Quantitative analysis method for compound state proportion in traditional Chinese medicine solution

Abstract

The invention relates to a quantitative analysis method for a composite state proportion in a traditional Chinese medicine solution, which quantitatively calculates the composition proportion of a traditional Chinese medicine compound based on the correlation between the mass transfer process of a composite state membrane in the traditional Chinese medicine solution and the component state. The invention provides the method for analyzing the composite state of the acid-base components of the traditional Chinese medicine without being separated from the original solution environment, the method is helpful for analyzing the real existence state of the acid-base compound under the complex solution system of the traditional Chinese medicine, the technical bottleneck that the structure of the compound formed by self-assembly of the acid-base components in the traditional Chinese medicine solution cannot be analyzed is solved, and the analysis of the composite state of the acid-base components in the traditional Chinese medicine solution is facilitated. The method is helpful for quality control and efficacy uniformity evaluation of traditional Chinese medicine preparations in the fields of medicines, pharmacology and the like.

Description

Technical Field

[0001] This invention relates to a quantitative analysis method for the proportion of complex states in a traditional Chinese medicine solution. Specifically, it relates to the correlation between the membrane mass transfer process and the component state of complexes and molecular components in a traditional Chinese medicine solution, and to the quantitative calculation of the composition ratio of the complexes. This method belongs to the pharmaceutical field. Background Technology

[0002] A complex is a product formed by the reaction of two compounds, resulting in the exchange of their chemical compositions. A complex can also be simply understood as the recombination of molecules. In the production of traditional Chinese medicine, due to the diversity of components, complexes often manifest as combinations formed between two or more different molecules, such as acid-base complexes or other complexes. Qingwei Huanglian Wan (Clear Stomach Coptis Pill) is composed of 14 traditional Chinese medicines, including Coptis chinensis and Glycyrrhiza uralensis. The effective components are extracted by decocting the herbs together, followed by filtration, concentration, granulation, and tableting. The effective components of Coptis chinensis are alkaloids, while Glycyrrhiza uralensis contains components with a certain degree of weak acidity. When the two are decocted together, they form a complex of berberine hydrochloride and glycyrrhizic acid. Qualitative analysis of the chemical composition of the decoction before and after combining Coptis chinensis with Evodia rutaecarpa and determination of the dissolution rate of the main component, berberine, revealed that the dissolution rate of berberine in the decoction decreased after the two were combined. This is mainly due to the formation of a large molecular complex between berberine and other alkaloids in Coptis chinensis and flavonoids in Evodia rutaecarpa. Phenolic acids and alkaloids are common components in traditional Chinese medicine. During the pharmaceutical process, acid-base complex reactions occur, altering the state of these components in solution and causing changes in their physicochemical properties such as solubility, sublimation, and volatility. Specifically, these changes manifest as alterations in the laws governing mass transfer during the pharmaceutical process, thus affecting the quality uniformity and product stability of traditional Chinese medicine preparations. Therefore, clarifying the proportion of acid-base complexes during the pharmaceutical process of traditional Chinese medicine is crucial for improving the standardization of traditional Chinese medicine preparation production.

[0003] Although researchers have developed advanced methods and techniques for studying the interactions of components in traditional Chinese medicine (TCM), such as infrared spectrophotometry (IR), nuclear magnetic resonance spectroscopy (NMR), X-ray diffraction (XRD), and molecular dynamics (MD), these methods can only qualitatively infer the state of the compounds and cannot accurately analyze the interactions between components in acid-base complexes of TCM. In the analysis of acid-base complexes, the aforementioned techniques are limited to requiring the compounds to be removed from their original solution environment. However, at this point, the state of the components has fundamentally changed, and the binding sites of intermolecular complexation reactions have changed, making accurate evaluation difficult. Furthermore, these methods can only provide qualitative analysis of the state of existence and cannot interpret the micellar state of saponin components in complex solution systems.

[0004] Based on the intrinsic relationship between the state of a complex and its separation behavior, this study amplifies the differences in permeability coefficients of complexes during separation by adjusting the transmembrane process. It then fits the mass transfer rate coefficients directly related to the state of the complexes and uses the separation behavior of monomers in the same solution environment as a reference to quantitatively calculate the proportion of complexes in traditional Chinese medicine solutions. This analysis helps guide the process control of traditional Chinese medicine preparations.

