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目的 :建立一种简便、快速、准确的测定重组人白介素 - 2制品中十二烷基硫酸钠 (SDS)含量的方法。方法 :用过量的吖啶橙溶液与一系列倍比稀释的不同浓度的SDS结合 ,产生有色的SDS -吖啶橙复合物 ,应用SpectraMax 2 5 0多功能酶标仪进行比色测定 ,并用Softmax分析软件分析结果。结果 :优化后的吖啶橙染料法测定重组人白介素 - 2制品中的SDS含量 ,最小检测限达到 0 0 2 μg·μL-1。经过 10次测定 ,均值为 (0 5 3± 0 0 0 6 3) μg·μL-1(n =10 ) ,RSD值为 11 9% ;当SDS的含量在 0 0 2～ 0 6 4μg·μL-1以内时 ,二次方程 (y =- 3 96× 10 -5x2 +0 0 2 7x +0 0 18)线性良好 (r >0 99) ;精密度试验的RSD值均小于 8% ;当SDS浓度在 0 0 2～ 0 32 μg·μL-1测定范围内时 ,SDS在样品中的平均回收率为10 4 4% (n =10 )。结论 :该法重复性好 ,灵敏度高 ,省时简便 ,可作为重组人白介素 - 2制品中SDS含量测定的一种常规方法
Objective: To establish a simple, rapid and accurate method for the determination of sodium dodecyl sulfate (SDS) in recombinant human interleukin - 2 preparations. Methods: The excess amount of acridine orange solution was combined with different serial dilutions of different concentrations of SDS to produce a colored SDS - acridine orange complex. The colorimetric determination was performed on a SpectraMax 250 multifunctional microplate reader. Softmax Analysis of software analysis results. Results: The optimized acridine orange dye method for the determination of recombinant human interleukin - 2 product SDS content, the minimum detection limit reached 0 0 2 μg · μL-1. After 10 determinations, the mean value was (0 53 ± 0 0 0 6 3) μg · μL-1 (n = 10) and the RSD value was 11 9%. When the SDS content was between 0 0 2 and 0 6 4 μg · μL (Y = - 3 96 × 10 -5x2 +0 0 2 7x +0 0 18) was good (r> 0 99). The RSD values of precision test were less than 8% The average recoveries of SDS in the sample ranged from 104 to 4% (n = 10) at concentrations ranging from 0 to 32 μg · μL-1. Conclusion: This method has good repeatability, high sensitivity and simple time - saving. It can be used as a routine method for determination of SDS content in recombinant human interleukin - 2