CN110907555B - Fingerprint detection method for ethyl acetate part of ligusticum wallichii - Google Patents
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Abstract
The invention discloses a fingerprint detection method for ethyl acetate part of ligusticum wallichii, which adopts high performance liquid chromatography for detection. The detection method can simultaneously detect 4 components of senkyunolide I, senkyunolide H, senkyunolide A and ligustilide in the ligusticum wallichii, can control the quality of the effective part with the analgesic effect, is simple, convenient and reliable, perfects the quality evaluation system of the ligusticum wallichii, guarantees the clinical curative effect of the ligusticum wallichii, and has practical popularization and application values.
Description
Technical Field
The invention relates to the technical field of traditional Chinese medicine fingerprint spectrums, in particular to a fingerprint spectrum detection method for ethyl acetate part of ligusticum wallichii.
Background
Ligusticum wallichii is the dried rhizome of Ligusticum wallichii Hort of Umbelliferae, is a famous Chuan birth canal medicinal material, has the effects of activating blood and promoting qi circulation, and dispelling wind and relieving pain, is used for treating symptoms such as thoracic obstruction and cardiodynia, chest and hypochondriac pain, traumatic swelling and pain, irregular menstruation, amenorrhea and dysmenorrhea, abdominal mass and pain, headache, rheumatism and arthralgia, and is used as a good medicine for activating blood and relieving pain by doctors of different generations. Modern pharmacological research shows that the ligusticum wallichii has pharmacological activities of protecting heart and cerebral vessels, tranquilizing and relieving pain, resisting coagulation and the like, and is widely used for treating angina pectoris, ischemic brain injury, migraine and the like of coronary heart disease in clinic. Tujiani et al, the screening and research on effective parts of chuanxiong rhizome for treating migraine, disclose that the effective part of chuanxiong rhizome for analgesia is an ethyl acetate part, but there are also documents, such as Yangwei et al, the analysis of effective parts of chuanxiong rhizome for analgesia and blood circulation by orthogonal design method, report that the analgesic effect of chuanxiong rhizome ethyl acetate part is not good enough. Experiments prove that the analgesic effect of the ethyl acetate part of the ligusticum wallichii is obvious, and the completely opposite conclusion is that: the extraction process is improper, so that ligustilide with tranquilizing, analgesic, antipyretic and antiinflammatory effects in the ethyl acetate part of rhizoma Ligustici Chuanxiong and senkyunolide compound with antioxidant injury, antiinflammatory and analgesic, anticoagulant and platelet aggregation resisting, vasodilating and cytotoxic effects are lost, and the content is low.
At present, due to the lack of a quality detection method for the ethyl acetate part of the ligusticum wallichii, the difference of effective components in the analgesic effective part of the ligusticum wallichii obtained by experiments and production is obvious, so that the condition that the conclusion about the analgesic effective part of the ligusticum wallichii in the existing research report is inconsistent appears, the clinical application of the ethyl acetate part of the ligusticum wallichii is limited, and the further development of the ligusticum wallichii is not favorable.
Disclosure of Invention
In order to solve the problems, the invention provides a fingerprint detection method of a ligusticum wallichii ethyl acetate part, which adopts high performance liquid chromatography for detection and comprises the following steps:
1) preparation of a reference solution: dissolving ligustilide control in methanol to obtain control solution;
2) preparing a test solution: pulverizing rhizoma Ligustici Chuanxiong, extracting with ethanol, filtering, concentrating the filtrate to obtain extract, dissolving in water, extracting with ethyl acetate, drying the extractive solution, dissolving in methanol, and filtering;
3) respectively sucking the reference solution and the test solution, injecting into a high performance liquid chromatograph, and recording the chromatogram; the chromatographic conditions were as follows:
a chromatographic column: octadecylsilane chemically bonded silica is used as a filling agent; mobile phase: taking water as a mobile phase A and methanol as a mobile phase B; the gradient elution procedure was as follows:
further, the contrast solution in the step 1) contains 10-20 mu g of ligustilide per 1 ml.
Further, the reference substance in the step 1) also comprises senkyunolide I, senkyunolide H and/or senkyunolide A; each 1ml of the reference solution contains 20-30 μ g of senkyunolide I, 2-5 μ g of senkyunolide H, and 15-25 μ g of senkyunolide A.