[0005] Based on the above background, the present invention provides an analytical method for the complex state of acid-base components in traditional Chinese medicine without removing them from the original solution environment. This invention will help clarify the true existence state of acid-base complexes in complex solution systems of traditional Chinese medicine and facilitate better work in pharmacology and other fields. Summary of the Invention

[0006] Purpose of the invention: The purpose of this invention is to provide a quantitative analysis method for the proportion of complex states in traditional Chinese medicine solutions, which can be used to analyze the existence state of complexes formed by acid-base components in complex solution environments.

[0007] Another objective of this invention is to provide a complex state analysis method that can be used to analyze the autonomous packing patterns in traditional Chinese medicine solutions and to evaluate the quality uniformity and efficacy differences of the corresponding preparations.

[0008] Technical Solution: A quantitative analysis method for the proportion of complex components in a traditional Chinese medicine solution, comprising the following steps: a. Take the traditional Chinese medicine solution to be analyzed and determine the molecular weight W of the target component; b. Take the traditional Chinese medicine solution to be analyzed, centrifuge at 10000 rpm for 15 minutes to obtain the supernatant, and detect the concentration C1 of the target component; c. Take the supernatant from step b, and separate it using ultrafiltration membranes with molecular weight cutoffs of 1, 3, 5, 8, 10, and 30 kDa, respectively, adjusting the transmembrane pressure difference to 0.05, 0.10, 0.15, and 0.20 MPa, collecting the ultrafiltrate respectively, detecting the concentration C2 of the target component, and calculating the permeate rate of the target component = C2 / C1 × 100%; d. Plot the logarithmic function equations of the permeate rate of the target component and the molecular weight cutoff of the ultrafiltration membrane under a series of transmembrane pressure differences, with a correlation coefficient greater than 0.9, where the logarithmic coefficient in the logarithmic function equation is the permeability coefficient K; e. Calculate the linear correlation equation between the transmembrane pressure difference and the permeability coefficient K in step d, with the slope being the permeate coefficient K. f. Using the permeate rate of the target component in the ultrafiltrate in step c as an indicator, the ultrafiltrate produced by the 30 kDa ultrafiltrate membrane is separated by ultrafiltration membranes with molecular weight cutoffs of 1, 3, 5, 8, 10, and 30 kDa. The transmembrane pressure difference is adjusted to 0.05, 0.10, 0.15, and 0.20 MPa, and the ultrafiltrate is collected respectively. The concentration C3 of the target component is detected, and the permeate rate of the target component is calculated as C3 / C2×100%. g. The logarithmic function equations of the permeate rate of the target component and the molecular weight cutoff of the ultrafiltration membrane at the series of transmembrane pressure differences in step f are plotted. The correlation coefficient is greater than 0.9, where the logarithmic coefficient in the logarithmic function equation is the permeate permeate coefficient Kv. h. The linear correlation equation between the transmembrane pressure difference and the permeate coefficient Kv in step g is calculated. The slope is the mass transfer rate Tv, and the correlation coefficient of the equation is greater than 0.9. i. Based on the mass transfer rates in steps e and h, the proportion P of the complex state in the traditional Chinese medicine solution is calculated with reference to formula (1).

[0009]

[0010] The formation of the complex involves a complex reaction between phenolic acids and alkaloids.

[0011] A quantitative analysis method for the proportion of complex states in a traditional Chinese medicine solution can be used for the quantitative calculation of the proportion of complex states in aqueous solutions during the quality analysis of traditional Chinese medicine preparations.

[0012] Beneficial effects: (1) It solves the technical bottleneck that the complex structure formed by the self-assembly of acid and base components in the solution system of Chinese medicine with multiple components cannot be analyzed. (2) It does not destroy the component composition and solvent system, and realizes the quantitative calculation of the existence state of acid and base complex of Chinese medicine in the original solution environment. Detailed Implementation

[0013] The following examples further illustrate the above-mentioned content of the present invention in detail, but should not be construed as limiting the scope of the subject matter of the present invention to the following examples. All technologies implemented based on the above-mentioned content of the present invention fall within the scope of the present invention.

