• Title/Summary/Keyword: high performance liquid chromatography %28HPLC%29

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Measurement of Total Plasma Homocysteine in Patients with Chronic Renal Failure Using HPLC/FLD (만성신부전증환자에서 HPLC/FLD를 이용한 혈장 Homocysteine의 측정)

  • Kyung-Ok Lee;Bo-Kyung Kang;Hyung-Jin Roh;Kwang-Suk Ryoo;Jeong-Yeon Yoo;Man-Jeong Paik;Kang-Hyeob Lee
    • Biomedical Science Letters
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    • v.3 no.1
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    • pp.29-35
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    • 1997
  • Cardiovascular disease has been the leading cause of death among patients with chronic renal failure. Many reports have been described that homocysteine is one of the independent risk factor to the occulsive vascular disease. In this study, HPLC/FLD (high performance liquid chromatography-fluorescence detector) technique was used to measure homocysteine level in patients with chronic renal failure and normal control group. The detection limit and recovery of total plasma homocysteine using HPLC/FLD were 98.6$\pm$5.8% and 0.2 nmol/ml, respectively. The linearity of this method was established in concentration range of 2~50 nmol/ml (correlation coefficient=0.9997). The concentrations of total plasma homocysteine were 6.81$\pm$1.54 nmol/ml and 27.28$\pm$14.94 nmol/ml in normal control (n=20) and patient group (n=90), respectively (p<0.05). In this study, the HPLC/FLD method showed high sensitivity and reproducibility for a routine clinical laboratory testing. Moreover determination of homocysteine level in plasma might be useful for a biochemical marker for predicting the cardiovascular diseases and for monitoring of therapeutic effect of lowering homocysteine in patients with chronic renal failure.

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Quantitative Determination of Marker Compounds in the Extracts of Camellia sinensis L. Sub-branches (Residual Products) by HPLC (HPLC에 의한 차나무 잔가지(부산물)의 추출물 내 지표 성분의 정량분석)

  • Lee, Min Sung;Im, Hyeon Jeong;Jeong, Hea Seok;Cho, Hae Jin;Woo, Hyun Sim;Oh, Yu Jin;Lee, Soo In;Kim, Hyun Chul;Ahn, Kyung Wan;Kim, Yeong Su;Kim, Dae Wook
    • Korean Journal of Medicinal Crop Science
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    • v.27 no.1
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    • pp.24-29
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    • 2019
  • Background: Camellia sinensis L.(CS) is a perennial evergreen species of plant whose leaves are used to produce tea. In this plant species, the parts used are the leaves, sub-branch parts are thrown out. Methods and Results: Ethanol extract of sub-branch parts was used for isolation of major compounds by column chromatography. Structures were identified as caffeine (1), (-)-epicatechin (2) and (-)-epicatechin gallate (3) by interpretation of spectroscopic analysis, including $^1H$- and $^{13}C$-NMR. High-performance liquid chromatography (HPLC) method was used to compare the quantitative level of marker compounds in various extraction solvents of sub-branch parts of CS. The content of caffeine, (-)-epicatechin, and (-)-epicatechin gallate in 30% ethanol extract showed higher value with $3.28{\pm}0.57mg/g$, $5.53{\pm}0.88mg/g$, and $1.29{\pm}0.24mg/g$, respectively. Conclusions: These results indicated that not only leaves parts but also sub-branch, could be a good source for the functional material and pharmaceutical industry.

Membrane Lipids of a Marine Ciliate Protozoan Uronema marinum

  • Seo Jung Soo;Kim Ki Hong;Lee Hyung Ho;Chung Joon Ki
    • Fisheries and Aquatic Sciences
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    • v.6 no.3
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    • pp.155-159
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    • 2003
  • Lipid composition and fatty acid composition were characterized in the membrane of a marine ciliate protozoan (Uronema marinum). Phospholipids accounted for 70% of total lipid, and the remainder was neutral lipids. Total phospholipids were separated as phosphatidylcholine $(24.26\%)$, phosphatidylethanolamine $(22.21\%)$, phosphatidylinositol $(6.14\%)$, phosphatidyl­serne $(5.11\%)$, cardiolipin $(3.07\%)$ and unidentified phospholipids $(28.72\%)$ through high performance liquid chromatography (HPLC). Fatty acid composition of neutral lipids and phospholipids was determined by gas chromatography (GC), based solely on comparision of retention times. In neutral lipids, the most abundant fatty acid group was monounsaturated fatty acid $(48.3\% of total fatty acids)$ with oleic acid (18:1) and nervonic acid (24:1). Saturated fatty acids comprised $29.6\%$ of total fatty acids, with palmitic acid (16:0), stearic acid (18:0) ane myristic acid (14:0), and polyunsaturated fatty acid accounted for $33.0\%$ with $Di-homo-\gamma-linolenic$ acid (20:3) and linoleic acid (18:2). Wherease phospholipids predominantly contained the fatty acid group in the following order: polyunsaturated fatty acids $(52.7\%\;of\;total\;fatty\;acids)$ with linoleic acid (18:2) and $\gamma-linolenic$ acid (18:3) > monounsaturated fatty acids $(28.5\%\;of\;total\;fatty\;acids)$ with oleic acid (18:1) and palmitoleic acid (16:1) > saturated fatty acids $(25.5\%\;of\;total\;fatty\;acids)$ with palmitic acid (16:0), stearic acid (18:0) and myristic acid (14:0).

