CN116642973A - Method for evaluating quality uniformity among batches of salvia miltiorrhiza injection - Google Patents

Method for evaluating quality uniformity among batches of salvia miltiorrhiza injection Download PDF

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CN116642973A
CN116642973A CN202310617445.3A CN202310617445A CN116642973A CN 116642973 A CN116642973 A CN 116642973A CN 202310617445 A CN202310617445 A CN 202310617445A CN 116642973 A CN116642973 A CN 116642973A
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谭欢钦
李存玉
李明明
沈平平
崔辉
吴祝军
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JIANGSU SHENLONG PHARMACEUTICAL CO Ltd
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Abstract

The invention provides a quality uniformity control method based on the concept of 'existence state-component content', and constructs an estimation method applicable to the existence state of phenolic acid components in a red sage root injection according to the correlation of component states and filtering behaviors. According to the action rule of phenolic acid components and membrane materials, polytetrafluoroethylene materials are selected as separation layers of the ultrafiltration membrane, and the interface rejection effect with water-soluble phenolic acid components is enhanced, so that the sensitivity of the change between the existence state of the components and the separation coefficient is improved, and the accuracy of the calculation result is ensured.

Description

Method for evaluating quality uniformity among batches of salvia miltiorrhiza injection
Technical Field
The invention belongs to the technical field of medicines, and particularly relates to a method for evaluating the quality uniformity among batches of salvia miltiorrhiza injection, in particular to a method for evaluating the quality uniformity among batches of salvia miltiorrhiza injection constructed based on the existence state and the ingredient distribution characteristics of traditional Chinese medicine ingredients.
Background
The research, production and quality control of the traditional Chinese medicine preparation follow the basic principles of controllable quality, safety and effectiveness, and are subject to the complexity of chemical components of the traditional Chinese medicine preparation and more sources of variation of production units, and the uniformity of the quality of the product is still difficult to ensure by the process operation mode of traditional index detection, so that the quality of the products is different among the same prescription, different manufacturers and even different batches.
The traditional Chinese medicine preparation quality uniformity control method relies on all links of product manufacture such as design development, production control and logistics management, and the like, carries out the concept that the quality is derived from process control, carries out the production process monitoring on seed seedling breeding, planting base optimization, medicinal material harvesting, decoction piece processing and preparation processes of traditional Chinese medicine, and refers to index components, characteristics/fingerprints in quality standards, so that the transmission rule of the components is clear, intelligent control is carried out on key production links, and the controllability of the production process is improved, thereby guaranteeing the uniformity of the traditional Chinese medicine preparation quality.
However, the current quality uniformity evaluation method can only adjust the production parameters by visual component content data, but does not understand why the same batch of medicinal materials and the same production process parameters are difficult to produce preparations with high quality, so that root causes causing quality differences cannot be interpreted.
The salviae miltiorrhizae injection has the effects of promoting blood circulation to remove blood stasis, promoting blood circulation and nourishing heart, takes salvianolic acid B, rosmarinic acid, protocatechuic aldehyde and sodium salvianolic acid as main medicinal components, is also a quality control index mainly concerned in the production process, and is difficult to realize quantitative control of the component state in actual production by only qualitatively judging the component existence state through the component dissociation constant at present. The method of correlation fitting according to the nanofiltration mass transfer coefficient and concentration can only calculate molecular state, and is difficult to collect nanofiltration separation data for the solution environment with higher viscosity of the salvia miltiorrhiza injection, so that obvious limitations exist.
Disclosure of Invention
The invention provides a quality uniformity control method based on the concept of 'existence state-component content', and constructs an estimation method applicable to the existence state of phenolic acid components in a red sage root injection according to the correlation of component states and filtering behaviors. According to the action rule of phenolic acid components and membrane materials, polytetrafluoroethylene materials are selected as separation layers of the ultrafiltration membrane, and the interface rejection effect with water-soluble phenolic acid components is enhanced, so that the sensitivity of the change between the existence state of the components and the separation coefficient is improved, and the accuracy of the calculation result is ensured.