[0014] Example 1: Calculation of the complex proportion of berberine in the aqueous extract of Coptis chinensis-Scutellaria baicalensis pair

[0015] Weigh 5.0 kg each of Coptis chinensis and Scutellaria baicalensis, add 12 times the amount of water, mix and reflux for extraction twice, 1 hour each time, combine the extracts, and find that the molecular weight of berberine is W336.36 Da. Take the combined extract, centrifuge at 10000 rpm for 15 minutes, obtain the supernatant, and determine the concentration of berberine by high performance liquid chromatography (HPLC). The chromatographic conditions are as follows: Waters e2695 HPLC system, full-wavelength UV detector, Waters XTerra MS C. 18 The chromatographic column was prepared with acetonitrile (A)-0.05 mol / L (B) potassium dihydrogen phosphate solution (50:50) (0.4 g of sodium dodecyl sulfate was added per 100 ml, and the pH was adjusted to 4.0 with phosphoric acid). The flow rate was 1.0 mL / min, the column temperature was 35 ℃, and the detection wavelength was 347 nm. The concentration of berberine in the supernatant, C1, was calculated to be 6.07 mg / mL using the external standard one-point method.

[0016] Take the supernatant and use ultrafiltration membranes with molecular weight cutoffs of 1, 3, 5, 8, 10, and 30 kDa for separation. Adjust the transmembrane pressure difference to 0.05, 0.10, 0.15, and 0.20 MPa respectively. Collect the ultrafiltrate and detect the concentration C2 of the target component. Calculate the permeability of the target component = C2 / C1 × 100%. The results are shown in Table 1.

[0017] Table 1

[0018]

[0019] Logarithmic function equations were plotted for the permeate flow rate (Y) of the target component versus the molecular weight cutoff (X) of the ultrafiltration membrane under various transmembrane pressure differences: Pressure 0.05 MPa: Y = 0.1712 ln(X) + 0.3716, correlation coefficient 0.9611; Pressure 0.10 MPa: Y = 0.1859 ln(X) + 0.2981, correlation coefficient 0.9592; Pressure 0.15 MPa: Y = 0.1956 ln(X) + 0.2308, correlation coefficient 0.9652; Pressure 0.20 MPa: ... Y = 0.2102ln(X) + 0.1486, with a correlation coefficient of 0.9655; the permeability coefficients K corresponding to transmembrane pressure differences of 0.05, 0.10, 0.15, and 0.20 MPa are 0.1712, 0.1859, 0.1956, and 0.2102, respectively; the linear correlation equation between transmembrane pressure difference (Δp) and permeability coefficient K is calculated as: K = 0.2534Δp + 0.1591, with a slope of mass transfer rate T of 0.2534 and a correlation coefficient of 0.9939.

[0020] Using the permeate permeate of berberine in the ultrafiltrate as an indicator, ultrafiltrate from a 30 kDa ultrafiltration membrane was separated using ultrafiltration membranes with molecular weight cutoffs of 1, 3, 5, 8, 10, and 30 kDa. The transmembrane pressure difference was adjusted to 0.05, 0.10, 0.15, and 0.20 MPa, and the ultrafiltrate was collected. The concentration C3 of the target component was detected, and the permeate permeate of the target component was calculated as C3 / C2 × 100%. The results are shown in Table 2.

[0021] Table 2

[0022]

[0023]

[0024] Logarithmic function equations were plotted for the permeate flow rate (Y) of the target component versus the molecular weight cutoff (X) of the ultrafiltration membrane under various transmembrane pressure differences: Pressure 0.05 MPa: Y = 0.1323 ln(X) + 0.598, correlation coefficient 0.9366; Pressure 0.10 MPa: Y = 0.1401 ln(X) + 0.5434, correlation coefficient 0.9465; Pressure 0.15 MPa: Y = 0.1596 ln(X) + 0.4662, correlation coefficient 0.9287; Pressure 0.20 MPa: Y = 0.1788 ln(X) + 0.4662, correlation coefficient 0.9287; Pressure 0.20 MPa: Y = 0.1788 ln(X) + 0.598, correlation coefficient 0.1323 ln(X) + 0.598, correlation coefficient 0.598; Pressure ...; Pressure n(X)+0.3963, correlation coefficient 0.9311; the permeability coefficients Kv corresponding to transmembrane pressure differences of 0.05, 0.10, 0.15, and 0.20 MPa are 0.1323, 0.1401, 0.1596, and 0.1788, respectively; calculate the linear correlation equation between transmembrane pressure difference (Δp) and permeability coefficient Kv: Kv=0.318Δp+0.113, the slope is the mass transfer rate Tv is 0.318, and the correlation coefficient of the equation is 0.9696; calculate the proportion P of the complex state in the traditional Chinese medicine solution according to formula (1).

