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OBJECTIVE:To develop a method for simultaneous determination of carbamazepine,venlafaxine,rosiglitazone and nifedipine in human plasma.METHODS:UPLC-MS/MS method was adopted to determine plasma sample after precipitated with methanol (containing 0.1％ formic acid) using pioglitazone hydrochloride as internal standard.The determination was performed on Waters ACQUITY UPLC HSS C18 column with mobile phase consisted of aqueous solution containing 0.01％ formic acid-methanol containing 0.01％ formic acid (gradient elution) at the flow rate of 0.3 mL/min.The column temperature was 50 ℃,and sample size was 5 μL.ESI was used for positive ion scanning by multi reaction monitoring mode.The ion pairs for quantitative analysis were m/z 237.00 to 194.05 (carbamazepine),m/z 278.20 to 58.10 (venlafaxine),m/z 358.08 to 135.04 (rosiglitazone),m/z 347.15 to 315.17 (nifedipine),m/z 357.09 to 134.06 (internal standard).RESULTS:The liner ranges of carbamazepine,venlafaxine hydrochloride,rosiglitazone hydrochloride and nifedipine were 2.00-2 000.00 (r=0.995 9,n=5),2.12-2 120.00(r=0.990 5,n=5),2.00-2 000.00(r=0.991 5,n=5) and 2.04-1 020.00(r=0.991 0,n=5) ng/mL;the minimum detection limits were 0.200,0.106,0.100,1.020 ng/mL,respectively.RSDs of inter-day and intra-day were less than 15％.The absolute values of RE were less than 15％.The extraction recoveries were 65.66％-96.36％ (RSD％ ＜15％,n=5),and matrix effects ranged 80.99％-114.33％.The plasma concentrations of carbamazepine in 2 epileptic patients were (1 500.41 ± 169.11),(1 159.01 ±59.24) ng/mL(RSD were 11.27％,5.60％,n=3).The plasma concentrations of nifedipine in 2 hypertensive patients were (14.68 ±1.92),(21.18 ± 3.59)ng/mL (RSD were 16.98％,13.10％,n=3).CONCLUSIONS:The method is simple,specific,sensitive,accurate and suitable for the monitoring of plasma concentration and pharmacodynamic study of above drugs.
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Objective To investigate the inhibitory effect of ester catechin monomers-EGCG, GCG on the growth of human colon carcinoma cell line SW480 and its potential mechanism. Methods MTT assay, soft-agar colony formation test, Hoechst 33258 stain and flow cytometry analysis were used to determine the inhibitory effect of theasinesin(TS), and its monomers-EGCG, GCG on SW480 growth. Results EGCG, GCG and TS significantly inhibited the growth of human colon carcinoma cell line SW480 in dose-dependent manner, and their half inhibitory concentration (IC 50 ) was 108.88, 183.21, and 83.36?g?ml -1 , respectively. 24 hours after treatment with IC 50 of EGCG, GCG and TS, the colony formation rate of SW480 cells in the experimental group was obviously lower than that in the control group. Hoechst 33258 staining showed typical apoptotic features such as cell shrinkage, nuclear condensation and cell splinter in the experimental group. A subdiploid peak before G 0/G 1 phase in the experimental group was observed by flow cytometry. Conclusion In accord with TS, EGCG and GCG could inhibit the growth of colon carcinoma cell line SW480 cells, the mechanism of which may be related to inducing cell apoptosis.