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An LC-MS Method for Determination of Betamethasone in Tissue Engineering Skin and Application to Dermatopharmacokinetic Study

[ Vol. 16 , Issue. 1 ]

Author(s):

Lingjun Li, Mengli Zhang, Yuping Cao, Pengcheng Ma*, Jun Wei, Lei Tao and Kun Qian   Pages 92 - 99 ( 8 )

Abstract:


Background and Objectives: Tissue engineering skin is a three-dimensional skin substitute cultured in the gas-liquid interface using the immortalized keratinocytes (HaCaT cells). In this study, the preliminary metabolism of betamethasone dipropionate by tissue engineering skin was studied and the pharmacokinetics methodology was established using betamethasone dipropionate gel as the target drug.

Methods: The betamethasone dipropionate gel was applied on the tissue engineering skin after the skin was cultured. Then the medium (receiving liquid) and skin were taken on 0.25, 0.75, 1.75, 3, 5, 8, 12, 24, 36, 48 h time points. The betamethasone concentration in the medium and skin was determinated by the LC-MS method. Chromatographic analysis was conducted using isocratic elution on a C18 column (150 mm × 2.0 mm, 5 μm) in mobile phase consisting of methanol and water (70 : 30, v/v). The mobile phase was pumped at a flow rate of 0.2 mL/min.

Results: This method exhibited linearity within the concentration range of 0. 1 to 50 μg /mL of betamethasone. The LLOQ was 0. 1 μg /mL. The intra- and inter-day precisions of betamethasone in the blank medium were all less than 10.69 % (RSD, %), while in the blank, skin homogenates were all less than 13.96 % (RSD, %). As a result, the betamethasone concentration in the medium and skin could both be detected, which suggested that betamethasone dipropionate could be metabolized to betamethasone through the tissue engineering skin.

Conclusion: It was feasible to use tissue engineering skin as a model to study the dermatopharmacokinetics of topical betamethasone dipropionate gel. The research could build a foundation for the dermato-pharmacokinetic study approach.

Keywords:

Betamethasone dipropionate, tissue engineering skin, determination, dermatopharmacokinetic, LC-MS, methodology.

Affiliation:

Institute of Dermatology,Chinese Academy of Medical Sciences & Peking Union Medical College, No. 12, Jiangwangmiao Street, Nanjing, 210042, Institute of Dermatology,Chinese Academy of Medical Sciences & Peking Union Medical College, No. 12, Jiangwangmiao Street, Nanjing, 210042, Institute of Dermatology,Chinese Academy of Medical Sciences & Peking Union Medical College, No. 12, Jiangwangmiao Street, Nanjing, 210042, Institute of Dermatology,Chinese Academy of Medical Sciences & Peking Union Medical College, No. 12, Jiangwangmiao Street, Nanjing, 210042, Institute of Dermatology,Chinese Academy of Medical Sciences & Peking Union Medical College, No. 12, Jiangwangmiao Street, Nanjing, 210042, Institute of Dermatology,Chinese Academy of Medical Sciences & Peking Union Medical College, No. 12, Jiangwangmiao Street, Nanjing, 210042, Institute of Dermatology,Chinese Academy of Medical Sciences & Peking Union Medical College, No. 12, Jiangwangmiao Street, Nanjing, 210042



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