• 제목/요약/키워드: reverse phase high-performance liquid chromatography

검색결과 151건 처리시간 0.034초

호도기름의 Triglyceride 조성(組成)에 관한 연구(硏究) (Triglyceride Composition of Walnut Oil)

  • 천석조;박영호
    • 한국식품영양과학회지
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    • 제13권3호
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    • pp.263-267
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    • 1984
  • 호도(胡桃)기름의 triglyceride 조성(組成)을 밝히기 위하여 TLC로서, 호도(胡桃)기름의 triglyceride를 분리하고, 이것을 HPLC에 의하여 PN별(別)로 triglyceride를 분획(分劃)하였다. PN 별(別)로 분획(分劃)한 각(各) 획분(劃分)을 다시 GLC에 의하여 acyl 탄소수별(炭素數別)로 분획(分劃)하였으며, 또한 PN별(別) 각획분(各劃分)의 지방산조성(脂肪酸組成)을 GLC로 분석(分析)하였다. 이들 세가지 chromatography의 분석결과(分析結果)로 부터 호도(胡桃)기름의 triglyceride 조성(組成)을 산정(算定)하였다. 산정(算定)한 호도(胡桃)기름의 triglyceride는 10종류였으며, 이들 중 1% 이상을 차지하는 triglyceride를 들면 다음과 같다. $(C_{18:2},\;C_{18:2},\;C_{18:2};\;53.3%)$, $(C_{18:1},\;C_{18:2},\;C_{18:2};\;10.1%)$, $(C_{18:1},\;C_{18:1},\;C_{18:2};\;5.4%)$, $(C_{18:1},\;C_{18:2},\;C_{18:3};\;4.3%)$, $(C_{18:0},\;C_{18:2},\;C_{18:2};\;3.9%)$, $(C_{18:0},\;C_{18:1},\;C_{18:2};\;2.0%)$, $(C_{18:0},\;C_{18:2},\;C_{18:2};\;1.8%)$.

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먹장어 (Eptatretus burgeri)의 피부로부터 Bradykinin-Related Peptide의 정제 (Purification of a Bradykinin-Related Peptide from the Skin of Hagfish. Eptatretus burgeri)

  • 신미정;김은정;김찬희;고혜진;김인혜;류홍수;허민도;정준기;박남규
    • 한국수산과학회지
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    • 제36권1호
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    • pp.30-34
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    • 2003
  • Hagfish bradykinin (BK)-related peptide가 먹장어 (E. burgeri)의 피부추출물로부터 분리, 정제되어졌다. 분자량이 875.8 Da인 hagfish BK는 $C_{18}$ 역상 및 양이온교환 high performance liquid chromatography로 순수하게 정제되었다. 자동아미노산서열분석기와 MALDI-TOF mass spectroscopy에서 얻은 결과의 조합으로 hagfish BK의 일차구조는 Gly-Thr-Ala-Gly-Ile-Gly-Pro-Phe-Arg로 판명되었다. 이 아미노산서열을 mammalian BK와 비교하면 다섯 개의 아미노산의 치환을 $(Arg^1{\rightarrow}Gly,\;Pro^2{\rightarrow}Thr,\;Pro^3{\rightarrow}Ala,\;Phe^5{\rightarrow}Ile,\;Ser^6{\rightarrow}Gly)$ 포함한다. Hagfish BK는 먹장어 장관에 대해 수축활성을 나타냈으며, 약 $10^{-11}\;M$의 농도에서 역치반응이 나타났다.

Isolation, Purification and Characterization of Antioxidative Bioactive Elastin Peptides from Poultry Skin

