• Title/Summary/Keyword: high performance liquid chromatography

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Antioxidant and Antiaging Assays of Hibiscus sabdariffa Extract and Its Compounds

  • Widowati, Wahyu;Rani, Andani Puspita;Hamzah, R. Amir;Arumwardana, Seila;Afifah, Ervi;Kusuma, Hanna Sari W.;Rihibiha, Dwi Davidson;Nufus, Hayatun;Amalia, Annisa
    • Natural Product Sciences
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    • v.23 no.3
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    • pp.192-200
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    • 2017
  • Skin aging is a complex biological process due to intrinsic and extrinsic factors. Free radical oxidative is one of extrinsic factors that induce activation of collagenase, elastase and hyaluronidase. Natural product from plants has been used as antioxidant and antiaging. This study aimed to evaluate antioxidant and antiaging properties of Hibiscus sabdariffa extract (HSE) and its compounds including myricetin, ascorbic acid, and ${\beta}$ carotene. The phytochemical of H. sabdariffa was determined using modified Farnsworth method and presence of phenols, flavonoids and tannins were in moderate content, whereas triterpenoids and alkaloids were in low content. Total phenolic content performed using Folin-Ciocalteu method, was $23.85{\mu}gGAE/mg$. Quantitative analysis of myricetin, ${\beta}-carotene$, and ascorbic acid of HSE was performed with Ultra-High Performance Liquid Chromatography (UHPLC) that shows $78.23{\mu}g/mg$ myricetin, $0.034{\mu}g/mg$ ${\beta}-carotene$, whilst ascorbic acid was not detected. HSE has lower activity on DPPH ($IC_{50}=195.73{\mu}g/mL$) compared to ${\beta}-carotene$, the lowest in ABTS assay ($IC_{50}=74.58{\mu}g/mL$) and low activity in FRAP assay ($46.24{\mu}MFe(II)/{\mu}g\;$) compared to myricetin, ${\beta}-carotene$. Antiaging was measured through inhibitory activity of collagenase, elastase, and hyaluronidase. HSE had weakest collagenase inhibitory activity ($IC_{50}=750.33{\mu}g/mL$), elastase inhibitory activity ($103.83{\mu}g/mL$), hyaluronidase inhibitory activity ($IC_{50}=619.43{\mu}g/mL$) compared to myricetin, ${\beta}-carotene$, and ascorbic acid. HSE contain higher myricetin compared to ${\beta}-carotene$. HSE has moderate antioxidants and lowest antiaging activities. Myricetin is the most active both antioxidant and antiaging activities.

Estimation of Measurement Uncertainty for the HPLC Analysis of Deoxynivalenol in Wheat (밀에서 HPLC에 의한 데옥시니발레놀 분석의 측정불확도 산정)

  • Ok, Hyun-Ee;Chang, Hyun-Joo;Ahn, Jang-Hyuk;Cho, Jae-Young;Chun, Hyang-Sook
    • Korean Journal of Food Science and Technology
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    • v.41 no.3
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    • pp.258-264
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    • 2009
  • The principal objective of this study was to estimate the measurement uncertainty associated with determination of deoxynivalenol (DON), a mycotoxin generated by Fusarium strain, in food. In service of this goal, wheat as a food matrix was analyzed via high performance liquid chromatography-ultraviolet (HPLC-UV) detection using an immunoaffinity column for clean-up. The uncertainty sources in the measurement process were identified by sample weight, final volume, and sample concentration in extraction volume with components including standard stock solution, working standard solution, 5 standard solutions, calibration curve, matrix, and instrument. The expanded uncertainty for DON at a concentration of 300 ${\mu}g/kg$ was estimated as 71.62 ${\mu}g/kg$ using a coverage factor of two, which provides a confidence level of approximately 95%. The most influential component in the uncertainty sources was the recovery of the wheat matrix, followed by the calibration curve. These results indicate that all efforts may be directed toward reducing the uncertainties of the recovery of the wheat matrix and the calibration curve to obtain a reliable HPLC-UV method for DON analysis in wheat.