Further, the mass volume ratio of the ligusticum wallichii, the ethanol, the water, the ethyl acetate and the methanol in the step 2) is 0.5 g: 50 ml: 50 ml: 15 ml: 25 ml.
Further, the ethanol is 60-80% (v/v, ml/ml) ethanol, preferably 70% (v/v, ml/ml) ethanol.
Further, the ligusticum wallichii in the step 2) is crushed and sieved by a No. 4 sieve.
Further, the extraction in the step 2) is ultrasonic extraction, and the ultrasonic extraction time is 30 min.
Further, the concentration of the step 2) is reduced pressure concentration at 25 ℃.
Further, the drying in the step 2) is drying under reduced pressure at 25 ℃.
Further, the detection wavelength in the chromatographic conditions of step 2): 280nm, the injection volume of 20ul, the flow rate of 1ml/min and the column temperature of 25 ℃.
Further, 8 characteristic peaks are totally included in the fingerprint, the peak corresponding to the ligustilide reference substance is an S peak, the relative retention time of each characteristic peak and the S peak is calculated, and the relative retention time is within +/-3% of a specified value; the values were peak 1:0.421, peak 2:0.443, peak 3:0.535, peak 4:0.760, peak 5:0.912, peak 6:0.926, peak 7:1.000, and peak 8: 1.060.
The fingerprint detection method of the ethyl acetate part of the ligusticum wallichii can simultaneously detect 4 components including senkyunolide I, senkyunolide H, senkyunolide A and ligustilide in the ligusticum wallichii, and can also control the quality of the effective part with the analgesic effect.
Obviously, many modifications, substitutions, and variations are possible in light of the above teachings of the invention, without departing from the basic technical spirit of the invention, as defined by the following claims.
The present invention will be described in further detail with reference to the following examples. This should not be understood as limiting the scope of the above-described subject matter of the present invention to the following examples. All the technologies realized based on the above contents of the present invention belong to the scope of the present invention.
Drawings
FIG. 1 HPLC chromatogram of the mixed control (A) (1: senkyunolide I; 2: senkyunolide H; 3: senkyunolide A; 7: ligustilide) and sample (B)
FIG. 224 batch HPLC fingerprint of ethyl acetate part of Ligusticum wallichii
Detailed Description
The reagents, reagents and equipment used in the embodiment of the present invention are known products and obtained by purchasing commercially available products.
Example 1 fingerprint detection of Ethyl acetate fraction of Ligusticum wallichii of the invention
1) Preparation of a reference solution: accurately weighing senkyunolide I, senkyunolide H, senkyunolide A and ligustilide, respectively dissolving with methanol to obtain reference stock solutions with appropriate mass concentration, accurately weighing the stock solutions into the same volumetric flask, diluting with methanol to scale, and shaking to obtain mixed reference solutions with senkyunolide I, senkyunolide H, senkyunolide A and ligustilide mass concentrations of 0.0210mg/m L, 0.0038mg/m L, 0.0197mg/m L and 0.0137mg/m L respectively;
2) preparing a test solution: taking 0.5g of ligusticum wallichii sample powder (screened by a No. 4 sieve), precisely weighing, placing the powder in a conical flask with a plug, adding 50mL of 70% ethanol for soaking, weighing, ultrasonically extracting for half an hour, cooling, weighing again, complementing the lost weight with 70% ethanol, shaking up, taking supernatant, filtering, concentrating the filtrate at 25 ℃ under reduced pressure, and recovering ethanol to obtain extract; dispersing the concentrated extract with 50mL water, extracting with 15mL ethyl acetate, repeating twice, mixing the extractive solutions, drying at 25 deg.C under reduced pressure to obtain solid, dissolving with 25mL methanol, and filtering with microporous membrane;
3) respectively sucking the reference solution and the test solution, injecting into a high performance liquid chromatograph, wherein the chromatographic conditions are as follows:
a chromatographic column: GS-120-5-C18(250 mm. times.4.6 mm, 5 μm) column; detection wavelength: 280 nm; the sample injection volume is 20 mu L, the flow rate is 1ml/min, and the column temperature is 25 ℃; taking water as a mobile phase A and methanol as a mobile phase B; the gradient elution procedure was as follows:
4) the fingerprint spectrum of the ethyl acetate part of the ligusticum wallichii has 8 characteristic peaks, the peak corresponding to the ligustilide reference substance is an S peak, the relative retention time of each characteristic peak and the S peak is calculated, and the relative retention time is within +/-3% of a specified value; the values were peak 1:0.421, peak 2:0.443, peak 3:0.535, peak 4:0.760, peak 5:0.912, peak 6:0.926, peak 7:1.000, and peak 8: 1.060.