  • Nadalian, Mehdi;Kamaruzaman, Nurkhuzaiah;Yusop, Mohd Shakir Mohamad;Babji, Abdul Salam;Yusop, Salma Mohamad
    • 한국축산식품학회지
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    • 제39권6호
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    • pp.966-979
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    • 2019
  • Muscle-based by-products are often undervalued although commonly reported having a high amount of natural bioactive peptides. In this study, elastin was isolated from the protein of broiler hen skin while its hydrolysate was prepared using Elastase. Assessment of antioxidative properties of elastin-based hydrolysate (EBH) was based on three different assays; 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) radical, 2,2'-azinobis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) radical and metal chelating ability. The EBH was purified further using ultrafiltration, gel filtration and Reverse- Phase High-Performance Liquid Chromatography (RP-HPLC). The IC50 of ABTS radical activities for EBH were decreased as EBH further purified using ultrafiltration (EBH III; 0.66 mg/mL)>gel filtration (EB-II; 0.42 mg/mL)>RP-HPLC (EB-II4; 0.12 mg/mL). The sequential identification of the peptide was done by matrix-assisted laser desorption/ionization time-of-flight/time-of-flight mass spectrometry (MALDI-TOF/ TOF-MS) of the potent fractions obtained from RP-HPLC (EB-II4). The presence of hydrophobic amino acids (Val and Pro) in the peptide sequences could potentially contribute to the high antioxidant activity of EBH. The sequences GAHTGPRKPFKPR, GMPGFDVR and ADASVLPK were identified as antioxidant peptides. In conclusion, the antioxidative potential from poultry skin specifically from elastin is evident and can be explored to be used in many applications such as health and pharmaceutical purposes.

들깨기름의 Triglyceride 조성(組成)에 관한 연구 (Triglyceride Composition of Perilla Oil)

  • 박영호;김동수;천석조
    • 한국식품과학회지
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    • 제15권2호
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    • pp.164-169
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    • 1983
  • 들깨기름의 트리-글리세리드조성(組成)을 밝히기 위하여 TLC에 의하여 시료유(試料油)로부터 트리-글리세리드를 분리하고, 이것을 ${\mu}-Bondapak\;C_{18}$ column을 사용한 HPLC에 걸어 PN별(別)로 트리-글리세리드를 분획(分劃)하였으며, 각획분(各劃分)을 분취(分取)하여 GLC로 아실 탄소수별(炭素數別)로 분획(分劃)하였다. 또한 PN별(別) 획분(劃分)의 지방산조성(脂肪酸組成)을 GLC에 의하여 분석하였다. 이들 분석 결과로부터 들깨기름을 구성하는 트리-글리세리드조성(組成)을 산정(算定)하였는데 그 수(數)는 15종류이며, 그 중 주요(主要)한 트리-글리세리드 및 그 비율은 다음과 같다. 즉, $(C_{18:3},\;C_{18:3},\;C_{18:3},\;68.0%)$, $(C_{18:2},\;C_{18:3},\;C_{18:3},\;6.7%)$, $(C_{18:1},\;C_{18:2},\;C_{18:3},\;5.9%)$, $(C_{16:0},\;C_{18:3},\;C_{18:3},\;4.3%)$, $(C_{18:1},\;C_{18:2},\;C_{18:3},\;3.8%)$, $(C_{18:1},\;C_{18:1},\;C_{18:3},\;3.2%)$, ($C_{16:0},\;C_{18:2},\;C_{18:3},\;2.0%)$, $(C_{18:2},\;C_{18:2},\;C_{18:3},\;1.5%)$$(C_{16:0},\;C_{18:1},\;C_{18:3},\;1.0%)$ 등이다.

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Selective production of red azaphilone pigments in a Monascus purpureus mppDEG deletion mutant

  • Balakrishnan, Bijinu;Lim, Yoon Ji;Hwang, Seok Hyun;Lee, Doh Won;Park, Si-Hyung;Kwon, Hyung-Jin
    • Journal of Applied Biological Chemistry
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    • 제60권3호
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    • pp.249-256
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    • 2017
  • The Monascus azaphilone (MAz) pigment is a well-known food colorant that has yellow, orange and red components. The structures of the yellow and orange MAz differ by two hydride reductions, with yellow MAz being the reduced form. Orange MAz can be non-enzymatically converted to red MAz in the presence of amine derivatives. It was previously demonstrated that mppE and mppG are involved in the biosynthesis of yellow and orange MAz, respectively. However, ${\Delta}mppE$ and ${\Delta}mppG$ knockout mutants maintained residual production of yellow and orange MAz, respectively. In this study, we deleted the region encompassing mppD, mppE and mppG in M. purpureus and compared the phenotype of the resulting mutant (${\Delta}mppDEG$) with that of an mppD knockout mutant (${\Delta}mppD$). It was previously reported that the ${\Delta}mppD$ strain retained the ability to produce MAz but at approximately 10% of the level observed in the wildtype strain. A chemical analysis demonstrated that the ${\Delta}mppDEG$ strain was still capable of producing both yellow and orange MAz, suggesting the presence of minor MAz route(s) not involving mppE or mppG. Unexpectedly, the ${\Delta}mppDEG$ strain was observed to accumulate fast-eluting pigments in a reverse phase high-performance liquid chromatography analysis. A LC-MS analysis identified these pigments as ethanolamine derivatives of red MAz, which had been previously identified in an mppE knockout mutant that produces high amounts of orange MAz. Although the underlying mechanism is largely unknown, this study has yielded an M. purpureus strain that selectively accumulates red MAz.