Based on the background, the method for evaluating the quality batch-to-batch uniformity of the salvia miltiorrhiza injection is provided, the quality control method of the salvia miltiorrhiza injection is enriched, the rationality of the batch-to-batch uniformity evaluation of the traditional Chinese medicine preparation is further improved, and the production risk of traditional Chinese medicine enterprises is effectively reduced.
Therefore, the invention provides a method for evaluating quality uniformity among batches of salvia miltiorrhiza injection, which comprises the following steps:
a. taking the red sage root injection, adopting polytetrafluoroethylene ultrafiltration membrane, regulating the transmembrane pressure difference to be 0.02-0.2MPa, collecting membrane flux J1 under corresponding pressure, collecting corresponding ultrafiltrate, and calculating retention rate R1;
b. according to the concentration of the index components in the red sage root injection, respectively preparing mixed solutions of index component reference substances, regulating the pH value of the solutions to 2.0-3.0, adopting polytetrafluoroethylene ultrafiltration membranes, regulating the transmembrane pressure difference to 0.02-0.2MPa, collecting membrane flux J2 under corresponding pressure, collecting corresponding ultrafiltrate, and calculating the retention rate R2 of the index components;
c. according to the concentration of the index components in the red sage root injection, respectively preparing mixed solutions of the index component reference substances, regulating the pH value of the solutions to be 6.0-7.0, adopting polytetrafluoroethylene ultrafiltration membranes, regulating the transmembrane pressure difference to be 0.02-0.2MPa, collecting membrane flux J3 under corresponding pressure, collecting corresponding ultrafiltrate, and calculating the retention rate R3 of the index components;
d. calculating the separation coefficient S of the index components when the solution has the molecular weight cut-off of 1kDa, 3kDa, 5kDa, 8kDa and 10kDa in the steps a-c according to a formula ln [ J/R-J ] -ln [ DK/delta ] =J/S, wherein delta is the thickness of an ultrafiltration membrane separation layer, and DK is a mass transfer constant;
e. fitting linear equations of the separation coefficient S and the molecular weight cutoff M of the index components in the red sage root injection in the step a and the mixed solution of the reference substances with different pH values in the steps b-c respectively;
f. respectively collecting linear equation slope K of the index components in the red sage root injection in the step a and the mixed solution of the reference substances with different pH values in the steps b-c;
g. according to the slope values (K) fitted by the mixed solutions of different pH reference substances, the ionic state 1% (pH 2.0) and the ionic state 99% (pH 7.0) of the corresponding component states (Z), taking the ionic state proportion as an abscissa, taking the slope K as an ordinate to make a linear equation of two points, and respectively carrying the slopes of the linear equation of salvianolic acid B, protocatechuic aldehyde, salvianic acid and rosmarinic acid in the salvianic acid injection into the equation to calculate the proportion of the molecular state and the ionic state of the index component in the salvianic acid injection;
h. taking the ratio of the molecular state to the ionic state as a weighting coefficient, and multiplying the weighting coefficient by the index component characteristics in the salvia miltiorrhiza injection to obtain a uniformity evaluation index, thereby evaluating the quality uniformity of the salvia miltiorrhiza injection among batches.
Further, the ultrafiltration membrane is made of polytetrafluoroethylene with the molecular weight cutoff of 1KDa-10 KDa.
Further, in the steps a, b and c, the transmembrane pressure difference is adjusted to be 0.02, 0.05, 0.10, 0.15 and 0.20MPa.
Further, in the steps a, b, C, the retention rate (%) = (1-C) Ultrafiltrate solution /C Stock solution ) X 100%, where C Ultrafiltrate solution C as the concentration of the index component in the ultrafiltrate Stock solution Is the concentration of the index component in the red sage root injection sample.
Further, the index components are salvianolic acid B, protocatechuic aldehyde, salvianic acid A and rosmarinic acid.
Further, the separation coefficient S characterizes a performance parameter of the index component approaching and penetrating the membrane separation layer.
