The Application of Fluorospectrophotometry on the Determination of Serum Albumin and Medicine
|Keywords||Spectrofluorimetric methods protein Ciprofloxacin hydrochloride Fluorescein Eosin Sudan|
Proteins play an essential role in life process. To develope new techniques for protein determination is very important in biological science, clinical diagnosis, drugs and food analysis; Medicine analysis is an important part of medicine research. The development of medicine science requires more sensible and quick methods for the determination of drugs.Spectrofluorimetric methods have the advantages of high sensitivity and good selectivity, they are also simple and quick. Therefore they have been widely used in the determination of drugs and proteins. In this paper, new spectrofluorimetric methods for the determination of bovine serum albumin （BSA）, human serum albumin （HAS）, ciprofloxacin hydrocloride（CPFX） and Sudan I were Studied. It includes four chapters:Chapter one:Summarized the advances in the determination of serum albumins and drugs by Spectrofluorimetric methods.64 reference papers were cited.Chapter two:Reported a new Spectrofluorimetric methods for the determination of BSA and HSA.It was found that in the condition of pH=9.4, small amount of protein could make the fluorescence intensity of the fluorescein-Cu2+-cetyltrimethyl ammonium bromide （CTMAB） system quenched greatly. Based on this, a sensible, simple, quick, low cost method for protein determination was built. Under the optimum conditions, the quenched fluorescence intensity is in proportion to the concentration of bovine serum albumin （BSA） and human serum albumin （HSA） in the range of 0.15～2.5 mg/L and 0.1～2.0 mg/L respectively with detection limits （3S/K） of 0.11 mg/L for BSA and 0.08 mg/L for HSA. This method has been applied to the determination of protein in actual human serum sample with satisfactory results. The relative standard deviations were 2.9%-3.7%for 6 parallel determinations of HSA in human serum sample and the recovery of HSA was 92.5%-107.5%.Chapter three:A simple, quick, sensible method for the determination of ciprofloxacin hydrochloride was built. In the medium of pH= 8.2 Clark-Lubs buffer solution, with the existence of Tween-20, Cu（Ⅱ）, eosin and ciprofloxacin hydrochloride could formate ternary chelate that lead to the fluorescence of eosin-Cu（Ⅱ） quenched. Under the optimum condition, the quenching degree is proportional with the concentration of ciprofloxacin hydrochloride in the range of 0.4～4.0 mg/L. the detection limit of the method is 0.11 mg/L. this method was applied in the determination of ciprofloxacin hydrochloride in tablets with satisfactory result. The recovery was 89.6%-97.0%and the RSD for 6 paralell determination of the ciprofloxacin hydrochloride in tablets was 2.41%-3.72%.Chapter four:The isochromatism between the fluorescence of Acridine Yellow and the absorption of SudanⅠwas studied, atλex/λem= 280/495 nm, SudanⅠabsorbed the fluorescence of Acridine Yellow effectively in the medium of ethanol, which made the fluorescence of Acridine Yellow quenched, based on those the new way to determine the concentration of Sudan I was built. The range of determining concentration of Sudan I was 1.5×10-6 mol/L～3.5×10-5 mol/L. The measurement has been applied to the determination of Sudan I in capsicum powder and chili sauce. The relative standard deviation was 0.8%～2.7% for 6 parallel determination of Sudan I samples and the recovery of Sudan I was 89.5%～109.0%. The detection limit of the method was up to 6.4×10-7 mol/L.