Various species of the genus Capsicum contain in their fruits an intensively sharp and pungent substance, Capsaicin, which was first isolated in an almost pure state by Thresh. Containing the pungent principle, Capsicums are used extensively in food as a spice and in medicine as a rubefacient and carminative.
Numerous methods have been proposed for the isolation, the chemical structure and the quantitative determination of Capsaicin. Modifying the several methods described before, the Capsaicin contents in various species of the genus Capsicum were determined as follows.
(1) The isolation of pure Capsaicin was the essential first step for the determination of Capsaicin contents.
Powdered cayenne pepper was extracted with acetone. By the method of ether alkali partition extraction slightly modified at this laboratory and by the recrystallization with light petroleum ether that was repeated ten times, the pure crystalline Capsaicin was obtained. Using this Capsaicin, the standard absorption curve was drawn with Beckman spectrophotometer model DU for the quantitative determination of Capsaicin.
(2) The powdered sample was extracted in a Soxhlet extractor with ether-acetone solvent system (3 : 1) for 25 hours. Capsaicin in this ether-acetone extracts was efficiently separated in a pure state by paper partition chromatography using 58% methanol solution as developing agent. It was found that 58% methanol was one of the most valuable solvent to separate Capsaicin from impurities such as sterols, fatty acids, waxes and carotenoid pigments.
(3) The colorimetric method modifying the Schulte-Kruger´s method which consists of measuring the red color produced with diazobenzenesulphonic acid was used. Capsaicin in various species of the genus Capsicum was determined quantitatively with use of Beckman spectrophotometer model DU at 480 mμ.
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