L, split ratio of 3: 1, and carrier gas of N2Column flow 4mL/min, air flow 350mL/min, H2The flow rate was 35 mL/min.
⑤ Agilent 19095N-123 INNOWAX capillary column (30m × 530 μm,1 μm), initial column temperature of 170 deg.C, holding for 5min, raising to 180 deg.C at 10 deg.C/min, holding for 5min, sample inlet temperature of 220 deg.C, detector temperature of 250 deg.C, sample inlet amount of 1.0 μ L, split ratio of 3: 1, and carrier gas of N2Column flow 4mL/min, air flow 350mL/min, H2The flow rate was 35 mL/min.
⑥ Agilent 19095N-123 INNOWAX capillary column (30m × 0.53.53 mm,1 μm), initial column temperature of 170 deg.C, holding for 5min, raising to 190 deg.C at 10 deg.C/min, holding for 5min, sample inlet temperature of 220 deg.C, detector temperature of 250 deg.C, sample injection amount of 1.0 μ L, split ratio of 3: 1, and carrier gas of N2Column flow 4mL/min, air flow 350mL/min, H2The flow rate was 35 mL/min.
⑦ Agilent 19095N-123 INNOWAX capillary column (30m × 530 μm,1 μm), initial column temperature of 170 deg.C, holding for 5min, raising to 200 deg.C at 10 deg.C/min, holding for 5min, sample inlet temperature of 220 deg.C, detector temperature of 250 deg.C, sample inlet amount of 1.0 μ L, split ratio of 3: 1, and carrier gas of N2Column flow 4mL/min, air flow 350mL/min, H2The flow rate was 35 mL/min.
2.2 preparation of the liquid medicine
Precisely weighing appropriate amount of patchouli alcohol and Mentholum reference substances, placing in 25mL brown volumetric flask, adding n-hexane to obtain reference substance mixed solution containing 0.1mg of patchouli alcohol and 5mg of Mentholum per 1mL, and filtering with 0.22 μm microporous membrane.
Accurately weighing 2.0g of the capsule content of the Chinese medicinal composition, placing in a 150mL triangular conical flask, precisely adding 50mL of chloroform, sealing, ultrasonically treating for 60 minutes (power 250W, frequency 40kHz, keeping water temperature below 50 ℃), filtering, and collecting filtrate; washing residue in the conical flask with 30mL of trichloromethane, filtering, combining trichloromethane solutions, volatilizing the solvent at low temperature, transferring the residue into a 5mL brown volumetric flask with n-hexane, filtering with a 0.22 μm microporous membrane, and collecting the subsequent filtrate.
3. Results of the experiment
3.1 chromatographic Condition examination results
Precisely absorbing 1.0 μ L of sample solution, injecting into gas chromatograph, inspecting according to chromatographic conditions (i-c), and recording chromatogram, as shown in fig. 1-7.
The result shows that the chromatographic conditions of the first step and the second step are poor in separation effect; the chromatographic conditions (c) and (d) are late in peak time after 6 min; the chromatographic conditions are sixteenth and seventeenth peak time of patchouli alcohol. Comprehensive investigation results show that the peak time of the chromatographic condition is appropriate, the separation effect is good, and the chromatographic condition is finally selected.
Precisely sucking 1.0 μ L of the mixed solution of the reference substances, injecting into a gas chromatograph, inspecting according to the chromatographic conditions, and recording the chromatogram, as shown in FIG. 8.
3.2 methodological investigation
And (4) performing precision investigation on the same mixed standard solution, continuously injecting samples for 6 times, and recording the peak area of the chromatogram. The results show that the RSD of the peak areas of the menthol and the patchouli alcohol are 0.74 percent and 0.53 percent respectively, which indicates that the precision of the instrument is good.
