皮肤冻存对肉桂挥发性成分体外透皮特性的影响

    Effect of Skin Cryopreservation on in Vitro Transdermal Properties of Volatile Components from Cinnamomi Cortex

    • 摘要:
      目的  建立同时测定肉桂挥发油中香豆素、肉桂醇、肉桂酸、肉桂醛和邻甲氧基肉桂醛含量的HPLC方法,并比较离体皮肤冻存对这些成分体外透皮特性的影响。
      方法  采用Franz扩散池法对肉桂油进行体外透皮吸收试验,以冻存前后的小鼠离体皮肤做渗透屏障,同步用MTT法和CCK8法监测皮肤活力。采用Agilent C18色谱柱(4.6 mm×250 mm,5 μm),乙腈-0.2%磷酸(30∶70)为流动相,流速1 mL·min−1,检测波长为250 nm和278 nm,同时测定透皮过程中肉桂这5个挥发性成分的含量变化,统计分析其28 d内多点下的24 h渗透规律。
      结果  香豆素、肉桂醇、肉桂酸、肉桂醛和邻甲氧基肉桂醛等成分在该色谱条件下分离度好,进样量分别在0.0013~0.74140.0043~2.37990.0105~5.79740.0389~44.58300.0027~1.5224 μg线性关系良好(r>0.999)。离体皮肤在常用冻存液中保存28 d,5个成分24 h的渗透速率变化范围分别是1.64~6.43、4.88~22.84、3.76~60.40、197.53~274.39、8.20~12.60 μg·cm−2·h−1;冻存皮肤的活力前7 d下降明显,之后仍继续缓慢下降。
      结论  所建立的HPLC方法可简便、快捷地用于肉桂油中5个挥发性成分的含量测定,可为相关药物的研发提供参考。离体皮肤冻存28 d,期间其活力会持续下降,5个成分24 h的渗透速率变化规律不同,用CCK8法和MTT法监测皮肤活力变化的趋势相近。

       

      Abstract:
      OBJECTIVE  To develop an HPLC determination method of coumarin, cinnamyl alcohol, cinnamic acid, cinnamaldehyde and 2-methoxy cinnamaldehyde in cinnamon oil, and compare the effects of in vitro skin cryopreservation on the transdermal properties of these components.
      METHODS The transdermal absorption of cinnamon oil was examined using Franz diffusion cells with cryopreserved mouse skin as the permeation barrier. Simultaneously, skin vitality was monitored using MTT and CCK8 assays. An Agilent C18 chromatographic column(4.6 mm×250 mm, 5 μm) with acetonitrile-0.2% phosphoric acid(30∶70) as the mobile phase at a flow rate of 1 mL·min−1 was employed, detection was performed at 250 nm and 278 nm to simultaneously monitor the content changes of these 5 volatile components, the 24 h permeation regularities were analyzed over a period of 28 d.
      RESULTS  Excellent separation of coumarin, cinnamyl alcohol, cinnamic acid, cinnamaldehyde and 2-methoxy cinnamaldehyde was achieved under the chromatographic conditions, good linear relationships(r>0.999) were observed within the concentration ranges of 0.00130.7414, 0.00432.3799, 0.01055.7974, 0.038944.5830, 0.00271.5224 μg respectively. The range of 24 h permeation rate changes of the above components was as followed: 1.64−6.43, 4.88−22.84, 3.76−60.40, 197.53−274.39, 8.20−12.60 μg·cm−2·h−1. The vitality of cryopreservation skin decreased rapidly within the first 7 d then more gradually thereafter.
      CONCLUSION  The proposed HPLC method offers a rapid and convenient means for determining the 5 volatile components in cinnamon oil, providing valuable insights for drug development. The vitality of cryopreservation skin in vitro continued to decline over 28 d. The variation patterns of 24 h permeation rates of the five components are different. Both CCK8 and MTT assays demonstrated similar trends in monitoring skin vitality.

       

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