Further, in the step h, the characteristic of the index component refers to one or more of component content, characteristic spectrum and fingerprint spectrum.
The invention has at least the following beneficial effects:
(1) Based on the mode of ingredient state-ingredient content dual-index synergy, the method for evaluating the quality uniformity of the salvia miltiorrhiza injection among batches is improved.
(2) According to the strong interface repulsive effect between phenolic acid components in the red sage root injection and polytetrafluoroethylene materials, the sensitivity of the change between the existence state of the components and the separation coefficient is improved, and the accuracy of the calculation result is ensured.
(3) According to the ionic state and molecular state of the phenolic acid in the salvia miltiorrhiza injection, the method for evaluating the quality uniformity among batches of the salvia miltiorrhiza injection with the correlation between multi-state and multi-component content is further formed, and the quality control method of the salvia miltiorrhiza injection is enriched.
Compared with the traditional evaluation indexes such as fingerprint, the batch-to-batch similarity of the salvia miltiorrhiza injection is high, so that the fingerprint similarity is high, and the batch-to-batch difference is difficult to evaluate; according to the invention, the ratio of the molecular state and the ionic state of the index component is used as a weighting coefficient, and the calculated quality uniformity evaluation index can reflect the quality uniformity of the salvia miltiorrhiza injection among batches more comprehensively and objectively.
Detailed Description
The above-described matters of the present invention will be further described in detail by way of examples, but it should not be construed that the scope of the above-described subject matter of the present invention is limited to the following examples, and all techniques realized based on the above-described matters of the present invention are within the scope of the present invention.
Example 1
Example 1: calculation of existence state of phenolic acid component in red sage root injection
Taking radix Salviae Miltiorrhizae injection (batch No. 2021-1201, jiangsu Shenlong pharmaceutical Co., ltd.), adopting polytetrafluoroethylene ultrafiltration membrane with molecular weight cut-off (MWCO) of 1kDa, 3kDa, 5kDa, 8kDa, 10kDa, regulating transmembrane pressure difference to 0.02, 0.05, 0.10, 0.15, 0.20MPa, collecting membrane flux under corresponding pressure, collecting corresponding ultrafiltration liquid, and detecting salvianolic acid B, protocatechuic aldehyde, salvianic acid A and rosmarinic acid content in the stock solution and ultrafiltrate by adopting the following chromatographic conditions.
Chromatographic conditions: SCIENTIFIC C18 chromatography column (4.6 mm. Times.250 mm,5 μm); column temperature: 30 ℃; detection wavelength: 286nm; flow rate: 1mL/min; sample injection amount: 10. Mu.L; mobile phase: the methanol-1% acetic acid aqueous solution was eluted in a gradient, and the elution procedure is shown in Table 1.
TABLE 1 elution procedure
Time/min Methanol/% 1% aqueous acetic acid solution/%
0 10 90
22 30 70
50 55 45
60 10 90
Linear relationship: preparing a mixed reference substance solution with mass concentrations of salvianolic acid B, protocatechuic aldehyde, salvianic acid A and rosmarinic acid of 1.520, 0.362, 0.610 and 0.850mg/mL respectively, precisely sucking 0.10, 0.20, 0.50, 1.00 and 2.00mL of the mixed reference substance solution respectively, placing the mixed reference substance solution in a 5mL measuring flask, fixing the volume of a 50% methanol aqueous solution to a scale, detecting by an Agilent high performance liquid chromatograph 1260, taking the peak area as an ordinate (Y), taking the concentration of the reference substance solution as an abscissa (X), obtaining a linear regression equation,
the salvianolic acid B Y =16.16x+10.02, and the R2= 0.9991 have good linear relation within the range of 30.4-608 mu g/mL;
protocatechuic aldehyde y=6.91x+12.45, r2=0.9995, and the linear relationship is good in the range of 7.24-144.8 μg/mL;
tanshinol y=4.02x+7.11, r2= 0.9992, and the linear relation is good in the range of 12.2-244 μg/mL;
rosmarinic acid y=10.68x+13.37, r2=0.9990, and the linear relationship is good in the range of 17 to 340 μg/mL.