[0025]

[0026] The proportion of berberine in the aqueous extract of the Coptis chinensis-Scutellaria baicalensis pair was 79.69%.

[0027] Example 2: Calculation of the complex proportion of chlorogenic acid in the aqueous extract of Coptis chinensis-Lonicera japonica herb pair

[0028] Weigh 5.0 kg each of Coptis chinensis and Lonicera japonica, add 12 times the amount of water, mix and reflux for extraction twice, 1 hour each time, combine the extracts, and find the molecular weight of chlorogenic acid to be W354.31 Da; take the combined extract, centrifuge at 10000 rpm for 15 minutes, obtain the supernatant, and determine the concentration of chlorogenic acid by high performance liquid chromatography (HPLC). The chromatographic conditions are as follows: Waters e2695 HPLC system, full-wavelength UV detector, Agela Venusil XBP C 18 The chromatographic column was used with gradient elution of acetonitrile (A) and 0.1% phosphoric acid solution (B) (0-8 min, 86%-81% B; 8-14 min, 81% B; 14-34 min, 81%-69% B; 34-35 min, 69%-10% B), flow rate 0.7 mL / min, column temperature 25℃, and detection wavelength 327 nm. The concentration of chlorogenic acid C1 in the supernatant was calculated to be 6.12 mg / mL using the single-point external standard method.

[0029] Take the supernatant and use ultrafiltration membranes with molecular weight cutoffs of 1, 3, 5, 8, 10, and 30 kDa for separation. Adjust the transmembrane pressure difference to 0.05, 0.10, 0.15, and 0.20 MPa respectively, collect the ultrafiltrate, detect the concentration C2 of the target component, and calculate the permeability of the target component = C2 / C1 × 100%. The results are shown in Table 3.

[0030] Table 3

[0031]

[0032] Logarithmic function equations were plotted for the permeate flow rate (Y) of the target component versus the molecular weight cutoff (X) of the ultrafiltration membrane under various transmembrane pressure differences: Pressure 0.05 MPa: Y = 0.0964 ln(X) + 0.6665, correlation coefficient 0.962; Pressure 0.10 MPa: Y = 0.1082 ln(X) + 0.59, correlation coefficient 0.9499; Pressure 0.15 MPa: Y = 0.1185 ln(X) + 0.5194, correlation coefficient 0.9277; Pressure 0.20 MPa: ... Y = 0.124ln(X) + 0.4859, with a correlation coefficient of 0.9371; the permeability coefficients K corresponding to transmembrane pressure differences of 0.05, 0.10, 0.15, and 0.20 MPa are 0.0964, 0.1082, 0.1185, and 0.124, respectively; the linear correlation equation between transmembrane pressure difference (Δp) and permeability coefficient K is calculated as: K = 0.1862Δp + 0.0885, with a slope of mass transfer rate T of 0.1862 and a correlation coefficient of 0.9764.

[0033] Using the permeate of chlorogenic acid in the ultrafiltrate as an indicator, ultrafiltrate from a 30 kDa ultrafiltration membrane was separated using ultrafiltration membranes with molecular weight cutoffs of 1, 3, 5, 8, 10, and 30 kDa. The transmembrane pressure difference was adjusted to 0.05, 0.10, 0.15, and 0.20 MPa, and the ultrafiltrate was collected. The concentration C3 of the target component was detected, and the permeate of the target component was calculated as C3 / C2 × 100%. The results are shown in Table 4.

[0034] Table 4

[0035]

[0036] Logarithmic function equations were plotted for the permeate flow rate (Y) of the target component versus the molecular weight cutoff (X) of the ultrafiltration membrane under various transmembrane pressure differences: Pressure 0.05 MPa: Y = 0.076 ln(X) + 0.7348, correlation coefficient 0.9583; Pressure 0.10 MPa: Y = 0.0876 ln(X) + 0.6837, correlation coefficient 0.9492; Pressure 0.15 MPa: Y = 0.0928 ln(X) + 0.6562, correlation coefficient 0.9379; Pressure 0.20 MPa: Y = 0.1116 ln(X) + 0.7348, correlation coefficient 0.9583; Pressure 0.10 MPa: Y = 0.1116 ln(X) + 0.7348, correlation coefficient 0.7348 ... (X)+0.5679, correlation coefficient 0.9256; the permeability coefficients Kv corresponding to the transmembrane pressure difference of 0.05, 0.10, 0.15, and 0.20 MPa are 0.076, 0.0876, 0.0928, and 0.1116, respectively; calculate the linear correlation equation between the transmembrane pressure difference (Δp) and the permeability coefficient Kv: Kv=0.224Δp+0.064, the slope is the mass transfer rate Tv is 0.224, and the correlation coefficient of the equation is 0.9501; calculate the proportion P of chlorogenic acid complex in the traditional Chinese medicine solution according to formula (1).

