GC-MS测定血清胆固醇及其标志物前处理条件正交试验优化研究

采用正交试验优化GC-MS测定血清胆固醇及其标志物的样品前处理条件，并对最优试验方案进行方法学评价。采用L₁₆(211)正交设计对影响GC-MS测定血清胆固醇及其标志物前处理的3个关键步骤共7个因素——皂化(KOH-乙醇浓度、皂化温度和时间)、萃取(正己烷用量)和衍生化(衍生化温度、时间和用量)进行优化。得到最优前处理条件组合如下：KOH-乙醇溶液浓度为1 mol·L-1;皂化温度70 ℃;皂化时间60 min;萃取剂用量2 mL;衍生化温度70 ℃;衍生化时间60 min;衍生化试剂用量100 μL。所建方法准确性和精密性良好，方法学评价指标优于文献报道。

Orthogonal Experiment for Optimization of the Sample Pretreatment Conditions for the Serum Cholesterol and Markers Determination by GC-MS

The obj ective is to use orthogonal experiment to optimize the pretreatment on the determination of serum cholesterol and its markers by GC-MS.And then the method is evaluated in a methodological perspective.The methodis to Use L16 (211 )or-thogonal experiment design to observe the influence of three key steps,althogether seven factors of pretreatment,which are sa-ponification (KOH ethanol solution concentration,temperature and time),extraction (dose)and derivatization (temperature , time and dose).As for the results,the conditions of optimal pretreatment are as follows:the ethanol solution is 1 mol·L-1 KOH,the saponification temperature is 70 ℃;the saponification time is 60 min;the Solvent quantity is 2 mL;the derivatization temperature is 70 ℃;the derivatization time is 60 min,and the derivatization agent is 100μL.Through the optimization by or-thogonal design and methodological evaluation,the determination of serum cholesterol and its markers by GC-MS is excellent in terms of accuracy and precision,and methodological evaluation indexes are better than those reported in other papers.