1Department of Internal Medicine, Inha University College of Medicine, Inchon, Korea 2Department of Surgery, Inha University College of Medicine, Inchon, Korea
Corresponding author:
Myung Sun Joo,
ABSTRACT
Background: Methods used for gallstone analysis include X-ray diffraction analysis, infrared anaylsis and chemical analysis. We measured total cholesterol of gallstones with enzymatic method. This method was shown to be simple and rapid compared with the IR spectrophotometry. Method: 1. Gallstones were obtained from 30 patients' gallbladder. 2. Gallstones were crushed into powder and then dissolved in N-N, dimethylformamide and dimethyl sulfoxide (8:2 by volume) mixture. 3.20 μl of this solvent mixed with 2 ml of reagent mixture and then incubated at 37ºC for 10 minutes. Reagent mixture contains phosphate buffer 0.1 mol/L, 4-aminoantipyrin 0.8 mmol/L, ρ-hydroxy-benzensulfonate 20 mmol/L, esterase 150 U/L, peroxidase 25 KU/L and cholesterol oxidase 200 U/L. 4. The absorbance of these reaction mixtures were measured at 500 nm using spectrophotometer(Beckman-Du 650). 5. The cholesterol calibration curve was made by using standard cholesterols and reagent mixture. 6. The cholesterol concentrations of gallstones were determined using cholesterol standard curve and each absorbances. 7. This concentrations were compared with the values using intrared spectrophotometry (Nicolet Inst. corp. Omnic and Quick IR) method. Result: Total cholesterol concentrations of 30 gallstones by enzymatic method correlated with cholesterol in gallstones determined by IR spectrophotometry method. Conclusion: Enzymatic method for total cholesterol determination of gallstone showed useful, simple and rapid.
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