CN112362788B - Method for determining content of chlorogenic acid in radix stemonae tuberose medicinal material and application of method - Google Patents
Method for determining content of chlorogenic acid in radix stemonae tuberose medicinal material and application of method Download PDFInfo
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Abstract
The invention discloses a method for measuring the content of chlorogenic acid in a radix stemonae tuberose medicinal material and application thereof, belonging to the technical field of analysis, quality identification and standard control of traditional Chinese medicines. Detecting the content of chlorogenic acid in the radix Stemonae by high performance liquid chromatography, and quantitatively analyzing the radix Stemonae to realize the control of radix Stemonae; the method is simple to operate, high in analysis speed, accurate and good in repeatability, provides a scientific and reliable analysis method for analyzing components such as chlorogenic acid in the radix stemonae, and lays a foundation for perfecting the quality standard of the radix stemonae.
Description
Technical Field
The invention relates to a method for determining the content of chlorogenic acid in an Stemona tuberosa medicinal material and application thereof, belonging to the technical field of analysis, quality identification and standard control of traditional Chinese medicines.
Background
Stemona tuberosa is dried root tuber of Stemona tuberosa of Stemonaceae, which is harvested in spring and autumn, removed fibrous root, cleaned, placed in boiling water for slightly scalding or steaming until there is no white core, taken out, and dried. Has the effects of moistening lung, descending qi, relieving cough, killing parasite and killing louse; generally used for treating new chronic cough, phthisis cough and cough; it is externally applied to head louse, body louse, enterobiasis, and pudendal pruritus.
At present, the research on content index components in the stemona sessilifolia medicinal material is rarely carried out, such as: the content index components of radix Stemonae against the leaf are not specified in the Chinese pharmacopoeia (2015 edition). In 2019, 22.02.9, a patent document with publication number CN109374787A entitled "construction method and detection method of UPLC characteristic spectrum of radix stemonae japonicae", wherein chlorogenic acid, neochlorogenic acid and cryptochlorogenic acid are respectively used as reference substances to prepare reference substance solution; preparing a reference solution of a reference medicinal material by using a radix stemonae tuberose reference medicinal material; respectively taking a stemona tuberosa medicinal material and a standard decoction sample, and preparing a test solution and a standard decoction test solution; respectively sucking reference substance solution, test sample solution, and standard decoction test sample solution, injecting into ultra high performance liquid chromatograph, and measuring; comparing the test sample spectrum of the radix Stemonae, and the standard decoction sample test sample spectrum, and calibrating the characteristic peaks of 8 water-soluble components to obtain the UPLC characteristic spectrum of the radix Stemonae. The characteristic map can be used for qualitatively and quantitatively analyzing the quality of the radix stemonae tuberose medicinal material, can ensure the quality of the traditional decoction of the radix stemonae tuberose prepared by adopting the medicinal material, and is also suitable for detecting other preparations containing the radix stemonae tuberose. The method adopts an ultra-high performance liquid phase method and mainly uses a characteristic spectrum for qualitative analysis. The method adopts an ultra-high performance liquid phase method, mainly uses a characteristic spectrum to carry out qualitative analysis, and has higher requirements on experimental equipment.
Disclosure of Invention
The invention aims to solve the problems in the prior art and provides a method for measuring the content of chlorogenic acid in a radix stemonae sessilifoliae medicinal material and application thereof. In the technical scheme, the content of chlorogenic acid in the radix stemonae tuberose is detected by using a high performance liquid chromatography, and the radix stemonae tuberose is subjected to quantitative analysis, so that the control of the radix stemonae tuberose medicinal material is realized; the method is simple to operate, high in analysis speed, accurate and good in repeatability, provides a scientific and reliable analysis method for analyzing components such as chlorogenic acid in the radix stemonae, and lays a foundation for perfecting the quality standard of the radix stemonae.