[0037]

[0038] The proportion of chlorogenic acid complex in the aqueous extract of Coptis chinensis-Lonicera japonica was 83.13%.

[0039] Example 3: Calculation of the complex proportion of matrine in the aqueous extract of the Sophora flavescens-Lonicera japonica herb pair

[0040] Weigh 5.0 kg each of Sophora flavescens and Lonicera japonica, add 10 times the amount of water, mix and reflux for extraction twice, 1 hour each time, combine the extracts, and check the molecular weight of matrine (W248.36 Da). Centrifuge the combined extract at 10000 rpm for 15 minutes to obtain the supernatant. Detect the concentration of matrine using high-performance liquid chromatography (HPLC). Chromatographic conditions are as follows: Waters e2695 HPLC system, full-wavelength UV detector, Waters XTerra MS C. 18 The chromatographic column used acetonitrile-anhydrous ethanol-2% phosphoric acid solution (81:10:9) as the mobile phase, with a flow rate of 1.0 mL / min, a column temperature of 30℃, and a detection wavelength of 220 nm. The concentration of matrine C1 in the supernatant was calculated to be 4.46 mg / mL using the external standard one-point method.

[0041] Take the supernatant and use ultrafiltration membranes with molecular weight cutoffs of 1, 3, 5, 8, 10, and 30 kDa for separation. Adjust the transmembrane pressure difference to 0.05, 0.10, 0.15, and 0.20 MPa respectively. Collect the ultrafiltrate and detect the concentration C2 of the target component. Calculate the permeability of the target component = C2 / C1 × 100%. The results are shown in Table 5.

[0042] Table 5

[0043]

[0044] Logarithmic function equations were plotted for the permeate flow rate (Y) of the target component versus the molecular weight cutoff (X) of the ultrafiltration membrane under various transmembrane pressure differences. For pressure 0.05 MPa: Y = 0.1271 ln(X) + 0.4988, correlation coefficient 0.9122; for pressure 0.10 MPa: Y = 0.1314 ln(X) + 0.4392, correlation coefficient 0.9121; for pressure 0.15 MPa: Y = 0.1482 ln(X) + 0.3726, correlation coefficient 0.9292; for pressure 0.20 MPa: ... Pa: Y = 0.1503ln(X) + 0.3136, correlation coefficient 0.9576; the permeability coefficients K corresponding to transmembrane pressure differences of 0.05, 0.10, 0.15, and 0.20 MPa are 0.1271, 0.1314, 0.1482, and 0.1503, respectively; the linear correlation equation between transmembrane pressure difference (Δp) and permeability coefficient K is calculated as: K = 0.162Δp + 0.1177, the slope is the mass transfer rate T is 0.162, and the correlation coefficient of the equation is 0.9744.

[0045] Using the permeate of matrine in the ultrafiltrate as an indicator, ultrafiltrate from a 30 kDa ultrafiltration membrane was separated using ultrafiltration membranes with molecular weight cutoffs of 1, 3, 5, 8, 10, and 30 kDa. The transmembrane pressure difference was adjusted to 0.05, 0.10, 0.15, and 0.20 MPa, and the ultrafiltrate was collected. The concentration C3 of the target component was detected, and the permeate of the target component was calculated as C3 / C2 × 100%. The results are shown in Table 6.

