高效液相色谱法同时测定血浆中S-腺苷甲硫氨酸和S-腺苷同型半胱氨酸  

Measurement of S-adenosylmethionine and S-adenosylhomocysteine in plasma with high performance liquid chromatography

作　　者：赵世晶[1] 赛娜[1] 任大林[1] 徐蓓[1] 刘川[1] 黄国伟[1] 

机构地区：[1]天津医科大学公共卫生学院营养与食品卫生系,天津300070

出　　处：《卫生研究》2013年第1期110-113,共4页Journal of Hygiene Research

基　　金：国家自然科学基金(No.81130053);天津市高等学校科技发展基金计划项目(No.20110143)

摘　　要：目的建立同时测定血浆中S-腺苷甲硫氨酸(SAM)和S-腺苷同型半胱氨酸(SAH)的高效液相色谱分析方法,并对正常人血浆中的含量进行测定。方法样品用400g/L三氯乙酸处理,离心取上清液过膜,以50mmol/L NaH2PO4缓冲液(pH4.38)、庚烷磺酸钠和甲醇为流动相,选用Venusil MP-C18(4.6mm×250mm,5μm)色谱柱进行色谱分离,流速为1.0ml/min,柱温30℃,紫外检测波长254nm。结果 SAM和SAH在相应检测浓度范围内与其响应值呈良好的线性关系(r>0.9999);方法回收率在94.14%～102.44%,相对标准偏差在1.00%～8.10%。正常人血浆中SAM和SAH的含量分别为0.032～0.080mg/L和0.004～0.010mg/L。结论该方法用于血浆中SAM和SAH的检测简便快速,准确可靠。Objective To develop a method for determination of S- adenosylmethionine （SAM） and S-adenosylhomocysteine （SAH） simultaneously in plasma by reversed-phase high performance liquid chromatography （HPLC）. Methods The plasma were hydrolyzed with trichloroacetic acid 400g/L, then the hydrolysate was adjusted to centrifuged and filtrated with a 0.45p.m membrane. The supernatants were separated on Venusil MP-Cxs column（4.6mm 250mm,5μm） at 30℃with a mobile phase of 50mmol/L NaH2PO4 （ pH 4.38 ） , C7 H15 NaO3 S and methanol, a flow rate of 1.0ml/ min, and UV detection at 254nm. Results The SAM and SAH in the corresponding concentration range showed a good linear relation with its peak area, correlation coefficient r 〉 0. 9999, recovery was 94.14% - 102.44% , RSD was 1.00% - 8.10%. The content of SAM and SAH in plasma of healthy people was 0. 032 -0. 080mg/L and 0. 004 - 0. 010mg/L respectively. Conclusion The validated method is simple, rapid accurate and reliable to the determination of SAM and SAH in plasma. J

关 键 词：高效液相色谱法 S-腺苷甲硫氨酸 S-腺苷同型半胱氨酸 血浆 

分 类 号：R446.11[医药卫生—诊断学]