• Title/Summary/Keyword: HPLC-ELSD

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The ceramide contents of Saccharomyces cerevisiae in batch culture (Saccharomyces cerevisiae 의 회분식 배양에서 세라마이드의 생산)

  • Kim, Se-Kyung;Noh, Yong-Ho;Yun, Hyun-Shik
    • KSBB Journal
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    • v.23 no.5
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    • pp.449-451
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    • 2008
  • Ceramide has become a widely used ingredient in cosmetic and pharmaceutical industries, however, only a few yeast strains were investigated for the synthesis of ceramide and the concentration was very low. Ceramide is not only a core intermediate of sphingolipids but also an important modulator of many cellular events including apoptosis, cell cycle arrest, senescence, differentiation, and stress responses. In this study S.cerevisiae was grown in a batch culture and the cellular content of ceramide was measured at different growth phases. The ceramide content was highest at stationary phase and 2.01 mg ceramide/g cell was obtained.

조류(microalgae)에서 얻어진 DHA powder로부터의 유지 추출 및 특성 연구

  • 조은진;이기택
    • Proceedings of the Korean Society of Postharvest Science and Technology of Agricultural Products Conference
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    • 2003.10a
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    • pp.141.2-141
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    • 2003
  • 고도불포화지방산 중 EPA(eicosapentaenoic acid)와 DHA(docosahexaenoic acid)는 여러 가지 생리활성 효과를 나타내는 것으로 알려져 있으며, 최근 관심이 주목되고 있다. 본 실험에서는 특히 DHA의 함량이 높은 해양미생물, 즉 조류(microalgae)인 Schizochytrium sp.에서 얻어진 DHA powder로부터 두 가지 방법으로 유지를 추출한 뒤 그 성질을 비교.분석하였다. 먼저, 8$0^{\circ}C$, soxhlet extractor로부터 6시간 동안 추출하고, 55$^{\circ}C$, shaking water bath로부터 같은 시간 동안 추출한 결과, 각각 25.5, 24.0%의 수율을 나타내었다. GC에 의한 지방산 조성 분석 결과, soxhlet extractor에서 추출한 경우, palmitic acid, DHA가 각각 30.98, 30.04 mol%로 주요 지방산인 것으로 나타났으며, shaking water bath에서 추출된 유지는 그 함량이 각각 31.44, 34.78 mol%를 나타내었다. 또한 pancreatic lipase에 의한 가수분해를 통해 sn-2 위치의 지방산 조성 분석 결과, DHA가 sn-2 위치에 주로 함유되어있는 것으로 조사되었다. 한편, HPLC-ELSD system을 이용하여 중성지방의 구성 성분을 분석 결과, 유지의 99%이상이 TAG(triacylglycerol) 형태로 이루어져 있었으며, 소량의 MAG(monoacylglycerol) 존재를 확인할 수 있었다.

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Simultaneous determination of saikosaponin derivatives in Bupleurum falcatum by HPLC-ELSD analysis using different extraction methods

  • Choi, Jungwon;Kim, Juree;Kang, Sam Sik;Lee, Sanghyun
    • Journal of Applied Biological Chemistry
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    • v.64 no.1
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    • pp.57-61
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    • 2021
  • Saikosaponin derivatives such as saikosaponins A, B1, B2, B3, B4, C, and D present in Bupleurum falcatum were analyzed by a high performance liquid chromatograph equipped with an evaporative light scattering detector, using different extraction solvents (water and 70% ethanol). The samples were injected into a YMC Pack Pro C18 column and separated using a gradient elution system with a mobile phase composed of acetonitrile and water at a flow rate of 1.1 mL/min. The content of saikosaponin derivatives was higher in 70% ethanol extract than in water extract. This study provides an efficient analytical method for determining the optimal conditions for extraction of saikosaponin derivatives, which can be used as a basis for development of functional foods and pharmaceutical products from B. falcatum.

Optimization of Enzymatic Synthesis Condition of Structured Lipids by Response Surface Methodology (반응표면분석에 의한 기능성 유지의 효소적 합성 조건 최적화)

  • Cho, Eun-Jin;Lee, Jong-Ho;Lee, Ki-Teak
    • Korean Journal of Food Science and Technology
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    • v.36 no.4
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    • pp.531-536
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    • 2004
  • Synthesis conditions were optimized using response surface methodology for producing structured lipids (SL) by interesterification of DHA-enriched algae oil derived from microalgae, Schizochytrium sp. and corn oil. Reaction was performed fer 24 hr at $55^{\circ}C$ catalyzed by immobilized lipase from Rhizomucor miehei (RM IM) in shaking water bath. Major fatty acids of SL were palmitic (21.70 mol%), oleic (20.20 mol%), and linoleic (27.34 mol%) acids, and DHA (15.06 mol%). To separate newly synthesized SL-triglycerides (TG) species, HPLC with evaporative light scatting detector (ELSD) was used. Production conditions were optimized using central composite design with reaction temperature $(35-75^{\circ}C,\;X_1)$, reaction time $(2-42\;hr,\;X_2)$, and enzyme concentration $(2-14%,\;X_3)$ as variables. When variables were $70.28^{\circ}C\;(X_1),\;28.74\;hr\;(X_2),\;and\;11.30%\;(X_3)$, maximum content of selected three peaks of synthesized SL-TG species was predicted as 6.97 area%.