Analysis of Florfenicol in Meats (식육 중 항생제 florfenicol의 분석)

  • Kuk, Ju-Hee;Song, Young-Me;Bae, Min-Seok;Go, Myoung-Jin;Yoo, Myung-Sang;An, Eun-Suk;Park, Eun-Heui;Go, Myoung-Jin;Kang, Kil-Jin
    • Journal of Food Hygiene and Safety
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    • v.23 no.4
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    • pp.319-323
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    • 2008
  • Analytical method for determination of florfenicol was developed for estimate veterinary drug residue of unestablished MRLs in meat. The method was validated in correspondence with the CODEX guideline for florfenicol residue in meat. The samples mixed with sodium sulfate were extracted with ethyl acetate. After clean-up, the residue was dissolved in mobile phase and analyzed using high-performance liquid chromatography with fluorescence detector. The calibration curve showed good linearity($r^2=0.9997$) within the concentration range of $0.05{\sim}1.0\;mg/kg$. The limit of detection(LOD) and limit of quantification(LOQ) were validated at 0.012 and 0.039 mg/kg, respectively. The recoveries in fortified meat ranged from 85.6 to 95.6%($1.1{\sim}5.3%$ RSD) at the 0.05 to 0.4 spiking levels. We monitored 150 samples of meats that were purchased in Korea(Seoul, Busan, Daegu, Daejeon and Gwangju). Among tested samples, florfenicol was detected in 1 of pig at the level of 0.040 mg/kg, and below LOQ in 1 of cattle, 2 of pig and 2 of chicken. The residues of florfenicol in the tested samples were within the MRLs.

Penetration of Some Humectants into Shark Muscle (상어육(肉)중의 수분유지제(水分維持劑) 침투(浸透))

  • Lee, Jong-Soo;Koo, Jae-Keun;Cha, Yong-Jun;Lee, Eung-Ho
    • Korean Journal of Food Science and Technology
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    • v.16 no.4
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    • pp.475-481
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    • 1984
  • Estimation of the penetration rate of humectants has been considered to be important in effective control of food processing when intermediate moisture food is manufactured by the moist-infusion method. In this study, when shark (Isurus glaucus) muscle was soaked in four common humectants (sucrose, sorbitol, glycerol, and propylene glycol), the equation of their penetration rate was drawn as a function of time using high performance liquid chromatography analysis. Penetration rates increased with soaking temperatures and decreased inversely with molecular weights of humectants. The penetrated amounts for 10% humectant solution reached about equilibrium after soaking for 10 hours and for 30% humectant after soaking for about 7 hours. In consideration with the penetration rate of the sample soaked in 10% humectant and complex solution of each 10% humectant, little difference was found between them. When the sample was soaked in 10% humectant and 30% humectant, it seemed to be able to apply the following regression equation to estimate the penetrated amounts: M = a log (c.t)+ b where M = penetrated amounts; c = concentration of humectant; t= soaking time; a, b = constant and c.t should be within $10^3\;-\;4{\times}10^4$.

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Isolation of Ginsenoside${-Rh}_1$ and ${-Rh}_2$ by High Performance Liquid Chromatography (고속액체(高速液體) 크로마토그래피에 의(依)한 Ginsenoside ${-Rh}_1$${-Rh}_2$ 의 분리(分離))

  • Choi, Jin-Ho;Kim, Woo-Jung;Hong, Soon-Keun;Oh, Sung-Ki;Oura, Hikokichi
    • Korean Journal of Food Science and Technology
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    • v.13 no.1
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    • pp.57-66
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    • 1981
  • An effective method for isolation of the major components of ginseng saponin such as $ginsenoside-Rb_{1},\;-Rb_2,$ -Rc, -Rd, -Re and $-Rg_1$, and the minor components such as ginsenoside-Rf, $-Rg_2,\;and-Rh_1$, was developed and reported in previous papers (J. Korean Agr. Chem. Soc., 23(4), 199 and 206(1980) The conditions and procedures used for isolation and identification for ginsenosides described in the previous papers were not sufficient enough for clean separation of minor components, $ginsenoside-Rh_1,\;and-Rh_2$. In this work, modifications in extraction method and in mobile phase for HPLC were attempted. It was found that application of ethyl acetate extraction at $60^{\circ}C$ for 3 hr on crude saponin resulted in a removal of diol group saponin from crude saponin which made it possible for using higher portion of acetonitrile in mobile phase. The mixed solvents of acetonitrile : water (92 : 8 and 94 : 6) gave excellent resolution of $ginsenoside-Rh_1\;and\;-Rh_2$.

