CN116008445A - Quality control method for one-test-multiple evaluation of cleavers - Google Patents
Quality control method for one-test-multiple evaluation of cleavers Download PDFInfo
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- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
Abstract
The invention discloses a quality control method for a cleaver with one test and multiple evaluations, and belongs to the technical field of traditional Chinese medicine component detection. The method adopts high performance liquid chromatography, uses chlorogenic acid as an internal reference, establishes a one-measurement-multiple-evaluation method of the cleavers, and calculates the component content of the novel chlorogenic acid, the hidden chlorogenic acid, the isochlorogenic acid A and the isochlorogenic acid C in the cleavers by using relative correction factors between the chlorogenic acid and the novel chlorogenic acid, the hidden chlorogenic acid, the isochlorogenic acid A and the isochlorogenic acid C in the cleavers. The method has no obvious difference with the component content result obtained by the external standard method, and the precision, repeatability and stability of the method are less than 2 percent, which indicates that the method can be used for measuring the content of 5 components in the cleavers, solves the problem that expensive reference substances such as new chlorogenic acid, cryptochlorogenic acid and the like are difficult to obtain, is simple and convenient to operate, has low detection cost, accurate measurement result and good repeatability and stability, and lays an experimental foundation for the quality control of the cleavers.
Description
Technical Field
The invention belongs to the technical field of detection of traditional Chinese medicine components, and particularly relates to a quality control method for one-test-multiple evaluation of a cleaver.
Background
The Galium is derived from dry whole herb of Galium aparine Linn. Var. Echinospermam (Wallr.) cuf of Rubiaceae, also called as Lala vine, rubia tenuifolia, saw blade, blood-activating grass, etc., and the Yao medicine name is Dong Yangmi, is medicinal material used by Guangxi Yao nationality, and has pharmacological effects of clearing heat, detoxicating, removing toxicity, resisting cancer, promoting urination, detumescence, eliminating phlegm, relieving pain, etc. Modern pharmaceutical chemistry researches show that the cleavers contain alkaloid, volatile oil, anthraquinone, phenolic acid, flavonoid and other components, wherein most phenolic acid components have the effects of scavenging free radicals, resisting viruses and bacteria and exciting the central nervous system, and are main active components in the cleavers.
The chemical components of the traditional Chinese medicine are complex and various, the quality control modes of multiple indexes are larger for the types and the quantity of the reference substances, and the price of some reference substances is more expensive, so Wang Zhiming and the like provide a multi-index quality control mode for measuring multiple evaluations, namely the content of other components can be calculated by using relative correction factors only by the concentration and peak area of one reference substance. The one-test multi-evaluation method is already applied to the measurement of traditional Chinese medicines in the 2020 edition of Chinese pharmacopoeia, and is widely applied to the quality control research of the traditional Chinese medicines and the multiple indexes of the preparation thereof.
At present, the research report of a one-test-multiple-evaluation method of the cleavers is not yet seen. In order to more comprehensively and effectively control the quality of the medicinal materials in the cleavers, 5 organic acid components of chlorogenic acid, isochlorogenic acid A, isochlorogenic acid C, neochlorogenic acid and cryptochlorogenic acid in the cleavers are taken as index components, a multi-index quality control mode of one measurement and multiple evaluation is established, and data support and theoretical basis are provided for the quality control and research of the medicinal materials in the cleavers.
Disclosure of Invention
Aiming at the problems, the invention provides a method for controlling the quality of one-test-multiple-evaluation of a cleaver, which adopts a high-performance liquid chromatography method, takes low-cost and easily-obtained chlorogenic acid as an internal reference, establishes the one-test-multiple-evaluation method of the cleaver, and calculates the content of each organic acid component in the cleaver. The method is simple and convenient to operate, low in detection cost, accurate in measurement result and good in repeatability and stability.
The invention is realized by the following technical scheme:
a quality control method for testing and evaluating the number of cleavers in one step adopts high performance liquid chromatography, uses chlorogenic acid as an internal reference, establishes a testing and evaluating method for testing and evaluating the number of cleavers in one step, and calculates the component content of the novel chlorogenic acid, the hidden chlorogenic acid, the isochlorogenic acid A and the isochlorogenic acid C in the cleavers by using relative correction factors between the chlorogenic acid and the novel chlorogenic acid, the hidden chlorogenic acid, the isochlorogenic acid A and the isochlorogenic acid C in the cleavers.
