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

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。

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

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 0.45μm membrane.The supernatants were separated on Venusil MP-C18 column(4.6mm×250mm,5μm)at 30℃ with a mobile phase of 50mmol/L NaH2PO4(pH 4.38)、C7H15NaO3S 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.