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Determination of Isomaltooligosaccharides in Yoghurts by Using HPLC-ELSD (HPLC-ELSD를 이용한 발효유 제품 중의 Isomaltooligosaccharides 분석법 개발)

  • Ko, Jinhyouk;Lee, Moon-Seok;Kwak, Byung-Man;Ahn, Jang-Hyuk;Park, Jong-Su;Kwon, Joong-Ho
    • Food Science of Animal Resources
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    • v.33 no.3
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    • pp.417-424
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    • 2013
  • A rapid and simple analytical method for the determination of 9 isomaltooligosaccharides (IMO) species in yoghurts was developed using dispersive solid phase extraction (dSPE) clean-up technic and high performance liquid chromatography with evaporative light-scattering detector (HPLC-ELSD). In this study, 9 IMO were extracted from samples simply with chemical reagent using ISO22662 IDF198 method and additional dSPE clean-up. The optimum instrument conditions for the determination were used carbohydrate ES $5{\mu}$ column with gradient elution of water and acetonitrile and ELS detector. The linearity of this method was expressed as the correlation coefficient ($r^2$), the results of IMO 9 species were shown in 0.9999. LOD and LOQ were respectively 7.9-22.1 mg/kg, 25.9-72.8 mg/kg. The accuracy of intra- and inter-day measurements were in the range from $84.3{\pm}4.5$ to $104.9{\pm}6.5%$, and the preceision of the intra- and inter-day measurements were in the range from 0.8 to 7.7%. The recoveries were from $84.3{\pm}4.5$ to $104.9{\pm}6.5%$. The determination results of IMO 9 species for the 9 yoghurts circulated in the market were in the range from $0.317{\pm}0.007$ to $1.624{\pm}0.050$ g/100 g. The newly developed method is appropriate for the determination of IMO in yoghurts, is a rapid and simple method with excellent resolution in compared with previous method.

Changes in the Growth Characteristics and the Content of Decursin and Decursinol angelate in Angelica gigas Nakai by Mineral and Hormonal Agent Treatment (미네랄 및 호르몬 제제 처리에 따른 당귀의 생육특성 및 Decursin, Decursinol Angelate 함량 변화)

  • Lee, Sang Hoon;Yun, Hyeong Muk;Koo, Sung Cheol;Lee, Woo Moon;Chang, Jae Ki;Koo, Hyun Jeong;Chang, Kwang Jin;Kim, Yeon Bok
    • Korean Journal of Medicinal Crop Science
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    • v.26 no.3
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    • pp.227-232
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    • 2018
  • Background: Angelica gigas Nakai is a perennial herb belonging to the family Umbelliferae. Its roots are utilized in traditional medicine. The aim of this study was to increase the yield of and the content of two indicator components (decursin and decursinol angelate) in A. gigas Nakai. Methods and Results: The roots of A. gigas Nakai were harvested 4-times from late August to late October in 2017. Two agents (trace element-TE, and plant hormone-HM) were applied 4 times at intervals of 2 weeks. The content of the two indicator components were analyzed by high performance liquid chromatography. The HM treatment showed the greatest increase in underground part yield and root diameter. The content of the two indicator components in the control (non-treatment) group was the highest in the underground part, but was higher in the aerial parts in the agent treatment group. After treatment with the agents, the content of the indicator components tended to decrease in the underground part. However, the total content of the indicator components in the two agent treatment groups exceeded the level of 6% set by the Korean Pharmacopoeia. Conclusions: The highest underground part yield was found in the HM treatment group, while the highest content of decursin and decursinol angelate were found in the control group. This study provides basic information for yield improvement in A. gigas.

