Journal of the Korean Society of Food Science and Nutrition (한국식품영양과학회지)

The Korean Society of Food Science and Nutrition (한국식품영양과학회)
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Determination of Soluble Carbohydrates in Soybean Seeds Using High Performance Liquid Chromatography with Evaporative Light Scattering Detection

증기화광산란 검출기를 이용한 콩 함유 수용성 탄수화물의 분석

  • Kim, Gyeong-Ha (Dept. of Herbal Medicine Resource, Kangwon National University) ;
  • Hwang, Young-Sun (Dept. of Herbal Medicine Resource, Kangwon National University) ;
  • Ahn, Kyung-Geun (Dept. of Herbal Medicine Resource, Kangwon National University) ;
  • Kim, Gi-Ppeum (Dept. of Herbal Medicine Resource, Kangwon National University) ;
  • Kim, Min-Ji (Dept. of Herbal Medicine Resource, Kangwon National University) ;
  • Hong, Seung-Beom (Dept. of Herbal Medicine Resource, Kangwon National University) ;
  • Moon, Jung-Kyeong (National Institute of Crop Science, RDA) ;
  • Choung, Myoung-Gun (Dept. of Herbal Medicine Resource, Kangwon National University)
  • 김경하 (강원대학교 보건과학대학 생약자원개발학과) ;
  • 황영선 (강원대학교 보건과학대학 생약자원개발학과) ;
  • 안경근 (강원대학교 보건과학대학 생약자원개발학과) ;
  • 김기쁨 (강원대학교 보건과학대학 생약자원개발학과) ;
  • 김민지 (강원대학교 보건과학대학 생약자원개발학과) ;
  • 홍승범 (강원대학교 보건과학대학 생약자원개발학과) ;
  • 문중경 (농촌진흥청 국립식량과학원 전작과) ;
  • 정명근 (강원대학교 보건과학대학 생약자원개발학과)
  • Received : 2014.03.19
  • Accepted : 2014.06.02
  • Published : 2014.07.31
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Abstract

In the present study, a new analytical method was devised for the simultaneous determination of soluble carbohydrates in soybean seeds using high performance liquid chromatography/evaporative light scattering detection (HPLC/ELSD). The limit of quantification (LOQ) for soybean soluble carbohydrates ranged from 5.6~7.6 mg/kg using the HPLC/ELSD method and from 16.2~33.9 mg/kg using the high performance liquid chromatography/refractive index detection (HPLC/RID) method. Therefore, the HPLC/ELSD method was more sensitive than HPLC/RID. The precision values for retention time and peak area of the HPLC/ELSD method were evaluated by inter-day (n=5) and intra-day (n=10) assays using a standard solution. All precision values (CV<2.5%) for soybean soluble carbohydrates were acceptable and fulfilled international acceptance criteria. All linear calibration curves were obtained with a correlation coefficient of $R^2$ >0.999. The contents of soluble carbohydrates for the "Shingikong" (yellow soybean) and "Cheongjakong 3" (black soybean) samples were analyzed using the HPLC/RID and HPLC/ELSD methods. The difference in carbohydrate contents between the two detection methods was significant. Carbohydrate contents in the HPLC/ELSD method were higher than those in the HPLC/RID method. Overall, the HPLC/ELSD method showed satisfactory resolution with a favorable LOQ and reproducibility. Therefore, these results indicate that the HPLC/ELSD method may be applied to determine the contents of soluble carbohydrates in soybean seeds and related food stuffs.

콩 함유 수용성 탄수화물의 동시분석 시 증기화광산란검출법(HPLC/ELSD) 및 굴절율검출법(HPLC/RID)의 분석 효율성, 정밀성 및 재현성을 상호 비교, 평가하여 콩 함유 수용성 탄수화물의 효율적 분석방법을 제공하고자 하였다. HPLC/ELSD 분석법 적용 시 콩 함유 수용성 탄수화물의 정량한계(LOQ)를 확인한 결과 5.6~7.6 mg/kg 범위를 나타낸 반면, RID는 16.2~33.9 mg/kg 범위를 나타내어 HPLC/ELSD 분석법이 RID 분석법 대비 감도가 향상된 양상을 확인하였고, intra-day(n=10) 및 inter-day(n=5) 분석재현성 평가에서도 피크의 머무름 시간(Rt.) 및 면적(peak area) 모두 최대 2.5% 미만의 변이계수를 나타내어 높은 분석 재현성을 확인할 수 있었다. 또한 농도별 표준시약을 이용하여 검량식의 직선성을 확인한 결과 모든 성분이 고도의 직선성($R^2$ >0.999)이 유지되어 정량적 분석이 가능함을 확인하였다. 콩 함유 수용성 탄수화물의 함량을 HPLC/ELSD 및 RID 방법으로 상호 비교한 결과 HPLC/ELSD 분석법을 적용하였을 경우 RID 분석법 대비 각 수용성 탄수화물의 함량이 더 높게 평가됨을 확인할 수 있었다.

Keywords

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