Pharmacokinetic comparison between Tanreqing Capsules Substitute and Tanreqing Capsules in rats by LC-MS/MS / 中国中药杂志

Due to the limited resource of bear bile powder, the major raw material of Tanreqing Capsules(TRQ), cultured bear bile powder is used as a replacement to develop the Tanreqing Capsules Substitute(TRQS). An LC-MS/MS method was established in this study for simultaneous quantitation of 8 compounds from TRQS in rat plasma: tauroursodeoxycholic acid(TUDCA), taurocheno-deoxycholic acid(TCDCA), ursodeoxycholic acid(UDCA), chenodeoxycholic acid(CDCA), ferulic acid, wogonoside, baicalin, and forsythoside A. Thereby, the pharmacokinetic behaviors of TRQ and TRQS were evaluated. Concentration of endogenous compounds TUDCA, TCDCA, UDCA, and CDCA was determined with the stable isotope surrogate analytes: D4-TUDCA, D4-TCDCA, D4-UDCA, and D4-CDCA. Plasma samples were extracted by acetonitrile-induced protein precipitation. The LC conditions are as follows: Waters BEH C_(18) column(2.1 mm×100 mm, 1.7 μm), mobile phase of 10 mmol·L~(-1) ammonium formate aqueous solution(containing 0.01% formic acid) and acetonitrile-methanol mixture(1∶5). MS conditions are as below: multiple reaction monitoring(MRM), ESI~(+/-). Concentration of UDCA, CDCA, TUDCA, and TCDCA was corrected with a response factor, which is the ratio between the responses recorded for the surrogate and the authentic analyte at the equal concentration. Each of the plasma components showed good linearity(r > 0.995 1). Accuracy and precision met the criteria(inter-day RSD<7.0%, RE 89.98%-112.0%; intra-day RSD<12%, RE 90.41%-111.2%). The recovery was 64.83%-119.9% and matrix effect was 87.15%-113.8%. The validated method was applied for pharmacokinetic study of TRQS and TRQ(po, 0.94 g·kg~(-1)). There was no significant difference in C_(max) and AUC_(0-24 h) of baicalin, UDCA, TUDCA, and TCDCA between the two groups, indicating similar pharmacokinetic behaviors between TRQS and TRQ in rats.

Research on Change of Constituents of Bear Bile Powder During Production Process of Tanreqing Capsules by HPLC-MS/MS / 中国药学杂志

OBJECTIVE: To identify the chemical components of bile acids from bear bile powder, the intermediate of bear bile powder and Tanreqing capsules by HPLC-MS/MS. METHODS: All samples were extracted with 70% methanol(V/V), and an Ultimate XB C18 column (4.6 mm×250 mm, 5 μm) was employed for separation with acetonitrile-0.1% formic acid as mobile phase in gradient elution. The MS spectrum was acquired in both positive and negative ion mode using ESI ion source. The chemical components were identified by the second mass spectrometric pyrolysis fragments, chromatographic peak retention time and fragmentation regularity summarized from the reference standards and the available literature. RESULTS: A total of 33 compounds were successfully identified or tentatively predicted, and six chemical compounds including tauroursodeoxycholic acid, taurochenodeoxycholic acid, ursodeoxycholic acid, chenodeoxycholic acid, 7α-hydroxy-3-oxo-5β-cholanic acid and one unknown constituent were finally transferred to Tanreqing capsules through the intermediate of bear bile powder. Moreover, 21 new chemical compounds (major ingredients of free bile acids) were generated during the production process of the intermediate, and 19 components were also detected in Tanreqing capsules. CONCLUSION: The investigation of the change of constituents in bear bile powder during Tanreqing capsules production provides a basis for the quality control and evaluation of Tanreqing capsules during production process.

Simultaneous Determination of 7 Components in Tanreqing Capsules by HPLC / 中国药房

OBJECTIVE:To establish a method for the simultaneous determination of 7 components in Tanreqing capsules.METHODS:HPLC method was adopted.The determination was performed on Ultimate XB C18 column with mobile phase consisted of acetonitrile-0.2％ formic acid (gradient elution) at a flow rate of 0.8 mL/min.The detection wavelength was set at 325 nm,and column temperature was 35 ℃.The sample size was 10 μL.RESULTS:The linear ranges of chlorogenic acid,isoforsythiaside A,forsythiaside A,isochlorogenic acid B,isochlorogenic acid A,isochlorogenic acid C and baicalin were 0.025-0.5 μg(r=0.999 6),0.025-0.5 μg (r=0.999 7),0.050-1.0 μg (r=0.999 9),0.025-0.5 μg((r=0.999 7),0.025-0.5 μg ((r=0.999 6),0.025-0.5 μg (r=0.999 6),0.750-1.5 μg(r=0.999 9),respectively.The limit of quantitation was no more than 1.5 ng,and the limit of detection was 0.5 ng.RSDs of precision,stability and reproducibility tests were all lower than 3.0％.The recoveries were 95.28％-106.30％ (RSD ranged 0.97％-2.14％,n=9).CONCLUSIONS:The method is simple,precise,stable and reproducible,and can be used for simultaneous determination of 7 components in Tanreqing capsules.