Preferably, the method comprises the following steps:
(1) Preparation of a mixed control solution: precisely weighing chlorogenic acid, neochlorogenic acid, cryptochlorogenic acid, isochlorogenic acid A and isochlorogenic acid, dissolving with methanol, and fixing volume to obtain mixed reference solution;
(2) Preparation of test solution: precisely weighing appropriate amount of Galium Teneri powder, precisely adding methanol solution, soaking, weighing, refluxing, cooling to room temperature, weighing again, supplementing weight with methanol, filtering, centrifuging the filtrate, and collecting supernatant as sample solution;
(3) High performance liquid chromatography assay: respectively taking the mixed reference substance solution prepared in the step (1) and the sample solution prepared in the step (2), and injecting the mixed reference substance solution and the sample solution into a high performance liquid chromatograph for testing to obtain a high performance liquid chromatogram of the mixed reference substance solution and a high performance liquid chromatogram of the sample solution;
(4) Positioning chromatographic peaks of components to be detected: chlorogenic acid is used as an internal reference substance, and chromatographic peaks are positioned according to relative retention values between chlorogenic acid and new chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid A and isochlorogenic acid C in a high performance liquid chromatogram of the mixed reference substance solution;
(5) Calculation of the relative correction factor: calculating peak area of each chromatographic peak by using high performance liquid chromatogram of the mixed reference solution, and calculating relative correction factors between chlorogenic acid and new chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid A and isochlorogenic acid C by using chlorogenic acid as internal reference;
(6) Calculating the content of the components to be detected: and calculating the component content of the chlorogenic acid, the isochlorogenic acid A and the isochlorogenic acid C in the sample solution by using the relative correction factors between the chlorogenic acid and the chlorogenic acid, the isochlorogenic acid A and the isochlorogenic acid C in the mixed reference solution.
Preferably, in step (5), the calculation of the relative correction factor is: the peak areas of the novel chlorogenic acid, the hidden chlorogenic acid, the isochlorogenic acid A and the isochlorogenic acid C are calculated by taking chlorogenic acid as an internal reference, and the relative correction factors are calculated by a multipoint correction method.
Preferably, the multipoint correction method is according to formula f i/s ＝f i /f s ＝(C s A i )/(A s C i ) Calculating a relative correction factor;
wherein A is s Peak area of internal reference, C s A is the concentration of the internal reference substance i For the component to be measuredPeak area, C i For the concentration of the component to be measured, f i/s Is a relative correction factor.
In the step (5), the relative correction factor of chlorogenic acid and neochlorogenic acid is 1.024-1.055, the relative correction factor of chlorogenic acid and cryptochlorogenic acid is 0.921-0.931, the relative correction factor of chlorogenic acid and isochlorogenic acid A is 1.281-1.325, and the relative correction factor of chlorogenic acid and isochlorogenic acid C is 1.181-1.217.
As a preferred embodiment, in the step (4), the calculation formula of the relative retention value is: r is (r) i/s ＝T R(i) /T R(s) ；
Wherein r is i/s For relative retention value, T R(i) For retention time of the component to be measured, T R(s) Is the retention time of the internal reference.
In the step (4), the relative retention value of chlorogenic acid and neochlorogenic acid is 0.585-0.588, the relative retention value of chlorogenic acid and cryptochlorogenic acid is 1.113-1.114, the relative retention value of chlorogenic acid and isochlorogenic acid A is 2.182-2.200, and the relative retention value of chlorogenic acid and isochlorogenic acid C is 2.515-2.545.
As a preferable mode of the technical scheme, in the step (3), the measurement conditions of the high performance liquid chromatography are as follows: the chromatographic column is Guangzhou Fei Nigen X-Penyx C 18 A 4.6X105 mm,5 μm model chromatographic column; the mobile phase is methanol-0.1% formic acid aqueous solution, and gradient elution is carried out, wherein methanol is mobile phase A, and 0.1% formic acid aqueous solution is mobile phase B; the flow rate is 0.8-1.0 mL/min; the detection wavelength is 280-327 nm; column temperature is 25-30 ℃; the time is 60min, and the operation is 10min later; the sample injection amount is 8-10 mu L.
As a preferred embodiment, the specific procedure of the gradient elution is as follows: 0-10 min, 15-20% A; 10-33 min, 20-38% A; 33-35 min,38% A; 35-45 min, 38-50% A.
As a preferred embodiment, in the step (2), the preparation of the sample solution comprises the following specific steps: taking 0.5-1 g of cleaver powder, precisely weighing, placing in a conical flask, precisely adding 15-20 mL of 60% methanol solution, firstly soaking for 20-30 min, weighing, refluxing for 50-60 min, cooling to room temperature, weighing again, supplementing weight with 60% methanol, filtering, taking 2-4 mL of continuous filtrate, centrifuging at the rotating speed of 10000-13000 r/s for 10-15 min, and taking supernatant as a sample solution.