The beneficial effects of the present invention are illustrated by the following experimental examples:
experimental example 1
1 materials and methods
1.1 medicaments
Ligusticum wallichii, collected in main producing areas and extraprovince introduction areas of Sichuan province, is identified as Ligusticum wallichii Liuusticum Chuanxiong Hort by professor Ma Jie Ying of Chengdu Chinese medicine university, and the sample information is shown in Table 1.
TABLE 1 rhizoma Ligustici Chuanxiong sample information
1.2 reagents
Senkyunolide I (batch number: CHB170926), senkyunolide H (batch number: CHB161228), senkyunolide A (batch number: CHB160917) and ligustilide (batch number: CHB170224) are purchased from Chengdu Cromar Biotech limited company, and the purity is more than or equal to 98%;
methanol is chromatographically pure (Fisher company, USA), water is purified water, ethanol, and ethyl acetate reagent are chemically pure, and are provided by chemical reagent factory of Chengdu Kelong.
1.3 instruments
LC-2010A high performance liquid chromatograph (Shimadzu, Japan), SQP one ten thousandth analytical balance (Sideris scientific instruments Co., Germany), BT125D one ten thousandth analytical balance (Sideris scientific instruments Co., Germany), KQ-300 ultrasonic cleaner (ultrasonic Equipment Co., Kunshan), RE-5203 rotary evaporator (Shanghai subsrong Biochemical instruments factory), SHB-III circulating water type multipurpose vacuum pump (Zhengzhou Changcheng Kong Co., Ltd.), Allegra TM X-22R Centrifuge refrigerated Centrifuge (BECKMAN COULTER Co., USA), VARIOSKAN FLASH 2.4.3 full wavelength multifunctional reader (Thermo Corp., USA).
2. Method of producing a composite material
2.1 study of HPLC fingerprint of effective fraction of chuanxiong rhizome in relieving pain-ethyl acetate
2.1.1 chromatographic conditions
The chromatographic column is a GS-120-5-C18(250mm, 4.6mm and 5 mu m) column; the mobile phase is A (water) -B (methanol); flow rate: 1.0 mL/min; column temperature: 25 ℃; detection wavelength: 280 nm; sample introduction amount: 20 mu L of the solution; because the effective components of the ethyl acetate of the hemlock parsley are complex, the following 6 elution programs which can separate characteristic peaks are obtained by a plurality of mobile phase gradient elution screens:
①0～5min：30％B；5～10min：30％～55％B；10～35min：55％～65％B；35～45min：65％～85％B；
②0～10min：30％～60％B；10～35min：60％～65％B；35～40min：65％～85％B；40～50min：85％～85％B；
③0～10min：30％～60％B；10～15min：60％～65％B；15～20min：65％～85％B；20～25min：85％～85％B；
④0～10min：30％～60％B；10～15min：60％～65％B；15～20min：65％～85％B；20～30min：85％～85％B；
⑤0～15min：30％～65％B；15～40min：65％～75％B；40～50min：75％～85％B；
⑥0～5min：30％～30％B；5～10min：30％～55％B；10～35min：55％～65％B；35～45min：65％～85％B；
gradient elution program (II) is used for methodology investigation because the separation degree of senkyunolide compounds in the chromatogram obtained by gradient elution program (II) is poor and does not meet the requirement that the separation degree in the chromatographic systematic applicability test specified in Chinese pharmacopoeia is more than 1.5.
2.1.2 preparation of control solutions
Accurately weighing appropriate amount of senkyunolide I, senkyunolide H, senkyunolide A, and ligustilide, respectively dissolving in methanol, and making into reference stock solution with appropriate mass concentration. Precisely measuring the above stock solution to the same volumetric flask, adding methanol to dilute to scale, and shaking to obtain mixed reference solutions with mass concentrations of senkyunolide I, senkyunolide H, senkyunolide A and ligustilide of 0.0210mg/m L, 0.0038mg/m L, 0.0197mg/m L and 0.0137mg/m L respectively.
