摘要
目的:建立一种快速、灵敏和准确测定病人血浆中甲氨蝶呤浓度的方法。方法:取病人血浆样品0.5mL 加内标(替加氟400.0μg·mL^(-1))50μL,混匀,加7%高氯酸溶液0.5mL,涡旋1min,离心(15000r·min^(-1))10min,取上清液50μL进样。以 Agilent ZORBAX Eclipe-XDB C_(18)(4.6mm×150mm,5μm)为分析柱;流动相为40mmol·L^(-1)磷酸二氢钾缓冲液(pH=4.5)-乙腈(91:9);流速为1.0mL·min^(-1);检测波长为304nm;柱温为25℃。结果:色谱峰分离良好,无干扰。测定数据根据浓度分高低两段进行线性回归,线性方程分别为:Y=0.2914X-0.1593,r=0.9999(5.0～100.0μg·mL^(-1));Y=0.2581X-0.0005,r=0.9999(0.05～5.0μg·mL^(-1)),定量限为(0.045±0.0025)μg·mL^(-1),最低检测浓度为0.020μg·mL^(-1)。方法回收率为94.8%～106.6%(n=7),提取回收率为83.3%～89.6%(n=7),日内 RSD 为1.6%～5.7%(n=7),日间 RSD 为1.7%～5.5%(n=7)。结论:本方法准确灵敏,操作简便、快速,适用于甲氨蝶呤大剂量化疗时病人血药浓度的监测。
Objective：To develop a new HPLC method to determine the concentration of methotrexate in human plasma. Methods ：0. 5 mL plasma sample with the internal standard tegafur added was extracted with 7% perchloric acid. After centrifugation, the supernatant was injected for analysis by HPLC. Separation was achieved on an Agilent ZORBAX Eclipe - XDB Cls （4. 6 mm × 150 mm ,5 μm） column at 25 ℃ and detected at 304 nm. The mobile phase consisted of 40 mmol ·L^-1 potassium dihydrogen phosphate（ adjusted to pH 4. 5 with phosphoric acid）and acetonitrile（91： 9）at a flow rate of 1.0 mL ·min^-1. Results：The linear equation at higher concentrationwas Y = 0. 2914X -0. 1593 ,r =0. 9999（5.0 - 100. 0μg·mL^-1 ） ,while that at lower concentration was Y =0. 2581X - 0. 0005, r = 0. 9999 （ 0. 050 - 5.0 μg·mL^-1 ） ; The limit of detection was 0. 020 μg·mL^-1 （ S/N 〉 4） and The limit of qualiilcation was （0. 045 ± 0. 0025 ） μg·mL^-1 （ S/N〉 10） ; The method and extraction recovery was 94. 8% - 106. 6% and 83.3% -89.6% respectively（ n = 7 ）, within - day precision（RSD） was 1.6% -5.7% （n = 7 ） and between - day precision （RSD） was 1.7 % - 5.5 % （ n = 7 ）. Conclusion： The method is accurate, sensitive and convenient, and is suitable for the determination of methotrexate concentration in human plasma.
出处
《药物分析杂志》
CAS
CSCD
北大核心
2006年第5期598-601,共4页
Chinese Journal of Pharmaceutical Analysis
