Korean Journal of Food Science and Technology (한국식품과학회지)

Korean Society of Food Science and Technology (한국식품과학회)
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Rapid and Simple Analytical Method for Removing Patulin from Apple Juice Using Solid Phase Extraction

고체상 추출법(SPE: Solid Phase Extraction)을 이용한 국내 사과주스 중 Patulin 간편.신속 분석방법

  • Yim, Jong-Gab (Department of Food Science and Technology, Dongguk University) ;
  • Jang, Hae-Won (Department of Food Science and Technology, Dongguk University) ;
  • Lee, Kwang-Geun (Department of Food Science and Technology, Dongguk University)
  • Received : 2009.10.22
  • Accepted : 2010.01.22
  • Published : 2010.06.30
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Abstract

Patulin, a secondary metabolite of mold, is commonly found in rotten apples. Many countries regulate patulin at levels ranging from 30 to $50\;{\mu}g/L$. Most analytical methods for removing patulin from apple juice include liquid-liquid extraction (LLE), which is time and labor intensive. To replace the LLE method, a solid-phase extraction (SPE) method has been developed for apple juice and unfiltered apple juice. A portion of the test sample was applied to a macroporous copolymer cartridge and washed with 5 mL of 1% sodium bicarbonate, followed by 5 mL of 1% acetic acid. Patulin was eluted with 5 mL of 2% acetonitrile in anhydrous ethyl ether. The mobile phase was tetrahydrofuran in water (0.8:99.2) and was detected with a UV detector at 276 nm. Recoveries ranged from 95 to 101% in test samples, and the minimum detectable level was 30 ppb. Because this SPE method is fast, easy, reliable, and inexpensive, it could be applicable for companies or analytical agencies to analyze patulin concentrations in apple juice.

Patulin은 곰팡이의 이차 대사산물로 주로 썩은 사과의 병반에서 발견된다. 사과주스 중에 포함된 patulin에 대한 실험법은 LLE법을 이용한 방법이 대부분이다. 이를 대체하기 위해 SPE 법을 통한 사과주스 중 patulin 분석법을 본 연구에서 시행하였다. 시료인 사과주스를 다공성 중합체인 고체상 카트리지에 주입한 후 1% sodium bicarbonate 용액 5 mL와 1% acetic acid 5 mL를 차례로 흘려 방해물질 등을 제거해준 다음 정제가 끝난 카트리지를 상온에서 1시간 정도 방치한 후 ethyl acetate/ethyl ether 5 mL를 이용해 추출하였다. 추출된 용액을 질소 농축기를 이용해 완전 농축한 후 1% acetic acid 용액 0.5 mL를 넣고 혼합 후 syringe filter로 여과하여 amber vial에 보관하여 분석에 사용하였다. HPLC 분석에 사용된 이동상은 THF 8 mL를 1 L 정용 플라스크에 넣고 물로 채워 사용하였다. HPLC 분석조건은 이동상을 0.7 mL/min 유속으로 흘려 276 nm에서 흡광도를 측정하였고, 샘플주입은 $20\;{\mu}g/L$로 하였다. LOD는 30 ng/mL 수준이었다. 실험 결과 SPE 법은 기존의 식품공전시험법(LLE)보다 분석시간이 1/3로 줄어들었고, 적은 양의 용매만으로 추출 및 정제가 가능하였으며 실험과정이 간단하여 공전의 방식보다 손쉽게 분석할 수 있었다. 특히, strata-X를 통한 SPE 추출방법은 재연성과 회수율 또한 우수해 저 농도 분석에도 적합하였다. 이와 같이 기존 분석법에 비해 신속하고 간소하며 정확한 정량분석 및 우수한 회수율과 신뢰도를 얻을 수 있어 다량의 분석을 요하는 연구기관 및 단체에 매우 효율적일 것으로 사료된다.

Keywords

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