2.1.3 preparation of test solutions
Taking about 0.5g of rhizoma ligustici wallichii sample powder (screened by a No. 4 sieve), precisely weighing, placing in a conical flask with a plug, adding 50mL of 70% ethanol for infiltration, weighing, carrying out ultrasonic extraction for half an hour, cooling, weighing again, supplementing the lost weight with 70% ethanol, shaking up, taking supernatant, filtering, concentrating the filtrate at 25 ℃ under reduced pressure, and recovering ethanol. Dispersing the concentrated extract with 50mL water, extracting with 15mL ethyl acetate, repeating twice, mixing extractive solutions, concentrating under reduced pressure at 25 deg.C, dissolving with 25mL methanol, and filtering with microporous membrane.
2.1.4 methodology investigation
And (3) precision test: taking the same rhizoma ligustici wallichii sample test solution (C19), continuously injecting samples for 6 times according to the chromatographic condition under the item of 2.1.1, recording a chromatogram, and calculating the relative retention time and the relative peak area of each common peak, wherein the relative retention time RSD of each common peak is less than 1%, and the relative peak area RSD is less than 3%, which indicates that the instrument precision is good.
And (3) repeatability test: taking 6 parts of the same ligusticum wallichii sample (C19) in total, precisely weighing, preparing a test solution according to the method under the item 2.1.3, respectively carrying out sample injection analysis according to the chromatographic condition under the item 2.1.1, recording a chromatogram, calculating the relative retention time and the relative peak area of each common peak, wherein the RSD of each common peak is less than 1%, and the RSD of the relative peak area is less than 3%, so that the method has good repeatability.
And (3) stability test: taking the same rhizoma ligustici wallichii sample test solution (C19), respectively carrying out sample injection analysis for 0, 2, 4, 6, 8 and 16h according to the chromatographic condition under the item of 2.1.1, recording a chromatogram, calculating the relative retention time and the relative peak area of each common peak, wherein the RSD of each common peak is less than 1%, and the RSD of the relative peak area is less than 3%, which indicates that the test solution has good stability in 16 h.
2.1.5 sample determination
The samples of 24 batches were prepared as a test solution according to the method under item "2.1.3", and analyzed under chromatographic conditions under item "2.1.1", and chromatograms recorded for 45 min.
2.1.6 data analysis
Introducing HPLC chromatograms of 24 batches of rhizoma Ligustici Chuanxiong effective component samples into a similarity evaluation system (2004A) of Chinese medicinal chromatogram fingerprint for similarity evaluation. And (3) carrying out principal component analysis on common peaks in the fingerprint of 24 batches of ligusticum wallichii effective part samples by adopting SPSS 21.0 software.
3. Results
Comparing the chromatograms of the control and the sample with the ultraviolet spectrum, the chromatographic peak in the sample is confirmed, 4 chromatographic peaks are identified, and the result is shown in fig. 1.
Introducing HPLC chromatograms of ethyl acetate parts of 24 batches of ligusticum wallichii samples into a national pharmacopoeia committee-traditional Chinese medicine chromatogram fingerprint similarity evaluation system (2004A), setting a sample of S1 as a reference chromatogram, selecting a median by a reference chromatogram generation method, setting the width of a time window to be 0.5, finally determining 8 peaks as characteristic peaks through multipoint correction and automatic matching, and analyzing the chromatogram data to obtain a result shown in figure 2: calculating the relative retention time of each characteristic peak and S peak, wherein the relative retention time is within + -3% of the specified value; the values were peak 1:0.421, peak 2:0.443, peak 3:0.535, peak 4:0.760, peak 5:0.912, peak 6:0.926, peak 7:1.000, and peak 8: 1.060.