[0046] Table 6

[0047]

[0048] Logarithmic function equations were plotted for the permeate flow rate (Y) of the target component versus the molecular weight cutoff (X) of the ultrafiltration membrane under various transmembrane pressure differences: Pressure 0.05 MPa: Y = 0.0887 ln(X) + 0.6659, correlation coefficient 0.965; Pressure 0.10 MPa: Y = 0.10021n(X) + 0.6, correlation coefficient 0.9392; Pressure 0.15 MPa: Y = 0.1244 ln(X) + 0.4921, correlation coefficient 0.9208; Pressure 0.20 MPa: Y = 0.1494 ln(X) + 0.3 915, correlation coefficient 0.968; the permeability coefficients Kv corresponding to transmembrane pressure differences of 0.05, 0.10, 0.15, and 0.20 MPa are 0.0887, 0.1002, 0.1244, and 0.1494, respectively; calculate the linear correlation equation between transmembrane pressure difference (Δp) and permeability coefficient Kv: Kv=0.4126Δp+0.0641, the slope is the mass transfer rate Tv is 0.4126, and the correlation coefficient of the equation is 0.9759; calculate the proportion P of matrine complex in the extract of Sophora flavescens-Lonicera japonica according to formula (1).

[0049]

[0050] The proportion of chlorogenic acid complex in the aqueous extract of Sophora flavescens-honeysuckle pair was 39.26%, while the proportion of chlorogenic acid complex in the aqueous extract of Coptis chinensis-honeysuckle pair in Comparative Example 2 was 83.13%, indicating that the proportion of matrine forming acid-base complex in Sophora flavescens was weaker than that in Coptis chinensis alkaloids.

[0051] Example 4: Calculation of the complex proportion of berberine in Coptis chinensis aqueous extract

[0052] Weigh 5.0 kg of Coptis chinensis herb, add 12 times the amount of water, mix and reflux for extraction twice, 1 hour each time, combine the extracts, and find that the molecular weight of berberine is W336.36 Da. Take the combined extract, centrifuge at 10000 rpm for 15 minutes, obtain the supernatant, and determine the concentration of berberine by high performance liquid chromatography (HPLC). The chromatographic conditions are as follows: Waters e2695 HPLC system, full-wavelength UV detector, Waters XTerra MS C. 18 The chromatographic column was prepared with acetonitrile (A)-0.05 mol / L (B) potassium dihydrogen phosphate solution (50:50) (0.4 g of sodium dodecyl sulfate was added per 100 mL, and the pH was adjusted to 4.0 with phosphoric acid). The flow rate was 1.0 mL / min, the column temperature was 35 °C, and the detection wavelength was 347 nm. The concentration of berberine in the supernatant, C1, was calculated to be 21.07 mg / mL using the one-point external standard method.

[0053] Take the supernatant and use ultrafiltration membranes with molecular weight cutoffs of 1, 3, 5, 8, 10, and 30 kDa for separation. Adjust the transmembrane pressure difference to 0.05, 0.10, 0.15, and 0.20 MPa respectively. Collect the ultrafiltrate and detect the concentration C2 of the target component. Calculate the permeability of the target component = C2 / C1 × 100%. The results are shown in Table 7.

[0054] Table 7

[0055]

[0056] Logarithmic function equations were plotted for the permeate flow rate (Y) of the target component versus the molecular weight cutoff (X) of the ultrafiltration membrane under various transmembrane pressure differences: Pressure 0.05 MPa: Y = 0.0745 ln(X) + 0.7284, correlation coefficient 0.9182; Pressure 0.10 MPa: Y = 0.078 ln(X) + 0.7036, correlation coefficient 0.9194; Pressure 0.15 MPa: Y = 0.0799 ln(X) + 0.6816, correlation coefficient 0.9072; Pressure 0.20 MPa: ... Y = 0.0818ln(X) + 0.6394, with a correlation coefficient of 0.9154; the permeability coefficients K corresponding to transmembrane pressure differences of 0.05, 0.10, 0.15, and 0.20 MPa are 0.0745, 0.078, 0.0799, and 0.0818, respectively; the linear correlation equation between transmembrane pressure difference (Δp) and permeability coefficient K is calculated as: K = 0.0476Δp + 0.0726, with a slope of mass transfer rate T of 0.0476 and a correlation coefficient of 0.9736.

[0057] Using the permeate permeate of berberine in the ultrafiltrate as an indicator, ultrafiltrate from a 30 kDa ultrafiltration membrane was separated using ultrafiltration membranes with molecular weight cutoffs of 1, 3, 5, 8, 10, and 30 kDa. The transmembrane pressure difference was adjusted to 0.05, 0.10, 0.15, and 0.20 MPa, and the ultrafiltrate was collected. The concentration C3 of the target component was detected, and the permeate permeate of the target component was calculated as C3 / C2 × 100%. The results are shown in Table 8.