Quantitative determination of inosine 5'-monophosphate dehydrogenase activity in human peripheral blood mononuclear cells by ion-pair reversed-phase high-performance liquid chromatography (이온쌍 역상 HPLC를 이용한 인체 말초혈액단핵구에서 이노신 5'-일인산 탈수소효소 활성의 정량적 측정)

  • Shin, Hye-Jin;Kwon, Soon-Ho;Park, Ji-Myeong;Kwon, Soon-Hyo;Lee, Kyoung-Ryul;Kim, Young-Jin;Lee, Sang-Hoo
    • Analytical Science and Technology
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    • v.23 no.6
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    • pp.531-536
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    • 2010
  • A quantitative analytical method has been established for the measurement of inosine 5'-monophosphate dehydrogenase (IMPDH) activity in human peripheral blood mononuclear cells (PBMCs) by ion-pair reversed-phase high performance liquid chromatography equipped with ultraviolet detection (HPLC/UV). IMPDH is a ${\beta}$-nicotinamide adenine dinucleotide hydrate (NAD+)-dependent dehydrogenase in which the enzyme converts inosine 5'-monophosphate (IMP) into xanthosine 5'-monophosphate (XMP). Its activity was measured by quantifying a HPLC chromatogram corresponding to XMP produced during the incubation of lysed PBMCs with IMP as a substrate and $NAD^+$ as a coenzyme. XMP produced was detected at a wavelength of 260 nm. The mobile phase was composed of a mixture of 37 mM potassium dihydrogen phosphate containing 7 mM tetra-n-butylammonium hydrogen sulfate adjusted to pH 5.5 and methanol (85:15, v/v) with a flow rate of 1 mL/min. The calibration curve was linear ($r^2$=0.999999) in the range of $0.2-50.0\;{\mu}M$ and the limit of quantification (LOQ) was $0.2\;{\mu}M$. The intra- and inter-day precisions were between 0.88-1.47% and 0.85-5.24%, respectively. The intra- and inter-day accuracies were between 98.74-99.99% and 99.95-101.65%, respectively. IMPDH activity in 11 Korean healthy volunteers ranged from 18.29 to 36.60 nmol/h/mg protein (mean = $27.70{\pm}6.28\;nmol/h/mg$ protein).

Expression and Purification of Herpes Simplex Virus Type 1 Protease (Herpes Simplex Virus Type 1 Protease의 발현 및 분리 정제)

  • Bae, Pan-Kee;Paeng, Jin-Wook;Kim, Jee-Hyun;Kim, Hae-Soo;Paik, Sang-Gi;Chung, In-Kwon;Lee, Chong-Kyo
    • The Journal of Korean Society of Virology
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    • v.29 no.3
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    • pp.175-182
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    • 1999
  • An attractive target for anti-herpes chemotherapy is the herpes simplex virus 1 (HSV-1) protease encoded by the UL26 gene. HSV-1 protease is essential for DNA packaging and virus maturation. To perform high throughput for potent inhibitors, the efficient production of larger amounts of highly purified enzyme and protease activity assay method must be established. In this report, expression in E. coli and purification of the protease gene of HSV-1 strain F was investigated. The protease gene was cloned pET28, and the nucleotide sequence of protease catalytic domain of HSV-1 compared strain F with other strains (KOS and CL101). In these results the F strain was different in base sequence. However, the amino acid sequence was identifical. The HSV-1 protease was purified with His-tagged affinity column. The analysis of HSV-1 protease activity was performed by high performance liquid chromatography.