Similarity evaluation is carried out on the 24 batches of sample chromatograms and the comparison fingerprint by adopting a traditional Chinese medicine chromatogram fingerprint similarity evaluation system (2004A), and the result is shown in table 2. The similarity of the ethyl acetate parts of the ligusticum wallichii of 24 batches is 0.967-1.000, which shows that the chemical components of the ethyl acetate parts of the ligusticum wallichii of all the places tend to be consistent on the whole, but because the 24 batches of ligusticum wallichii samples have differences in the ecological environments of producing areas such as altitude, illumination, rainfall, soil and the like, the contents of the chemical components of the parts of the samples in different batches have certain differences.
TABLE 224 degree of similarity between fingerprint of ethyl acetate part of rhizoma Ligustici Chuanxiong and reference fingerprint
KMO statistics and Bartlett tests were performed on 8 common peaks from 24 samples and the results indicated that principal component analysis could be performed. A total of 4 principal components were obtained by principal component analysis, and the cumulative contribution rate was 83.952%. 4 principal component scores in 24 samples are calculated, and the 24 samples are comprehensively judged through an individual comprehensive judgment function (C), and the judgment results are shown in a table 3. As can be seen from the table, the total evaluation score of 24 samples was-0.584 to 1.145, and the quality of the samples could be evaluated and controlled based on the evaluation score.
TABLE 324 batches of the principal Components and the comprehensive evaluation scores of Ligusticum chuanxiong
The experimental result shows that the invention can be used for controlling the quality of the ethyl acetate part of the ligusticum wallichii by detecting the 8 peaks, if the 8 peaks exist, the ethyl acetate part of the ligusticum wallichii can be judged, and if the 8 peaks do not exist, the ethyl acetate part of the ligusticum wallichii is judged not to be the ethyl acetate part of the ligusticum wallichii or the ethyl acetate part of the ligusticum wallichii with unqualified quality; simultaneously, the content of 4 ingredients including senkyunolide I, senkyunolide H, senkyunolide A and ligustilide can be simultaneously detected.
In conclusion, the ethyl acetate extraction parts in the extraction parts of the ligusticum wallichii play a role in analgesia, and HPLC fingerprint researches on the ethyl acetate effective parts of the ligusticum wallichii in different producing areas show that the chemical component types in the ethyl acetate effective parts of the ligusticum wallichii in the producing areas are basically consistent, but the contents are different. The detection method of the invention provides a basis for the quality control of the ethyl acetate effective part of the ligusticum wallichii, perfects the quality evaluation system of the ligusticum wallichii, ensures the clinical curative effect of the ligusticum wallichii, and has practical popularization and application values.
Claims (10)
1. A fingerprint detection method for ethyl acetate part of rhizoma ligustici wallichii is characterized by comprising the following steps: the detection is carried out by adopting a high performance liquid chromatography, and the method comprises the following steps:
1) preparation of a reference solution: dissolving ligustilide, senkyunolide I, senkyunolide H and senkyunolide A in methanol to obtain reference solution;
2) preparing a test solution: pulverizing rhizoma Ligustici Chuanxiong, extracting with ethanol, filtering, concentrating the filtrate to obtain extract, dissolving in water, extracting with ethyl acetate, drying the extractive solution, dissolving in methanol, and filtering;
3) respectively sucking the reference solution and the test solution, injecting into a high performance liquid chromatograph, and recording the chromatogram; the chromatographic conditions were as follows:
a chromatographic column: octadecylsilane chemically bonded silica is used as a filling agent; mobile phase: taking water as a mobile phase A and methanol as a mobile phase B; the gradient elution procedure was as follows:
。
2. The detection method according to claim 1, characterized in that: in the step 1), each 1ml of the reference solution contains 10-20 mu g of ligustilide.
3. The detection method according to claim 1, characterized in that: step 1) every 1ml of the reference substance solution contains 20-30 μ g of senkyunolide I, H2-5 μ g of senkyunolide and A15-25 μ g of senkyunolide A.
4. The detection method according to claim 1, characterized in that: step 2), the mass volume ratio of the ligusticum wallichii to the ethanol to the water to the ethyl acetate to the methanol is 0.5 g: 50 ml: 50 ml: 15 ml: 25 ml.
5. The detection method according to claim 1 or 4, characterized in that: the ethanol is 60-80% v/v, ml/ml ethanol.
6. The detection method according to claim 1, characterized in that: and step 2), crushing the ligusticum wallichii and screening the ligusticum wallichii by a No. 4 sieve.
