对乙酰氨基酚论文：三厂家对乙酰氨基酚口服制剂生物等效性与体内外.doc

对乙酰氨基酚论文：三厂家对乙酰氨基酚口服制剂生物等效性与体内外相关性研究【中文摘要】建立对乙酰氨基酚血浓度测定方法；以健康志愿者为研究对象,对体内外释放存在差异的不同厂家对乙酰氨基酚口服制剂进行人体生物等效性研究,评价以体外释放试验作为口服制剂质量标准的可行性,以及体外溶出存在差异产品,以体外溶出试验代替体内吸收实验的可能性；评价体外释放试验与体内吸收的相关性及对乙酰氨基酚片的质量,为其临床合理应用、质量评价提供依据。方法：实验采用单剂量三周期三交叉实验设计。健康志愿者12名,男性,随机分为3组,分别于试验当日晨空腹一次口服安徽联谊药业股份有限公司(A)、上海华源安徽仁济制药有限公司(B)、中美史克制药有限公司(R)对乙酰氨基酚片1片(规格0.5g),于药前和药后0.25、0.5、0.75、1.0、1.5、2.0、2.5、3.0、4.0、7.0、10.0、15.0h取肘静脉血4mL,4000r/min离心l0min,取血浆于-20贮存,备测。清洗期为7天,7天、14天后各组交叉服药,进行第二、三周期试验。以咖啡因为内标,采用高效液相色谱法测定对乙酰氨基酚经时血浓度,色谱柱为Diamonsil(钻石)C18(5m,2504.6mm)；流动相为乙腈-水(1783,VV),用前经0.45m微孔滤膜过滤并经超声脱气；流速1.OmLmin-1；柱温室温；检测波长237nm；进样量10L，保留时间定性,内标法定量。根据所测对乙酰氨基酚血浓度-时间数据,应用DAS 2.0实用药代动力学程序计算主要药代动力学参数(t1/2,AUC0-t,AUCo0-),Cmax和Tmax,对主要药代动力学参数进行方差分析、双单侧t检验和(1-2)置信区间分析,评价生物等效性。参照中国药典口服制剂体外溶出试验要求,进行三制剂体外溶出度试验,计算三制剂的体外溶出参数T50和Td,评价体外释放与体内吸收的相关性。结果：对乙酰氨基酚在0.0225gmL-1范围内线性关系良好(r=0.9966)。当信噪比2(SN2)时,血浆最低定量限为0.02g-mL-1,符合试验要求。低、中、高浓度(含对乙酰氨基酚0.05,1和20gmL-l)质控样本绝对回收率均满足试验要求。对乙酰氨基酚A、B、R片口服给药后对乙酰氨基酚药动学参数t12分别为(2.4110.506)h、(2.8490.554)h和(2.7940.543)h,Tmax为(1.3750.598)h、(0.9791.003)h和(0.9790.432)h；AUC0-15为(27.24310.866)gmL-1,(27.6438.008)g-mL-1h和(26.7557.005)gmL-1h;AUC0为27.68010.944)gml-1h, (28.3638.156)gml-1h和(27.4637.289)mL-1h。对乙酰氨基酚A、B、R片体外溶出参数T5o分别为35.221min、3.33Omin和2.530min,Td为46.828min,4.590 min和3.620min。结论：1.多因素方差分析显示,A、B、R三制剂口服三交叉试验对乙酰氨基酚主要药代动力学参数AUC和Cmax药物间及周期间无显著性差异,但个体间存在统计学差异。进一步双单侧t检验结果符合生物等效性的统计学要求,符合生物等效的假设, A、B、R三种对乙酰氨基酚制剂具有生物等效性,为生物等效制剂。2.A、B、R三种对乙酰氨基酚片体外溶出度与体内吸收相关性分析结果显不,Cmax,Tmax与T50,Td均有较好相关性,可在一定程度上以体外溶出速度预测体内吸收速度和程度。但三片剂体外溶出差异较大,B和R片剂在30min内基本溶出100%，而A片剂30min时仅溶出40%左右。体内吸收(hTmax表示)也存在差异,即A片剂Tmax为1.3750.598h,B和R片剂Tmax分别为0.9791.003h和0.9790.432h。【英文摘要】:To evaluate the bioequivalence and to compare the differences of absorption in vitro and distribution in vivo of Acetaminophen tablets from three different manufacturers, for the possible applications of bioequivalence as quality method of oral preparations; To evaluate the inherent quality of acetaminophen tablets, to provide guideline for its reasonable clinical application and quality evaluation.Methods:Health volunteers were involved in a single dose,3-period cross-over test. 12 healthy male volunteers were divided into 3 groups randomly, and orally given Acetaminophen Tablets A (Anhui Lanyi Pharmaceutical Co., LTD), Tablets B (Shanhai huayuananhuirenji Pharmaceutical Co., LTD), Tablets R (Tianjin Smith Kline) 0.5g, respectively.4ml elbow vein blood were collected before and 0.25,0.5, 0.75,1,1.5,2,2.5,3,4,7,10,15.0h after administration. All blood were heparinized and centrifuged at 4000rpm for 10 minutes, and plasma was separated and stored at-20for analysis. Volunteers were taken Tables A, B or R after one or two washing period (7 days or 14 days), in second- or