Determination of differential markers of Modified Huanglian Ointment and its skin protective effect
OBJECTIVE To establish the high performance liquid chromatography(HPLC)fingerprint of Modified Huan-glian Ointment,screen for differential markers with chemical pattern recognition,detect them quantitatively and examine its skin protective effect.METHODS The similarity of 21 batches of Modified Huanglian Ointment was examined by HPLC fingerprint.With chemical pattern recognition and a variable importance projection(VIP)value＞1 as a standard,differential biomarkers affecting the quality of Modified Huanglian Ointment were screened.The quality of Modified Huanglian Ointment was evaluated by content determination and its skin protective effect examined by an inhibition of Modified Huanglian Ointment on tyrosinase and melanin production in B16F10 cells.RESULTS The similarity of 21 batches of samples(S1-S21)was＞0.950 in HPLC finger-print of Modified Huanglian Ointment.Among 17 common peaks,8 peaks were phellodendron hydrochloride,sesamol,ferulic acid,coptis hydrochloride,berberine hydrochloride,palmatine hydrochloride,curcumin and sesamin.OPLS-DA results showed that peak 3(phellodendrine hydrochloride),peak 5(ferulic acid),peak 15(curcumin)and peak 16(sesamin)were differential markers affecting the quality of Modified Huanglian Ointment.And their contents were 0.021 0-0.028 7,0.006 7-0.010 0,0.021 5-0.025 8 and 0.209 5-0.235 5 mg·g-1 respectively.Moreover,the inhibitory rate of Modified Huanglian Ointment on tyrosinase and melanin was 100.33％and 59.80％respectively.CONCLUSION The established fingerprint and content deter-mination provide a reliable theoretical basis for quality evaluation of Modified Huanglian Ointment.It provides a scientific basis for quality control for industrial production.Modified Huanglian Ointment has some skin protection effect and it provides references for its clinical applications.
traditional Chinese medicine ointmentcontent determinationquality evaluationskin protection
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