高效液相色谱法测定质粒pUDKH中曲拉通X-100的含量

Quantitative analysis of Triton X-100 in plasmid pUDKH by high performance liquid chromatography

目的检测质粒pUDKH最终产品中去污剂曲拉通X 10 0 (TritonX 10 0 )的残留含量。方法用高效液相色谱法测定TritonX 10 0残留含量 ,色谱柱为C18柱 ,流动相为乙腈 水 (70∶30 ) ,流速为 1.0ml/min ,进样量 10 μl,检测波长 2 2 3nm。结果平均回收率为 10 0 .19% ,日内精密度为 4 .36 % ,日间精密度为 4 .17% ,TritonX 10 0在 1.2 5～2 0 μg/ml浓度范围内呈线性关系。pUDKH产品中的TritonX 10 0含量在 2 .2 7～ 3.0 0 μg/ml之间。TritonX 10 0的最低检测限可达 1.0 μg/ml。 结论此法有良好的准确度与精密度 ,供试品不需预处理 ,不受其它成分的干扰。

PurposeIn the process of plasmid pUDKH production, Triton X-100 was used as a virus inactivated solvent. The residual Triton X-100 concentration must be determined in final products.MethodsA high performance liquid chromatography was used. The system included a Hypersil silica column, eluted with acetonitrile -H 2O(70∶30). The flow rate was 1.0 ml/min. ResultsThe detection limit was low (1.0 μg/ml) with detection at 223nm. The calibration linear range was between 1.25 μg/ml and 20 μg/ml.ConclusionThe method was rapid. It did not need sample pretreatment. The residual Triton X-100 in our final pUDKH products was determined at 2.27～3.00 μg/ml level.