third-period. An HPLC method was used to determine the concentration of acetaminophen in plasma. Diamonsil (diamond) C18 column (5m,2504.6mm) with acetonitrile-water (17/83, V/V) as mobile phase at flow rate of 1.0mLmin-1 was used to separate acetaminophen in human plasma. The column temperature was maintained at 25, and the detection wavelength was 237nm. Acetaminophen concentration were determined at above chromatography condition and its main pharmacokinetic parameters such as t1/2, AUC0-t, AUC0-, were calculated by software of DAS 2.0. Analysis of variance, t-test and (1-2) confidence interval were used to evaluate the bioequivalence.The dissolution tests of acetaminophen tablets were conducted according to Guidelines affiliated to Chinese Pharmacopoeia and T50 and Td of acetaminophen tablets were calculated for the evaluation of the correlation between dissolution in vitro and absorption in vivo.Results:Acetaminophen was linear in the range of 0.0225g-mL-1 (r=0.9966). The Limit of quantification was 0.02g-mL-1 with the signal to noise ratio2 (S/N2). The recoveries of acetaminophen at 0.05,1 and 20g-mL-1 met the requirements. The main acetaminophen pharmacokinetics parameters after oral administration of Acetaminophen Tablet A, B and R were as follows, t1/2 (2.4110.506) h,(2.8490.554) h and (2.7940.543) h, Tmax (1.3750.598) h,(0.9791.003) h and (0.9790.432)h; AUC015 (27.24310.866)g-mL-1, (27.6438.008)g-mL-1h and (26.7557.005)g-mL-1h; AUC0(27.68010.944)g-mL-1h, (28.3638.156)g-mL-1h and (27.4637.289)g-mL-1h. The dissolution parameters of Acetaminophen Tablet A, B and R were as follows, T5035.221min,3.330min and 2.530min, Td 46.828min,4.590min and 3.620min.Conclusion:1. Multiple factor analysis of variance showed that main pharmacokinetic parameters such as AUC and Cmax, of Acetaminophen Tablets A, B and R had no significant differences between preparations and periods, but there were significant differences between individuals. Further bioequivalence of t-test met the statistical requirements and complied with the bio-equivalent hypothesis, and three acetaminophen A, B, R tablets were bioequivalent.2. The dissolution in vitro of acetaminophen A, B, R tablets and absorption in vivo were well correlated in Cmax, Tmax and T50, Td, which indicated that in a certain extent, dissolution speed in vitro could reflect the absorption rate and degree in vivo.Acetaminophen Tablets B and R were dissoluted to 100% within 15 min, but Tablet A was dissoluted to only 40% in 30 min. here are differences, Tmax of tablet A in vivo was (1.3750.598) h, tablets B and R were (0.9791.003) h and (0.9790.432) h, respectively.【关键词】对乙酰氨基酚 咖啡因 生物等效性 体内外相关性 HPLC【英文关键词】acetaminophen caffeine bioequivalence in vitro-in vivo correlation HPLC【