And according to the retention (%) = (1-C) Ultrafiltrate solution /C Stock solution ) X 100%, where C Ultrafiltrate solution C as the concentration of the index component in the ultrafiltrate Stock solution Is the concentration of the index component in the red sage root injection sample. Flux data and retention data for the membrane pore sizes of the series are shown in table 2.
TABLE 2 flux data for series of membrane pore sizes, rejection
TABLE 3 rejection of index Components at different rejection pressures and molecular weights
According to the concentration of salvianolic acid B, protocatechuic aldehyde, salvianic acid A and rosmarinic acid (0.15 mg/ml of salvianolic acid B, 0.22mg/ml of protocatechuic aldehyde, 0.60mg/ml of salvianic acid A and 0.20mg/ml of rosmarinic acid) in the salvianic acid A injection, preparing a mixed solution of salvianolic acid B, protocatechuic aldehyde, salvianic acid A and rosmarinic acid reference substances, regulating the pH value of the solution to be 2.0, adopting polytetrafluoroethylene ultrafiltration membranes with interception molecular weights of 1kDa, 3kDa, 5kDa, 8kDa and 10kDa, regulating the transmembrane pressure difference to be 0.02, 0.05, 0.10, 0.15 and 0.20MPa, collecting membrane flux under corresponding pressure, and collecting corresponding ultrafiltrate, calculating the interception rate of components, and obtaining data as shown in Table 5.
TABLE 4 Membrane flux at the corresponding pressures
TABLE 5 rejection of indicated components at various rejection pressures and molecular weights
Preparing a mixed solution of salvianolic acid B, protocatechuic aldehyde, salvianic acid A and rosmarinic acid reference substances according to the concentration of the salvianolic acid B, protocatechuic aldehyde, salvianic acid A and rosmarinic acid in the red sage root injection, regulating the pH value of the solution to 7.0, adopting polytetrafluoroethylene ultrafiltration membranes with the interception molecular weights of 0.02, 0.05, 0.10, 0.15 and 0.20MPa, regulating the transmembrane pressure difference to 0.02, 0.05, 0.10, 0.15 and 0.20MPa, collecting membrane flux under corresponding pressure, collecting corresponding ultrafiltrate, calculating the component interception rate, and collecting the result as shown in table 7.
TABLE 6 Membrane flux at the corresponding pressures
TABLE 7 rejection of index Components at different rejection pressures and molecular weights
According to the correlation of the membrane flux J, the separation coefficient S and the retention rate R, the separation coefficient S of the salvianolic acid B, protocatechuic aldehyde, salvianic acid and rosmarinic acid when the retention molecular weight of the salvianic acid injection is 1kDa, 3kDa, 5kDa, 8kDa and 10kDa is calculated according to the formula ln [ J/R-J ] -ln [ DK/delta ] =J/S, and the result is shown in Table 8.
TABLE 8 separation coefficient S of index components
The separation coefficient S of salvianolic acid B, protocatechuic aldehyde, salvianic acid A and rosmarinic acid at the molecular weight cut-off of 1kDa, 3kDa, 5kDa, 8kDa and 10kDa was calculated for the mixed control solution (pH 2.0), and the results are shown in Table 9.
TABLE 9 separation coefficient S of index components
The separation coefficient S of salvianolic acid B, protocatechuic aldehyde, salvianic acid A and rosmarinic acid at the molecular weight cut-off of 1kDa, 3kDa, 5kDa, 8kDa and 10kDa was calculated for the mixed control solution (pH 7.0), and the results are shown in Table 10.
TABLE 10 separation coefficient S of index components
The linear equation of the separation coefficient S and the molecular weight cut-off (M) of salvianolic acid B, protocatechuic aldehyde, salvianic acid A and rosmarinic acid in the mixed solution of the salvianolic acid B injection and the reference substances with different pH values are respectively fitted, the correlation coefficients are all more than 0.9, and the results are shown in Table 11.