And (3) observing linear relation, precisely sucking 0.2, 0.4, 0.8, 1.0, 1.5 and 2.0 mu L of the mixed reference substance solution, injecting the mixed reference substance solution into an ultra-high performance liquid chromatograph, measuring, and recording chromatographic peak areas. The peak area was plotted as the ordinate (Y) and the amount of sample (. mu.g) as the abscissa (X) to prepare a standard curve. The linear equations of mingmbh and patchouli alcohol are respectively: Y-19.864X-0.8172 (r-0.9991), and Y-10.438X-8.5637 (r-0.9993). The result shows that the linear relationship between patchouli alcohol (20-200 ng) and menthol (1-10 mug) is good.
And (3) taking the same test solution, respectively injecting samples for determination within 0, 2, 4, 8, 16 and 24 hours after preparation, recording peak areas of respective chromatographic peaks, and calculating RSD of the chromatographic peak areas of the menthol and the patchouli alcohol. The results show that the RSDs of the chromatographic peak areas of menthol and patchouli alcohol within 24h are 1.44% and 1.71%, respectively, indicating that the stability of the test solution is good within 24 h.
And (4) repeatedly inspecting, namely taking 2.0g of the same batch of samples (S40) and 6 parts in total, precisely weighing, preparing a test solution for measurement, recording the area of each chromatographic peak and calculating the content. As a result, the average contents of menthol and patchouli alcohol in 6 test samples are 15.54mg/g and 0.251 mg/g; indicating that the repeatability of the method is good.
Accuracy examination 1.0g and 6 parts of capsule content with known content in the same batch (S40) are respectively put into a 150mL triangular conical flask, 15.6mg and 0.25mg of menthol and patchouli alcohol standard substances are respectively added, 50mL of trichloromethane is precisely added, and a test solution is prepared and tested by a sealing plug. The results show that the recovery rates of the menthol and the patchouli alcohol are respectively 95.17 percent and 95.39 percent, and the method has better accuracy.
3.3 content determination of 2 volatile components in the Chinese medicinal composition Capsule
Taking different batches of Chinese medicinal composition capsules, preparing a test solution according to a method 2.2, measuring and calculating the content by a method 2.1, and the result is shown in table 2.
TABLE 2 measurement results of menthol and patchouli alcohol content
The measurement results show that the content difference of volatile components of menthol and patchouli alcohol in the traditional Chinese medicine composition capsules of different batches is large, and the variation coefficients are 14% and 16% respectively. According to the dosage of the menthol in the prescription, the menthol should theoretically contain 21.42mg/g of the menthol in each capsule, but the actual measurement result shows that the content of the menthol is in the range of 11.80(S33) to 20.49(S11) mg/g, which is obviously lower than the theoretical value. According to the dosage of the patchouli in the prescription, each traditional Chinese medicine composition capsule theoretically contains 0.243mg/g of patchouli alcohol, but the actual measurement result shows that the content range of the patchouli alcohol is between 0.12(S1) and 0.24(S13) mg/g, which is obviously lower than the theoretical value. The contents of the components of the high-temperature, high-humidity and illuminated samples are obviously reduced, so that the menthol can be considered to be added by adopting an inclusion process in the preparation process; to avoid losses during storage, high temperatures, high humidity and light exposure should be avoided during storage. The results show that the reasonability of the process and the storage condition have certain influence on the quality consistency and stability of the traditional Chinese medicine composition capsules.
Example 2
The formulation and preparation method are the same as in example 1.
The detection method comprises the following steps:
chromatographic conditions are Agilent 19095N-123 INNOWAX capillary column (30m × 530 μm,1 μm), initial column temperature of 150 deg.C, holding time of 8min, raising to 200 deg.C at 10 deg.C/min, holding time of 5min, sample inlet temperature of 220 deg.C, detector temperature of 250 deg.C, sample injection amount of 1.0 μ L, split ratio of 3: 1, and carrier gas of N2Column flow 4mL/min, air flow 350mL/min, H2The flow rate was 35 mL/min.
Preparation of mixed control solution: precisely weighing appropriate amount of patchouli alcohol and Mentholum reference substances, placing in 25mL brown volumetric flask, adding n-hexane to obtain reference substance mixed solution containing 0.1mg of patchouli alcohol and 5mg of Mentholum per 1mL, and filtering with 0.22 μm microporous membrane.
