• Title/Summary/Keyword: analysis by HPLC

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Evaluation of Enzymatic Method using an Automated Chemistry Analyzers for Homocysteine Measurement

  • Shim, Moon-Jung
    • Korean Journal of Clinical Laboratory Science
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    • v.39 no.3
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    • pp.196-200
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    • 2007
  • In clinical practice, homocysteine has gained popularity because its elevated values are strongly associated with an increased risk of cardiovascular disease. More recently, a new enzymatic colorimetric assay for homocysteine in biological sample, suitable for automated clinical analyzers, has been proposed. To evaluate one of these enzymatic methods and compare the results obtained with this method with those of an immunoenzymatic method, thirty-two samples were analyzed for total homocysteine by HiSens$^{(R)}$ homocysteine reagent on the automated chemistry analyzers TBA 200FR and compared to the widely used immunoenzymatic method ADVIA Centaur. In TBA 200FR, the within-run CVs of two control materials were 3.23% and 0.92%, respectively; the between run CVs were 4.58% and 2.55%, respectively. And in ADVIA 1650, the within-run CVs were 6.81% and 0.99%, respectively; the between run CVs were 9.0% and 3.9%, respectively. The recovery for homocysteine was 100% ($60.8{\mu}mol/L$), 99.1% ($48.64{\mu}mol/L$), 96.3% ($36.48{\mu}mol/L$), 96.1% ($24.32{\mu}mol/L$), and 92.1% ($12.16{\mu}mol/L$). The regression equation of TBA 200FR vs. ADVIA Centaur was y=0.9095x-2.5086 (r=0.9632). And the regression equation for the ADVIA 1650 chemistry vs. Immulite 2000 was y=0.8418x + 0.3207 (r=0.9625). In conclusion, this enzymatic method using automated chemistry analyzer for homocysteine assay shows acceptable analytical performance. I suggest that this assay will be suitable for routine analysis.

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Optimal Extract Methods of Antioxidant Compounds from Coat of Grape Dreg (포도부산물인 과피로부터 항산화 물질 최적 추출방법 확립)

  • Yoo, Mi-Ae;Chung, Hae-Kyung;Kang, Myung-Hwa
    • Korean Journal of Food Science and Technology
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    • v.36 no.1
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    • pp.134-140
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    • 2004
  • Optimal extraction method for antioxidant compounds from coat of grape dreg was established. Extracts were prepared with ethanol solutions containing phosphoric, formic, acetic, HCl, TFA, and citric acids. Antioxidant compounds of grape coat were determined by HPLC analysis and evaluated for antioxidizing effects using in vitro model system. Peonidin-3-glucoside content was the highest in 0.1% HCl-added ethanol extract. The extract prepared from pure ethanol without organic acids showed the highest content of cyanidin-3-glucoside among samples tested. Resveratrol and quercetin contents, the most important antioxidants, were highest in 0,1% HCl-added extract. Electron-donating ability was high in 0.1% acetic acid-added extract. SOD-like activities were 95.08% and 94.39% in 0.1% formic and 0.1% citric acid extracts, respectively. Inhibitory effects on peroxidation of egg yolk lecithin were observed in phosphoric (60.51%), citric (59,27%), and formic acid (56,77%) extracts. Hydrogen radical-scavenging activity was 59.47% in 0,1% HCl extract. Results suggest addition of 0.1% HCl in ethanol solution affords the highest yield in antioxidant compounds and antioxidant activities.

Application as a Cosmeceutical Ingredient of Euryale ferox Seed Extract (가시연꽃 종자 추출물의 화장품 원료로서의 특성)

  • Choo, Soo-Jin;Kim, Young-Hee;Ryoo, In-Ja;Xu, Guang-Hua;Yoo, Ick-Dong
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.35 no.4
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    • pp.309-315
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    • 2009
  • In our search for the natural cosmetic ingredients, we found that Euryale ferox seed extract exhibited the strong antioxidative activity. Five active compounds were isolated from the ethyl acetate extract through various chromatographic methods and their structures were determined by NMR and MS spectral analysis. These compounds were identified as fucosterol (1), 3-(4-hydroxy-3-methoxybenzyl)-4-[(7'R),5'-dihydroxy-3'-methoxybenzyl]tetrahydrofuran (2), resorcinol (3), pyrogallol (4) and 4-O-methylgallic acid (5).We evaluated the antioxidative, antielastase activities and melanogenesis inhibitory effects of these compounds. The $SC_{50}$ values of compounds 2 ~ 5 for free radical scavenging activity were $17.0\;{\sim}\;100.2\;{\mu}M$ and especially compounds 4 and 5 were 6-fold more effective than ferulic acid as a positive control. And compounds 2 ~ 4 inhibited human neutrophil elastase with $IC_{50}$ values of $18.8\;{\sim}\;78.2\;{\mu}M$ and compound 3 also inhibited melanin synthesis in B16F10 melanoma cells with an $IC_{50}$ value of $492.8\;{\mu}M$. These results suggest that Euryale ferox extract having a lot of various active ingredients may be useful as a natural multi-functioning agent.