[0058] Table 8

[0059]

[0060] Logarithmic function equations were plotted for the permeate flow rate (Y) of the target component versus the molecular weight cutoff (X) of the ultrafiltration membrane under various transmembrane pressure differences: Pressure 0.05 MPa: Y = 0.0659 ln(X) + 0.7651, correlation coefficient 0.9113; Pressure 0.10 MPa: Y = 0.0749 ln(X) + 0.725, correlation coefficient 0.9221; Pressure 0.15 MPa: Y = 0.0811 ln(X) + 0.682 7. Correlation coefficient 0.9555; Pressure 0.20 MPa: Y = 0.0924 ln(X) + 0.6222, correlation coefficient 0.9967; Permeability coefficients Kv corresponding to transmembrane pressure differences of 0.05, 0.10, 0.15, and 0.20 MPa are 0.0659, 0.0749, 0.0811, and 0.0924, respectively; Calculate the linear correlation equation between transmembrane pressure difference (Δp) and permeability coefficient Kv: Kv ＝ =0.1714Δp+0.0572, the slope is the mass transfer rate Tv is 0.1714, and the correlation coefficient of the equation is 0.988; the proportion of berberine complex in Coptis chinensis extract P is calculated with reference to equation (1).

[0061]

[0062] The proportion of berberine in the aqueous extract of Coptis chinensis was 27.77%, indicating that berberine in the extract may have undergone a complex reaction with tannins, amino acids and other components in Coptis chinensis. However, the proportion of the complex state was lower than that of 79.69% in the Coptis chinensis-Scutellaria baicalensis pair in Example 1.

Claims

1. A method for quantitative analysis of the proportion of complex states in a traditional Chinese medicine solution, characterized in that, Includes the following steps: a. Take the solution of the Chinese medicine to be analyzed and check the molecular weight W of the target component; b. Take the Chinese medicine solution to be analyzed, centrifuge at 10,000 rpm for 15 minutes, obtain the supernatant, and detect the concentration C1 of the target component; c. Take the supernatant from step b, and use ultrafiltration membranes with molecular weight cutoffs of 1, 3, 5, 8, 10, and 30 kDa for separation. Adjust the transmembrane pressure difference to 0.05, 0.10, 0.15, and 0.20 MPa respectively, collect the ultrafiltrate, detect the concentration C2 of the target component, and calculate the permeability of the target component = C2 / C1 × 100%. d. Plot a series of logarithmic function equations for the permeate rate of the target component and the molecular weight cut-off of the ultrafiltration membrane under different transmembrane pressures, with a correlation coefficient greater than 0.9, where the logarithmic coefficient in the logarithmic function equation is the permeability coefficient K; e. Calculate the linear correlation equation between the transmembrane pressure difference and the permeability coefficient K in step d, with the slope being the mass transfer rate T, and the correlation coefficient of the equation being greater than 0.9; f. Using the permeation rate of the target component in the ultrafiltrate from step c as an indicator, ultrafiltrate generated by a 30 kDa ultrafiltration membrane is separated using ultrafiltration membranes with molecular weight cutoffs of 1, 3, 5, 8, 10, and 30 kDa. The transmembrane pressure difference is adjusted to 0.05, 0.10, 0.15, and 0.20 MPa, and the ultrafiltrate is collected respectively. The concentration C3 of the target component is detected, and the permeation rate of the target component is calculated as C3 / C2 × 100%. g. Plot the logarithmic function equations of the permeate rate of the target component and the molecular weight cut-off of the ultrafiltration membrane under the series of transmembrane pressure differences in step f, with a correlation coefficient greater than 0.9, where the logarithmic coefficient in the logarithmic function equation is the permeate coefficient Kv of the ultrafiltrate; h. Calculate the linear correlation equation between the transmembrane pressure difference and the permeability coefficient Kv in step g, with the slope being the mass transfer rate Tv, and the correlation coefficient of the equation being greater than 0.9; i. Based on the mass transfer rates in steps e and h, calculate the proportion P of the complex state in the traditional Chinese medicine solution according to equation (1).

2. The method for quantitative analysis of the proportion of complex states in a traditional Chinese medicine solution according to claim 1, characterized in that, The formation of the complex involves a complex reaction between phenolic acids and alkaloids.

3. The method for quantitative analysis of the proportion of complex states in a traditional Chinese medicine solution according to claim 1, characterized in that, It can be used for the quantitative calculation of the proportion of complex states in aqueous solutions during the quality analysis of traditional Chinese medicine preparations.

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