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Objective:To establish the determination method for berberine hydrochloride in Shenshe ointments. Methods: The quantitative analysis of berberine hydrochloride in Shenshe osintments was carried out by HPLC with a Kromsal C18 chromatographic column (250 mm × 4. 6 mm, 5 μm), the mobile phase was acetonitrile-0. 1% phosphoric acid (49∶51) (adding 0. 2g sodium dode-cylsulphate into 100 ml solution) with the flow rate of 1. 0 ml·min-1 and the detection wavelength of 320nm, the temperature of col-umn was room temperature, and the injection volume was 20 μl. Results: The linear range was 0. 059 2-0. 296 0 g·L-1 with good correlation(r=0. 999 4). The average recovery was 99. 80% and RSD was 0. 24%(n=6). Conclusion: The established method is accurate, specific and reproducible, and suitable for the determination of berberine hydrochloride in Shenshe ointments.
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Objective To establish a content determination method for paeoniflorin in qisheng capsule. Methods The quantitative analysis of paeoniflorin in qisheng Capsule was carried out by high-performance liquid chromatography ( HPLC) . The chromatographic separation was achieved by using a Kromasil C18 chromatographic column (4. 6 mm×250 mm,5 μm) with a mobile phase consisting of methanol,water (1585) at a flow rate of 1. 0 mL·min-1 and 230 nm detection wavelength. Results The linear range was 2. 5-12. 5 μg·mL-1( r =0. 999 9). The average recovery and RSD of the method were 99. 97%and 0. 94%. Conclusion The method is accurate,specific,reproducible,which can effectively be used in quality control of paeoniflorin in qisheng capsule.
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Objective To investigate the role of different culture densities of embryoid bodies (EBs) in cardiac dif-ferentiation of mouse embryonic stem cells (ESCs). Methods The mouse ESCs were cultured in hanging drops for 3 days, followed by another 2 days for suspension culture to form EBs. Suspended EBs of different densities (60 or 120 EBs/60 mm tissue culture dish) were transferred onto tissue culture plates. The cardiac specific troponin T (TnT) was detected by immu-nocytochemistry. The percentage of beating EBs was calculated at different time points. The mRNA expression of cardiac spe-cific transcriptional factors Nkx2.5, GATA4 and cardiac specific proteinsβ-MHC and ANF were detected by RT-PCR. The protein expressions of TnT and p38 were detected by Western-blot assay. Results Spontaneously beating EBs were posi-tively stained with TnT. There were significantly higher percentage of beating EBs, higher gene expression levels of Nkx 2.5, GATA4,β-MHC and ANF and higher protein expression of TnT in high density culture group than those of low density cul-ture group (P＜0.05). Furthermore, there was significantly higher activity of p38 pathway in high density culture group than that of low density culture group. And the percentages of beating EBs and TnT protein expression were decreased by p 38 pathway inhibitor SB203580. Conclusion The culture density of EBs is important in regulating the cardiac differentiation of ESCs. The high cell-density density culture of EBs enhances the cardiac differentiation of ESCs, which might be mediated by p38 signaling pathway.
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OBJECTIVE: To study the bioequivalence of tegaserod maleate dispersible tablets in healthy volunteers.METHODS: A single oral dose of 6mg test or reference preparations of tegaserod maleate was given to 22 healthy volunteers in a randomized crossover study.The plasma concentrations of tegaserod were determined by LC/MS/MS assay.RESULTS: The main pharmacokinetic parameters of test and reference products were as follows: tmax(0.86? 0.22) and(1. 01? 0.24) h;Cmax(2.21? 0.69) and(2.05? 0.64) ng? mL1;AUC0～ 17(6.35? 2.48) and(6.47? 1.99) ng? h? mL-1,AUC0～ ∞(6.69? 2.59) and(6.70? 2.03) ng? h? mL-1,respectively.The relative bioavailability of test to reference preparation was(98.2? 22.1) %.CONCLUSION: The reference preparation and the test preparation are bioequivalent.