高效液相色谱紫外检测法同时测定小鼠红细胞中S-腺苷甲硫氨酸和S-腺苷同型半胱氨酸  

Simultaneous Determination of S-Adenosylmethionine and S-Adenosylhomocysteine in Red Blood Cells of Mice by High Performance Liquid Chromatography with Ultraviolet Detection

作　　者：唐秀芳[1] 王箭[1] 张晓清[1] 高茹菲[2] 丁敏[1] 

机构地区：[1]重庆医科大学检验医学院,临床检验诊断学教育部重点实验室,重庆400016 [2]重庆医科大学生殖生物学研究室,重庆400016

出　　处：《分析化学》2013年第4期570-574,共5页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金(No.30973195)资助项目

摘　　要：建立了高效液相色谱紫外检测同时测定红细胞中S-腺苷甲硫氨酸(SAM)和S-腺苷同型半胱氨酸(SAH)的方法。采用ZORBA×Eclipse×DB-C8色谱柱(150 mm×4.6 mm,5μm),以含3 mmol/L庚烷磺酸钠的40 mmol/L磷酸钠缓冲溶液(pH 3.72)-甲醇(95∶5,V/V)为流动相,流速0.9 mL/min,柱温35℃,紫外检测波长设置:0～8.00 min,450 nm;8.01～16.00 min,260 nm。SAH与SAM的线性范围分别为0.25～3.0 mg/L和0.50～10 mg/L,检出限分别为0.15和0.07 mg/L。方法日内和日间相对标准偏差(RSD)分别小于6.5%和7.4%,平均回收率为80.9%～116.2%。利用本方法对19例叶酸缺乏孕小鼠和11例健康对照孕小鼠红细胞进行测定。结果表明:叶酸缺乏组SAH明显高于健康对照组,而SAM两组间无显著差异。A high performance liquid chromatographic method with ultraviolet（UV） detection was established to determine S-adenosylmethionine（SAM） and S-adenosylhomocysteine（SAH） in red blood cells simultaneously.A ZORBA×Eclipse×DB-C8 column（150 mm×4.6 mm,5μm）was used in the analysis.The temperature was set at 35 ℃.The separation was carried out using the mobile phase consisting of 3 mmol/L sodium heptanesulfonate in 40 mmol/L sodium phosphate buffer（pH 3.72）-methanol（95 ∶ 5,V/V） at a flow rate of 0.9 mL/min.The eluates were monitored by the ultraviolet detection setting at 450 nm from 0 to 8.00 min and at 260 nm from 8.01 to 16.00 min.The linearities were from 0.25 mg/L to 3 mg/L for SAH,0.5 mg/L to 10 mg/L for SAM.The detection limit was 0.15 mg/L for SAH,and 0.07 mg/L for SAM.The relative standard deviations of the intra-day and inter-day precision were less than 6.5% and 7.4%,respectively.The mean recoveries were from 80.9% to 116.2%.Red blood cell samples of nineteen folate deficiency and eleven healthy pregnant mice were tested.The results demonstrated that the SAH in the red blood cells from the folate deficiency group was significantly higher than that of the healthy group,while no significant difference was found in the concentration of SAM between the two groups.

关 键 词：高效液相色谱 S-腺苷甲硫氨酸 S-腺苷同型半胱氨酸 红细胞 

分 类 号：Q26[生物学—细胞生物学] O657.3[理学—分析化学]