注射用糜蛋白酶的比活与纯度研究

目的分别建立一种适用于注射用糜蛋白酶比活测定的方法以及RP-HPLC纯度测定方法。方法采用凯氏定氮法测定注射用糜蛋白酶中的总蛋白质含量，再结合效价计算出比活。纯度采用Grace214TPC4色谱柱（25Ommx4．6mm，5um），以0．1％三氟乙酸水溶液和0．09％三氟乙酸乙腈为流动相进行梯度洗脱，流速1．0m1／min，检测波长214nm，柱温30℃。由凯氏定氮法得到的比活结果与采用RP-HPLC法测得的纯度结果呈显著正相关（r=0．940，p〈0．01，n=5）。结果两种方法重复性、精密度和加样回收率均良好，可用于注射用糜蛋白的质控。结论对于多组分的生化药物，在主成分能与杂质分离的情况下，可采用纯度法来控制投料质量，而对于成分复杂者，则可考虑用比活代替纯度来进行质量控制。

Study on the specific activity and purity determination of Chymotrypsin for Injection

The methods for the specific activity and purity determination of Chymotrypsin for Injection were respectivelyestablished.To determinate the total protein content of Chymotrypsin for Injection by Kjeldahl determination of nitrogen,the specific activity was calculated by potency and total protein content.The RP-HPLC analysis of purity was performed on a Grace 214TP C4（250mm x 4.6ram, 5 um）columnusing gradient elution with mobile phase of 0.1% trifluoroacetie acid in water and 0.09% trifluoroacetic acid in acetonitrile .The assay was carried out at a flow rate of 1.0ml/min at 30~ with the UV detection at 214nm. The specific activity of Chymotrypsin for Injection showed positive correlation with its purity （r=0.940, P〈0.01,n=5） .The repeatability, accuracy and average recovery of both two methods are satisfying,they can be used for the quality control of Chymotrypsin for Injection.