Phospholipids Isolation from Squid Viscera Residues After Supercritical Carbon Dioxide Extraction (오징어 내장의 초임계 이산화탄소 추출 잔류물로부터 인지질의 분리)

  • U, Pyoung-Ook;Chun, Byung-Soo
    • Korean Chemical Engineering Research
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    • v.48 no.6
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    • pp.741-746
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    • 2010
  • Phospholipids were recovered from squid viscera residues by ethanol extraction after supercritical carbon dioxide($SCO_2$) extraction and from squid viscera was not processed $SCO_2$ by various organic solvent extraction. $SCO_2$ extraction were performed at $45^{\circ}C$ and 20 MPa for removal of non polar lipid molecules from freeze dried squid viscera sample. Phospholipids were extracted from freeze dried squid viscera sample by chloroform, hexane, methanol, and ethanol and from $SCO_2$extracted squid viscera sample by ethanol. The pH was fixed at 5.7 for all phospholipids extraction conditions. Phospholipid classes were analyzed by HPLC equipped with evaporative light scattering detector (ELSD). Phosphatidyl choline(PC) extracted by ethanol from $SCO_2$ extracted residues was higher than that of extracted by ethanol from squid viscera. But phosphatidyl ethanolamine(PE) and phosphatidic acid(PA) were extracted higher percentage in raw squid viscera. The fatty acid compositions in phospholipids extract by ethanol extract from $SCO_2$ extracted residues were analyzed by gas chromatography(GC). Docosahexanoic acid(DHA) was found in highest percentage in phospholipid extract.

Effect of Expression of Genes in the Sphingolipid Synthesis Pathway on the Biosynthesis of Ceramide in Saccharomyces cerevisiae

  • Kim, Se-Kyung;Noh, Yong-Ho;Koo, Ja-Ryong;Yun, Hyun-Shik
    • Journal of Microbiology and Biotechnology
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    • v.20 no.2
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    • pp.356-362
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    • 2010
  • Ceramide is important not only for the maintenance of the barrier function of the skin but also for the water-binding capacity of the stratum corneum. Although the exact role of ceramide in the human skin is not fully understood, ceramide has become a widely used ingredient in cosmetic and pharmaceutical industries. Compared with other microorganisms, yeast is more suitable for the production of ceramide because yeast grows fast and is non-toxic. However, production of ceramide from yeast has not been widely studied and most work in this area has been carried out using Saccharomyces cerevisiae. Regulating the genes that are involved in sphingolipid synthesis is necessary to increase ceramide production. In this study, we investigated the effect of the genes involved in the synthesis of ceramide, lcb1, lcb2, tsc10, lac1, lag1, and sur2, on ceramide production levels. The genes were cloned into pYES2 high copy number vectors. S. cerevisiae was cultivated on YPDG medium at $30^{\circ}C$. Ceramide was purified from the cell extracts by solvent extraction and the ceramide content was analyzed by HPLC using ELSD. The maximum production of ceramide (9.8 mg ceramide/g cell) was obtained when the tsc10 gene was amplified by the pYES2 vector. Real-time RT-PCR analysis showed that the increase in ceramide content was proportional to the increase in the tsc10 gene expression level, which was 4.56 times higher than that of the control strain.

Optimization of Demucilage Process of Opuntia ficus-indica var. saboten Fruit using High Hydrostatic Pressure Enzyme Dissolution (고압 효소 액화 장치를 이용한 백년초 점질물 분해 공정의 최적화)

  • Im, Sungbin;Lee, Hyungjae;Shim, Jae-Yong;Kim, Tae-Rahk;Kim, Dae-Ok
    • KSBB Journal
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    • v.30 no.2
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    • pp.63-68
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    • 2015
  • This study aimed to develop and optimize a demucilaging process of Opuntia ficus-indica var. saboten (OFI) fruit to increase its usability as functional food ingredient and food additive. Viscozyme and Novozym 33095 as multienzyme complex having a broad spectrum of carbohydrases and pectolytic enzymes, respectively, were used in enzymatic dissolution along with high hydrostatic pressure liquefaction. To optimize the liquefaction process using high hydrostatic pressure liquefying extractor, response surface methodology with 3-factor central composite design was employed with reaction factors such as temperatures (25, 32, 40, 48, and $55^{\circ}C$), pressures (20, 40, 60, 80, and 100 MPa), and times (15, 30, 45, 60, and 75 min). At optimum conditions ($25^{\circ}C$, 100 MPa, and 58.275 min) for high hydrostatic pressure liquefaction process, the processed OFI fruit juice was predicted to have viscosity at 2.917 poise, partly due to the release of free sugars such as fructose and glucose detected using HPLC-ELSA system. The results above suggests that the OFI fruit juice with decreased viscosity may be used for various manufacturing processes of food, beverage, ice cream, and cosmetics.