7. The detection method according to claim 1, characterized in that: the extraction in the step 2) is ultrasonic extraction, and the ultrasonic extraction time is 30 min.
8. The detection method according to claim 1, characterized in that: step 2) concentrating under reduced pressure at 25 ℃; the drying is carried out at 25 ℃ under reduced pressure.
9. The detection method according to claim 1, characterized in that: step 3) detection wavelength in chromatographic conditions: 280nm, the injection volume is 20 mu L, the flow rate is 1ml/min, and the column temperature is 25 ℃.
10. The detection method according to any one of claims 1 to 9, characterized in that: the fingerprint contains 8 characteristic peaks, the peak corresponding to ligustilide reference is S peak, the relative retention time of each characteristic peak and S peak is calculated, and the relative retention time is within +/-3% of the specified value; the values were peak 1:0.421, peak 2:0.443, peak 3:0.535, peak 4:0.760, peak 5:0.912, peak 6:0.926, peak 7:1.000, and peak 8: 1.060.
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1707260A (en) * | 2005-05-11 | 2005-12-14 | 沈阳药科大学 | Angelica sinensis and ligusticum wallichii Chinese medicinal crop and establishing method for its preparation finger print and standard fingerprint |
| CN101015552A (en) * | 2006-12-29 | 2007-08-15 | 天津大学 | Ligustilide extract and its preparing process and application |
| CN101226177A (en) * | 2008-01-29 | 2008-07-23 | 南方医科大学 | Method for identification of Szechwan Ligusticum wallichii |
| CN102716160A (en) * | 2012-06-28 | 2012-10-10 | 天津大学 | Hemlock parsley phthalide type ingredient effective part as well as preparation method and application of hemlock parsley phthalide type ingredient effective part |
| CN104597153A (en) * | 2014-12-30 | 2015-05-06 | 上海现代中医药股份有限公司 | Assay method of ligusticum wallichii fingerprint |
| CN104713956A (en) * | 2014-12-30 | 2015-06-17 | 上海现代中医药股份有限公司 | Method for determining fingerprint chromatography of radix astragali and ligusticum wallichii extract products |
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|---|---|---|---|---|
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Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1707260A (en) * | 2005-05-11 | 2005-12-14 | 沈阳药科大学 | Angelica sinensis and ligusticum wallichii Chinese medicinal crop and establishing method for its preparation finger print and standard fingerprint |
| CN101015552A (en) * | 2006-12-29 | 2007-08-15 | 天津大学 | Ligustilide extract and its preparing process and application |
| CN101226177A (en) * | 2008-01-29 | 2008-07-23 | 南方医科大学 | Method for identification of Szechwan Ligusticum wallichii |
| CN102716160A (en) * | 2012-06-28 | 2012-10-10 | 天津大学 | Hemlock parsley phthalide type ingredient effective part as well as preparation method and application of hemlock parsley phthalide type ingredient effective part |
| CN104597153A (en) * | 2014-12-30 | 2015-05-06 | 上海现代中医药股份有限公司 | Assay method of ligusticum wallichii fingerprint |
| CN104713956A (en) * | 2014-12-30 | 2015-06-17 | 上海现代中医药股份有限公司 | Method for determining fingerprint chromatography of radix astragali and ligusticum wallichii extract products |
Non-Patent Citations (5)
| Title |
|---|
| Isolation and identification of anti-inflammatory constituents from Ligusticum chuanxiong and their underlying mechanisms of action on microglia;Terry C.T.Or等;《Neuropharmacology》;20110531;第60卷(第6期);823-831 * |
| Simultaneous quantification of 12 bioactive components of Ligusticum chuanxiong Hort. by high-performance liquid chromatography;Ru Yan等;《Journal of Pharmaceutical and Biomedical Analysis》;20041128;87-95 * |
| 不同溶剂对川芎药材中有效成分提取效果的影响;杨学东 等;《中国中药杂志》;20120731;第37卷(第13期);1942-1945 * |
| 川芎中的化学成分的抗炎活性及其质量控制的研究;孙存霞 等;《医药论坛杂志》;20150731;第36卷(第7期);58-60 * |
| 川芎药材的高效液相色谱质量控制方法研究;丁明玉 等;《药物分析杂志》;20041231;第24卷(第4期);351-353 * |
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