盐酸阿莫罗芬乳膏体外猪皮渗透贮留量的HPLC测定方法
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摘要
目的:研究盐酸阿莫罗芬乳膏体外经猪皮渗透贮留量的HPLC测定方法。方法:采用巴马小香猪离体猪皮和改良的Franz扩散池进行体外渗透试验。采用90%乙腈作为提取溶剂,酮康唑作为内标,色谱柱为Diamonsil-C18柱(150 mm×4.6 mm,5μm),流动相为甲醇-磷酸二氢铵溶液(磷酸调节p H值至3.0)(70∶30),流速为1.0 m L·min-1,检测波长为214 nm,柱温为30℃。结果:盐酸阿莫罗芬的浓度在1.13~8.05μg·m L-1范围内与峰面积线性关系良好,低、中、高浓度平均回收率分别为(105.48±2.58)%,(99.46±1.79)%和(101.41±2.00)%,精密度良好(RSD为1.36%)。定量限和检测限浓度分别为6.73和2.71 ng·m L-1。结论:该方法专属性好、准确度高、重现性好,可应用于盐酸阿莫罗芬乳膏的皮肤生物等效性研究。
Objective: To establish an HPLC method for the in vitro determination of retention of amorolfine hydrochloride cream via pig skin penetration. Methods: The penetration experiments on in vitro pig skin were performed by modified Franz's cell using Bama miniature pig skin. After being extracted with 90% acetonitrile solution,samples were separated by HPLC method on a Diamonsil C18column( 150 mm × 4. 6 mm,5 μm) with mobile phases of methanol-ammonium dihydrogen phosphate solution( 70 ∶ 30,adjusted p H to 3. 0 with phosphoric acid)and ketoconazole as an internal standard. The flow rate was 1. 0 m L·min- 1. The detection wavelength was 214 nm.Results: Calibration curve for amorolfine hydrochloride was linear in the range of 1. 13 ~ 8. 05 μg·m L- 1. The average recoveries of low,medium and high concentrations were( 105. 48 ± 2. 58) %,( 99. 46 ± 1. 79) % and( 101. 41 ± 2. 00) %,respectively. Precision was good( RSD = 1. 36%). The limit of quantification and detection was 6. 73 and 2. 71 ng·m L- 1,respectively. Conclusion: The method has good specificity,high accuracy,and good reproducibility. Thus it can be used for the skin bioequibalence study related to amorolfine hydrochloride cream.
Objective: To establish an HPLC method for the in vitro determination of retention of amorolfine hydrochloride cream via pig skin penetration. Methods: The penetration experiments on in vitro pig skin were performed by modified Franz's cell using Bama miniature pig skin. After being extracted with 90% acetonitrile solution,samples were separated by HPLC method on a Diamonsil C18column( 150 mm × 4. 6 mm,5 μm) with mobile phases of methanol-ammonium dihydrogen phosphate solution( 70 ∶ 30,adjusted p H to 3. 0 with phosphoric acid)and ketoconazole as an internal standard. The flow rate was 1. 0 m L·min- 1. The detection wavelength was 214 nm.Results: Calibration curve for amorolfine hydrochloride was linear in the range of 1. 13 ~ 8. 05 μg·m L- 1. The average recoveries of low,medium and high concentrations were( 105. 48 ± 2. 58) %,( 99. 46 ± 1. 79) % and( 101. 41 ± 2. 00) %,respectively. Precision was good( RSD = 1. 36%). The limit of quantification and detection was 6. 73 and 2. 71 ng·m L- 1,respectively. Conclusion: The method has good specificity,high accuracy,and good reproducibility. Thus it can be used for the skin bioequibalence study related to amorolfine hydrochloride cream.
引文
[1]TABARA K,SZEWCZYK AE,BIENIAS W,et al.Amorolfine vs.ciclopirox-lacquers for the treatment of onychomycosis[J].Postep Derm Alergol,2015,32(1):40-45.
[2]王成刚,余立.HPLC法测定盐酸阿莫罗芬搽剂的含量[J].中国抗生素杂志,2000,25(3):226-227.
[3]GAO Y,LI L,ZHANG JJ,et al.Simultaneous determination of triacetin,acetic ether,butyl acetate and amorolfine hydrochloride in amorolfine liniment by HPLC[J].Pak J Pharm Sci,2012,25(2):389-394.
[4]田璐,张蜀,林华庆,等.经皮给药制剂体外透皮扩散试验方法的研究进展[J].中国药房,2012,23(29):2761-2763.
[5]DANIELA K,JAMES DB,JIM ER,et al.Modeling the effect of experimental variables on the in vitro permeation of six model compounds across porcine skin[J].Int J Pharm,2013,443(1-2):58-67.
[6]MICHAEL HQ,ULLA H,BO K,et al.Evaluation of Gottingenminipig skin for transdermal in vitro permeation studies[J].Eur J Pharm Sci,2000,11(1):59-68.
[7]LIU Y,CHEN JY,SHANG HT,et al.Light microscopic,electron microscopic,and immunohistochemical comparison of Bamaminipig(Sus scrofa domestica)and human skin[J].Comp Med,2010,60(2):142-148.
[8]祁少芳,高丽琼,舒文娟,等.盐酸阿莫罗芬在水溶液中的降解动力学研究[J].药学进展,2010,34(11):518-522.
[9]张肖玲,张蜀,邓红,等.盐酸多塞平的经皮渗透性比较研究[J].中国新药杂志,2013,22(4):477-481.
[10]罗荣,张新华,周临,等.盐酸布替萘芬乳膏的离体透皮吸收研究[J].中国新药杂志,2012,21(3):310-323.
[2]王成刚,余立.HPLC法测定盐酸阿莫罗芬搽剂的含量[J].中国抗生素杂志,2000,25(3):226-227.
[3]GAO Y,LI L,ZHANG JJ,et al.Simultaneous determination of triacetin,acetic ether,butyl acetate and amorolfine hydrochloride in amorolfine liniment by HPLC[J].Pak J Pharm Sci,2012,25(2):389-394.
[4]田璐,张蜀,林华庆,等.经皮给药制剂体外透皮扩散试验方法的研究进展[J].中国药房,2012,23(29):2761-2763.
[5]DANIELA K,JAMES DB,JIM ER,et al.Modeling the effect of experimental variables on the in vitro permeation of six model compounds across porcine skin[J].Int J Pharm,2013,443(1-2):58-67.
[6]MICHAEL HQ,ULLA H,BO K,et al.Evaluation of Gottingenminipig skin for transdermal in vitro permeation studies[J].Eur J Pharm Sci,2000,11(1):59-68.
[7]LIU Y,CHEN JY,SHANG HT,et al.Light microscopic,electron microscopic,and immunohistochemical comparison of Bamaminipig(Sus scrofa domestica)and human skin[J].Comp Med,2010,60(2):142-148.
[8]祁少芳,高丽琼,舒文娟,等.盐酸阿莫罗芬在水溶液中的降解动力学研究[J].药学进展,2010,34(11):518-522.
[9]张肖玲,张蜀,邓红,等.盐酸多塞平的经皮渗透性比较研究[J].中国新药杂志,2013,22(4):477-481.
[10]罗荣,张新华,周临,等.盐酸布替萘芬乳膏的离体透皮吸收研究[J].中国新药杂志,2012,21(3):310-323.