Wu Tianfeng, Zhao Shijing, Li Wen, Ma Fei, Zhao Jian'gang, Huang Guowei
Wei Sheng Yan Jiu. 2015 Jul;44(4):632-5.
To develop a method for determination of S-adenosylmethionine (SAM) and S-adenosylhomocysteine (SAH) simultaneously in mice liver by reversed-phase high performance liquid chromatography (HPLC).
Mice livers were weighed, emulsified quickly by handheld homogenate, four times the volume of 0.6 mol/L HCLO4 added for ultrasonic, and the hydrolysate was adjusted to centrifuged and filtrated with a membrane. The supernatants were separatedonVenusil MP-C18 column (250 mm x 4.6 mm, 5 µm) at 30 °C with a mobile phase of 50 mmol/L NaH2PO4, 10 mmol/L C7H15NaO3S and methanol, a flow rate of 1.0 ml/min and UV detection at 254 nm.
The SAM and SAH in the corresponding concentration range showed a good linear relation with its peak area, correlation coefficient ( r > 0. 9990 ) , recovery was 92.20%-101.38%, RSD was 2.88%-6.78%. The average within-day precision of SAM and SAH was 4.14% and 3.71%, and the average day to day precision was 7.51% and 9.54%. The content of SAM and SAH in mice liver was 3.14-6.09 mg/L (31.44-60.98 nmol/g wet weight) and 1.29-3.10 mg/L (13.38-32.17 nmol/g wet weight) respectively.
The validated method is simple, rapid accurate and reliable to the determination of SAM and SAH in mice liver.
建立一种反相高效液相色谱法（HPLC）同时测定小鼠肝脏中S-腺苷甲硫氨酸（SAM）和S-腺苷高半胱氨酸（SAH）的方法。
称取小鼠肝脏，用手持匀浆器快速乳化，加入4倍体积的0.6 mol/L高氯酸进行超声处理，将水解产物调节后离心并用膜过滤。上清液在Venusil MP-C18柱（250 mm×4.6 mm，5 µm）上于30℃分离，流动相为50 mmol/L磷酸二氢钠、10 mmol/L庚烷磺酸钠和甲醇，流速为1.0 ml/min，在254 nm处进行紫外检测。
SAM和SAH在相应浓度范围内与其峰面积呈良好的线性关系，相关系数（r＞0.9990），回收率为92.20% - 101.38%，相对标准偏差（RSD）为2.88% - 6.78%。SAM和SAH的日内平均精密度分别为4.14%和3.71%，日间平均精密度分别为7.51%和9.54%。小鼠肝脏中SAM和SAH的含量分别为3.14 - 6.09 mg/L（31.44 - 60.98 nmol/g湿重）和1.29 - 3.10 mg/L（13.38 - 32.17 nmol/g湿重）。
该验证方法用于测定小鼠肝脏中SAM和SAH简单、快速、准确且可靠。
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