Birmingham B K, Greene D S
J Pharm Sci. 1983 Nov;72(11):1306-9. doi: 10.1002/jps.2600721118.
An assay for the separation and quantification of folinic acid in serum was developed using high-performance liquid chromatography with electrochemical detection. Folinic acid was extracted from serum using a C18 minicolumn treated with dibasic ammonium phosphate. The drug was eluted from this column with methanol, which was evaporated under a nitrogen stream at 50 degrees. The mobile phase, pH 3.5 ammonium phosphate buffer-methanol-acetonitrile (93:4:3), was pumped at a flow rate of 3.0 ml/min. The recovery of folinic acid added to human serum was 101.11 +/- 8.5% (mean +/- SD). A plot of folinic acid peak height as a function of concentration was linear over the range of 2.5 X 10(-7) to 2.5 X 10(-6) M. Neither methotrexate nor other reduced folates interfered with the analysis of folinic acid. Sample preparation and analysis can be completed within 2 min of sample collection.
开发了一种使用高效液相色谱-电化学检测法分离和定量血清中亚叶酸的分析方法。使用经磷酸氢二铵处理的C18微型柱从血清中提取亚叶酸。用甲醇将药物从该柱上洗脱下来，在50℃氮气流下蒸发甲醇。以3.0 ml/min的流速泵送pH 3.5的磷酸铵缓冲液-甲醇-乙腈（93:4:3）流动相。添加到人体血清中的亚叶酸回收率为101.11±8.5%（平均值±标准差）。在2.5×10⁻⁷至2.5×10⁻⁶ M范围内，亚叶酸峰高与浓度的关系图呈线性。甲氨蝶呤和其他还原型叶酸均不干扰亚叶酸的分析。样品制备和分析可在采集样品后2分钟内完成。
