HPLC法测定人红细胞中的S-腺苷同型半胱氨酸和S-腺苷甲硫氨酸  

HPLC determination of S-adenosylhomocysteine and S-adenosylmethionine in human red blood cells

作　　者：杜望春[1] 杨涛[2] 周海峰[2] 沈杰[2] 

机构地区：[1]上海健康医学院,上海201318 [2]复旦大学附属华东医院药剂科,上海200040

出　　处：《中南药学》2015年第9期979-982,共4页Central South Pharmacy

基　　金：上海市高等职业教育重点专业建设[No.FY(14)700-A5-1-13;No.FY(15)700-A6-1-04]

摘　　要：目的建立HPLC法测定人红细胞中S-腺苷同型半胱氨酸（SAH）和S-腺苷甲硫氨酸（SAM）浓度的方法。方法人红细胞经同体积醋酸盐缓冲液稀释后,经10%高氯酸蛋白沉淀后直接进样。采用Athena C18 WP柱（150 mm×4.6 mm,3μm）,柱温35℃,流动相为含5 mmol·L-1庚烷磺酸钠的50 mmol·L-1 Na H2PO4缓冲液（pH 4.0）-甲醇（80：20,v/v）,流速：1.0 m L·min-1,检测波长254 nm。采用外标法定量。结果SAH和SAM分别在0.025-10 mg·L-1和0.05-20 mg·L-1内线性关系良好,SAH和SAM的日内、日间RSD均〈12%,加样回收率在95.5%-106.3%,样品提取后在24 h内稳定性良好。结论该法快速、简便、准确,适用于SAH和SAM的体内分析检测。Objective To establish an HPLC method to determine the concentration of S-adenosylhomocysteine（SAH） and S-adenosylmethionine（SAM） in human red blood cells（RBCs）. Methods Based on protein precipitation, SAH and SAM in the RBCs samples were determined by HPLC with ultraviolet detection and the detector wavelength was 254 nm. HPLC analytical column was Athena C18 WP（150 mm×4.6 mm, 3 μm） at 35 ℃. The separation was carried out with the mobile phase of 5 mmol·L- 1 sodium heptanesulfonate in 50 mmol·L- 1 sodium phosphate buffer（pH= 4.0）-methanol（80 ：20, v/v） at 1.0 m L·min- 1. The assay utilized a quantification method of external standard.Results Calibration curves were linear for SAH and SAM at 0.025- 10 mg·L- 1 and 0.05- 20 mg·L- 1, respectively. The precision of within-day and between-day were good. Conclusion The method can be used to determine the concentrations of SAH and SAM in human RBCs.

关 键 词：高效液相色谱法 S-腺苷同型半胱氨酸 S-腺苷甲硫氨酸 人红细胞 

分 类 号：Q26[生物学—细胞生物学]