目的 建立聚山梨酯20、40、60和80中甲醛和乙醛杂质的含量测定方法,并研究样品中甲醛和乙醛总含量与过氧化值的相关性。方法 样品与2, 4-二硝基苯肼(DNPH)在室温反应60 min,生成甲醛-DNPH和乙醛-DNPH,采用HPLC进行检测;照2015年版《中国药典》通则0713测定样品的过氧化值。结果 本法专属性好,目标峰与相邻杂质峰完全分离;甲醛和乙醛分别在0.1~35.0 μg·mL-1和0.2~50.0 μg·mL-1内线性关系良好(r甲醛=0.999 8、r乙醛=0.999 6);加标平均回收率为88.6%~103.5%,相对标准偏差(RSD)为1.2%~2.6%;日内精密度和日间精密度RSD均小于6%;甲醛的检测限为0.03 μg·mL-1,定量限为0.1 μg·mL-1;乙醛的检测限为0.06 μg·mL-1,定量限为0.2 μg·mL-1。样品中甲醛含量为0~593.1 μg·g-1,乙醛含量为0~740.5 μg·g-1;大多数样品的过氧化值小于10。结论 建立的HPLC柱前衍生化法适用于聚山梨酯类辅料中醛类杂质的含量测定;过氧化值与甲醛和乙醛总含量之间相关性差,建议在聚山梨酯系列药用辅料质量标准中补充醛类检查项目。
Abstract
OBJECTIVE To develop a quantitative determination method of formaldehyde and acetaldehyde and study the correlation between aldehydes and peroxide value in polysorbate 20, 40, 60 and 80.METHODS Formaldehyde and acetaldehyde in polysorbates reacted with 2,4-dinitrophenylhydrazine(DNPH) for 60 min at room temperature and stable phenylhydrazone compounds,formaldehyde-DNPH and acetaldehyde-DNPH, were obtained, respectively.The two phenylhydrazone compounds were simultaneously detected by HPLC to calculate the contents of formaldehyde and acetaldehyde in polysorates. The peroxide value of polysorbates was also determined according to chapter <0713> of Chinese Pharmacopoeia 2015. RESULTS The HPLC method for quantitative determination of aldehydes in polysorates had good specificity. The resolutions of formaldehyde-DNPH and acetaldehyde-DNPH were 2.0 and 2.9, respectively. There was good linearity in the range of 0.1-35.0 μg·mL-1 for formaldehyde(r=0.999 8) and 0.2-50.0 μg·mL-1 for acetaldehyde(r=0.999 6). The average spiked recoveries were in the range of 88.6%-103.5% and their relative standard deviations(RSDs) were in the range of 1.2%-2.6%.The RSDs of intra-day and inter-day precision were not more than 6.0%.The limits of detection and quantitation were 0.03 and 0.1 μg·mL-1 for formaldehyde and 0.06 and 0.2 μg·mL-1 for acetaldehyde, respectively. The contents of aldehydes in polysorbates from different manufacturers were 0-593.1 μg·g-1 for formaldehyde and 0-740.5 μg·g-1 for acetaldehyde, however, the peroxide values of most polysorbate products were less than 10. CONCLUSION The pre-column derivation HPLC method is suitable for assay of trace aldehydes in polysorbates. Moreover, the weak correlation between aldehydes and peroxide value indicated that the latter testing item is not enough to control the risk of aldehydes in polysorbates. Therefore, aldehydes as well as quantitative determination method are suggested to be included in the monographs of polysorbates to improve their quality standard.
关键词
聚山梨酯 /
甲醛 /
乙醛 /
高效液相色谱法 /
过氧化值
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Key words
polysorbate /
formaldehyde /
acetaldehyde /
HPLC /
peroxide value
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中图分类号：
R917
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参考文献
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脚注
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基金
中国食品药品检定研究院中青年研究发展基金项目资助(2017A2)
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