Compared with the prior art, the invention has the advantages that:
1. the invention adopts high performance liquid chromatography, takes low-cost and easily available chlorogenic acid as an internal reference, establishes a multi-evaluation method for the cleavers, and calculates the component content of the novel chlorogenic acid, the hidden chlorogenic acid, the isochlorogenic acid A and the isochlorogenic acid C in the cleavers by using relative correction factors between the chlorogenic acid and the novel chlorogenic acid, the hidden chlorogenic acid, the isochlorogenic acid A and the isochlorogenic acid C in the cleavers. Meanwhile, the content of 5 organic acids in the cleavers is measured by an external standard method, and the values obtained by the two methods are compared, so that no significant difference exists, and the method has the advantages that the precision, the repeatability and the stability of RSD (reactive species decomposition) are less than 2%, so that the method can be used for measuring the content of 5 organic acids in the cleavers, solves the problem that expensive reference substances such as new chlorogenic acid, cryptochlorogenic acid and the like are difficult to obtain, and lays an experimental foundation for the quality control of the cleavers.
2. The method selects chlorogenic acid as an internal reference to perform one-measurement-multiple-evaluation method for determining the content of organic acid in the cleavers, the content of the chlorogenic acid in the cleavers is high, the reference substances are more common, and more expensive reference substances such as newer chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid A and the like are easy to obtain, and the price is substantial, so that the time cost and the detection cost can be saved by taking the chlorogenic acid as the internal reference, and the method is rapid and accurate, and can more comprehensively and effectively control the quality of the cleavers.
3. According to the invention, chlorogenic acid is used as an internal reference, relative correction factors between the chlorogenic acid and the novel chlorogenic acid, the cryptochlorogenic acid, the isochlorogenic acid A and the isochlorogenic acid C in the cleavers are calculated, and the novel chlorogenic acid, the cryptochlorogenic acid, the isochlorogenic acid A and the isochlorogenic acid C in the cleavers are calculated by using the relative correction factors, so that the detection cost and the detection time are greatly reduced, the quantitative detection method of the cleavers is simplified, the efficiency is improved, and the method has important significance on quality control of the cleavers.
4. According to the invention, the results of the one-test multi-evaluation method and the external standard method are compared, the content results of the two methods are not significantly different, and the RSD of the precision, repeatability and stability of the method are less than 2%, so that the method can be used for effectively measuring the content of chlorogenic acid, neochlorogenic acid, cryptochlorogenic acid, isochlorogenic acid A and isochlorogenic acid C5 organic acids in the medicinal materials of the cleavers.
5. The invention establishes a method for detecting the cleavers by one test and multiple evaluations, can more comprehensively and effectively control the quality of the cleavers, and avoids the problems of high cost, high detection cost, complex operation and the like of partial reference substances in the multi-component content measurement.
Drawings
Fig. 1 is a structural diagram of chlorogenic acid, neochlorogenic acid, cryptochlorogenic acid, isochlorogenic acid a and isochlorogenic acid C5 components, in fig. 1: a-chlorogenic acid; b-neochlorogenic acid; c-cryptochlorogenic acid; d-isochlorogenic acid A; e-isochlorogenic acid C.
Fig. 2 is a high performance liquid chromatogram of the sample solution and the mixed control solution of example 1, fig. 2: a-high performance liquid chromatogram of the sample solution; b-high performance liquid chromatogram of the mixed reference substance solution; 1-neochlorogenic acid; 2-chlorogenic acid; 3-cryptochlorogenic acid; 4-isochlorogenic acid A; 5-isochlorogenic acid C.
FIG. 3 is a linear diagram of the novel chlorogenic acid of example 1.
FIG. 4 is a linear diagram of chlorogenic acid in example 1.
FIG. 5 is a linear graph of cryptochlorogenic acid in example 1.
FIG. 6 is a linear diagram of isochlorogenic acid A in example 1.
FIG. 7 is a linear diagram of isochlorogenic acid C in example 1.
Detailed Description
The present invention is further illustrated by the following examples, which are only intended to illustrate the present invention and not to limit the scope of the present invention.
Example 1
1. Experimental materials
1.1 instruments
The instruments used in the present invention are shown in the following tables 1-1, all of which are commercially available
TABLE 1-1 instrument information
1.2 reagents
The apparatus used in the present invention is shown in the following tables 1-2, all of which are commercially available
TABLE 1-2 reagent information
1.3 medicinal materials
Cleavers were collected in Wu Mingou Yilingcun, nanning city peak, guest city Jin Xiuxian, gui Lin Shi May you be city, fengshan city, fenghuang city, luzhai county, liuzhou, fengshan county Qiao Yin village. The dried whole herb was identified by teacher in Guangxi university of Chinese medicine Wei Songji as Galium aparine Linn. Var. Echinospermam (Wallr.) cuf.
2. Experimental methods and results
2.1 chromatographic conditions
Guangzhou phenanthridine X-Penyx C 18 Chromatographic column (4.6 mm. Times.250 mm,5 μm), mobile phase: methanol-0.1% formic acid water, eluting (see Table 2-1), 1.0mL/min flow rate, 327nm detection wavelength, 30deg.C column temperature, and 10 μl sample injection amount. Under the above chromatographic conditions, the chromatographic peak separation effect of 5 index components in the cleavers is better, and the figure is shown in the figure 2. The molecular structural formula of each component is shown in figure 1.