Preparation of a test solution: accurately weighing 2.0g of the capsule content of the Chinese medicinal composition, placing in a 150mL conical flask, precisely adding 50mL of methanol, sealing, shaking for 60 min, filtering, and collecting the filtrate; washing the residue in the conical flask with 30mL of methanol, filtering, combining methanol solutions, volatilizing the solvent at low temperature, transferring the residue into a 5mL brown volumetric flask with n-hexane, filtering with a 0.22 μm microporous membrane, and collecting the subsequent filtrate.
And (3) determination: precisely sucking 1.0 μ L of each of the test solution and the mixed reference solution, injecting into a gas chromatograph, and recording chromatogram.
Example 3
The formulation and preparation method are the same as in example 1.
The detection method comprises the following steps:
chromatographic conditions are Agilent 19095N-123 INNOWAX capillary column (30m × 530 μm,1 μm), initial column temperature 180 deg.C, holding for 3min, raising to 200 deg.C at 8 deg.C/min, holding for 5min, sample inlet temperature 220 deg.C, detector temperature 250 deg.C, sample injection amount 1.0 μ L, split ratio 3: 1, and carrier gas N2Column flow 4mL/min, air flow 350mL/min, H2The flow rate was 35 mL/min.
Preparation of mixed control solution: precisely weighing appropriate amount of patchouli alcohol and Mentholum reference substances, placing in 25mL brown volumetric flask, adding n-hexane to obtain reference substance mixed solution containing 0.1mg of patchouli alcohol and 5mg of Mentholum per 1mL, and filtering with 0.22 μm microporous membrane.
Preparation of a test solution: accurately weighing 2.0g of the capsule content of the Chinese medicinal composition, placing in a 150mL conical flask, precisely adding 50mL of acetone, sealing, extracting under reflux for 60 min, filtering, and collecting the filtrate; washing the residue in the conical flask with 30mL of acetone, filtering, combining acetone solutions, volatilizing the solvent at low temperature, transferring the residue into a 5mL brown volumetric flask with n-hexane, filtering with a 0.22 μm microporous membrane, and collecting the subsequent filtrate.
And (3) determination: precisely sucking 1.0 μ L of each of the test solution and the mixed reference solution, injecting into a gas chromatograph, and recording chromatogram.
Example 4
The formulation and preparation method are the same as in example 1.
The detection method comprises the following steps:
chromatographic conditions are Agilent 19095N-123 INNOWAX capillary column (30m × 530 μm,1 μm), initial column temperature 160 deg.C, holding for 6min, raising to 200 deg.C at 15 deg.C/min, holding for 5min, sample inlet temperature 220 deg.C, detector temperature 250 deg.C, sample injection amount 1.0 μ L, split ratio 3: 1, and carrier gas N2Column flow 4mL/min, air flow 350mL/min, H2The flow rate was 35 mL/min.
Preparation of mixed control solution: precisely weighing appropriate amount of patchouli alcohol and Mentholum reference substances, placing in 25mL brown volumetric flask, adding n-hexane to obtain reference substance mixed solution containing 0.1mg of patchouli alcohol and 5mg of Mentholum per 1mL, and filtering with 0.22 μm microporous membrane.
Preparation of a test solution: accurately weighing 2.0g of the capsule content of the Chinese medicinal composition, placing in a 150mL conical flask, precisely adding 50mL of methanol, sealing, shaking for 60 min, filtering, and collecting the filtrate; washing the residue in the conical flask with 30mL of methanol, filtering, combining methanol solutions, volatilizing the solvent at low temperature, transferring the residue into a 5mL brown volumetric flask with n-hexane, filtering with a 0.22 μm microporous membrane, and collecting the subsequent filtrate.
And (3) determination: precisely sucking 1.0 μ L of each of the test solution and the mixed reference solution, injecting into a gas chromatograph, and recording chromatogram.
Example 5
The formulation and preparation method are the same as in example 1.
