Abstract
Objective To develop a high performance liquid chromatographic(HPLC)method for the determination of riboflavin in hepatic tissue.Methods The hepatic samples were grinded uniformly with homogenizer.The homogenate was then treated with 0.1 mol/L HCL and 10%trypsin hydrolysis,and analyzed after fihration.An Atlantis C18 column(150 mm x4.6 mm,5μm)was used.The mobile phase consisted of 35%methanol and 65%5 mmol/L ammonium acetate solution.The flow rate was 1.0 ml/min.The spectrophotofluorometer was set at a wavelength of 450 nm for excitation and 520 nm for emission.Results A good linear correlation between riboflavin concentration(from 1.0 ug/mL to 200 ng/mL)and fluorescence intensity was established.The detection limit was 1.0 ng/mL.The standard substance of riboflavin and the samples in single-day and multi-day relative standard deviations were in the range of 2.83%-3.76%and 0.71%-2.17%.respectively.The recoveries for riboflavin in hepatic tissue were in the ranges of 98.6%-101.1%.Conclusion The established HPLC method is quick,accurate,and sensitive,and can be applied to the determination of riboflavin in hepatic tissue. Key words: High performance liquid chromatography; Hepatic tissue; Riboflavin
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