0.99)與峰面積呈良好的線性關(guān)系。結(jié)論 建立了重組人IL-12注射液中泊洛沙姆188含量檢測的高效液相色譜法,并對建立的分析方法進(jìn)行專屬性、準(zhǔn)確度、精密度、檢測限、定量限、線性、范圍和耐用性的驗(yàn)證,測定了3批重組人IL-12注射液中的泊洛沙姆188的量。;Objective To develop a HPLC method for potency determination of poloxamer 188 in recombinant human IL-12 injection. Methods A Waters Alliance 2695 system with a sepax poly RP-100 column was used for separation and subsequent detection by an evaporative light scattering detector(ELSD); 0.1%TFA-water and 0.1%TFA-acetonitrile were used as eluent A and B, and gradient elution conditions were as follows:0-5 min, 50%A→50%A; 5-6 min, 50%A→5%A; 6-12 min, 5%A→5%A; 12-13 min, 5%A→50%A; 13-20 min, 50%A→50%A. The flow rate was 1.0 mL/min; the column was maintained at 30℃. Results The RSD of retention time and peak area for poloxamer 188 were 0.19% and 1.7%, respectively (n=6); the recovery rates at low (50%), middle (100%) and high (150%) dose were 93.3%, 93.1% and 93.8%, and the average was 93.4%; the potency of poloxamer 188 between 1.5 μg and 9.0 μg was linearly related to peak area. Conclusions A HPLC for potency determination of poloxamer 188 in recombinant IL-12 injection was successfully developed and validated in specificity, accuracy, precion, limit of detection (LOD), limit of quantitation (LoQ), linearity, detection range and robustness. The developed method was finally applied to potency determination of poloxamer 188 in three recombinant IL-12 injections."/>