Determination of Preservatives in Pharmaceuticals and Personal Care Products (의약품 및 개인위생·생활용품 중 보존제 함유량 분석)

  • Kim, Boyoung;Lee, Joohee;Kim, Seonyeob;Lee, Eunji;Choi, Chorong;Kho, Younglim
    • Journal of Environmental Health Sciences
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    • v.42 no.1
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    • pp.53-60
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    • 2016
  • Objectives: Preservatives are commonly used in pharmaceuticals, cosmetics and other products to extend the expiration date and prevent the growth of microorganisms. Preservatives are generally effective in controlling mold and inhibiting yeast growth, and against a wide range of bacterial attacks as well. They also adversely affect the quality of sperm and cause precocious puberty in children. This study was performed to analyze seven preservatives used in pharmaceuticals and personal care products. Methods: Five kinds of pharmaceuticals and personal care products (PPCPs) were examined for analysis with a high performance liquid chromatography-diode array detector. Each sample was homogenized and the targeted compounds were extracted with methanol. The suspended particulate was removed by syringe filter. Next, the sample was injected into an HPLC system. The separation of the seven preservatives was achieved with a C18 column and gradient mode. The accuracies were between 73% and 120% and precision was lower than 11.58% (RSD). Results: All of the calibration curves showed good linearity with a coefficient of determination ($r^2$) over 0.999. Among the PPCP samples, the detection rate of preservatives was 32.5% for pharmaceuticals, 44.8% for toothpaste, 76.9% for mouthwash, 40.0% for body lotion and 56.0% for wet tissues. The average concentrations of the preservatives in PPCPs were BA 1141.0 mg/kg, MP 709.8 mg/kg, EP 624.9 mg/kg, PP 216.9 mg/kg, BP 167.8 mg/kg, and TCS 538.2 mg/kg. The most frequently detected preservatives in pharmaceuticals and personal care products were BA, MP and PP. The concentrations of preservatives exceeded Korean regulatory standards in 11 samples of medicines, three of mouthwash and two of body lotion. Conclusion: We found that most of the PPCP samples contained various preservatives. It is necessary to identify which preservatives were used and to determine the level of preservatives in PPCPs and to assess the health risk to susceptible populations such as children.

Biodistribution of 99mTc Labeled Integrin Antagonist

  • Jang, Beom-Su;Park, Seung-Hee;Shin, In Soo;Maeng, Jin-Soo;Paik, Chang H.
    • Toxicological Research
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    • v.29 no.1
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    • pp.21-25
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    • 2013
  • The selective targeting of an integrin ${\alpha}_v{\beta}_3$ receptor using radioligands may enable the assessment of angiogenesis and integrin ${\alpha}_v{\beta}_3$ receptor status in tumors. The aim of this research was to label a peptidomimetic integrin ${\alpha}_v{\beta}_3$ antagonist (PIA) with $^{99m}Tc(CO)_3$ and to test its receptor targeting properties in nude mice bearing receptor-positive tumors. PIA was reacted with tris-succinimidyl aminotriacetate (TSAT) (20 mM) as a PIA per TSAT. The product, PIA-aminodiacetic acid (ADA), was radiolabeled with $[^{99m}Tc(CO)_3(H_2O)_3]^{+1}$, and purified sequentially on a Sep-Pak C-18 cartridge followed by a Sep-Pak QMA anion exchange cartridge. Using gradient C-18 reverse-phase HPLC, the radiochemical purity of $^{99m}Tc(CO)_3$-ADA-PIA (retention time, 10.5 min) was confirmed to be > 95%. Biodistribution analysis was performed in nude mice (n = 5 per time point) bearing receptor-positive M21 human melanoma xenografts. The mice were administered $^{99m}Tc(CO)_3$-ADA-PIA intravenously. The animals were euthanized at 0.33, 1, and 2 hr after injection for the biodistribution study. A separate group of mice were also co-injected with 200 ${\mu}g$ of PIA and euthanized at 1 hr to quantify tumor uptake. $^{99m}Tc(CO)_3$-ADA-PIA was stable in phosphate buffer for 21 hr, but at 3 and 6 hr, 7.9 and 11.5% of the radioactivity was lost as histidine, respectively. In tumor bearing mice, $^{99m}Tc(CO)_3$-ADA-PIA accumulated rapidly in a receptor-positive tumor with a peak uptake at 20 min, and rapid clearance from blood occurring primarily through the hepatobiliary system. At 20 min, the tumor-to-blood ratio was 1.8. At 1 hr, the tumor uptake was 0.47% injected dose (ID)/g, but decreased to 0.12% ID/g when co-injected with an excess amount of PIA, indicating that accumulation was receptor mediated. These results demonstrate successful $^{99m}TC$ labeling of a peptidomimetic integrin antagonist that accumulated in a tumor via receptor-specific binding. However, tumor uptake was very low because of low blood concentrations that likely resulted from rapid uptake of the agent into the hepatobiliary system. This study suggests that for $^{99m}Tc(CO)_3$-ADA-PIA to be useful as a tumor detection agent, it will be necessary to improve receptor binding affinity and increase the hydrophilicity of the product to minimize rapid hepatobiliary uptake.

