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Objective To investigate the effect of compatibility on brain tissue distribution in rat of corydaline, tetrahydropalmatine, protopine, imperatorin and isoimperatorin in Corydalis Rhizoma and Angelicae Dahuricae Radix, and to provide further experimental evidence for confirming the Q-marker of Yuanhu Zhitong Prescription. Methods After oral administration of Corydalis Rhizoma, Angelicae Dahuricae Radix, and Yuanhu Zhitong Prescription extracts respectively, the concentration of five compounds in brain tissue samples at different time points was determined by UPLC-MS/MS, and the effect of compatibility on brain tissue distribution was further evaluated by kinetic parameters. Results Compared with single-herb group, AUC and Cmax of corydaline, tetrahydropalmatine and protopine in Yuanhu Zhitong Prescription group were increased significantly, and MRT of imperatorin and isoimperatorin were prolonged at the same time. Conclusion Corydalis Rhizoma and Angelicae Dahuricae Radix exist synergistic effect, the compatibility of these two herbs promote brain tissue distribution of corydaline, tetrahydropalmatine, protopine, imperatorin and isoimperatorin, which can be used as Q-marker of Yuanhu Zhitong Prescription.
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Objective To determine the technological conditions for the purification of the total alkaloid from the Corydalis Rhizoma (CR) by macroporous adsorption resin. Methods Total alkaloids of CR were determined by acid dye colorimetry, palmatine hydrochloride, dehydrocorydaline, tetrahydropalmatine, and corydaline were determined by HPLC. Six macroporous adsorption resins were investigated with the absorption rates, elution rates, and the content of the total alkaloid and four alkaloids of CR by static and dynamic adsorbing experiments. The purification process conditions of the total alkaloid of CR were optimized by the loading amount and volume flow of sample, the type and volume of the impurity removal of solvent and elution solvent and so on. The stability of the purification process was investigated by 5, 10 times enlargement. Results D141 type macroporous adsorption resin was the best choice for the purification of the total alkaloid from CR, the optimized parameters were as follows: Drug concentration was 0.6 g/mL of medicinal material and was added to the D141 macroporous resin column that the ratio of diameter to height range from 1:5 to 1:9 at a flow rate of 2 BV/h to 2 BV, 1.3 BV of purified water was used to remove impurities, and then 6 BV 95% ethanol was used as eluent at a flow rate of 2 BV/h. The purity of the total alkaloid of CR was up to 68.19% after purification, and the content of palmatine hydrochloride, dehydrocorydaline, tetrahydropalmatine, and corydaline was 1.95%, 11.74%, 4.93%, and 6.36%, respectively. The purity of the total alkaloid of the CR can reach more than 65% by 5 times and 10 times enlargement. Conclusion The purity of the total alkaloids can reach more than 65% after verification test, and the transfer rate of total alkaloids and four alkaloid monomers of CR can reach more than 85%, indicating that D141 macroporous adsorption resin can effectively purify total alkaloids from CR, and can be applied to industrial production.
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Objective: To prepare the molecularly imprinted polymer (MIPs) of L-tetrahydropalmatine (L-THP) by the molecular imprinting technique and study on solid-phase extraction. Methods: Using L-THP as template, methyl acrylic acid (MAA) as functional monmer, and ehtylene glycol dimethacrylate (EDMA) as a cross-linking agent to prepare the L-THP-MIPs. A test was conducted to investigate the selectivity and the specificity of solid-phase extraction. Results: The experiment showed that the MIPs had the specific adsorption to L-THP, but did not have the specific adsorption to corydaline the structural analogue with L-THP. Conclusion: The L-THP-MIPs have a good selectivity and the specificity of L-THP.
