동물성 식품에서의 효소 가수분해에 의한 비오틴 추출
Enzymatic hydrolysis for biotin extraction from animal-derived foods
0
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
HPLC를 이용하여 효소처리 및 추출방법을 달리하여 여러 가지 식품군의 biotin 함량을 분석하였다. 동물성 식품의 육류(beef, foreshank), 어패류(mackerel), 난류(yolk, raw) 중 각 1종을 선정하여 분석을 진행하였으며 모두 pepsin 처리를 진행한 고압멸균 추출법에서 유의적으로 높은 biotin 함량을 보였다. 추후 실험에서는 육류, 어패류, 난류에 대해 pepsin 처리 및 고압멸균기를 이용한 biotin 추출법을 이용하였다. 실험에 사용한 육류 시료 중 소고기의 앞다리(foreshank), 목심(chuck), 사태(shank), 닭고기의 다리(leg)는 pepsin 처리에 따라 biotin이 유의적으로 증가하였으나, 나머지 시료에서는 유의적인 차이가 나타나지 않았다. 어패류의 고등어(mackerel), 갈치(hair tail), 문어(giant octopus), 꼬막(cockle)은 pepsin 처리로 인하여 biotin의 함량이 높게 나타났다. 난류의 경우, 닭의 난황에서는 효소처리에 따른 biotin 함량이 유의적으로 증가하였으나, 닭의 전란과 메추리의 전란에서는 효소처리에 따라 biotin 함량이 증가하지 않았다. 본 연구에서 사용된 biotin 추출법 및 분석법 검증을 진행하였다. Biotin 표준품과 SRM 1849a의 retention time이 일치하는 것을 알 수 있었으며, biotin 표준품의 7개 농도를 분석하여 상관계수(R2)가 0.999 이상으로 나타났다. 또한 정확성 및 정밀성 검증은 AOAC(2002)에서 제시한 범위에 충족하는 것을 확인하였다. 검출한계 및 정량한계는 0.009 μg/100 g과 0.024 μg/100 g으로 나타나 미량으로 존재하는 biotin을 우수하게 검출할 것으로 사료된다. 분석법 검증을 통해 본 연구에서 이용된 biotin 추출 및 분석방법의 신뢰성을 확보할 수 있었으며, 식품 중 미량 존재하는 biotin을 추출하는데 적용 가능할 것으로 판단된다. 따라서 다양한 식품에서 biotin을 효과적으로 추출하기 위해서는 각 식품의 matrix에 따른 효소처리가 필요하며, 이는 식품 중 함유된 biotin의 영양정보를 구축하는 데 활용될 수 있을 것으로 기대된다.
Abstract
Biotin exists as free or covalently bound to protein through a lysine residue. Therefore, the extraction of free biotin form is difficult in foodstuffs. Thus, the aim of this study was to investigate the extraction method of biotin in animal-derived foods by enzymatic hydrolysis (pepsin) using autoclave extraction or sonication extraction. Biotin contents of animal origin foods such as meat (beef, foreshank), fish and shells (mackerel), and egg (yolk, raw) were significantly higher in the autoclave extraction method with pepsin treatment. Therefore, these methods were selected for further applications in this study. As a result, pepsin-assisted extraction methods showed significantly higher contents of biotin compared with non-enzyme-treated samples in most of meat, fish, shellfish, and egg. The performed extraction methods in this study were validated according to AOAC guideline. An excellent linearity of biotin standard was observed over R2=0.999. All analytical methods for biotin showed the acceptable accuracy (97-101% recovery) and the precision (1-4% RSD for the repeatability, 0-4% RSD for the reproducibility). The limits of detection and quantification for biotin analysis were 0.009 and 0.024 μg/100 g, respectively. These results may be useful for providing reliable data about the biotin contents in commonly consumed foods in Korea.