TABLE 11 Linear equation of separation coefficient S and molecular weight cut-off (M) for index components
The linear equation slopes of salvianolic acid B, protocatechuic aldehyde, salvianic acid and rosmarinic acid in the mixed solution of the red sage root injection and the different pH reference substances are respectively collected, the linear equation (which is calculated by taking the ionic state proportion as the abscissa and taking the slope K as the ordinate and using the excel two-point linear relation) of two points is made according to the slope value (K) fitted by the two pH reference substances, the ionic state proportion and the molecular state proportion of the corresponding component state (Z) are calculated according to the ionic state 1% (pH 2.0) and the ionic state 99% (pH 7.0), and the slopes of four components in the red sage root injection are brought into the linear equation, and the results are shown in Table 12.
Table 12
Composition of the components Linear equation Ionic state ratio% Molecular state ratio%
Salvianolic acid B Z＝-11.31K+110.31 48.46 51.54
Protocatechuic aldehyde Z＝-15.08K+114.08 35.81 64.19
Salvianic acid A Z＝-21.86K+120.86 5.89 94.11
Rosmarinic acid Z＝-16.04K+115.04 28.27 71.73
Wherein the ion state ratio of the components in the red sage root injection of batches 2021-1201 is respectively 48.46% of salvianolic acid B, 35.81% of protocatechuic aldehyde, 5.89% of tanshinol and 28.27% of rosmarinic acid.
Example 2: uniformity analysis of red sage root injection among different batches
The ionic ratios of the components in the red sage root injections of batches 2021-1101, 2021-1110, 2021-1201, 2021-1208 were examined by the same technical means as in example 1, and the results are shown in Table 13.
TABLE 13
The uniformity evaluation indexes of the four components are respectively compared and averaged with the uniformity evaluation indexes of the batches 2021-1110 by taking the ionic state proportion as a weighting coefficient and multiplying the ionic state proportion with the concentration of the components in the solution to obtain uniformity evaluation indexes, and the uniformity evaluation indexes are respectively 0.912, 1.000, 0.904 and 0.961.
Example 3: evaluation of blood lipid regulating effect of red sage root injection in different batches
Animals: male ICR mice, SPF grade, 70, body mass (20+ -2) g, supplied by the university of Nanjing laboratory animal center. The test animal was approved by SCXK 2020-0005, and the license plate number was SYXK 2018-0049. The raising environment is ventilated and dry, quiet, sufficient in light and free to take drinking water. The experimental process meets the ethical requirements of experimental animals of Nanjing university of traditional Chinese medicine, and the trial and batch number is 410983211100040072.
Drug and reagent: the injection comprises Saviae Miltiorrhizae radix injection (Jiangsu Shenlong pharmaceutical Co., ltd., lot numbers 20211101, 20211201), saviae Miltiorrhizae radix injection (Shenwei pharmaceutical Co., ltd., lot number SW-202210-1), fenofibrate capsule (French Li Bofu Ni pharmaceutical Co., lot number H20200105).
The method comprises the following steps: ICR 70 mice were filled with 20mL/kg of a high-fat emulsion (containing 15g of lard, 6g of cholesterol, 2g of sodium cholate, 0.2g of propylthiouracil, 4mL of Tween-80, distilled water to a volume of 100mL, reference to the lipid emulsion preparation method) per day. On day 21 of molding, 10 mice were randomly sampled, blood was collected, and the TG, CHO, LDL-C content of serum was detected, all three of which were significantly elevated as successful molding. The 60 mice after successful molding were randomly divided into model group, non-specific group, 20211101 group, 20211201 group, SW-202210-1 group, 10 mice each. 10 mice that were not subjected to the molding treatment were used as a blank group.
After grouping, the model group and each administration group continued to administer the high-fat emulsion every morning, the model group was intraperitoneally injected with physiological saline every afternoon, and each administration group was interfered with the corresponding drug every afternoon, and was administered at a clinically equivalent dose of 1-fold for 70kg adults. The human injection dose of the red sage root injection is 20mL per day, the converted mouse intraperitoneal injection dose is 2.5mL/kg per day, the oral dose of the fenofibrate capsule is 200mg per day, and the converted mouse gastric lavage dose is 26mg/kg per day. The blank group was infused with saline daily in the morning and intraperitoneally injected with saline in the afternoon. Each group was continuously intervened for 21 days.