TABLE 2-1 gradient elution procedure
2.2 examination of the preparation method of the sample solution
2.2.1 extraction solvent investigation
By looking at different extraction solvents, i.e. different concentrations of methanol solutions (35%, 60%, 85%). Under the same extraction method, the cleavers are extracted by different solvents for 60min, cooled to room temperature, fully shaken up to supplement the lost weight, filtered, and the filtrate is centrifuged (13000 r/s) for 10min, and the supernatant is taken for measurement, and the results are shown in Table 2-2. The method shows that 60% of methanol is best in extraction, the extraction is more complete, and the content of each component is higher; when the extraction solvent is ethanol, the chromatographic peak and the impurity peak of the sample are not separated, and the separation degree is unqualified.
Table 2-2 different extraction solvent assay results (n=2)
2.2.2 extraction method investigation
The extraction and reflux were examined using 60% methanol as the extraction solvent, 2 extraction methods. The content of each corresponding ingredient was calculated, and the results are shown in tables 2 to 3. The results show that the reflux extraction method is better than the ultrasonic extraction method, the content of chlorogenic acid and cryptochlorogenic acid in the reflux extraction is higher than that in the ultrasonic extraction, and the extraction is more sufficient, so that the reflux extraction is selected as the extraction method for preparing the test sample.
Tables 2-3 determination results (n=3) of different extraction methods
2.2.3 extraction time investigation
1g (3 parts each) of cleavers is taken, precisely weighed, precisely added with 60% methanol, extracted under reflux for 30min, 60min and 90min respectively, taken out and cooled to room temperature, the lost weight is complemented, uniformly shaken, filtered, and the filtrate is centrifuged (13000 r/s) for 10min, and the supernatant of the solution is taken for measurement, and the results are shown in tables 2-4. The results show that the optimal extraction time is 60min, since the content of each component is the highest and the extraction can be completed when the reflux time is 60min.
Tables 2-4 measurement results of different extraction times (n=2)
2.2.4 solvent quantity investigation
1g (3 parts each) of cleaver powder is taken, precisely weighed, 20mL, 25mL and 50mL of 60% methanol solution are respectively and precisely added, soaked for 30min, weighed, extracted under reflux for 60min, weighed again after being cooled, complemented by 60% methanol, filtered, and the obtained filtrate is taken in a 2mL centrifuge tube, centrifuged for 10min, injected with sample, and the measurement results are shown in tables 2-5. The results show that when the solvent amount is 20mL, the content of the extracted components such as the chlorogenic acid is relatively high.
Tables 2-5 different solvent amount measurement results (n=2)
In summary, the preparation method of the sample comprises the following steps: 20mL of 60% methanol was added thereto, and the mixture was extracted under reflux for 1 hour.
2.3 preparation of Mixed control solution
Accurately weighing appropriate amounts of reference substances of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid A and isochlorogenic acid C respectively, placing into a 10mL volumetric flask, dissolving with chromatographic methanol, and fixing volume. 1.111mg/mL of neochlorogenic acid, 1.743mg/mL of chlorogenic acid, 0.931mg/mL of cryptochlorogenic acid, 0.949mg/mL of isochlorogenic acid A and 0.958mg/mL of isochlorogenic acid C are prepared. Respectively precisely sucking 1mL, 6mL, 1mL and 0.3mL of each stock solution, placing into a 50mL volumetric flask, adding chromatographic methanol to fix volume to scale marks to obtain mixed reference substance solution, wherein the concentrations of the chlorogenic acid, the cryptochlorogenic acid, the isochlorogenic acid A and the isochlorogenic acid C are respectively 0.02222mg/mL, 0.2092mg/mL, 0.01862mg/mL, 0.01898mg/mL and 0.005748mg/mL.
2.4 preparation of sample solutions
About 1g of cleaver powder is taken, precisely weighed, placed in a 25mL conical flask, and precisely added with 20mL of 60% methanol solution. Soaking for 30min, weighing, refluxing for 60min, cooling to room temperature, weighing again, supplementing with 60% methanol, filtering, centrifuging (13000 r/s) for 10min with 2mL of the filtrate, and collecting supernatant as sample solution.
2.5 methodology investigation
2.5.1 Linear relationship test
Precisely sucking 0.5mL, 1mL, 2mL, 4mL, 5mL, 7.5mL and 10mL of the mixed reference substance solution under the item "2.3", respectively placing the mixed reference substance solution into a 10mL volumetric flask, adding chromatographic methanol for constant volume, and preparing 7 mixed reference substance solutions with different concentrations. Measured according to the chromatographic conditions under item "2.1". A standard curve was drawn with the concentrations C (mg/mL) of the respective component controls as the abscissa (X) and the peak areas (A) as the ordinate (Y). The results are shown in tables 2-6, FIGS. 3, 4, 5, 6, and 7. The results show that chlorogenic acid, novel chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid A and isochlorogenic acid C have good linear relationship in the concentration ranges of 0.01046-0.2092 mg/g, 0.01111-0.02222 mg/g, 0.000931-0.01862 mg/g, 0.000949-0.01898 mg/g and 0.0002874-0.005748 mg/g respectively.