In order to achieve the technical purpose, the following technical scheme is proposed:
a method for measuring the content of chlorogenic acid in an Stemona tuberosa medicinal material comprises the following steps:
A. chromatographic conditions and system suitability tests include:
a chromatographic column: octadecylsilane chemically bonded silica is used as a filler, the column length is 250mm, the inner diameter is 4.6mm, and the particle size is 5 mu m;
column temperature of the chromatographic column: 35 ℃;
mobile phase: acetonitrile is taken as a mobile phase A, and 0.4% phosphoric acid solution is taken as a mobile phase B;
flow rate of mobile phase: 1.0mL/min;
sample introduction amount: 10 mu L of the solution;
wavelength: 325nm;
the number of theoretical plates is not less than 5000 calculated according to the peak of chlorogenic acid;
the flow mobile phase is eluted according to the gradient program:
B. preparation of control solutions: precisely weighing chlorogenic acid reference substance, adding methanol, and making into solution containing 40 μ g per 1 mL;
C. preparation of a test solution: sieving the powder with a third sieve, weighing 0.5g, adding 25mL of methanol, sealing, weighing, ultrasonically treating (power 600W, frequency 40 kHz) for 30min, cooling, weighing again, supplementing the weight loss with methanol, shaking, filtering, and collecting the filtrate;
D. and (3) determination: precisely sucking the reference solution and the sample solution, respectively, injecting into a liquid chromatograph, and measuring.
Preferably, the methanol concentration is 50%.
Based on the technical scheme, the method for measuring the content of chlorogenic acid in the radix stemonae sessilifoliae medicinal material is applied to quality control of the radix stemonae sessilifoliae medicinal material for quantitative analysis.
By adopting the technical scheme, the beneficial technical effects brought are as follows:
1) In the invention, a standard curve is drawn by adopting a common high performance liquid method, a specific reference substance type, a sample solution extraction and matched chromatographic conditions to obtain a linear relation between the concentration of chlorogenic acid and a corresponding peak area to obtain the content of chlorogenic acid in the radix stemonae tuberosa medicinal material, and the method is applied to quantitative analysis of the radix stemonae tuberosa medicinal material to realize quality control of the radix stemonae tuberosa medicinal material;
meanwhile, a scientific and reliable analysis method is provided for analyzing components such as chlorogenic acid in the radix stemonae tuberosa medicinal material, and a foundation is laid for perfecting the quality standard of the traditional Chinese medicine radix stemonae tuberosa;
2) The technical scheme of the invention (such as: using a common high performance liquid chromatograph) compared with the prior art (such as: an ultra-high performance liquid chromatograph is adopted in CN 109374787A), the economic effect is better;
3) In the chromatographic condition, 0.4% phosphoric acid solution is adopted as the mobile phase B, the related pH value is lower, the detection of the target detection object chlorogenic acid is more stable under the condition, and the stability of the detection method and the accuracy of the detection result are further effectively ensured.
Drawings
FIG. 1 is a graph of the UV absorption spectrum of a chlorogenic acid control in example 4;
FIG. 2 is a chromatogram of different column temperatures in example 4;
FIG. 3 is a chromatogram of different flow rate surveys in example 4;
FIG. 4 is a chromatogram of the experimental investigation result of the specificity in example 5;
FIG. 5 is a standard curve diagram of chlorogenic acid as a reference in example 5.
Detailed Description
In the following, the technical solutions in the embodiments of the present invention are clearly and completely described, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
In the following examples, reference is made to an apparatus comprising:
high performance liquid chromatograph: waters e2698-2996 model HPLC, agilent 1260 model HPLC;
an electronic balance: ME204E/02, MS205D M, XPE26 (Mettler-Tollido instruments, inc.);
an ultra-pure water machine: cell type 1810A (Shanghai Mohler scientific instruments, inc.);
an ultrasonic cleaner: KQ5200DB type (600W, 40KHz; ultrasonic instruments, inc. of Kunshan);
and (3) chromatographic column: agilent ZORBTX SB-Aq 4.6X 250mm, ZORBTX Eclipse Plus C18.6X 250mm, agilent 5 HC-C18 (2) 4.6X 250mm.
In the following examples, the reagents involved included:
methanol (chromatographically pure), acetonitrile (chromatographically pure), phosphoric acid (chromatographically pure), water (ultrapure water), and other reagents are analytically pure;
in the following examples, the test articles referred to include:
stemona tuberosa medicinal material batch number: 010029-1906001, XLS201908241, XLS201908242, XLS201909897, XLS201909898, XLS201909899, XLS201909900, XLS201909902, XLS201909903, XLS201910157, XLS201910158, XLS201910159, XLS201910160, XLS201910161, XLS201910162, XLS201910163 and XLS201910164.
