HPLC测定小儿复方氨基酸注射液中甲硫氨酸的有关物质

Determination of related substances of methionine in Pediatric compound amino acid injection

目的采用HPLC法测定小儿复方氨基酸注射液中甲硫氨酸的有关物质,探讨市售制剂包装的合理性。方法采用HILIC色谱柱(250 mm×4.6 mm,5μm),流动相A为0.01 mol·L^(-1)磷酸二氢铵溶液-乙腈(50∶50),流动相B为乙腈,梯度洗脱,柱温30℃,检测波长200 nm,进样量10μL。结果甲硫氨酸降解产生的主要有关物质为甲硫氨酸亚砜,各强制降解条件下均未产生甲硫氨酸砜,其检出量可以有效评价甲硫氨酸的降解情况,甲硫氨酸亚砜与样品中其他组分峰能完全分离;0.5~106.9μg·mL^(-1)甲硫氨酸亚砜与峰面积呈良好的线性关系(r=0.9999);回收率为98.81%~100.62%;重复性与进样精密度均符合要求;测定了7个生产厂家的135批样品,甲硫氨酸亚砜的含量为甲硫氨酸标示量的0.02%~0.71%,相同厂家生产的不同批次样品中,甲硫氨酸亚砜的含量差异较大,推测其生产工艺存在不稳定性。市售聚丙烯安瓿包装具有明显的透氧性,不加外层塑料包装在空气中长期放置会严重影响产品的质量。结论所用方法可测定小儿复方氨基酸注射液中甲硫氨酸的降解产物甲硫氨酸亚砜,实现产品质量的控制,同时为制剂包装的选择提供了依据。

OBJECTIVE To determine the related substances of methionine in Pediatric compound amino acid injection(PCAAI)by HPLC,and to discuss the rationality of the current commercial preparation packaging.METHODS HILIC column(250 mm×4.6 mm,5μm)was used.The mobile phase A was 0.01 mol·L^(-1) ammonium dihydrogen phosphate solution-acetonitrile(50∶50),the mobile phase B was acetonitrile,and the related substances were detected with gradient elution.Column temperature was 30℃,the injection volume was 10μL and the detection wavelength was at 200 nm.RESULTS The main related substance produced by the degradation of methionine is methionine sulfoxide,and no methionine sulfone is produced under the forced degradation conditions.The detection amount of methionine sulfoxide can effectively evaluate the degradation of methionine,and methionine sulfoxide can be completely separated from other peaks in the sample.The linear relationship of methionine sulfoxide was good in the range of 0.5^(-1)06.9μg·mL^(-1)(r=0.9999);the recovery rate was between 98.81%and 100.62%;according to the repeatability and injection precision met the requirements;according to the determination of 135 batches of samples from 7 manufacturers,the content of methionine sulfoxide was within the range of 0.02%to 0.71%of the labelled amount of methionine.Among different batches of samples produced by the same manufacturer,the content of methionine sulfoxide varies greatly,it was speculated that the production process is unstable.In the study,it was also found that the commercially available polypropylene ampoule bottle packaging had obvious oxygen permeability,and long-term storage in the air without the outer plastic packaging will seriously affected the product quality.CONCLUSION The degradation product of methionine in PCAAI,which can be determined and quality controlled by the established method,and the basis for the selection of preparation packaging is also provided.