Seasonal Variation of Primary Productivity and Pigment of Phytoplankton Community Structure in the Seomjin Estuary (섬진강 하구역의 계절별 일차생산력 및 식물플랑크톤 색소 조성 변화)

  • Min, Jun-Oh;Ha, Sun-Yong;Chung, Mi-Hee;Choi, Bo-Hyung;Lee, Yeon-Jung;Youn, Seok-Hyun;Yoon, Won-Duk;Lee, Jae-Seong;Shin, Kyung-Hoon
    • Korean Journal of Ecology and Environment
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    • v.45 no.2
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    • pp.139-149
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    • 2012
  • Four in situ incubation experiments were conducted in May, August and November 2009, as well as February 2010 to determine the seasonal primary productivity and the community structure of phytoplankton at the Seomjin estuary. The primary production of phytoplankton ranged from 9 to 3560 mgC $m^{-2}\;d^{-1}$. Primary productivity was the highest in the summer season (August), which was influenced by improved optical and temperature conditions of the water, as well as the supply of nutrients derived from its surrounding watershed. Particularly, the upper station (SJ-1, SJ-2) of Seom-jin estuary showed a higher productivity, as a result of inflow of input nutrients originated from the terrestrial source. The fucoxanthin, as an index pigment of diatoms showed the highest concentration (0.74~9.51 ${\mu}g\;L^{-1}$) at all stations, occupying 30~80% to total Chl a concentrations. The phytoplankton species composition determination, using a microscope showed similar results to the pigment analysis, which indicated diatom Skeletonema costatum, as the dominant species. The primary productivity in Seom-jin estuary indicates temporally and spatially large variation, according to different environmental conditions. Also, Skeletonema costatum has euryhaline features with relatively higher contribution.

Preparation and Physicochemical Properties of Soluble Dietary Fiber Extracts from Soymilk Residue at High Temperature (두유박 수용성 식이섬유의 고온 추출물 제조와 이화학적 특성구명)

  • Park, Chun-Ho;Kim, Hyun-Jung;Moon, Tae-Wha
    • Korean Journal of Food Science and Technology
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    • v.29 no.4
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    • pp.648-656
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    • 1997
  • Thermal treatment of soymilk residue was carried out at 140, 150, and $160^{\circ}C$ for possible use as a raw material for dietary fiber, and some physicochemical properties of the extracts were investigated. Soluble dietary fiber(SDF) content of the extracts prepared under optimal conditions was more than 30% suggesting the conversion of insoluble dietary fiber to SDF. The main sugar components of the extracts were glucose, galactose, and arabinose. Analysis of the moelcular weight distribution by high performance size exclusion chromatography revealed that the proportion of high molecular weight fraction decreased and that of middle-sized polymer increased as the extraction temperature increased. The viscosity of aqueous solution of the extracts decreased with an increase in extraction temperature, but showed no trend as pH changed. The solubility increased with extraction temperature showing the highest at $160^{\circ}C$. The extract at $140^{\circ}C$ had the biggest calcium-binding capacity, which correlated with the changes in viscosity.

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Pharmacokinetic and Bioequivalence Study of Zolpidem Tartate in Healthy Volunteers