In the following examples, reference controls include:
chlorogenic acid (China institute for food and drug testing, batch number: 110752-201817, content in 99.8%);
example 1
A method for measuring the content of chlorogenic acid in a radix stemonae stemona medicinal material comprises the following steps:
A. chromatographic conditions and system suitability tests include:
and (3) chromatographic column: octadecylsilane chemically bonded silica is used as a filler, the column length is 250mm, the inner diameter is 4.6mm, and the particle size is 5 mu m;
column temperature of the chromatographic column: 35 ℃;
mobile phase: acetonitrile is taken as a mobile phase A, and 0.4% phosphoric acid solution is taken as a mobile phase B;
flow rate of mobile phase: 1.0mL/min;
sample introduction amount: 10 mu L of the solution;
wavelength: 325nm;
the number of theoretical plates is not less than 5000 calculated according to chlorogenic acid peak;
the flow mobile phase was as shown in table 1 below following a gradient elution procedure:
B. preparation of control solutions: precisely weighing chlorogenic acid reference substance, adding methanol, and making into solution containing 40 μ g per 1 mL;
C. preparation of a test solution: sieving the powder with a third sieve, weighing 0.5g, adding 25mL of methanol, sealing, weighing, ultrasonically treating (power 600W, frequency 40 kHz) for 30min, cooling, weighing again, supplementing the weight loss with methanol, shaking, filtering, and collecting the filtrate;
D. and (3) determination: precisely sucking the reference solution and the sample solution, respectively, injecting into a liquid chromatograph, and measuring.
Wherein, in steps B and C, the methanol concentration is 50 percent.
TABLE 1 gradient elution
| Time (min) | Mobile phase A (%) | Mobile phase B (%) |
| 0～10 | 7 | 93 |
| 10～12 | 7→5 | 93→95 |
| 12～35 | 5 | 95 |
| 35～40 | 5→13 | 95→87 |
| 40～50 | 13 | 87 |
Example 2
Based on example 1, the method for determining the content of chlorogenic acid in the stemona tuberosa is applied to quality control of the stemona tuberosa for quantitative analysis.
Example 3
Based on examples 1-2, this example will examine the kind of extraction solvent, extraction method, extraction time, and material ratio in the preparation of the sample solution, and further describe the present technical solution.
1. Investigation of extraction solvent species
Weighing the powder (batch number: 010029-1906001), sieving with a third sieve, weighing 0.5g (ten parts), placing into a conical flask with a plug, adding 25mL of water, 25mL of 50% methanol, 25mL of 70% methanol, 25mL of methanol and 25mL of 50% ethanol, respectively, sealing, weighing, ultrasonically extracting (power 600W, frequency 40 kHz) for 30min, cooling, weighing again, supplementing the lost weight with the corresponding extraction solvent, shaking, filtering, taking the subsequent filtrate, and measuring to obtain the results shown in the following table 2.
TABLE 2 analysis results of different extraction solvents
The results show that: when 50% methanol, 70% methanol and 50% ethanol are used as extraction solvents, the extraction rate is basically the same; and finally, determining that 50% methanol is used as an extraction solvent of the radix stemonae.
2. Examination of extraction methods
Weighing the powder (lot number: 010029-1906001), sieving with a third sieve, weighing 0.5g (four parts), placing in a conical flask with a plug, adding 25mL of 50% methanol, sealing the plug, weighing, ultrasonically extracting (power 600W, frequency 40 kHz), heating and refluxing for 30min, cooling, weighing again, supplementing the lost weight with 50% methanol, shaking up, filtering, taking the subsequent filtrate, and then measuring, wherein the obtained results are shown in the following table 3.
TABLE 3 analysis results of different extraction methods
The results show that: the chlorogenic acid obtained by ultrasonic extraction and reflux extraction is respectively 0.163% and 0.166%, and the chlorogenic acid content has no significant difference in the two extraction modes, so that a simple ultrasonic extraction mode is selected as the extraction mode of the radix stemonae japonicae medicinal material.
3. Extraction time review
Weighing the powder (lot number: 010029-1906001), sieving with a third sieve, weighing 0.5g (six parts), placing into a conical flask with a plug, adding 25mL of 50% methanol, sealing the plug, weighing, ultrasonically extracting (power 600W, frequency 40 kHz) for 20min, 30min and 40min, cooling, weighing again, supplementing the lost weight with 50% methanol, shaking up, filtering, taking the subsequent filtrate, and then measuring to obtain the results shown in the following table 4.