The mice are fasted without water for 12 hours, the fasting body mass is measured, after the last administration for 2 hours, the anesthesia is carried out by intraperitoneal injection of pentobarbital sodium, the blood is taken from the eyeground venous plexus, the centrifugation (with the centrifugal radius of 8 cm) is carried out for 10 minutes at 3000r/min, and the serum is separated for standby. Serum TG, CHO, HDL-C, LDL-C was detected using a fully automated biochemical analyzer.
Statistical methods: statistical analysis was performed using SPSS Statistics 22.0 software. The measurement data is represented by mean value +/-variance, and accords with normal distribution, the comparison between two groups adopts t test, and the comparison between multiple groups adopts single factor variance analysis; non-parametric testing was used for those that did not meet normal distributions. The difference of P < 0.05 is statistically significant. The results are shown in Table 14.
TABLE 14
Group of TG CHO HDL-C LDL-C
Blank group 1.26±0.25 2.92±0.53 6.97±1.81 0.41±0.16
Model group 2.04±0.76* 3.57±0.80* 5.81±1.14* 0.64±0.22*
Non-special group 1.43±0.37# 2.47±0.69# 6.35±1.04 0.42±0.14#
20211101 group 1.58±0.46# 2.86±0.30# 6.42±0.49 0.52±0.08#
20211201 group 1.52±0.39# 2.55±0.26# 6.33±0.57 0.47±0.10#
SW-202210-group 1 1.64±0.55# 2.98±0.45# 6.56±0.60 0.59±0.13#
Note that: p < 0.05 compared to the blank; compared to the model group, #P < 0.05.
Comparing the red sage root injection of different manufacturers and batches in table 14, compared with blank group, the contents of TG, CHO and LDL-C in model group are obviously raised (P < 0.05), and compared with model group, the contents of CHO and LDL-C in each administration group are obviously lowered (P < 0.05), and the contents of TG in non-special group are obviously lowered (P < 0.05); the HDL-C content of each administration group is compared with that of the model group, and the difference has no statistical significance (P is more than 0.05); the difference between the TG, CHO, LDL-C contents of 20211101 group, 20211201 group and SW-202210-1 group was statistically significant (P < 0.05) compared with the model group. Compared with the two batches 20211101 and 20211201 of Jiangsu shenlong pharmaceutical industry limited company, the influence on the blood lipid level is relatively close, the four index components of salvianolic acid B, protocatechuic aldehyde, salvianic acid and rosmarinic acid in the two batches are relatively consistent, the reason for the influence difference is possibly related to the existence state of the components, and the difference of SW-202210-1 is possibly the result of the combined action of the component content difference and the existence state.
Example 4: homogeneity evaluation of red sage root injection produced by different manufacturers
Using the same technical procedure as in example 1, 3 manufacturers, two lots of each manufacturer, were collected and labeled SL-20221101, SL-20221201, SW-202210-1, SW-202212-1, SH-2023001, SH-2023005, respectively, for the ionic ratios of the components in the red sage root injections, and the results are shown in Table 15.
TABLE 15
From the difference in the existence state of the components between different factories and batches in table 13, the uniformity evaluation indexes are obtained by multiplying the ionic state proportion as a weighting coefficient by the concentration of the components in the solution, and the uniformity evaluation indexes of the four components are respectively compared and averaged with the uniformity evaluation indexes of the batch SL-20221101 by taking the batch SL-20221101 as a reference, so that the uniformity is respectively 1.000, 0.964, 0.730, 0.845, 0.782 and 0.457.
The fingerprint spectrum evaluation is carried out on the batches, the results are all larger than 0.85, the batch-to-batch uniformity is difficult to evaluate, but the quality uniformity control method based on the correlation concept of the existence state and the component content can reflect the quality difference among batches of different manufacturers more according to the correlation of the component state and the filtering behavior.