Table 2-6 5 Linear relation of ingredients
2.5.2 precision test
The mixed reference substance solution under the item "2.3" is precisely sucked, and the sample is continuously injected for 6 times according to the chromatographic condition under the item "2.1", so as to determine the peak area. The peak areas RSD were calculated to be 0.26%,0.69%,0.91%,0.49%,0.62%, respectively. The precision of the instrument was shown to be good, and the data are shown in tables 2-7.
Tables 2-7 precision test results (n=6)
2.5.3 stability test
About 1g of cleaver powder is taken and precisely weighed. Extracting with 60% methanol under reflux for 60min with 20mL. 10 mu L of the sample solution is precisely sucked, and the sample solution is injected into a chromatograph at 0, 2, 4, 8, 12 and 24 hours respectively, and is measured according to the chromatographic condition under the item "2.1". As a result, the RSD values of the components of the cleavers were 0.14%,0.21%,0.52%,0.96% and 1.72%, respectively. The stability of the sample solution over 24 hours was shown to be good, and the results are shown in tables 2-8.
Tables 2-8 stability test results (n=6)
2.5.4 repeatability test
6 parts of the same place of production of the cleavers are taken, each about 1g of the cleaver powder is precisely weighed. Prepared as under "2.4", and peak areas were measured by continuously feeding 6 parts as under "2.1". The peak area RSD is calculated. The results are shown in tables 2-9. The contents of the components are 0.5157mg/g,3.849mg/g,0.4276mg/g,0.06304mg/g and 0.01637mg/g, and the RSD is 0.75%,1.04%,1.64%,1.66% and 1.60% respectively. The results show good reproducibility of the process and are shown in tables 2-9.
Tables 2-9 repeatability test results (n=6)
2.5.5 sample recovery test
6 parts of cleavers powder with known 5 component contents are weighed, each part is 0.5g, each part is precisely weighed, new chlorogenic acid (1.111 mg/mL), chlorogenic acid (2.061 mg/mL), cryptochlorogenic acid (0.931 mg/mL), isochlorogenic acid A (0.949 mg/mL) and isochlorogenic acid C (0.958 mg/mL) are precisely weighed respectively, 2mL, 9mL, 2mL, 0.3mL and 0.1mL of stock solution of each reference substance are put into a 200mL volumetric flask, and 60% methanol is added to fix the volume to a scale mark, so that mixed reference substance mother solution is obtained. Precisely sucking 20mL of the mother liquor of the mixed reference substance, heating and refluxing for extraction for 60min, cooling and supplementing weight, filtering and centrifuging for 10min, taking the supernatant of the solution, feeding the supernatant into an automatic sample injection bottle, measuring according to the chromatographic condition under the item of 2.1, and respectively calculating the sample adding recovery rate and RSD. The results are shown in tables 2-10, tables 2-11, tables 2-12, tables 2-13, and tables 2-14. The RSD of each component is less than 3%, which indicates that the sample recovery rate of the method is qualified.
Tables 2-10 results of sample recovery of chlorogenic acid (n=6)
Tables 2-11 chlorogenic acid sample recovery results (n=6)
Tables 2-12 recovery results of cryptochlorogenic acid sample (n=6)
Tables 2-13 results of recovery of isochlorogenic acid A (n=6)
Tables 2-14 results of recovery of isochlorogenic acid C (n=6)
2.6 calculation of relative correction factor
The relative correction factor calculation formula: f (f) i/s ＝f i /f s ＝(C s A i )/(A s C i ). The calculation formula for the concentration can be deduced from the calculation formula: c (C) i ＝(C s ×A i )/(f i/s ×A s ). A in the two formulas s Peak area of internal reference substance, C s Is the concentration of an internal reference substance; a is that i For the peak area of a certain component to be measured, C i For a certain concentration of the component to be measured, f i/s Is a relative correction factor. The experiment calculates f by a plurality of concentration points i/s Average value as quantitative f i/s Values.
Precisely sucking the reference substance solution under the item "2.5.1", sequentially injecting into chromatograph for determination, and recording chromatographic peak area. The relative correction factors for the other 4 components were calculated with chlorogenic acid. The results are shown in tables 2-15. The results showed that the relative correction factors obtained for the different concentrations were substantially identical, with RSD values of less than 2%.