Proximate, Free Sugar, Fatty acids Composition and Anthocyanins of Saekso 2 Corn Kernels (옥수수 색소 2호 품종의 일반성분, 유리당, 지방산 및 안토시아닌 색소의 함량)

  • Lee, Ki Yeon;Kim, Tae hee;Lim, Sang Hyun;Park, Jong yeol;Kim, Kyung Hee;Ahn, Mun Seob;Kim, Hee Yeon
    • Journal of Food Hygiene and Safety
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    • v.31 no.5
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    • pp.335-341
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    • 2016
  • This study was performed to provide basic data of Saekso 2 corn kernels. Proximate composition, free sugars, fatty acids and anthocyanin content of Saekso 2 corn kernels were analyzed. Proximate composition of dried Saekso 2 corn kernels were represented 8.84% moisture, 1.44% crude ash, 5.46% crude lipid, 10.31% crude protein. Free sugars composition by HPLC/ELSD showed that sucrose (1.00%), glucose (0.63%), maltose (0.52%), fructose (0.44%) were present. The composition of fatty acids in Saekso 2 corn kernels was analyzed by GC/FID. 11 species of fatty acids were analyzed in Saekso 2 corn kernels. The ratio of saturated fatty acids and unsaturated fatty acids 16.09 : 83.91. Content of linoleic acid was the highest in fatty acids. The total anthocyanin content in Saekso 2 corn kernels was 0.24% by UV/Vis. Anthocyanin components separated and quantified using HPLC/MS/MS. Cyanidin 3-O-glucoside chloride (C-3-G), pelargonidin 3-O-glucoside chloride (Pg-3-G) and peonidin 3-O-glucoside chloride (Pn-3-G) of anthocyanin were analyzed in Saekso 2 corn kernels.

Determination of Fructooligosaccharides and Raffinose in Infant Formula by High Performance Liquid Chromatography with Evaporative Light Scattering Detector (HPLC-ELSC를 이용한 조제분유 중 fructooligosaccharides 및 raffinose 분석)

  • Shin, Man-Sub;Park, Jae-Woo;Cho, Mi-Ran;Song, Sung-Ok;Kim, Chun-Sun;Choi, Chun-Bae;Lee, Seoung-Won;Lee, Ki-Woong;Chang, Chi-Hoon;Kwak, Byung-Man
    • Korean Journal of Food Science and Technology
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    • v.38 no.6
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    • pp.725-729
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    • 2006
  • A method was developed for the determination of fructooligosaccharides and raffinose contents in infant formula. The samples were extracted and analyzed by liquid chromatography equipped with carbohydrate column and evaporative light scattering detector. The mobile phase used for the gradient mode was water-acetonitrile, at a flow rate of 1.0mL/min. The method showed a mean recovery of 95-99%, the relative standard deviation obtained in the precision study was 0.774-8.982%, the quantification and detection limits were 25-50mg/L.

Proximate, Mineral and Sugar Composition of Rehmannia glutinosa by Cultivars (품종별 지황의 일반성분, 무기질 및 당 조성)

  • Oh, Hye-Lim;Kim, Na-Yeon;Lee, Kun-Jong;Yang, Kee-Heun;Doh, Eun-Soo;Song, Mi-Ran;Park, Jong-Yoon;Kim, Mee-Ree
    • Journal of the East Asian Society of Dietary Life
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    • v.22 no.3
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    • pp.365-370
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    • 2012
  • Proximate composition, reducing sugar, and mineral content of several cultivars of Rehmannia glutinosa were analyzed. the moisture and soluble solid content of fresh Rehmannia glutinosa ('Korea', 'Kokang', 'Sewon 10', 'Sewon 11' and 'Jihwang 1') were 74.6~78.4% and $19.6{\sim}22.4^{\circ}Brix$, respectively. Proximate composition of dried Rehmannia glutinosa ranged from 82.91~86.94% carbohydrate, 3.38~5.70% crude protein, 2.5~3.0% crude ash and 3.47~3.80% fiber. Sugar composition by HPLC/ELSD showed that sucrose (4.49~7.75 g/100 g), raffinose (2.96~4.78 g/100 g) and stachyose (42.36~45.87 g/100 g) were present, whereas monosaccharides were not detected in 5 cultivars of dried Rehmannia glutinosa. Mineral compositions of dried Rehmannia glutinosa by ICP-AES were Ca (639.9~782.0 ppm), Fe (128.5~634.9 ppm), Na (119~150 ppm), K (6,639.1~10,448.0 ppm), Mg (372.2~981.8 ppm) and Zn (8.8~474.2 ppm). However, Co and Mo were not detected. Among 5 cultivars of Rehmannia glutinosa, 'Kokang' and 'Sewon 10' contain higher amounts of Fe, Ca, K, Mg, Cu and Zn than the other cultivars.