홍어 껍질 콜라겐 펩타이드의 지표성분 밸리데이션 및 생리활성
Validation of an analytical method for the quantification of a marker compound and determination of its biological activities in skate skin collagen peptides
0
조회
0
다운로드
3
인용
0
참고문헌
키워드
초록
In this study, we developed and validated a high-performance liquid chromatography/photodiode array method for the quantification of glycine- proline-hydroxyproline (GPH) as a marker compound in skate skin collagen peptides (SCPs). The accurate molecular mass and amino acid sequence of this marker were determined using a QTOF mass spectrometer. Chromatographic separations were conducted using a 95:5 isocratic mobile phase of 0.1% trifluoroacetic acid (A) and acetonitrile/methanol (1:4 v/v, B) at a flow rate of 0.3 mL/min; the separated compounds were detected at 214 nm using a Jupiter® 4 μm Proteo 90Å column. The method was validated for specificity, linearity, limit of detection (LOD), limit of quantitation (LOQ), accuracy, and precision. It was found to be linear in the range of 1.0-50.0 μg/mL, with a good correlation coefficient (R2=0.9999) and excellent specificity. The LOD and LOQ were 0.07 and 0.22 μg/mL, respectively. A recovery study determined the accuracy of the method, with an average recovery of 103.76%, 100.35% and 103.97% of the marker at 10, 15 and 25 μg/mL, respectively, and a precision study showed a relative standard deviation of less than 2%. Additionally, the DPPH and ABTS radical scavenging activities of the SCPs increased in a concentration-dependent manner, and UVprotective effect was confirmed by human keratinocyte (HaCaT cell) viability. Our study thus provides the foundation for developing functional foods or cosmetics using SCPs.
Abstract
In this study, we developed and validated a high-performance liquid chromatography/photodiode array method for the quantification of glycine- proline-hydroxyproline (GPH) as a marker compound in skate skin collagen peptides (SCPs). The accurate molecular mass and amino acid sequence of this marker were determined using a QTOF mass spectrometer. Chromatographic separations were conducted using a 95:5 isocratic mobile phase of 0.1% trifluoroacetic acid (A) and acetonitrile/methanol (1:4 v/v, B) at a flow rate of 0.3 mL/min; the separated compounds were detected at 214 nm using a Jupiter® 4 μm Proteo 90Å column. The method was validated for specificity, linearity, limit of detection (LOD), limit of quantitation (LOQ), accuracy, and precision. It was found to be linear in the range of 1.0-50.0 μg/mL, with a good correlation coefficient (R2=0.9999) and excellent specificity. The LOD and LOQ were 0.07 and 0.22 μg/mL, respectively. A recovery study determined the accuracy of the method, with an average recovery of 103.76%, 100.35% and 103.97% of the marker at 10, 15 and 25 μg/mL, respectively, and a precision study showed a relative standard deviation of less than 2%. Additionally, the DPPH and ABTS radical scavenging activities of the SCPs increased in a concentration-dependent manner, and UVprotective effect was confirmed by human keratinocyte (HaCaT cell) viability. Our study thus provides the foundation for developing functional foods or cosmetics using SCPs.
참고문헌 (37)
- 안소언, "수산부산물의 발생·이용 실태 평가 및 해양바이오 산업화 방안", Ocean and Polar Research, 43(3), pp.149~164 (2021), 한국해양과학기술원[DOI]
- Official Methods of Analysis, Guidelines for Standard Method Performance Requirements Appendix F, "AOAC", AOAC International, 6~9 (2019)
- 백장미, "저이용 홍어 가공 부산물을 이용한 고기능성 물질의 생산기술개발에 관한 연구(1) - 홍어연골을 이용한 콘드로이틴 황산 소재개발", 한국환경과학회지, 25(5), pp.645~654 (2016), 한국환경과학회[DOI]
- Bailey AJ, "Connective Tissue in Meat and Meat Products", pp.1~24 (1989), Elsevier Applied Science
- Cho SH, "The effect of processing conditions on the properties of gelatin from skate (Raja kenojei ) skin", Food Hydrocoll, 20, pp.810~816 (2006)
- 최명락, "홍어 숙성과 기능성", 생명과학회지, 13(5), pp.675~683 (2003), 한국생명과학회