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A Survey of the Presence of Aflatoxins in Food (식품 중 아플라톡신 오염도 조사)

  • Park, Min-Jung;Yoon, Mi-Hye;Hong, Hae-Geun;Joe, Tae-Suk;Lee, In-Sook;Park, Jeong-Hwa;Ko, Hoan-Uk
    • Journal of Food Hygiene and Safety
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    • v.23 no.2
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    • pp.108-112
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    • 2008
  • A survey of total aflatoxin levels was conducted on 158 samples (nuts, fermented foods and their processed products) collected in local markets in Gyeonggi-do and Domestic Internet Site. The total aflatoxins were quantified by the immunoaffinity column clean-up method followed by high performance liquid chromatography (HPLC)-fluorescence detector (FLD). Aflatoxins were found in 45(28.5%) samples including 34 nuts and nut products, 7 soybean pastes, 1 meju, 1 bean product and 2 corn snacks with a range of $0.02{\sim}3.96\;{\mu}g/kg$. These results show that the contamination level of aflatoxin in foods consumed in Korea is low compared with the standard in Korea Food Code($10\;{\mu}g/kg$ as aflatoxin $B_1$). Aflatoxin $B_1$ content was increased in peanuts and com snacks during storage but it was decreased in doenjang (soybean paste).

Analysis of Bioconversion Components of Fermentation Hwangryunhaedok-tang (발효 황련해독탕의 생물 전환 성분분석)

  • Lee, Kwang Jin;Lee, BoHyoung;Jung, Pil Mun;Lian, Chun;Ma, Jin Yeul
    • YAKHAK HOEJI
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    • v.57 no.4
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    • pp.293-298
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    • 2013
  • Hwangryunhaedok-tang (HRT) is a traditional herbal medicine, which has been known as a useful prescription for anti-biotic, anti-inflammatory, anti-oxidative and immunosuppressive activity. In this study, the variation in the amount of eight bioactive components of Hwangryunhaedok-tang (HRT) and its fermentation HRT with Lactobacillus casei KFRI 127, Lactobacillus curvatus KFRI 166 and Lactobacillus confuses KFRI 227 was investigated via high-performance liquid chromatography coupled with diode array detection (HPLC-DAD). Simultaneous qualitative and quantitative analysis of eight bioactive components; geniposide, genipin, baicalin, wogonoside, palmatine, berberine, baicalein and wogonin was achieved by comparing their retention times ($t_R$) and UV spectra with those of the standard components. All calibration curve of standard components showed good linearity ($r^2$ >0.979). As a result, the geniposide amount was $15.52{\pm}0.19{\mu}/mg$ that as a main components in HRT. The wogonoside was decreased by 29.28~58.35% with Lactobacillus casei KFRI 127 and L. confuses KFRI 227 ($3.17{\pm}0.31{\mu}g/mg$ and $3.55{\pm}0.13{\mu}g/mg$) compared with the original HRT ($5.02{\pm}0.14{\mu}g/mg$). Otherwise wogonin was increased by 16.28~41.86% with Lactobacillus casei KFRI 127 and L. confuses KFRI 227 ($0.61{\pm}0.01{\mu}g/mg$ and $0.50{\pm}0.02{\mu}g/mg$) compared with the original HRT($0.43{\pm}0.00{\mu}g/mg$). HRT fermented with L. casei KFRI 127 and L. confuses KFRI 227 were evaluated as creating the changes in wogonoside to that aglycon wogonine. In the fermented HRT using Lactobacillus acidophilus KFRI 166, the genipin was only detected, among 3 species of fermentation strains. Thus, these results considered that the strains 166 were exhibited the remarkable changes in genipin.

Changes of Saponin and β-Glucan Content on the Cultured Ginseng with Mushroom Mycelia (버섯균사체로 배양된 인삼 Saponin과 β-Glucan 함량 변화)

  • Joung, Eun-Mi;Hwang, In-Guk;Lee, Hyeon-Yong;Jeong, Jae-Hyun;Yu, Kwang-Won;Jeong, Heon-Sang
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.8
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    • pp.1084-1089
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    • 2009
  • This study investigated the changes of saponin and $\beta$-glucan content on the cultured ginseng with mushroom mycelia of Phellinus linteus (PL), Ganoderma lucidum (GL), and Hericium erinaceum (HE). Cultured ginsengs with mushroom mycelia were extracted with 80% ethanol, fractionated with n-buthanol, and analysed for ginsenosides by high performance liquid chromatography (HPLC). Crude saponin content of raw ginseng was 4.11% (d.b) but cultured ginseng with mushroom mycelia of PL, GL, and HE were increased to 6.74, 6.77 and 6.23% (d.b), respectively. Ginsenoside-Rd, among the 12 ginsenosides which were available for analysis, was remarkably increased to 13.61, 24.26, and 32.69 mg/g, respectively (raw ginseng: 0.80 mg/g). The $\beta$-glucan content of cultured ginseng with mushroom mycelia of PL, GL, and HE were decreased to 8.85, 5.51 and 5.46% rather than mushroom mycelia of 29.14, 19.44, and 23.39% (d.b), respectively.