Claims (7)

1. A method for evaluating quality uniformity among batches of salvia miltiorrhiza injection, comprising the following steps:
a. taking the red sage root injection, adopting polytetrafluoroethylene ultrafiltration membrane, regulating the transmembrane pressure difference to be 0.02-0.2MPa, collecting membrane flux J1 under corresponding pressure, collecting corresponding ultrafiltrate, and calculating retention rate R1;
b. according to the concentration of the index components in the red sage root injection, respectively preparing mixed solutions of index component reference substances, regulating the pH value of the solutions to 2.0-3.0, adopting polytetrafluoroethylene ultrafiltration membranes, regulating the transmembrane pressure difference to 0.02-0.2MPa, collecting membrane flux J2 under corresponding pressure, collecting corresponding ultrafiltrate, and calculating the retention rate R2 of the index components;
c. according to the concentration of the index components in the red sage root injection, respectively preparing mixed solutions of the index component reference substances, regulating the pH value of the solutions to be 6.0-7.0, adopting polytetrafluoroethylene ultrafiltration membranes, regulating the transmembrane pressure difference to be 0.02-0.2MPa, collecting membrane flux J3 under corresponding pressure, collecting corresponding ultrafiltrate, and calculating the retention rate R3 of the index components;
d. calculating the separation coefficient S of the index components when the solution has the molecular weight cut-off of 1kDa, 3kDa, 5kDa, 8kDa and 10kDa in the steps a-c according to a formula ln [ J/R-J ] -ln [ DK/delta ] =J/S, wherein delta is the thickness of an ultrafiltration membrane separation layer, and DK is a mass transfer constant;
e. fitting linear equations of the separation coefficient S and the molecular weight cutoff M of the index components in the red sage root injection in the step a and the mixed solution of the reference substances with different pH values in the steps b-c respectively;
f. respectively collecting linear equation slope K of the index components in the red sage root injection in the step a and the mixed solution of the reference substances with different pH values in the steps b-c;
g. according to the slope values (K) fitted by the mixed solutions of different pH reference substances, the ionic state 1% (pH 2.0) and the ionic state 99% (pH 7.0) of the corresponding component states (Z), taking the ionic state proportion as an abscissa, taking the slope K as an ordinate to make a linear equation of two points, and respectively carrying the slopes of the linear equation of salvianolic acid B, protocatechuic aldehyde, salvianic acid and rosmarinic acid in the salvianic acid injection into the equation to calculate the proportion of the molecular state and the ionic state of the index component in the salvianic acid injection;
h. taking the ratio of the molecular state to the ionic state as a weighting coefficient, and multiplying the weighting coefficient by the index component characteristics in the salvia miltiorrhiza injection to obtain a uniformity evaluation index, thereby evaluating the quality uniformity of the salvia miltiorrhiza injection among batches.
2. The evaluation method according to claim 1, wherein the ultrafiltration membrane is a polytetrafluoroethylene ultrafiltration membrane having a molecular weight cut-off of 1kDa to 10 kDa.
3. The method according to claim 1, wherein in the steps a, b, and c, the transmembrane pressure difference is adjusted to 0.02, 0.05, 0.10, 0.15, and 0.20MPa.
4. The evaluation method according to claim 1, wherein in the steps a, b, C, the retention rate (%) = (1-C Ultrafiltrate solution /C Stock solution ) X 100%, where C Ultrafiltrate solution C as the concentration of the index component in the ultrafiltrate Stock solution Is the concentration of the index component in the red sage root injection sample.
5. The evaluation method according to claim 1, wherein the index component is salvianolic acid B, protocatechuic aldehyde, tanshinol and rosmarinic acid.
6. The evaluation method according to claim 1, wherein the separation coefficient S characterizes a performance parameter of the index component approaching and penetrating the membrane separation layer.
7. The evaluation method according to claim 1, wherein in the step h, the characteristic of the index component means one or more of component content, characteristic spectrum, fingerprint spectrum.
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