.hplc-ms/ms Method For The Determination Of Human Plasma Aquin Calcium Concentration In Human Plasma In Pharmacokinetic Studies

Objective To establish a RP-HPLC-MS/MS method for the determinati on of Acamprosate Calcium(ACS) in Human plasma.Methods API 3000 LC-MS/MS system and Gemini C₁₈ analysis colu mn (50 mm×3.0 mm, 3μm) were used. The protein of plasma samp le was precipitated with acetonitrile . After evaporation of the supernatan t, the residue was dissolved in mobile phase, washed with dichlorometh ane and injected onto the column. The mobile phase of 10 mmol.L^-1 a mmonium acetate- methanol(95:5, adjust pH 7.4 with ammonia water)was p umped at 0.2 mL.min^-1 through the column.Results The retention time for ACS is 2.4 min, The standard curve was linear over the concentration range of 2.0～2048μg.L^-1. The lowes t concentration of detection in plasma was 2.0μg.L^-1. The method reco very was 93 %～104 %; the intra-day RSD less than 4.0%; inter-day RSD less than 11.6 %.Conclusion This method was found to be simple, rapid, sensitive and accurate for determination of ACS in human plasma.Part 2 A human pharmacokinetics research of Acamprosate CalciumObjective Study Pharmacokinetics that the ACS tablets.Methods A randomized cross-over study design was adopted in the si ngle oral study. The plasma concentration of ACS were determined by HPLC-MS/MS method and the data were analyzed automatically with DAS 2.0 pharmacokinetic program. The pharmacokinetic parameters wer e analyzed by SPSS 10.5 statistic software.Results The main pharmacokinetic parameters of single oral 333, 666, 1332 mg were as follows, respectively. C_max were 134.8±103.9μg·L^-1, 297.5±188.1μg·L^-1 and 385.4±155.7μg·L^-1, T_max were 7.50±2.58 h, 7.42±2.15 h and 8.083±3.118 h. AUC_0-t were 1771.5±323.1μg L^-1·h, 3708.9±1194.9μg L^-1·h, 6421.2±2485.7μg L^-1·h, T_1/2 were 9.96±4.69 h, 10.97±8.60 h and 11.78±6.15 h; MRT_0～t, VRT_0～t, V_1/F, CL/F and T_lag were 15.65±2.19 h～17.60±2.47 h, 104.48±44.14 h²ï½ž119.37±43.39 h², 3069.9±1343.5 L～3248.3±1360.7 L, 225.2±99.2 L·h^-1～211.7±74.7 L·h^-1 and 2.42±1.14 h～2.87±0.65 h. The main pharmacokinetic parameters of standard multiple oral dose and multiple oral dose with meal were as follows, respectively. AUCss, AUC_0-t, C_max, C_min, C_av, DF were 3126.0±948.0μg L^-1·h and 2187.3±1164.4μg L^-1·h, 13513±5340μg L^-1·h and 9163±4832μg L^-1·h, 564.1±138.6μg·L^-1 and 423.2±216.1μg·L^-1, 419.0±156.7μg·L^-1 and 300.1±164.6μg·L^-1, 390.8±118.5μg·L^-1 and 273.4±145.6μg·L^-1,0.393±0.425 and 0.459±0.301, T_max, T_1/2, MRT_0-t, VRT_0-t, V_1/F, CL/F were 8.75±4.05 h and 8.30±5.96 h, 13.60±10.60 h and 8.74±3.35 h, 17.14±3.28 h and 17.15±2.71 h, 128.8±47.7 h² and 114.8±25.7 h², 943.3±513.7 L and 1478.8±1185.9 L, 60.9±36.0 L·h^-1 and 125.3±89.0 L·h^-1.±Conclusion Single oral dose followed open one compartment model. There was no significant difference between male and female volunteers in pharmacokinetic parameters except 666mg CL/F, among different doses in pharmacokinetic parameters except AUC, C_z, C_max. The relative bioavailability of multiple oral doses with meal was 67.8 % of standard multiple oral dose. The C_max of multiple oral doses with meal was 75.0% of standard multiple oral doses. There was no significant difference between C_av(moring) and C_av(evening) of multiple oral doses with meal.