高效液相色谱法测定人体血浆中甲氨蝶呤浓度的改进

目的 改进高效液相色谱（HPLC）法测定人体血浆中甲氨蝶呤浓度的方法.方法 观察HPLC法测定人体血浆中甲氨蝶呤浓度常规方法和改进方法对色谱峰和出峰时间的影响.常规方法以Symmetryshield RD 18柱（150 mm ×3.9 mm,5.0 μm）为色谱柱,流动相为磷酸盐缓冲液（pH =7.2）-甲醇（88∶12）,流速为1.0 mL/min,柱温为35℃,Waters486型可变波长检测器,检测波长为302 nm;改进方法流动相改为磷酸盐缓冲液（pH =7.2）-乙腈（88∶12）,其他均与常规方法相同.结果 两种方法比较,改进方法色谱峰杂质峰少,分离度好,且较常规方法出峰时间缩短（P＜0.05）.结论 改进方法准确可靠,杂质峰少,分离度好,检测时间短,更适用于甲氨蝶呤的血药浓度监测.

Improvement of HPLC Determination of Methotrexate Concentration in Human Plasma

Objective To improve the high performance liquid chromatography（HPLC） method for determining the methotrexate concentra- tion in human plasma. Methods The influences of the conventional and improvement HPLC Method for determining the plasma methotrex- ate concentration on the chromatographic peak and its appearance time were observed. The conventional method： the Symmetrshield RD 18 column（150 mm ×3.9 mm, 5.0 μm） was used. The mobile phase consisted of 0. 01 mL/min phosphate buffer（pH =7.2）- methanol（88 ： 12）. The flow rate was 1.0 mL/min and the column temperature was 35 ℃. The detective wave length was 302 nm. The improved method： the mobile phase was changed to phosphate buffer（pH=7.2）-acetonitriler（88：12） and the others were the same as the conventional method. Results Compared with the conventional method, the improved method had few impurity peaks in the chromatographic peaks, the good separation degree and the short peak appearance time（ P〈 0. 05）. Conslusion The improved method is accurate and reliable with the advantages of few impurity peaks, good separation degree and short detection time and more suitable for the monitoring the methotrexate concentration in human plasma.