TABLE 4 analysis results of different extraction times
The results show that: the extraction time has no significant influence on the extraction efficiency, wherein the measurement result is slightly high at 30min, so the extraction time is selected to be 30min.
4. Investigating material ratio
Weighing the powder (lot number: 010029-1906001), sieving with a third sieve, weighing 0.5g (six parts), placing into a conical flask with a stopper, adding 50% methanol 10mL, 25mL, 50mL, sealing the stopper, weighing, ultrasonically extracting (power 600W, frequency 40 kHz) for 30min, cooling, weighing again, supplementing the lost weight with 50% methanol, shaking up, filtering, taking the subsequent filtrate, and measuring to obtain the results shown in Table 5 below.
TABLE 5 analysis results of different sample weights
The results show that: the extraction efficiency of the chlorogenic acid by the material ratio has no obvious influence, so that the solvent addition amount is 25mL based on the consideration of cost, pollution discharge and the like.
Example 4
This embodiment will be further described by examining chromatographic conditions (wavelength, column temperature, flow rate) in the measurement based on examples 1 to 3.
1. Wavelength investigation
Collecting the chlorogenic acid reference solution with full-wavelength spectrum, and analyzing the spectrogram to determine that the optimum detection wavelength for measuring chlorogenic acid content in radix Stemonae is 325nm (shown in FIG. 1).
2. Investigation of column temperature
The column temperatures were set to 25 ℃, 30 ℃ and 35 ℃ respectively, and the chromatographic conditions were the same as in example 1 except for the above measurement. The results of evaluation using the degree of separation of chlorogenic acid, the number of theoretical plates, tailing factor, and the like are shown in table 6 below and fig. 2.
TABLE 6 analysis results of different column temperatures
| Serial number | Column temperature (. Degree. C.) | Retention time | Number of theoretical plate | Tailing factor | Degree of separation |
| 1 | 25 | 43.971 | 298297 | 0.97 | 1.19 |
| 2 | 30 | 42.617 | 122797 | 0.99 | 1.52 |
| 3 | 35 | 38.699 | 10243 | 0.92 | 1.77 |
The results show that: when the column temperature is 35 ℃, the peak type is good, so that the column temperature is selected to be 35 ℃.
3. Investigation of flow velocity
The flow rates were set to 0.8mL/min, 1.0mL/min, and 1.2mL/min, respectively, and the other chromatographic conditions were the same as in example 1, and the results are shown in Table 7 below and FIG. 3, in which the degree of separation of the chlorogenic acid chromatographic peak, the number of theoretical plates, and the tailing factor were used as evaluation indices.
TABLE 7 analysis results of different flow rates
| Serial number | Flow rate (mL/min) | Retention time | Number of theoretical plate | Tailing factor | Degree of separation |
| 1 | 0.8 | 44.919 | 326574 | 0.90 | 1.66 |
| 2 | 1.0 | 38.556 | 10166 | 0.90 | 1.77 |
| 3 | 1.2 | 30.780 | 8629 | 0.93 | 1.64 |
The results show that when the flow rates are 0.8mL/min, 1.0mL/min and 1.2mL/min, the peak shape, separation and the like of the chromatogram have no significant effect, and under the integration of other factors, the flow rate is 1.0mL/min.
4. Chromatographic conditions and System suitability test results
Octadecylsilane chemically bonded silica is used as a filler (the column length is 250mm, the inner diameter is 4.6mm, and the particle size is 5 μm); acetonitrile is taken as a mobile phase A, 0.4% phosphoric acid solution is taken as a mobile phase B, and gradient elution is carried out according to the specification in the following table 8; the detection wavelength is 325nm; the column temperature was 35 ℃; the flow rate was 1.0ml/min. The number of theoretical plates is not less than 5000 calculated according to chlorogenic acid peak.
TABLE 8 gradient elution
Example 5
Based on examples 1-4, the following methodological studies were also performed in this example to further illustrate the present technical solution.