The detection method comprises the following steps:
chromatographic conditions are Agilent 19095N-123 INNOWAX capillary column (30m × 0.53mm,1 μm), initial column temperature of 170 deg.C, holding for 5min, raising to 190 deg.C at 10 deg.C/min, holding for 5min, sample inlet temperature of 220 deg.C, detector temperature of 250 deg.C, sample injection amount of 1.0 μ L, split ratio of 3: 1, and carrier gas of N2Column flow 4mL/min, air flow 350mL/min, H2The flow rate is 35mL/min。
Preparation of mixed control solution: precisely weighing appropriate amount of patchouli alcohol and Mentholum reference substances, placing in 25mL brown volumetric flask, adding n-hexane to obtain reference substance mixed solution containing 0.1mg of patchouli alcohol and 5mg of Mentholum per 1mL, and filtering with 0.22 μm microporous membrane.
Preparation of a test solution: accurately weighing 2.0g of the capsule content of the Chinese medicinal composition, placing in a 150mL triangular conical flask, precisely adding 50mL of dichloromethane, sealing, ultrasonically extracting for 60 minutes, filtering, and collecting the filtrate; washing the residue in the Erlenmeyer flask with 30mL of dichloromethane, filtering, combining dichloromethane solutions, volatilizing the solvent at low temperature, transferring the residue into a 5mL brown volumetric flask with n-hexane, filtering with a 0.22 μm microporous membrane, and taking the subsequent filtrate to obtain the final product.
And (3) determination: precisely sucking 1.0 μ L of each of the test solution and the mixed reference solution, injecting into a gas chromatograph, and recording chromatogram.
Claims (4)
1. A detection method of a traditional Chinese medicine composition is characterized by comprising the following steps:
step a, preparing a test solution: adding chloroform into the Chinese medicinal composition, and extracting to obtain a test solution;
step b, preparation of a reference substance solution: dissolving reference substances patchouli alcohol and menthol in n-hexane to obtain reference substance solution;
step c, detecting, namely respectively injecting the reference substance solution and the test substance solution into a gas chromatograph, and determining according to the gas chromatography, wherein the chromatographic conditions comprise that a chromatographic column is an INNOWAX capillary column with the specification of 30m × 0.53mm and 1 micron, the initial column temperature is 170 ℃, the initial column temperature is kept for 5min, the temperature is increased to 180 ℃ at the speed of 10 ℃/min, the initial column temperature is kept for 5min, the injection port temperature is 220-230 ℃, the FID detector temperature is 250-260 ℃, the split ratio is (3-10): 1, and the carrier gas is N2；
The traditional Chinese medicine composition comprises the following raw materials: 200-300 parts of fructus forsythiae, 200-300 parts of honeysuckle, 50-100 parts of fried ephedra, 50-100 parts of fried bitter almond, 200-300 parts of gypsum, 200-300 parts of isatis root, 200-300 parts of rhizoma dryopteris crassirhizomae, 200-300 parts of houttuynia cordata, 50-100 parts of pogostemon cablin, 20-70 parts of rheum officinale, 50-100 parts of rhodiola rosea, 5-10 parts of menthol and 50-100 parts of liquorice.
2. The detection method of claim 1, wherein the Chinese medicinal composition comprises the following raw materials: 255 parts of fructus forsythiae, 255 parts of honeysuckle, 85 parts of roasted ephedra, 85 parts of fried bitter almond, 255 parts of gypsum, 255 parts of isatis root, 255 parts of male fern rhizome, 255 parts of heartleaf houttuynia herb, 85 parts of cablin potchouli herb, 51 parts of rhubarb, 85 parts of rhodiola rosea, 7.5 parts of menthol and 85 parts of liquorice.
3. The detection method according to claim 1 or 2, wherein the concentration of the sample solution in the step a is 10-20 mg/mL.
4. The detection method according to claim 1, wherein the concentrations of patchouli alcohol and menthol in the control solution in the step b are 0.1-0.2 mg/mL and 3-10 mg/mL, respectively.
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| PCT/CN2018/110117 WO2019072245A1 (en) | 2017-10-13 | 2018-10-12 | Traditional chinese medicine composition gas chromatography testing method |
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