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

作　　者：郭润姿 黄言 孙澳 于继云 王宇 GUO Runzi;HUANG Yan;SUN Ao;YU Jiyun;WANG Yu(Gu'an Dingtai Haigui Biotechnology Co.,Ltd.,Langfang,Hebei,China 065500)

机构地区：[1]固安鼎泰海规生物科技有限公司,河北廊坊065500

出　　处：《中国药业》2024年第1期64-67,共4页China Pharmaceuticals

基　　金：河北省科学技术厅重点研发计划项目[20372404D]。

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。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,6μm),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 10μL.Results The linear range of mannitol was 0.02-5.00 mg/mL(R2=1.0000,n=6).The limit of quantification(LOQ)was 13.37μg/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 

分 类 号：R927.2[医药卫生—药学] R979.1