Analytical Methods for the Isolation of Dehydrotomatine and ${\alpha}$-Tomatine in Tomato Fruits by Use of Alumina Column Chromatography and High-Performance Liquid Chromatography

Alumina Column Chromatography와 HPLC에 의한 토마토의 Dehydrotomatine 및 ${\alpha}$-Tomatine 단리방법 연구

  • Choi, Suk-Hyun (Dept. of Food Service Industry, Seowon University) ;
  • Kim, Hyen-Ryung (Dept. of Foodservice Management & Cuisine, Gyeongju University) ;
  • Lee, Jin-Shik (Dept. of Food Service Industry, Uiduk University)
  • 최석현 (서원대학교 외식산업학과) ;
  • 김현룡 (경주대학교 외식.조리학과) ;
  • 이진식 (위덕대학교 외식산업학과)
  • Received : 2010.10.21
  • Accepted : 2010.12.11
  • Published : 2010.12.01

Abstract

Tomato fruits(Lycoperisicon esculentum) synthesize the glycoalkaloids dehydrotomatine and ${\alpha}$-tomatine, possibly as defense against bacteria, fungi and insects. We developed a new effective method to prepare and purify dehydrotomatine and ${\alpha}$-tomatine that exists in tomato fruits using alumina column chromatography and high performance liquid chromatography (HPLC). The tomato glycoalkaloids(TGA) in tomato was extracted with 2% acetic acid, and then precipitated with ammonium hydroxide(pH=10.5). The dry precipitate substance was applied on alumina column, and then fractionated with water saturated n-butylalcohol. The TGA(Fr. No. 26~36) were collected and dried under reduced pressure. The TGA was performed on a reverse phase HPLC(Inertsil ODS-2, $5\;{\mu}m$), eluted with acetonitrile/20mM $KH_2PO_4$(24:76, v/v) at 208 nm. Two peaks were detected on HPLC, and individual peak was collected by repeating HPLC. Furthermore, to confirm the identity dehydrotomatine and ${\alpha}$-tomatine, each peak isolated was hydrolyzed with 1N HCl into sugar and aglycone tomatidine. The sugars were converted to trimethylsilyl ester derivatives. The nature and molar ratios of sugars were identified by gas-liquid chromatography(GLC) and the aglycone by high-performance liquid chromatography(HPLC). The first peak (Rt=17.5 min) eluted from HPLC was identified as dehydrotomatine, and second peak(Rt=21.0 min) was as ${\alpha}$-tomatine. This technique has been used effectively to prepare and isolate dehydrotomatine and ${\alpha}$-tomatine from tomato fruits.

Keywords

References

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