참고문헌 (27)
- AOAC., "AOAC Guidelines for Single Laboratory Validation of Chemical Methods for Dietary Supplements and Botanicals", Association of Official Agricultural Chemists International, ~ (2002)
- Blake CJ, "Analytical procedures for water-soluble vitamins in foods and dietary supplements : A review", Anal Bioanal Chem, 389, pp.63~76 (2007)
- Blanco A, "Meidcal biochemistry", pp.251~273 (2017), Acdemic Press
- Chemat F, "Ultrasound-assisted extraction in food analysis", Handbook of food analysis instruments, 11, pp.85~103 (2008)
- Dhakal S, "Thermal and high-pressure treatment stability of egg-white avidin in aqueous solution", J Food Process Eng, 43, pp.e13481~ (2020)
- Eitenmiller RR, "Vitamin Analysis for the Health and Food Science", pp.535~560 (2007), CRC Press Taylor & Francis Group
- Gropper SS, "Advanced Nutrition and Human Metabolism", pp.335~341 (2018), Cengage Learning
- Holler U, "Quantification of biotin in feed, food, tablets, and premixes using HPLC-MS", MS, 2006, pp.831, 8~16
- Hoppner K, "Pantothenic acid and biotin retention in cooked legumes", J Food Sci, 58, pp.1084~1085 (1993)
- Joseph G, "Determination of biotin by liquid chromatography coupled with immunoaffinity column cleanup extraction : Single-laboratory validation, first action", J AOAC Int, 99, pp.1110~1112 (2016)
- KNS, "Dietary Reference Intakes for Koreans 2015", pp.45~46 (2015), The Korean Nutrition Society
- Kwon JH, "Monitoring of biotin content in frequently consumed foods in Korea", J Korean Soc Food Cult, 35, pp.278~284 (2020)
- Kwon JH, "Biotin analysis in selected agricultural foods consumed in Korea", J Korea Soc Food Sci Nutr, 48, pp.1366~1372 (2019)
- Lahely S, "Determination of biotin in foods by high-performance liquid chromatography with post-column derivatization and fluorimetric detection", Food Chem, 65, pp.253~258 (1999)
- Machado FF, "Solubility and density of egg white proteins : Effect of pH and saline concentration", LWT-Food Sci Technol, 40, pp.1304~1307 (2007)
- "Ministry of education, culture, science, and technology"[URL]
- Mun GJ, "Validation of pepsin-assisted extraction and immunoaffinity-HPLC", DAD analysis for vitamin B12 in seafood, 2018, pp.47, 168~175
- Pacheco-Alvarez D, "Biotin in metabolism and its relationship to human disease", Arch Med Res, 33, pp.439~447 (2002)
- Pakin C, "Fluorimetric determination of pantothenic acid in foods by liquid chromatography with post-column derivatization", J Chromatogr A, 1035, pp.87~95 (2004)
- Rural Development Administration, "Korean Food Composition Table. 10th revision", Rural Development. Administration, ~ (2021)
- Rural Development Administration, "Korean Food Composition Table. 9th revision", Rural Development. Administration, ~ (2016)
- Said HM, "Biotin : Biochemical, Physiological and Clinical Aspects", Subcell Biochem, 56, pp.1~19 (2012)
- Shin YW, "Establishment of biotin analysis by LC-MS", MS method in infant milk formulas, 2016, pp.31, 327~334
- Sponton OE, "Protein nanovehicles produced from egg white. Part 1 : Effect of pH and heat treatment time on particle size and binding capacity", Food Hydrocolloids, 73, pp.67~73 (2017)
- Teaguel AM, "Biotin is stable in frozen foods", FASEB J, 18, pp.A143~ (2004)
- White 3rd HB, "Role of avidin and other biotin-binding proteins in the deposition and distribution of biotin in chicken eggs. Discovery of a new biotin-binding protein", Biochem J, 241, pp.677~684 (1987)
- Wolf R, "Rapid quantification of biotin in human skin extracts after dermal application using high-performance liquid chromatography-electrospray mass spectrometry", Anal Commun, 34, pp.335~337 (1997)