摘要
目的:建立一种氨基固相萃取-HPLC法测定艾地骨化醇软胶囊中3个同分异构体的有关物质。方法:采用氨基固相萃取法,经乙酸乙酯-正己烷洗脱,无水乙醇提取,提取液经减压蒸干后用乙腈复溶,经ODS色谱柱(250 mm×4.6 mm, 5μm)分析,以水-乙腈为流动相,梯度洗脱,流速为1.0 mL·min^(-1),进样量为50μL,采用紫外检测器,在检测波长265 nm处检测有关物质。结果:固相萃取前处理可有效去除空白油干扰,艾地骨化醇与各杂质均可基线分离;在低温4℃条件下考察溶液稳定性,24 h时供试品溶液中各杂质面积百分比为98.6%~102.3%;杂质对照品溶液中,24 h时Tachy杂质、前体、艾地骨化醇和Trans杂质的面积百分比分别为101.2%、96.9%、98.5%和96.2%;艾地骨化醇、trans杂质、前体和tachy杂质的定量限分别为0.107、0.102、0.128、0.063μg·mL^(-1),检测限分别为0.054、0.051、0.064、0.031μg·mL^(-1);四者质量浓度分别在0.1~6.3μg·mL^(-1)、0.1~1.2μg·mL^(-1)、0.1~5.6μg·mL^(-1)和0.1~1.1μg·mL^(-1)范围内与峰面积呈良好线性关系,相关系数(n=5)分别为0.999 9、1.000、1.000和0.999 4;tachy杂质、前体和trans杂质回收率(n=6)分别为106.6%、88.6%和102.8%,RSD(n=6)分别为6.7%、3.1%和2.1%。3批样品含前体5.0%、5.0%和5.1%,杂质tachy、杂质Trans和未知单杂均未检出。结论:该方法操作简单,经济实用,安全性高,专属性强,重复性好,准确度高,为艾地骨化醇软胶囊及类似油性基质的药物质量研究提供可靠依据。
Objective:To establish an amino solid-phase extraction-HPLC method for the determination of three related substances in eldecalcitol soft capsules.Methods:The eldecalcitol soft capsules were extracted by an amino solid-phase extraction method,followed by eluting with ethyl acetate:n-hexane and anhydrous ethanol.The residue was then dissolved with acetonitrile and water after vacuum evaporation.An ODS column(250 mm×4.6 mm,5μm)was used.Water-acetonitrile with gradient elution was employed.The flow rate was 1.0 mL·min^(-1),the injection volume was 50μL,and the detection wavelength was set at 265 nm.Results:Interference from blank excipients was eliminated with pretreatment.Baseline separation and good selectivity of eldecalcitol and other impurities was achieved.The stability of the test and reference solution was investigated at 4℃.The area change percentage of the impurities in the test solution were 98.6%-102.3%at 24 h.The area change percentage of the impurities in the reference solution were 101.2%,96.9%,98.5%and 96.2%at 24 h,respectively.The quantitation limits of eldecalcitol,trans impurity,pre-eldecalcitol and tachy impurity were 0.107μg·mL^(-1),0.102μg·mL^(-1),0.128μg·mL^(-1)and 0.063μg·mL^(-1),respectively.The detection limits were 0.054μg·mL^(-1),0.051μg·mL^(-1),0.064μg·mL^(-1),0.031μg·mL^(-1),respectively.The linear ranges of 4 components were 0.1-6.3μg·mL^(-),0.1-1.2μg·mL^(-),0.1-5.6μg·mL^(-)and 0.1-1.1μg·mL^(-1),respectively.The correlation coefficients(n=5)were 0.9999,1.000,1.000 and 0.9994,respectively.The average recoveries(n=6)of tachy impurity,pre-eldecalcitol and trans impurity were 106.6%,88.6%and 102.8%with RSDs of 6.7%,3.1%and 2.1%,respectively.In the three batches of samples,no impurities were detected except pre-eldecalcitol.The contents of pre-eldecalcitol were 5.0%,5.0%and 5.1%,respectively.Conclusion:The method is simple to operate,economical and practical with high safety,strong specificity,good repeatability and high accuracy.The method provides a reliable
作者
李洁
王清亭
张丽敏
张佳鸿
刘振飞
陈阳生
LI Jie;WANG Qing-ting;ZHANG Li-min;ZHANG Jia-hong;LIU Zhen-fei;CHEN Yang-sheng(CP Pharmaceutical Qingdao Co.,Ltd.,Qingdao 266000,China)
出处
《药物分析杂志》
CAS
CSCD
北大核心
2024年第6期1009-1016,共8页
Chinese Journal of Pharmaceutical Analysis
基金
泰山产业领军人才工程资助(tscx202211121)。
关键词
固相萃取
艾地骨化醇软胶囊
油性基质
活性维生素D
同分异构体
有关物质
solid-phase extraction
eldecalcitol soft capsules
oily substrate
activated vitamin D
structural isomer
related substances