Tables 2-15 relative correction factors
2.6.1 Effect of different chromatography columns on the relative correction factor
Investigation of different chromatographic columns [ Guangzhou Fei Nigen X-Penyx C 18 (4.6×250mm，5μm)；Eclipse XDB C 18 (4.6×250mm，55μm)；Acclaim120 C 18 (4.6×250mm，5μm)]For f i/s The effect of the values, the results are shown in tables 2-16, RSD<2, showing that the relative correction factors have good durability under different chromatographic columns。
Tables 2-16 determination of relative correction factors for different chromatographic columns
2.7 positioning of chromatographic peaks to be measured
The experiment is to locate chromatographic peaks by the ratio of the retention time of the component to be detected to the retention time of chlorogenic acid as an internal reference. The calculation formula is as follows: r is (r) i/s ＝T R(i) /T R(s) (wherein r i/s For relative retention value, T R(i) For retention time of the component to be measured, T R(s) Retention time for internal reference
The 7 mixed reference solutions under the item "2.3" were precisely aspirated, and the chromatographic peak area was recorded as determined under the chromatographic conditions under the item "2.1". As a result, the relative retention values among the components to be detected are not greatly different, and RSD is less than 5%, so that the method can be used for chromatographic peak positioning. The results are shown in tables 2-17.
Tables 2 to 17 relative retention values
2.8 comparison of the results of the external Standard method and the one-test-multiple evaluation method
Taking about 1g of the powder of the cleavers in different producing areas, precisely weighing, precisely adding 20mL of 60% methanol, carrying out reflux extraction for 60min, measuring the peak area of a sample according to the chromatographic condition under the item of 2.1, calculating the content of corresponding components, calculating other components except chlorogenic acid by using the obtained relative correction factors, comparing the results obtained by the two methods by taking the relative error as a parameter, and obtaining the results with the relative error of less than 5% by the methods shown in tables 2-18 and tables 2-19, wherein the established method has good credibility, and shows that the one-measurement-multiple evaluation method and the external standard method can be used for measuring the content of active components in the cleavers.
Tables 2-18 external standard method and one-test-multiple-evaluation method measurement results (n=2)
Table 2-19 relative error of external standard method and one-test-multiple evaluation method
3. Discussion of the invention
The cleavers obtained by the experiment of the invention are sourced from different places, and the picking periods are different, and the content of active ingredients in the herbs is different. And the picked medicinal materials are placed for a long time, so that the chlorogenic acid content is reduced, and isomers of the medicinal materials are mutually converted. According to the experiment, different extraction solvents such as 35% methanol, 60% methanol and 85% methanol are inspected, and as a result, the extraction effect of each component is optimal when 60% methanol is used as the extraction solvent, and the extraction of the effective components is incomplete, the chromatographic peak separation effect is poor, part of the chromatographic peak separation effect is bimodal, and the peak type is poor when ethanol is used for extraction.
The experiment of the invention examines different mobile phase systems such as methanol-0.1% formic acid, methanol-0.05% glacial acetic acid water solution and methanol-0.1% phosphoric acid: when methanol-0.05% glacial acetic acid is used as a mobile phase, the chromatographic peak separation effect of the chlorogenic acid is poor, shoulder peaks appear, and the peak shape is unsightly; when methanol-0.1% formic acid is used as a mobile phase, the chromatographic peak separation effect of each component is good, and the peak area is relatively high; methanol-0.1% phosphoric acid separated the components well. Formic acid is a volatile acid, however, which can improve chromatographic column and instrument lifetime, so methanol-0.1% formic acid is selected as the mobile phase.
The medicine contains chlorogenic acid, new chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid A and isochlorogenic acid C, and are isomers, wherein the content of chlorogenic acid is high, and compared with the expensive comparison products such as new chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid A and the like, the comparison products are more common, and the price is substantial, so that the invention selects chlorogenic acid as an internal reference to carry out one-measurement and multi-evaluation study on the cleavers. The method for calculating the relative correction factor by experiments adopts a multi-point correction method, namely, an average value obtained by calculating a plurality of different concentration points is taken as the relative correction factor for quantification, and the method also adopts a slope correction method, wherein according to a regression equation Y=ax+b, X=Y-b)/a=Y/a-b/a, b/a is negligible when a/b is greater than 100, and the slope of the regression equation is the relative correction factor. The method for positioning the chromatographic peak to be detected has a relative retention value and a retention time difference method, and the experiment adopts the relative retention value method which is more stable and has smaller fluctuation than the retention time difference method.
4. Conclusion(s)
The experiment of the invention adopts a high performance liquid chromatograph to measure, and uses chlorogenic acid as an internal reference, a multi-evaluation method of a cleaver is established, and the component content of the novel chlorogenic acid, the hidden chlorogenic acid, the isochlorogenic acid A and the isochlorogenic acid C in the cleaver is calculated by using relative correction factors between the chlorogenic acid and the novel chlorogenic acid, the hidden chlorogenic acid, the isochlorogenic acid A and the isochlorogenic acid C in the cleaver. Meanwhile, the content of 5 organic acids in the cleavers is measured by an external standard method, and the values obtained by the two methods are compared, so that no significant difference exists, and the method has the advantages that the precision, the repeatability and the stability of RSD (reactive species decomposition) are less than 2%, so that the method can be used for measuring the content of 5 organic acids in the cleavers, solves the problem that expensive reference substances such as new chlorogenic acid, cryptochlorogenic acid and the like are difficult to obtain, and lays an experimental foundation for the quality control of the cleavers.