1. Specificity experiments
1. Preparation of control solutions: adding 50% methanol into chlorogenic acid reference substance to obtain 40 μ g solution per 1 mL;
2. preparation of a test solution: sieving the powder with a third sieve, weighing 0.5g, adding 50% methanol 25mL, sealing, weighing, ultrasonic processing (power 600W, frequency 40 kHz) for 30min, cooling, weighing again, supplementing the weight loss with 50% methanol, shaking, filtering, and collecting the filtrate;
3. preparation of negative control solution: transferring 10mL of 50% methanol into a conical flask with a plug, performing ultrasonic treatment (power 600W and frequency 40 kHz) for 30min, and filtering to obtain a subsequent filtrate;
the obtained control solution, test solution and negative control solution were respectively injected into HPLC for measurement, and the obtained results are shown in fig. 4, which indicates that: the negative control solution chromatogram has no interference to the measurement of the peak to be measured, which shows that the method has good specificity.
2. System adaptability survey
The control solution was sampled five times continuously, the peak area of chlorogenic acid was recorded, and the RSD value was calculated, and the results are shown in table 9 below.
TABLE 9 System Adaptation results
The results show that: the peak area RSD value of the chlorogenic acid reference substance is 0.7%, and the system adaptability of the method is good.
3. Linear relation
1mL, 2mL, 3mL, 4mL, 5mL and 6mL of chlorogenic acid reference substance solution (166.7864 mu g/mL) are respectively put into a 10mL volumetric flask for constant volume to prepare a series of chlorogenic acid reference substance solutions with gradient concentrations; then, injecting into a high performance liquid chromatograph, analyzing to obtain a peak area, drawing a standard curve with the chlorogenic acid reference substance concentration (μ g/mL) as an abscissa X and the peak area as an ordinate Y, and calculating to obtain a linear regression equation, wherein the obtained results are shown in table 10 and fig. 5 below.
TABLE 10 chlorogenic acid Standard Curve analysis results
| Numbering | Concentration (X, μ g/ml) | Peak area (Y) |
| 1 | 16.67864 | 417.3063 |
| 2 | 33.35728 | 873.3271 |
| 3 | 50.03592 | 1290.7212 |
| 4 | 66.71456 | 1787.6584 |
| 5 | 83.39320 | 2280.9661 |
| 6 | 100.07184 | 2699.8623 |
The results show that: the standard curve of chlorogenic acid is Y =27.63x-54.95, R 2 =0.999, which shows that the chlorogenic acid concentration in the sample is in the range of 16.68-100.07 mug/mL, and the linear relation is good.
4. Stability survey
The chromatographic peak areas of chlorogenic acid were measured at 0h, 2h, 6h, 8h, 16h, and 24h in the same sample (batch No.: 010029-1906001) solution, and the results are shown in Table 11 below.
TABLE 11 examination of stability of chlorogenic acid
The results show that: under the experimental condition, the RSD value of the chlorogenic acid peak area is 0.2%, and the test solution has good stability within 24 hours.
5. Repeatability survey
The same test sample (batch number: 010029-1906001) 0.5g, six portions were taken, the test sample solution was prepared by the same operator according to the method in example 1, the content of chlorogenic acid in the six portions of test sample was measured, and the results are shown in table 12 below.
TABLE 12 results of repeated experiments
The results show that: the RSD value of the chlorogenic acid content in the test sample is 0.9%, and the method has good repeatability.
6. Intermediate precision investigation
The same test sample (lot number: 010029-1906001) was taken in an amount of 0.5g and twelve portions, and solutions of the test sample were prepared by different operators (A, B) at different times (I, II) according to the method described in example 1, and measured by a Waters e2698-2996 high performance liquid chromatograph and an Agilent 1260 high performance liquid chromatograph, respectively, as shown in Table 13 below.
TABLE 13 results of intermediate precision examination
The results show that: when different operators use different instruments to detect the test sample at different time, the RSD value of the chlorogenic acid content is 1.2%, and the method has good intermediate precision.
7. Accuracy survey
Taking 0.25g of a test sample (batch number: 010029-1906001, chlorogenic acid content: 0.170%) with known chlorogenic acid content, and adding a certain amount of chlorogenic acid reference substance (purity: 96.8%) into six parts, respectively, preparing and measuring a test sample solution according to the method in example 1, and calculating the recovery rate, wherein the calculation formula is as follows:
TABLE 14 chlorogenic acid sample recovery results
The result shows that the content recovery rate of the chlorogenic acid is 95.7-98.8%, the average value is 97.4%, and the method has good accuracy.