곰보배추에 함유된 6-Hydroxyluteolin 7-O-Glucoside 분석을 위한 HPLC/DAD 분석법 밸리데이션 (HPLC/DAD Method Validation of 6-Hydroxyluteolin 7-O-Glucoside Analysis from Salvia plebeia)

  • 이학동;바헤;최중원;김주리;유아람;배민정;이상현
    • 생약학회지
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    • 제52권3호
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    • pp.186-191
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    • 2021
  • We developed a method to identify and quantify 6-hydroxyluteolin 7-O-glucoside in the powder of Salvia plebeia (PS) using high-performance liquid chromatography coupled with diode array detector (HPLC/DAD) and equipped with reverse-phase INNO C18 column. The analytical method was optimized and validated using novel parameters. The obtained values for the limits of detection and quantification were 3.60 and 10.90 ㎍/mL, respectively. Calibration curve showed good linearity in the concentration range tested (0.00625-0.1 mg/mL, r2 = 1.0000), high accuracy (96.2-101.4%), and precision values (RSD ≤ 0.27%). Our analysis support the use of our method for accurately identifying and quantifying 6-hydroxyluteolin 7-O-glucoside from PS in routine analyses and large-scale extraction processes for content determination.

산지별 밤꿀에 함유된 Kynurenic Acid의 정량 분석과 분석법 검증 (Quantitative Analysis of Kynurenic Acid in Chestnut Honey from Different Regions and Method Validation)

  • 김주리;김도윤;이상현
    • 생약학회지
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    • 제53권2호
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    • pp.111-118
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    • 2022
  • Chestnut honey is a sweet dark-colored honey with a distinct bitter aftertaste. It contains numerous phenolic compounds and alkaloids and is noted for its antioxidant and anti-inflammatory activities. However, it has been established that there are differences in the composition and activity of chestnut honey constituents depending on the region of origin, the sources of which warrant further research. In this study, we analyzed the kynurenic acid (KA) contents in chestnut honey produced in nine different regions in Korea, using high-performance liquid chromatography in conjunction with ultraviolet detection, and validated the analytical method developed. Use of a reverse-phase column and detection at a wavelength of 240 nm were found to be optimal for the detection of KA. Similar evaluation of an optimal method for extracting KA from chestnut honey revealed that extraction using 10% EtOH at 20 times the sample volume over a 6 h period was the most suitable for obtaining a high content of KA. Among the nine regional chestnut honeys assessed, KA content was found to be highest in the "Gongju" sample (1.14 mg/g), followed by that in the "Cheongdo" and "Damyang" samples. Validation of the KA analytical method revealed a good analyte linearity, with a correlation coefficient (r2) of 0.9995, an accuracy of between 92.37% and 107.35%, and good precision (RSD ≤ 1.05%). Our findings in this study, based on a validated quantitative analytical method for KA, could make an important contribution to establishing a data profiling procedure for characterizing chestnut honeys produced in different regions, and may also provide basic data for the identification of functional honey.