The foregoing description of the preferred embodiments of the invention is not intended to be limiting, but rather is to cover all modifications, equivalents, and alternatives falling within the spirit and principles of the invention.
Claims (10)
1. A quality control method for one-test-multiple-evaluation of a cleaver is characterized in that a high-performance liquid chromatography method is adopted, chlorogenic acid is used as an internal reference, one-test-multiple-evaluation method of the cleaver is established, and the component content of the novel chlorogenic acid, the hidden chlorogenic acid, the isochlorogenic acid A and the isochlorogenic acid C in the cleaver is calculated by using relative correction factors between the chlorogenic acid and the novel chlorogenic acid, the hidden chlorogenic acid, the isochlorogenic acid A and the isochlorogenic acid C in the cleaver.
2. The method for quality control of a cleaver assay according to claim 1, characterized in that it comprises the following steps:
(1) Preparation of a mixed control solution: precisely weighing chlorogenic acid, neochlorogenic acid, cryptochlorogenic acid, isochlorogenic acid A and isochlorogenic acid, dissolving with methanol, and fixing volume to obtain mixed reference solution;
(2) Preparation of test solution: precisely weighing appropriate amount of Galium Teneri powder, precisely adding methanol solution, soaking, weighing, refluxing, cooling to room temperature, weighing again, supplementing weight with methanol, filtering, centrifuging the filtrate, and collecting supernatant as sample solution;
(3) High performance liquid chromatography assay: respectively taking the mixed reference substance solution prepared in the step (1) and the sample solution prepared in the step (2), and injecting the mixed reference substance solution and the sample solution into a high performance liquid chromatograph for testing to obtain a high performance liquid chromatogram of the mixed reference substance solution and a high performance liquid chromatogram of the sample solution;
(4) Positioning chromatographic peaks of components to be detected: chlorogenic acid is used as an internal reference substance, and chromatographic peaks are positioned according to relative retention values between chlorogenic acid and new chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid A and isochlorogenic acid C in a high performance liquid chromatogram of the mixed reference substance solution;
(5) Calculation of the relative correction factor: calculating peak area of each chromatographic peak by using high performance liquid chromatogram of the mixed reference solution, and calculating relative correction factors between chlorogenic acid and new chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid A and isochlorogenic acid C by using chlorogenic acid as internal reference;
(6) Calculating the content of the components to be detected: and calculating the component content of the chlorogenic acid, the isochlorogenic acid A and the isochlorogenic acid C in the sample solution by using the relative correction factors between the chlorogenic acid and the chlorogenic acid, the isochlorogenic acid A and the isochlorogenic acid C in the mixed reference solution.
3. The method for quality control of a cleaver assay of claim 2, wherein in step (5), the relative correction factor is calculated as: the peak areas of the novel chlorogenic acid, the hidden chlorogenic acid, the isochlorogenic acid A and the isochlorogenic acid C are calculated by taking chlorogenic acid as an internal reference, and the relative correction factors are calculated by a multipoint correction method.
4. The method for quality control of a Galium-Teum-assay of claim 3, wherein said multipoint correction method is according to formula f i/s ＝f i /f s ＝(C s A i )/(A s C i ) Calculating a relative correction factor;
wherein A is s Peak area of internal reference, C s A is the concentration of the internal reference substance i For the peak area of the component to be measured, C i For the concentration of the component to be measured, f i/s Is a relative correction factor.
5. The method for quality control of a cleaver assay according to claim 3, wherein in step (5), the relative correction factor of chlorogenic acid and neochlorogenic acid is 1.024-1.055, the relative correction factor of chlorogenic acid and cryptochlorogenic acid is 0.921-0.931, the relative correction factor of chlorogenic acid and isochlorogenic acid A is 1.281-1.325, and the relative correction factor of chlorogenic acid and isochlorogenic acid C is 1.181-1.217.
6. The method for quality control of a cleaver assay of claim 2, wherein in step (4), the relative retention value is calculated according to the formula: r is (r) i/s ＝T R(i) /T R(s) ；
Wherein r is i/s For relative retention value, T R(i) For retention time of the component to be measured, T R(s) Is the retention time of the internal reference.
7. The method for quality control of a cleaver assay according to claim 6, wherein in step (4), the relative retention value of chlorogenic acid and neochlorogenic acid is 0.585-0.588, the relative retention value of chlorogenic acid and cryptochlorogenic acid is 1.113-1.114, the relative retention value of chlorogenic acid and isochlorogenic acid A is 2.182-2.200, and the relative retention value of chlorogenic acid and isochlorogenic acid C is 2.515-2.545.
