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建立高效液相色谱法测定门冬酰胺酶含量的方法。采用TSKgel G3000SW色谱柱(7.5 mm×300mm,10μm,TOSOH),以0.lmol/L磷酸盐缓冲液(p H 6.7)(取磷酸二氢钠6.0g、磷酸氢二钠20.2g,加水900ml,用磷酸或氢氧化钠溶液调节p H值至6.7,加水至1000ml),检测波长为280 nm,流速0.6 m L/min,柱温为室温,进样量为20μL。结果显示,门冬酰胺酶效价在100~300IU/m L范围内线性关系良好,线性回归方程:Y=1.7834C-1.8667(R2=0.999),平均回收率为102.36%(RSD=0.4045%)。可见,高效液相色谱法简便、灵敏、重复性好,可作为门冬酰胺酶效价测定方法。
To establish a HPLC method for the determination of asparaginase content. Using a TSKgel G3000SW column (7.5 mm × 300 mm, 10 μm, TOSOH), 0.1 mol / L phosphate buffer (p H 6.7) (taking sodium dihydrogen phosphate 6.0 g, disodium hydrogen phosphate 20.2 g, Adjust the pH value to 6.7 with phosphoric acid or sodium hydroxide solution and add water to 1000 ml). The detection wavelength is 280 nm and the flow rate is 0.6 m L / min. The column temperature is room temperature and the injection volume is 20 μL. The results showed that the asparaginase titer was linear in the range of 100-300 IU / mL with a linear regression equation of Y = 1.7834C-1.8667 (R2 = 0.999) with an average recovery of 102.36% (RSD = 0.4045%). . Visible, high performance liquid chromatography is simple, sensitive, reproducible, and can be used as asparaginase titer method.