Colorimetric determination of sulfite in "kanpyo" (dried gourd shavings) and "konnyakuseiko" (devil's-tongue fine powder) using sulfite oxidase and catalase

Abstract

A simple and convenient method for colorimetric determination of sulfite in foods based on its conversion to formaldehyde with sulfite oxidase and catalase was developed. Sulfite in a sample was extracted with water and then diluted with methanol. One mL of sample solution containing about 5-10 micrograms of sulfite was taken into a test tube with a ground-glass stopper, and 3 mL of 0.04 mol/L borate buffer (pH 8.7), 1 mL of 0.4% 3-methyl-2-benzothiazolinone hydrazone (MBTH) solution, 2,000 units of catalase solution and 1.0 units of sulfite oxidase were added. The mixture was incubated for 35 minutes at 37 degrees C. Then 0.15 mL of 1 mol/L hydrochloric acid and 5 mL of 0.2% iron(III) nitrate solution were added. The reaction mixture was transferred to a measuring flask after standing for 5 minutes at room temperature, and diluted to 20 mL with methanol. The absorbance of this solution was measured using a spectrophotometer at the wavelength of 635 nm. The calibration curve prepared with sodium sulfite showed linearity between 0 to 16 micrograms/mL as sulfur dioxide. The recoveries of sulfite in "Kanpyo" (dried gourd shavings) and "Konnyaku-seiko" (devil's-tongue fine powder) by the proposed method were 97-104%, and the coefficients of variation were below 6%. The sulfite values in these foods determined by the proposed method were reasonably consistent with those obtained by the bubbling distillation-alkaline titration method.