同位素稀释气相色谱质谱法测定血清总胆固醇  

Determination of serum cholesterol by isotope dilution gas chromatography mass spectrometry

作　　者：张江涛[1] 胡翠华[1] 张传宝[1] 赵海舰[1] 马嵘[1] 谢洁红[1] 汪静[1] 董军 王抒 申子瑜[1] 陈文祥[1] 

机构地区：[1]卫生部北京医院卫生部临床检验中心,100730 [2]老年医学研究所生化室

出　　处：《中华检验医学杂志》2007年第3期260-263,共4页Chinese Journal of Laboratory Medicine

基　　金：863计划资助项目(2006AA020909)

摘　　要：目的建立一种新的同位素稀释-气相色谱-质谱(ID/GC/MS)测定人血清总胆固醇的方法。方法取一定量的血清样品与[3,4-^(13)C_2]-胆固醇内标溶液充分混匀,水解并提取溶液中的胆固醇后,用 N,O-二-(甲基硅烷)三氟乙酰胺(BSTFA)衍生胆固醇为三甲基硅烷醚;气相色谱-四极杆质谱(GC/MS)选择检测目标离子。检测标准溶液和血清样品的 m/z 368和 m/z 370的离子强度并积分,校正标准溶液中天然同位素胆固醇对内标的影响,将校正后的胆固醇和内标的 m/z 368和 m/z370面积比对胆固醇标准溶液浓度做线性回归。用拟合的回归方程定量血清样品胆固醇浓度。结果建立的 ID/GC/MS 测定胆固醇的方法平均批内变异系数0.04%～0.81%。分析美国国家标准物质与技术研究所(NIST)2个浓度水平的标准血清 SRM1951a,相对偏差分别是0.19%和0.90%。结论建立的 ID/GC/MS 测定胆固醇的方法操作简单、精密,可以用做血清胆固醇的准确定量。Objective To develop a new isotope dilution gas chromatography mass spectrometry method （ID/GV/MS） for the measurement of serum cholesterol. Methods Serum was mixed with an isotope labeled internal standard （[ 3,4-13 C ]-cholesterol） and treated with alcoholic sodium hydroxide to hydrolyze cholesterol ester to cholesterol. Cholesterol and internal standard was extracted and derived by N, O-Bis（trimethylsflyl） trifluoroacetamide to trimethylsilyl ethers. The derivation products were analyzed by capillary column GC combined with electron impact MS using scan and selected ion monitor （SIM） modes. Signals of cholesterol internal standard were corrected for the contributions from cholesterol and the signal ratio of cholesterol to internal standard for the calibrators were linearly regressed against cholesterol concentrations. The resulted regression equation was used for the calculation of serum cholesterol concentrations. Results The new ID/GC/MS method showed a mean within-run coefficient variance （CV） of 0. 04% -0. 81%. Comparison with two levels of standard reference material （SRM1951a） of National Institute of Standards and Technology （ NIST ） displayed a bias of 0. 19% and 0.90% respectively. Conduslon A time-gaining ID/MS method has been established that is highly precise and accurate and can be used for the measurement of serum cholesterol.

关 键 词：脂蛋白类 胆固醇 气相色谱-质谱 血清 

分 类 号：R446.1[医药卫生—诊断学]