Establishment and application of fluorescence quantitative analysis method for transdermal absorption of different oligosaccharides in vitro
Objective The quantitative analysis method of fluorescein isothiocyanate(FITC)of different oligosaccharides in cream was established,and the transdermal ability of oligosaccharides was investigated in vitro.Methods Using Franz diffusion cell,the FITC fluorescence quantitative analysis method of different oligosaccharides was established by using Bama pig skin as the carrier and referring to the verification guidelines for quantitative analysis methods of biological samples in Chinese Pharmacopoeia(2020 edition,volume Ⅳ,9012 general rules),and the transdermal content of different oligosaccharides in cream was determined.Results A fluorescence quantitative analysis method for the transdermal absorption of chitosan oligosaccharide(COS),alginate oligosaccharide(AOS)and glucan oligosaccharide(GOS)in cream was established.The results showed that the mass concentration of FITC-fluorescence in and under the skin had a good linear relationship with fluorescence intensity,of which FITC-COS was 0.92-45.83 μg/mL,FITC-GOS was 0.71-42.44 μg/mL,and FITC-AOS was 0.28-27.85 μg/mL(in the skin)and 0.28-16.71 μg/mL(under the skin),and R2＞0.99.The accuracy,precision and stability of the method were in line with the requirements of the guiding principles.Conclusion The quantitative analysis method of FITC-fluorescence of different oligosaccharides in cream was established,and the results showed that the retention amount in the skin and the subcutaneous penetration amount was chitosan oligosaccharide＞alginate oligosaccharide＞glucan oligosaccharide.The retention amount of chitosan oligosaccharide in the skin was(35.81±2.09)μg/cm2,and the subcutaneous penetration amount was(118.72±6.91)μg/cm2.
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