개량된 방법에 의한 사람혈소판으로부터 TGF-$\beta$ 1의 분리 (Purification of TGF-$\beta$ 1 from Human Platelets by an Improved Method)

  • 신충건;김상국;문병조;김평현;전계택;남상욱;김장환;이종원
    • KSBB Journal
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    • 제14권1호
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    • pp.9-16
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    • 1999
  • Transforming growth factor $\beta$1(TGF-$\beta$1)은 여러 가지 생물학적 활성을 가지는 관계로 의학적 치료제로서 사용될 가능성이 크다. 본 연구에서는 혈소판추출, 젤여과, 양이온교환 크로마토그래피 및 역상 HPLC등 네 단계의 정제과정으로 이루어져 있는 정제공정을 이용한 TGF-$\beta$1을 값싸고 효울적으로 정제하였다. 이 과정을 거쳐 최종적으로 얻어진 TGF-$\beta$1은 비환원조건하에서 SDS-PAGE를 행한 결과 구매된 TGF-$\beta$1 표준품과 일치한 위치에서 한 개의 band가 관찰되어 순수하다는 것을 확인하였으며 또한 이것이 Westem blot를 통하여 TGF-$\beta$1 항체와 결합하는 것으로부터 TGF-$\beta$1임을 확인하였다 또한, mink lung epithelial cell line 을 이용한 성장저해 실험을 통해 정제된 TGF-$\beta$1이 구매되TGF-$\beta$1 표준품보다 조금 높은 활성을 가지는 것을 확인하였다 최종적으로 농축혈소판 10단위로부터 약 3.7$\mu$g의 정제된 TGF-$\beta$1이 얻어져 그 최종수율은 약 21%였다.

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Evaluation of the Digestibility of Korean Hanwoo Beef Cuts Using the in vitro Physicochemical Upper Gastrointestinal System

  • Jeon, Ji-Hye;Yoo, Michelle;Jung, Tae-Hwan;Jeon, Woo-Min;Han, Kyoung-Sik
    • 한국축산식품학회지
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    • 제37권5호
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    • pp.682-689
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    • 2017
  • The aim of this study was to investigate the digestibility of different Korean Hanwoo beef cuts using an in vitro digestion model, in vitro physicochemical upper gastrointestinal system (IPUGS). The four most commonly consumed cuts - tenderloin, sirloin, brisket and flank, and bottom round - were chosen for this study. Beef samples (75 g) were cooked and ingested into IPUGS, which was composed of mouth, esophagus, and stomach, thereby simulating the digestion conditions of humans. Digested samples were collected every 15 min for 4 h of simulation and their pH monitored. Samples were visualized under a scanning electron microscope (SEM) to examine changes in the smoothness of the surface after digestion. Analysis of the amino acid composition and molecular weight (MW) of peptides was performed using reverse-phase high performance liquid chromatography and sodium dodecyl sulfate polyacrylamide gel electrophoresis, respectively. Following proteolysis by the gastric pepsin, beef proteins were digested into peptides. The amount of peptides with higher MW decreased over the course of digestion. SEM results revealed that the surface of the digested samples became visibly smoother. Total indispensable and dispensable amino acids were the highest for the bottom round cut prior to digestion simulation. However, the total amount of indispensable amino acids were maximum for the tenderloin cut after digestion. These results may provide guidelines for the elderly population to choose easily digestible meat cuts and products to improve their nutritional and health status.

Evaluation of Haemagglutinin Content by RP-HPLC to Generate Pandemic Influenza Vaccine

  • Kang, Hyunkyung;Roh, Hang Sik;Song, Hyemin;Lee, Kwangmoon;Chung, Seung-Tae;Ban, Sang-ja;Mo, In Pil;An, Beum-Soo;Ahn, Chi-Young
    • Toxicological Research
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    • 제32권4호
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    • pp.269-274
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    • 2016
  • The potency of influenza vaccine is determined based on its hemagglutinin (HA) content. In general, single radial immunodiffusion (SRID) assay has been utilized as the standard method to measure HA content. However, preparation of reagents for SRID such as antigen and antibody takes approximately 2~3 months, which causes delays in the development of influenza vaccine. Therefore, quantification of HA content by other alternative methods is required. In this study, we measured HA contents of H1N1 antigen and H1N1 influenza vaccine by reverse phase-high performance liquid chromatography (RP-HPLC) methods. The presence of HA1 and HA2 was investigated by silver staining and Western blot assay. In addition, accuracy and repeatability of HA measurement by RP-HPLC were evaluated. Comparison of HA concentration by SRID and RP-HPLC revealed a precise correlation between the two methods. Our results suggest that RP-HPLC assay can replace SRID in the event of a pandemic flu outbreak for rapid vaccine development.