  • Park, Jun-Sung;Myung, Ja-Hye;Wang, Hun-Sik;Koo, Ja-Seong;Cho, Won-Kyung;Cha, Kwang-Ho;Park, Hee-Jun;Kim, Min-So;Kim, Jeong-Soo;Hwang, Sung-Joo
    • Journal of Pharmaceutical Investigation
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    • v.41 no.3
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    • pp.191-196
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    • 2011
  • In this study simple and sensitive high performance liquid chromatographic method using a commercially available column, was developed and validated for the determination of zolpidem tartrate in human plasma. The developed method with suitable validation was applied to a bioequivalence study of two different kinds of zolpidem tartrate. Two different formulations containing 10 mg of zolpidem tartate (CAS : 99294-93-6) were compared in 24 healthy male volunteers in order to compare the bioavailability and prove the bioequivalence. The study was performed in an open, single dose randomized, 2-sequence, cross-over design in 24 healthy male volunteers with a one-week washout period. Blood samples for pharmacokinetic profiling were drawn at selected times during 12 h. The mean $AUC_{0-12h}$, $C_{max}$, $T_{max}$ and $T_{1/2}$ were $676.6{\pm}223.4$ $ng{\cdot}h{\cdot}mL^{-1}$, $177.4{\pm}34.2$ $ng{\cdot}mL^{-1}$, and $0.8{\pm}0.4$ and $3.5{\pm}2.1$, respectively, for the test formulations, and $640.7{\pm}186.6$ $ng{\cdot}h{\cdot}mL^{-1}$, $193.0{\pm}64.5$ $ng{\cdot}mL^{-1}$, and $0.9{\pm}0.4$ and $2.7{\pm}0.9$, respectively, for the reference formulation. Both primary target parameters $AUC_{0-12h}$ and $C_{max}$ were log-transformed and tested parametrically by analysis of variance (ANOVA). 90% confidence intervals of $AUC_{0-12h}$ and $C_{max}$ were in the range of acceptable limits of bioequivalence (80-125%). Based on these results, the two formulations of zolpidem tartate are considered to be bioequivalent.

Extraction Conditions of Radical Scavenging Caffeoylquinic Acids from Gomchui (Ligularia fischeri) Tea (곰취차로부터 라디칼 소거능을 갖는 Caffeoylquinic Acid류 화합물의 추출조건)

  • Kim, Sang-Min;Kang, Suk-Woo;Um, Byung-Hun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.3
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    • pp.399-405
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    • 2010
  • After Gomchui tea was prepared from leaves of Ligularia fischeri (Ledeb.) Turcz by blanching method, the antioxidant activity of major compounds in Gomchui tea was assessed. On-line HPLC-ABTS analysis revealed that caffeoylquinic acids (chlorogenic acids), such as 5-O-caffeoylquinic acid (5-CQA), 3,4-di-O-caffeoylquinic acid (3,4-DCQA), 3,5-di-O-caffeoylquinic acid (3,5-DCQA) and 4,5-di-O-caffeoylquinic acid (4,5-DCQA), were the major antioxidant compounds in Gomchui tea. The extraction efficiency of these compounds were examined in the various conditions such as extraction temperature, time and solvent. The results demonstrated that the extraction amount with water increased in proportion to extraction time (1~10 min) and temperature ($8{\sim}80^{\circ}C$). These active compounds were also extracted with water even at $8^{\circ}C$ (60% of $80^{\circ}C$), indicating that water is very good extraction solvent for extraction of these antioxidant constituents. However, the extraction efficiency of these compounds decreased when ethanol percentage in water increased. The extraction efficiency between Gomchui powder (no blanching) and tea was significantly different, and 60% of total antioxidant compounds in tea was removed from fresh leaves into water in blanching process, especially 3,5-DCQA (over 90%). Meanwhile, the sonication method didn't affect the extraction of these compounds in all solvents. These results suggest that Gomchui tea can be a good candidate for the tea beneficial to human health.

Quantification and Varietal Variation of Rutin in Mulberry Fruits (뽕나무 계통별 오디함유 rutin 분석 및 품종변이)

  • Kim Hyun-bok;Kim Sun-lim
    • Journal of Sericultural and Entomological Science
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    • v.46 no.1
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    • pp.1-5
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    • 2004
  • Rutin has many beneficial effects on human health. It was established that rutin antagonizes the increase of capillary fragility associated with hemorrhagic disease, reduces high blood pressure, decreases the permeability of the vessels, and has an antiedema effect, reduces the risk of arteriosclerosis, and shows antioxidant activity. Rutin analysis with mulberry fruits was carried out by high-performance liquid chromatography (HPLC) to reveal the possibility of source of functional food. As a result, mean content of fifty accessions was 0.14±0.050% DW. Among the tested fifty accessions, 'Sabangso' was showed the highest rutin in 0.29%DW, whereas 'Simseol' was the lowest content in 0.05% DW. Also, we researched the change of rutin content according to harvested date. The rutin contents of earlier harvested groups were higher than later harvested groups. Simultaneously, fruity characteristics as well as rutin content were researched and analyzed to select the functional mulberry varieties for the production of fruit. From the six accessions which were contained high rutin content, we selected three suitable varieties such as 'Ficus', 'Kangsun', and 'Palcheongsipyung'.