8. Chromatographic column durability investigation
Comparing the influence of different chromatographic columns on the determination of the content of chlorogenic acid in the stemona sessilifolia, respectively examining the chromatographic columns of Agilent ZORBTX SB-Aq 4.6X 250mm, ZORBTX Eclipse Plus C18.6X 250mm and Agilent 5 HC-C18 (2) 4.6X 250mm, taking the stemona sessilifolia (batch number: 010029-1906001) for determination, and comparing the other chromatographic conditions with the example 1, the results are shown in the following table 15.
TABLE 15 durability examination results
The results show that: the RSD value of the chlorogenic acid content in the stemona medicinal material is 0.5 percent, and the method has good durability.
Example 6
Based on examples 1-5, this example further illustrates the technical solution by verifying the content of chlorogenic acid in the stemona tuberosa medicinal material.
Seventeen batches of radix stemonae tuberose medicinal materials are used as test samples, test sample solutions are prepared and measured by the method in example 1, peak areas are recorded, and the content of chlorogenic acid is calculated, and the results are shown in the following table 16.
TABLE 16 measurement results of seventeen batches of radix Stemonae
| Numbering | Batch number | Content (%) |
| 1 | 010029-1906001 | 0.165 |
| 2 | XLS201908241 | 0.107 |
| 3 | XLS201908242 | 0.258 |
| 4 | XLS201909897 | 0.102 |
| 5 | XLS201909898 | 0.171 |
| 6 | XLS201909899 | 0.179 |
| 7 | XLS201909900 | 0.186 |
| 8 | XLS201909902 | 0.182 |
| 9 | XLS201909903 | 0.122 |
| 10 | XLS201910157 | 0.113 |
| 11 | XLS201910158 | 1.191 |
| 12 | XLS201910159 | 0.193 |
| 13 | XLS201910160 | 0.122 |
| 14 | XLS201910161 | 0.115 |
| 15 | XLS201910162 | 0.178 |
| 16 | XLS201910163 | 0.109 |
| 17 | XLS201910164 | 0.223 |
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.
Claims (1)
1. A quality control method for radix stemonae tuberose medicinal material based on chlorogenic acid content is characterized by comprising the following steps:
A. chromatographic conditions and System suitability test
And (3) chromatographic column: octadecylsilane chemically bonded silica is used as a filler, the column length is 250mm, the inner diameter is 4.6mm, and the particle size is 5 mu m;
column temperature of the chromatographic column: 35 ℃;
mobile phase: acetonitrile is taken as a mobile phase A, and 0.4% phosphoric acid solution is taken as a mobile phase B;
flow rate of mobile phase: 1.0mL/min;
sample introduction amount: 10 mu L of the solution;
wavelength: 325nm;
the number of theoretical plates is not less than 5000 calculated according to the peak of chlorogenic acid;
the mobile phase is eluted according to the gradient program:
0-10 min, the mobile phase A is 7%, and the mobile phase B is 93%;
10-12 min, the mobile phase A is 7 → 5%, and the mobile phase B is 93 → 95%;
12-35 min, the mobile phase A is 5%, and the mobile phase B is 95%;
35-40 min, the mobile phase A is 5 → 13%, and the mobile phase B is 95 → 87%;
40-50 min, the mobile phase A is 13 percent, and the mobile phase B is 87 percent;
B. preparation of control solutions
Precisely weighing chlorogenic acid reference substance, and adding 50% methanol solution to obtain solution containing 40 μ g per 1 mL;
C. preparation of test solution
Sieving the powder with a third sieve, weighing 0.5g, adding 25mL of 50% methanol solution, sealing, weighing, ultrasonic treating for 30min, cooling, weighing again, supplementing the weight loss with methanol, shaking, filtering, and collecting the filtrate;
wherein, in the ultrasonic treatment, the power is 600W, and the frequency is 40kHz;
D. measurement of
Precisely sucking the reference solution and the sample solution, respectively, injecting into a liquid chromatograph, and measuring;
then, the method is applied to the quality control of the stemona tuberosa medicinal material for quantitative analysis.
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| CN111007171B (en) * | 2019-12-12 | 2022-09-09 | 哈尔滨市康隆药业有限责任公司 | Construction method and identification method of stemona tuberosa fingerprint |
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