固相萃取人体血液中安定条件的统计学筛选与响应面优化

Optimization of solid phase extraction conditions of diazepam from human plasma using statistical screening and response surface method

采用Plackett-Buman(P-B)法和中心复合设计(Central Composite Design,简称CCD)对影响固相萃取安定的6个因素进行筛选优化。P-B实验设计与统计学分析表明:pH、上样速度、洗脱液用量是影响回收率的3个关键因素。以回收率为响应目标,对3因素进行中心复合设计,并经响应面法优化分析得到影响回收率的二阶模型,确定了安定萃取实验的最优操作条件:pH10.20,上样速度0.67 mL/min,洗脱液用量2.60 mL,实测回收率达到91.26%。在0.10～10.00μg/mL的范围内本方法线性良好(R2>0.99),检测限为0.07μg/mL,日内和日间相对标准偏差(RSD)<10%,准确度(RE)<±6.0%。

Plackett-Burman（P-B） design and Central Composite Design（CCD） were applied to optimize 6 factors for the extraction diazepam by solid phase extraction.The P-B design and its following statistical analysis indicate that the pH,sample speed,eluent dosage were the 3 key factors which significantly influenced recovery of diazepam as the response objective,and a quadratic model was obtained by CCD for the 3 factors by analysis of response surface optimization.Under the optimal extraction conditions,pH 10.20,sample speed of 0.67 mL/min and eluent dosage of 2.60 mL,the recovery of diazepam reached 91.26%.The calibration curves were linear over the concentration range of 0.10～10.00 μg/mL（R2〉0.99） and the lower limit of quantification（LOQ） was 0.07 μg/mL.The intra-and inter-day precision（RSD） were less than 10% and accuracy（RE） was less than±6.0%.