高效液相色谱-示差折光法测定NMM抗肿瘤DNA疫苗中甘露醇含量
Content Determination of Mannitol in NMM Anti-Tumor DNA Vaccine by HPLC-RID
郭润姿 1黄言 1孙澳 1于继云 1王宇1
作者信息
- 1. 固安鼎泰海规生物科技有限公司,河北 廊坊 065500
- 折叠
摘要
目的 建立测定NMM抗肿瘤DNA疫苗中甘露醇含量的高效液相色谱-示差折光法.方法 检测器为示差折光检测器,色谱柱为SUGAR SC1011 阳离子钙型交换柱(300 mm×8 mm,6µm),流动相为纯化水,流速为 1.0 mL/min,柱温为 80℃,检测池温度为37℃,进样量为 10µL.结果 甘露醇的质量浓度在 0.02～5.00 mg/mL(R2 = 1.000 0,n = 6)范围内与峰面积线性关系良好;定量限为 13.37µg/mL;精密度、重复性、稳定性试验结果的RSD均小于 2.0%;加样回收率为 97.76%,RSD为 0.97%(n = 9).3 批小试样品和中试样品中甘露醇的含量分别为 20.02,19.95 mg/mL.结论 该方法操作简便、重复性好、结果准确,可用于NMM抗肿瘤DNA疫苗中甘露醇的含量测定.
Abstract
Objective To establish a high-performance liquid chromatography differential refractive index detector(HPLC-RID)method for the content determination of mannitol in NMM anti-tumor DNA vaccines.Methods RID was adopted,the chromatographic column was SUGAR SC1011 cation calcium exchange column(300 mm×8 mm,6 µm),the mobile phase was purified water,the flow rate was 1.0 mL/min,the column temperature was 80℃,the temperature of detection cell was 37℃,and the injection volume was 10 µL.Results The linear range of mannitol was 0.02-5.00 mg/mL(R2 = 1.000 0,n = 6).The limit of quantification(LOQ)was 13.37 µg/mL.The RSDs of precision,repeatability,and stability test results were lower than 2.0%.The recovery rate of mannitol was 97.76%with an RSD of 0.97%(n = 9).The contents of mannitol in three batches of small-test and pilot-test samples were 20.02 mg/mL and 19.95 mg/mL,respectively.Conclusion The method is simple,accurate and specific,which can be used for the content determination of mannitol in NMM anti-tumor DNA vaccines.
关键词
NMM抗肿瘤DNA疫苗/重组生物制品/甘露醇/高效液相色谱法/示差折光检测器/含量测定Key words
NMM anti-tumor DNA vaccine/recombinant biological products/mannitol/HPLC/RID/content determination引用本文复制引用
基金项目
河北省科技厅重点研发项目(20372404D)
出版年
2024
国家科技期刊平台
NSTL
万方数据