- Cui X, "Western blot analysis of type Ⅰ, Ⅲ, Ⅴ, Ⅵ collagen after laser epithelial keratomileusis and photorefractive keratectomy in cornea of rabbits", Yan Ke Xue Bao, 21, pp.141~148 (2005)
- Devita L, "The bioactivity prediction of peptides from tuna skin collagen using integrated method combining in vitro and in silico", Foods, 10, pp.2739~ (2021)
- Ding D, "Isolation and identification of and antioxidant collagen peptide from skipjack tuna (Katsuwonus pelamis ) bone", RSC Adv, 9, pp.27032~27041 (2019)
- Hong H, "Preparation of low-molecular-weight, collagen hydrolysates (peptides): Current progress, challenges, and future perspectives", Food Chem, 301, pp.125222~ (2019)
- Hwang JH, "Purification and characterization of molecular species of collagen in the skin of skate (Raja kenojei )", Food Chem, 100, pp.921~925 (2007)
- Iwai K, "Identification of food-derived collagen peptides in human blood after oral ingestion of gelatin hydrolysates", J Agric Food Chem, 53, pp.6531~6536 (2005)
- 정갑섭, "http:", 10.5762, pp.KAIS.2016.17.6.1, 한국산학기술학회논문지[DOI]
- 정갑섭, "홍어 콜라겐 펩타이드의 산화적 스트레스 완화를 통한 항염증효과", 한국응용과학기술학회지, 35(4), pp.1369~1378 (2018), 한국응용과학기술학회[DOI]
- Jerome SP, "Identification of collagen fibrils in scleroderma skin", J Invest Dermatol, 90, pp.48~54 (1988)
- 강건희, "홍어껍질 추출물의 추출특성과 유지 산화억제 효과 및 콜라겐겔 제조", 한국산학기술학회논문지, 13(11), pp.5637~5645 (2012), 한국산학기술학회[DOI]
- 김애향, "콜라겐 트리펩타이드를 함유한 콜라겐 가수분해물의 피부 광노화 예방 효과", 한국식품과학회지, 51(5), pp.466~472 (2019), 한국식품과학회[DOI]
- 김애향, "콜라겐 트리펩타이드를 고함량으로 함유하는콜라겐 가수분해물의 피부 보습 효과", 한국식품과학회지, 50(4), pp.420~429 (2018), 한국식품과학회[DOI]
- 김동환, "광어껍질 펩신 가수분해물 유래 콜라게네이즈 저해 펩타이드의 서열분석과 분자도킹", 한국식품영양과학회지, 51(4), pp.322~333 (2022), 한국식품영양과학회[DOI]
- 김정기, "콜라겐 펩타이드의 피부 장벽 보호 효과", 한국식품과학회지, 43(4), pp.458~463 (2011), 한국식품과학회
- Kleinnijenhuis AJ, "Identification of collagen 1α3 in teleost fish species and typical collision induced internal fragmentations", Food Chem X, 14, pp.100333~ (2022)
- Kumazawa Y, "A rapid and simple LC-MS method using collagen marker peptides for identification of the animal source of leather", J Agric Food Chem, 64, pp.6051~6057 (2016)
- 권민철, "동해 오징어 (Todarodes pacificus) 껍질 유래 콜라겐의 UV 보호능 및 미백효과", 한국수산과학회지, 41(1), pp.7~12 (2008), 한국수산과학회
- 권민철, "불가사리(Asterias amurensis) 콜라겐 유래 저분자 펩타이드의 피부주름 억제활성", 한국식품과학회지, 39(6), pp.625~629 (2007), 한국식품과학회
- 이현정, "홍어 껍질 유래 콜라겐의 지방대사 조절을 통한 간 조직 지질축적 억제 효과", 한국식품영양과학회지, 47(3), pp.235~242 (2018), 한국식품영양과학회
- 이지해, "AP 콜라겐 효소분해 펩타이드에 의한 자외선 손상으로부터 피부세포 보호 및 광노화 억제 효능", 한국식품영양과학회지, 50(2), pp.119~127 (2021), 한국식품영양과학회
- Lee JK, "Characterization of β-secretase inhibitory peptide purified from skate skin protein hydrolysate", Eur Food Res Technol, 240, pp.129~136 (2015)
- 이미진, "어류비늘에서 추출한 콜라겐펩타이드의 제조 및 유효성 분석", 한국응용과학기술학회지, 26(4), pp.457~466 (2009), 한국응용과학기술학회
- "Guidelines on analytical method validation for pharmaceuticals, etc" (2015)[URL]
- "Certified functional ingredient for health/functional food" (2016)[URL]
- "Guidelines for the standard procedure for preparing test methods for food, etc" (2016)[URL]
- "Guidelines for preparation of submission data for approval of functional ingredient for health/functional food" (2022)[URL]
- Ngo DH, "Angitensin - I converting enzyme inhibitory peptides from antihypertensive skate (Okamejei kenojei ) skin gelatin hydrolysate in spontaneously hypertensive rats", Food Chem, 174, pp.37~43 (2015)
- Ohara H, "Collagenderived dipeptide, proline-hydroxyproline, stimulates cell proliferation and hyaluronic acid synthesis in cultured human dermal fibroblasts", J Dermatol, 37, pp.330~338 (2010)
- 박성하, "홍어류(Dipturus chilensis ) 껍질로부터 분리 정제된 collagenase-1 저해 펩타이드의 특성", 한국수산과학회지, 44(5), pp.456~463 (2011), 한국수산과학회
- Yamamoto S, "Absorption and urinary excretion of peptides after collagen tripeptide ingestion in humans", Biol Pharm Bull, 39, pp.428~434 (2016)
- 양수진, "명태 껍질 유래 콜라겐의 분자량에 따른 이화학적 특성 및 생리활성", 한국식품영양과학회지, 43(10), pp.1535~1542 (2014), 한국식품영양과학회