Journal of Food Hygiene and Safety (한국식품위생안전성학회지)

The Korean Society of Food Hygiene and Safety (한국식품위생안전성학회)
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Development of Method for Determining 4-Hydroxy-L-isoleucine in Health Functional Foods by HPLC

건강기능식품에서 HPLC를 이용한 4-Hydroxy-L-isoleucine 분석법 연구

  • Lee, Se-Yun (Nutrition and Functional Food Research Team, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety) ;
  • Jeong, Hee-Sun (Nutrition and Functional Food Research Team, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety) ;
  • Hu, Soo-Jung (Nutrition and Functional Food Research Team, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety) ;
  • Lee, Hye-Young (Nutrition and Functional Food Research Team, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety) ;
  • Oh, Jae-Myoung (Nutrition and Functional Food Research Team, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety)
  • 이세윤 (식품의약품안전처 식품의약품안전평가원 영양기능연구팀) ;
  • 정희선 (식품의약품안전처 식품의약품안전평가원 영양기능연구팀) ;
  • 허수정 (식품의약품안전처 식품의약품안전평가원 영양기능연구팀) ;
  • 이혜영 (식품의약품안전처 식품의약품안전평가원 영양기능연구팀) ;
  • 오재명 (식품의약품안전처 식품의약품안전평가원 영양기능연구팀)
  • Received : 2020.03.13
  • Accepted : 2020.05.07
  • Published : 2020.06.30
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Abstract

This study was conducted to develop a method of analysis for 4-hydroxy-L-isoleucine in the seed extract of fenugreek (Trigonella foenum graecum), a health functional food that contains dietary fiber. The analytical method for 4-hydroxy-L-isoleucine was derived with O-phthaldialdehyde reagent (OPA) and determined by HPLC-PDA. The method was performed on a Capcell Pak C18 UG 120 column (4.6×250 mm, 5 ㎛) in isocratic elution mode using disodium phosphate and acetonitrile. The validation of the developed analytical method was conducted by evaluating several parameters; selectivity, linearity, limit of detection (LOD), limit of quantification (LOQ), accuracy and repeatability. Excellent linearity (R2=0.999) was observed for 4-hydroxy-L-isoleucine in the concentration range (5-100 ㎍/mL). Observed recoveries of these compounds were found to be between 91.7 and 96.4%. Precision was between 0.2 and 2.4% relative standard deviation (%RSD).

본 연구는 건강기능식품 호로파종자식이섬유 중 4-hydroxyL-isoleucine에 대한 분석법을 개발하는 연구이다. 최적분석 조건을 확립하기 위해 시료 채취량, 전처리 용매 및 이동상 용매 조건을 비교 검토하였으며, HPLC-PDA를 이용하여 시료 중의 4-hydroxy-L-isoleucine를 분석하였다. 분석 시 사용한 컬럼은 Capcell Pack UG120 C18 (Shiseido, 4.6×250 mm, 5 ㎛)이며, 유도체 시약으로 OPA를 선정하였다. 확립된 시험법에 대해 특이성, 직선성, 검출한계, 정량한계, 정확성, 정밀성 등의 밸리데이션을 수행하였다. 5-100 ㎍/mL 농도에서 결정계수(R2) 0.999 이상으로 높은 직선성을 확인하였다. 또한, 회수율은 91.7-96.4%이었고, 정밀성은 0.5-1.1%의 상대표준편차(%RSD)를 확인하였다. 개발된 시험법은 호로파종자추출물 중 4-hydroxy-L-isoleucine 분석을 위한 시험법으로 활용되기에 적합한 것으로 판단된다.

Keywords

References

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