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Objective: To establish an HPLC method for simultaneous assay of seven main active constituents (protopine, coptisine, palmatine, dehydrocorydaline, D-tetrahydro jatrorrhizine, tetrahydropalmatine, and corydaline) in Corydalis Rhizoma, and on this basis, establishing a methodology of quantitative analysis on multi-components by single marker (QAMS) to validate the feasibility of method and technical adaptability of quality control applications for Corydalis Rhizoma. Methods: Taking seven components in Corydalis Rhizoma as indicators, two correction methods were used to establish the relative correction factor (fk/s) between each component and tetrahydropalmatine. Then the correction factor was used to calculate the amount of each component in Corydalis Rhizoma and finally to achieve this method. In the meantime, the external standard method was used to measure the above seven components to compare the difference between the calculated and measured value of the two fk/s, and to validate the correctness and adaptability of QAMS. Results: The methodology of QAMS which was used to evaluate the seven kinds of alkaloids in Corydalis Rhizoma was established; There was no significant difference between the data calculated by QAMS in different columns and instruments and the values measured by the external standard method. Conclusion: The QAMS method for measuring the components of protopine, coptisine, palmatine, dehydrocorydaline, D-tetrahydro jatrorrhizine, tetrahydropalmatine, and corydaline in Corydalis Rhizoma is reliable and accurate, it could be used to control the quality of crude drugs and herbal pieces of Corydalis Rhizoma.
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Objective: To establish HPLC method for the simultaneous determination of protopine, palmatine hydrochloride, berberine hydrochloride, dehydrocorydaline, tetrahydropalmatine, tetrahydroberberine, and corydaline in Cuyanhusuo Granule. Methods: The analysis was performed on Ultimate AQ-C18 column (250 mm×4.6 mm, 5 μm) by gradient elution of acetonitrile-0.1% phosphoric acid (adjust to pH 6.0 with triethylamine) (10:90). The UV detection wavelength was set at 280 nm and the flow rate was 1.0 mL/min. Results: The linear ranges of protopine, palmatine hydrochloride, berberine hydrochloride, dehydrocorydaline, tetrahydropalmatine, tetrahydroberberine, and corydaline were 6.8-119.0 (r=0.9999), 24.38-426.65 (r=0.9999), 8.88-155.40 (r=0.9999), 77.66-1359.05 (r=0.9999), 41.4-724.5 (r=0.9999), 6.70-117.25 (r=0.9999), and 25.50-446.25 ng (r=0.9999). The average recoveries (n=6) were 98.2% (RSD=2.0%), 99.6% (RSD=2.8%), 100.2% (RSD=1.3%), 99.0% (RSD=2.2%), 100.8% (RSD=2.6%), 98.7% (RSD=2.5%), and 97.7% (RSD=2.2%), respectively. Conclusion: This method is simple and rapid, and can be used for the quality control of Cuyanhusuo Granule with satisfactory separation and repeatability.
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Objective: To establish a method of HPLC for simultaneous determination of six ingredients in Yuanhu Zhitong Dripping Pills. Methods: Separation was performed on a Diamonsil C18 column (250 mm × 4.6 mm, 5 μm); Mobile phase was acetonitrile (A)-0.04% ammonium acetate (acetic acid tone pH 4.0) (B), gradient elution: 0-10 min, 20% A, 10-25 min, 20%-25% A, 25-70 min, 25%-75% A; flow rate was 1 mL/min; column temperature was 35 ℃; detection wavelength was 280 nm; and injection volume was 10 μL. The contents of dl-tetrahydropalmatine, palmatine, dehydrocorydaline, d-corydaline, imperatorin, and isoimperation were determined. Results: The linear ranges were 14.94-149.40 μg/mL for dl-tetrahydropalmatine, 1.97-19.72 μg/mL for palmatine, 15.94-159.36 μg/mL for dehydrocorydaline, 9.72-97.20 μg/mL for d-corydaline, 2.82-28.17 μg/mL for imperatorin, and 0.61-6.06 μg/mL for isoimperation, respectively, with the correlation r > 0.999 0. The extraction recoveries varied from 95% to 105% (RSD varied from 1.82% to 2.99%). The contents of dl-tetrahydropalmatine, palmatine, dehydrocorydaline, d-corydaline, imperatorin, and isoimperation was 0.368-0.483, 0.058-0.082, 0.532-0.766, 0.315-0.459, 0.145-0.212, and 0.030-0.053 mg/g, respectively. Conclusion: The method is accurate and reliable, which can be used for the quality control of Yuanhu Zhitong Dripping Pills.