目的: 建立苦参软膏体外释放评价方法,比较苦参软膏工艺变更前后的体外释放行为一致性。方法: 采用改良Franz扩散池建立了软膏剂体外释放测定方法,以2020年版《中华人民共和国药典》苦参软膏含量测定项下的高效液相色谱方法为基础进行优化,建立苦参软膏释放量测定方法,评价不同处方工艺苦参软膏体外释放一致性。苦参软膏制剂主要释放氧化苦参碱、槐定碱、苦参碱和槐果碱4种活性成分。色谱柱采用Shim-pack GIS C18柱 (100 mm×4.6 mm,5 μm),流动相 A 为乙腈,B为0.1%磷酸溶液(三乙胺调节pH至8.0),梯度洗脱,检测波长为210 nm,柱温为 35 ℃,进样量为 100 μL,流速为2.0 mL·min-1。结果: 6批次苦参软膏样品均在18 h完全释放,释放量均>85%,工艺变更前后3批样品的释放速率分别为4 058.9、4 137.8、3 873.6 μg·cm-2·h-1/2和4 177.9、4 333.9、4 389.1 μg·cm-2·h-1/2,工艺变更前后样品释放速率比值的90%标准置信区间为89.80%~102.94%,落在FDA一致性规定的75%~133.33%限度范围内;工艺变更前后样品累积释放曲线的f2相似因子为69.9,满足50≤f2≤100。结论: 所建方法符合苦参软膏体外释放评价要求,处方工艺变更前后样品体外释放等效。
Objective: To develop an evaluation method of in vitro release test for Kushen ointment and investigate the consistency of release behavior in pre-and post-manufacturing change of Kushen ointment. Methods: The modified Franz diffusion cell was employed for the in vitro release test of ointments. The dissolution was determined by HPLC method based on the test of Kushen ointment in Chinese Pharmacopoeia (2020 edition). The consistency of release behavior of Kushen ointment varied with the formulation and preparation process was evaluated. The released drug substances included four active ingredients oxymatrine, sophoridine, matrine and sophocarpine. Chromatographic separation was performed on a Shim-pack GIS C18 (100 mm×4.6 mm, 5 μm) column using a mobile phase of acetonitrile (A) and 0.1% phosphoric acid solution (adjusted to pH 8.0 with triethylamine) (B) with a gradient elution. The detection wavelength was 210 nm, the column temperature was 35 ℃, the injection volume was 100 μL, and the flow rate was 2.0 mL·min-1. Results: All 6 batches of Kushen ointment samples were completely released at 18 h and the released amount were more than 85%. The in vitro release rate of 3 batches of Kushen ointment samples in pre-manufacturing change were 4 058.9, 4 137.8 and 3 873.6 μg·cm-2·h-1/2, respectively. The in vitro release rate of 3 batches of Kushen ointment samples in post-manufacturing change were 4 177.9, 4 333.9 and 4 389.1 μg·cm-2·h-1/2, respectively. The 90% confidence interval of the in vitro release rate of Kushen ointment samples in pre-and post-manufacturing change was 89.80%-102.94% within the quality consistency range from 75% to 133.33% defined by the FDA guideline. The cumulative release curve similarity factor f2 between pre- and post-manufacturing change of Kushen ointment was 69.9 within the range from 50 to 100. Conclusion: The release behavior of Kushen ointment variation with the formulation and preparation process was demonstrated to be quality consistent through the evaluation method of in vitro release test for Kushen ointment and met the requirements.
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