8. The method for quality control of a cleaver assay of claim 2, wherein in step (3), the conditions for high performance liquid chromatography are: the chromatographic column is Guangzhou Fei Nigen X-Penyx C 18 A 4.6X105 mm,5 μm model chromatographic column; the mobile phase is methanol-0.1% formic acid aqueous solution, and gradient elution is carried out, wherein methanol is mobile phase A, and 0.1% formic acid aqueous solution is mobile phase B; the flow rate is 0.8-1.0 mL/min; the detection wavelength is 280-327 nm; column temperature is 25-30 ℃; the time is 60min, and the operation is 10min later; the sample injection amount is 8-10 mu L.
9. The method for quality control of a cleaver assay of claim 8, wherein the gradient elution is performed by the following steps: 0-10 min, 15-20% A; 10-33 min, 20-38% A; 33-35 min,38% A; 35-45 min, 38-50% A.
10. The method for quality control of a cleaver assay of claim 2, wherein in step (2), the sample solution is prepared by the following steps: taking 0.5-1 g of cleaver powder, precisely weighing, placing in a conical flask, precisely adding 15-20 mL of 60% methanol solution, firstly soaking for 20-30 min, weighing, refluxing for 50-60 min, cooling to room temperature, weighing again, supplementing weight with 60% methanol, filtering, taking 2-4 mL of continuous filtrate, centrifuging at the rotating speed of 10000-13000 r/s for 10-15 min, and taking supernatant as a sample solution.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1161868A1 (en) * | 2000-06-07 | 2001-12-12 | Aventis Cropscience S.A. | New herbicidal compositions |
| CN108473484A (en) * | 2015-10-01 | 2018-08-31 | 赛诺米克斯公司 | It can be used as the compound of TRPM8 conditioning agents |
| CN110297061A (en) * | 2019-07-25 | 2019-10-01 | 广西中医药大学 | Survey the methods for commenting method measurement Chinese ixeris herb Content of Chlorogenic Acid, caffeic acid and galuteolin content using one more |
| EP3572078A1 (en) * | 2018-05-24 | 2019-11-27 | ETH Zurich | Tomm6-interacting extracts and compounds for use in the treatment and prophylaxis of nervous system diseases |
| EP3888461A1 (en) * | 2020-04-03 | 2021-10-06 | Bayer AG | Non-aqueous dispersions comprising inhibitors of hydroxyphenylpyruvate-dioxygenase and citric acid |
-
2022
- 2022-08-18 CN CN202210992632.5A patent/CN116008445B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1161868A1 (en) * | 2000-06-07 | 2001-12-12 | Aventis Cropscience S.A. | New herbicidal compositions |
| CN108473484A (en) * | 2015-10-01 | 2018-08-31 | 赛诺米克斯公司 | It can be used as the compound of TRPM8 conditioning agents |
| EP3572078A1 (en) * | 2018-05-24 | 2019-11-27 | ETH Zurich | Tomm6-interacting extracts and compounds for use in the treatment and prophylaxis of nervous system diseases |
| CN110297061A (en) * | 2019-07-25 | 2019-10-01 | 广西中医药大学 | Survey the methods for commenting method measurement Chinese ixeris herb Content of Chlorogenic Acid, caffeic acid and galuteolin content using one more |
| EP3888461A1 (en) * | 2020-04-03 | 2021-10-06 | Bayer AG | Non-aqueous dispersions comprising inhibitors of hydroxyphenylpyruvate-dioxygenase and citric acid |
Non-Patent Citations (5)
| Title |
|---|
| LI JIANCHEN: "Research on quality control method of Galii Aparinis Herba", 《HEBEI JOURNAL OF INDUSTRIAL SCIENCE & TECHNOLOGY》, vol. 34, no. 3, 1 May 2017 (2017-05-01), pages 162 - 6 * |
| LI JIANCHEN: "Research on quality control method of Galii Aparinis Herba", 《HEBEI JOURNAL OF INDUSTRIAL SCIENCE & TECHNOLOGY》, vol. 34, no. 3, 31 May 2017 (2017-05-31), pages 162 - 6 * |
| 农新维: "壮药毛冬青配方颗粒一测多评的定量研究", 《中国现代应用药学》, vol. 34, no. 8, 30 August 2017 (2017-08-30), pages 1118 - 1121 * |
| 陈剑: "高效液相法色谱法测定猪殃殃中绿原酸含量", 《广州化工》, vol. 42, no. 24, 13 December 2014 (2014-12-13), pages 116 - 117 * |
| 陈剑: "高效液相法色谱法测定猪殃殃中绿原酸含量", 《广州化工》, vol. 42, no. 24, 30 December 2014 (2014-12-30), pages 116 - 117 * |
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