邻苯二甲醛柱前衍生化-HPLC测定复方氨基酸注射液中蛋氨酸亚砜的含量

Determination of Methionine Sulfoxide in Compound Amino Acid Injection by HPLC with OPA Pre-column Derivatization

关键词:  邻苯二甲醛  柱前衍生  高效液相色谱  复方氨基酸注射液  蛋氨酸亚砜

Key words:  O-phthaladehyde(OPA)  pre-column derivatization  HPLC  Compound Amino Acid injection  methionine sulfoxide

摘要:

目的 建立测定复方氨基酸注射液中蛋氨酸亚砜含量的方法。方法 采用邻苯二甲醛（O-phthaladehyde，OPA）柱前衍生HPLC。色谱柱为Agilent ZORBAX Eclipse AAA C₁₈柱（150 mm×4.6 mm，5µm），检测波长为338 nm，流动相A为0.04 mol·L^-1磷酸二氢钠溶液（调节pH至7.8），流动相B为乙腈-甲醇-水（45：45：10），梯度洗脱，流速为2.0 ml·min^-1，柱温为40℃，供试品溶液与OPA衍生剂及硼酸盐缓冲液按设定程序自动混匀后进样。结果 蛋氨酸亚砜的质量浓度线性范围为0.000 5~0.050 0 mg·mL^-1（R=0.999 9）；定量限为0.250 5 μg·mL^-1，检测限为0.083 5 μg·mL^-1；精密度、稳定性、重复性试验的RSD<3.0%；平均回收率为99.1%（RSD=1.33%，n=9）。结论 该方法操作简便，可用于复方氨基酸注射液中蛋氨酸亚砜的含量测定。

Abstract:

OBJECTIVE To establish a method for the content determination of methionine sulfoxide in Compound Amino Acid injection. METHODS O-phthaladehyde(OPA) pre-column derivatization HPLC was performed on the column of Agilent ZORBAX Eclipse AAA C₁₈(150 mm×4.6 mm, 5 µm) with mobile phase A 0.04 mol·L^-1 sodium dihydrogen phosphate solution (adjust pH to 7.8), mobile phase B acetonitrile-methanol-water (45:45:10)(gradient elution) at the flow rate of 2.0 ml·min^-1, the detection wavelength was 338 nm and the column temperature was 40℃. The sample solution was automatic mixed with OPA agent and borate buffer according to set procedure. RESULTS The linear range of methionine sulfoxide was 0.000 5-0.050 0 mg·mL^-1(R=0.999 9). The limit of quantitation was 0.250 5 μg·mL^-1, and the limit of detection was 0.083 5 μg·mL^-1. RSDs of precision, stability and repetitive test were all <3.0%. The average recovery was 99.1%(RSD=1.33%, n=9). CONCLUSION The method is simple, rapid and suitable for the content determination of methionine sulfoxide in Compound Amino Acid injection.