目的: 建立克立硼罗软膏体外透皮方法,为克立硼罗软膏生物等效性提供参考依据。方法: 采用HPLC法测定克立硼罗软膏渗透量,采用垂直式的Franz扩散池法研究软膏体外释放特性。采用Thermo BDS Hypersil-C18（150 mm×4.6 mm,5 μm）色谱柱,以0.1%磷酸溶液-乙腈（60∶40）为流动相,流速1.5 mL · min-1,柱温30 ℃,检测波长254 nm,进样量10 μL。结果: 样品色谱图中克立硼罗峰形良好;阴性样品色谱图中在克立硼罗位置未出峰,不干扰测定;克立硼罗质量浓度在0.026~21.02 μg · mL-1的范围内线性关系良好（r≥0.999）;重复性6份试验中,将自制品与参比制剂最大渗透速率（Jmax）的均值进行比较,比值为0.94;同时对Jmax进行置信区间计算,考察90%置信区间为87.9%~102.0%;各样品质量守恒百分比为94.1%~101.1%（n=12）;对照品溶液室温放置26 h,峰面积的RSD为0.44%,自制品及参比制剂分别于室温、32 ℃水浴放置26 h,峰面积的RSD为0.78%~1.1%,说明溶液稳定性较好。对3批样品检测,自制品和参比制剂的Jmax均值的比值均在0.92~0.97范围内,同时对Jmax进行置信区间计算,考察90%置信区间为84.8%~100.7%。结论: 该方法操作简单、便捷,方法选择性高,专属性强,准确度较高。
Objective: To establish an in vitro permeation method for crisaborole ointment and to provide a reference for the bioequivalence of crisaborole ointment. Methods: The HPLC method was used to determine the permeability of crisaborole ointment, and the vertical Franz diffusion pool method was used to study the release characteristics of the ointment in vitro. The chromatographic column was Thermo BDS Hypersil-C18(150 mm×4.6 mm, 5 μm), with 0.1% phosphoric acid solution-acetonitrile (60∶40) as the mobile phase, flow rate of 1.5 mL · min-1, column temperature of 30 ℃, detection wavelength of 254 nm, and injection volume of 10 μL. Results: The peak shape of crisaborole in the chromatogram of the test sample was good. There was no peak at the position of crisaborole in the negative sample chromatogram, which did not interfere with the determination. The linear relationship of crisaborole was good within the range of 0.026-21.02 μg · mL-1 (r≥0.999). In six repeated experiments, the mean maximum transmittance (Jmax) of the self-made product and the reference formulation were compared (odds ratio: 0.94). Simultaneously calculate the confidence interval for Jmax, with a 90% confidence interval ranging from 87.9% to 102.0%. The mass conservation percentage of each sample was 94.1%-101.1% (n=12). The reference solution was left at room temperature for 26 h, with a peak area RSD of 0.44%. The self-made product and reference preparation were left in a water bath at room temperature and 32 ℃ for 26 h, with peak area RSD ranging from 0.78% to 1.1%, indicating good stability of the solution. For the three batches of samples tested, the ratio of the mean Jmax values of the self-made product and the reference formulation was within the range of 0.92-0.97. Simultaneously calculate the confidence interval for Jmax, with a 90% confidence interval ranging from 84.8%-100.7%. Conclusion: This method is simple and convenient to operate, with high selectivity, strong exclusivity and high accuracy.
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