HPLC法测定复方氨基酸注射液(18AA)中焦谷氨酸的含量

目的: 建立HPLC法测定复方氨基酸注射液(18AA)中焦谷氨酸的含量。方法: 采用Agilent ZORBAX SB C₁₈色谱柱(250 mm×4.6 mm,5 μm),以乙腈-0.01 mol·L^-1磷酸氢二铵溶液(pH 1.6)为流动相梯度洗脱,流速1.0 mL·min^-1,柱温15 ℃,检测波长205 nm,进样量20 μL。结果: 焦谷氨酸峰与样品中其他组分峰的分离度良好,溶液在24 h内稳定;测定33家生产企业158批样品,焦谷氨酸含量在4.4~97.9 mg·L^-1之间,不同企业不同规格之间数据差异较大。5%规格的含量平均值约为22 mg·L^-1,12%规格的含量平均值约为63 mg·L^-1。结论: 经方法学验证,本法可以有效地测定复方氨基酸注射液(18AA)中焦谷氨酸的含量。不同来源不同规格的复方氨基酸注射液(18AA)中焦谷氨酸的含量差异较大。

Objective: To establish an HPLC method for determination of pyroglutamic acid in compound amino acid injection(18AA). Methods: The determination was performed on an Agilent ZORBAX SB C₁₈ column(250 mm×4.6 mm,5 μm),with a mobile phase consisting of mixture of acetonitrile and 0.01 mol·L^-1 diammonium hydrogen phosphate buffer(pH 1.6)by gradient elution. The column temperature was set at 15 ℃ with a flow rate of 1.0 mL·min^-1. The detection wavelength was 205 nm. A 20 μL aliquot was injected into HPLC for analysis. Results: The resolution of peaks of pyroglutamic acid and other compounds was good. The solution was stable in 24 hours. The content of pyroglutamic acid in samples was from 4.4 mg·L^-1to 97.9 mg·L^-1. The difference in specifications of content of pyroglutamic acid from different producers was significant. The average content was 22 mg·L^-1in the samples of 5% specification,and 63 mg·L^-1in the samples of 12% specification. Conclusion: The method is validated to be useful for determination of content of pyroglutamic acid in compound amino acid injection(18AA). The contents of pyroglutamic acid in compound amino acid injection(18AA)from different sources and of different specifications are significantly different.

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