Antioxidant and α-Glucosidase Inhibitory Activities of the Extracts of Aster koraiensis Leaves (국내산 벌개미취 잎 추출물의 α-glucosidase 억제능 및 항산화 활성 평가)

  • Lee, Tae Gu;Hyun, Soo Wang;Lee, Ik Soo;Park, Bong Kyun;Kim, Jin Sook;Kim, Chan Sik
    • Korean Journal of Medicinal Crop Science
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    • v.26 no.5
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    • pp.382-390
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    • 2018
  • Background: The plant Aster koraiensis has long been used as an ingredient in folk medicine. It has been reported that Aster koraiensis extract (AKE) prevents the progression of diabetes-induced retinopathy and nephropathy. However, although these beneficial effects of AKE on diabetes complications have been identified, the antidiabetic effects of AKE have not yet been completely investigated and quantified. In the present study, the glucose-lowering and antioxidant effects of aqueous and ethanolic AKEs were evaluated. Methods and Results: The glucose-lowering effects of aqueous and ethanolic (30%-, 50%-, and 80%-ethanol) AKEs were investigated via ${\alpha}$-glucosidase inhibitory assays. The mode of inhibition by AKEs on ${\alpha}$-glucosidase was identified through kinetic analysis. The total antioxidant capacity of each of the 4 AKEs was evaluated by assessing their conversion rate of $Cu^{2+}$ to $Cu^+$. The content of chlorogenic acid and 3,5-di-O-caffeoylquinic acid, the bioactive compounds in AKE, in each extract were analyzed by high performance liquid chromatography (HPLC). The AKEs showed potent ${\alpha}$-glucosidase inhibitory activity with mixed inhibition mode, and significant antioxidant capacity. Conclusions: These results of this study suggested that the AKEs tested had ${\alpha}$-glucosidase inhibitory and antioxidant effects. Among the extracts, the 80% ethanol extract showed the most significant ${\alpha}$-glucosidase inhibitory activity, with a half maximal inhibitory concentration ($IC_{50}$ value) of $1.65{\pm}0.36mg/m{\ell}$ and a half maximal effective concentration ($EC_{50}$ value) for its antioxidant activity of $0.42{\pm}0.10mg/m{\ell}$. It can therefore be used as a source of therapeutic agents to treat diabetes patients.

Comparison of Anti-oxidative and Anti-inflammatory Effect of Atractylodes Interspecific Hybrid Cultivar Roots (삽주 육성품종 뿌리의 항산화 및 항염증 효과 비교)

  • Jeong, Hyeon Soo;Jeong, Jin Tae;Lee, Jeong Hoon;Park, Chun Geon;Choi, Je Hun;Jang, Gwi Yeong;Kim, Jang Wook;Chang, Jae Ki;Kim, Dong Hwi;Lee, Seung Eun
    • Korean Journal of Medicinal Crop Science
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    • v.26 no.5
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    • pp.391-400
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    • 2018
  • Background: Atractylodes radix is a well-known medicinal crop having many physiological effects. This study was conducted to select useful Atractylodes japonica ${\times}$ Atractylodes macrocephala (AJM) cultivars by comparing anti-oxidative and anti-inflammatory efficacies. Methods and Results: Seven extracts from AJM cultivars were used to treat lipopolysacchride (LPS)-treated BV2 cells, and the effects on cell viability and inhibition on reactive oxygen species (ROS) and nitric oxide (NO) production were analyzed. In vitro scavenging activities of 2,2-diphenyl-1-picrylhydrazyl (DPPH) and peroxynitrite ($NOO^-$) radicals were also investigated. Contents of total phenol, atractylenolide I, and atractylenolide III in the AJM extracts were measured using high performance liquid chromatography (HPLC) or spectrophotometry. The experiments show that none of the seven extracts was cytotoxic above 89.2% at $20-250{\mu}g/m{\ell}$. Extracts of Gowon, Dawon, Sangchul, and Huchul inhibited ROS generation in a dose-dependent manner, and Sangchul extract showed the highest inhibition on ROS production. All the AJM extracts showed effective inhibitory activity after on NO release in the LPS-treated BV2 cells, and Sangchul extract showed the highest activity. Sangchul extract had the most potent scavenging activities for $NOO^-$ and had some DPPH radical scavenging effect. Sangchul extract also had the highest content at total phenol and atractylenolide I content. Atractylenolide III was not detected in the AJM extracts. Conclusions: The results suggested that Sangchul was the most useful anti-oxidative and anti-inflammatory resource among the AJM cultivars.

Bioequivalence of LANIDIEM® Tablet 4 mg to Vaxar® Tablet 4 mg(Lacidipine 4 mg) (박사르®정 4 밀리그램(라시디핀 4 mg)에 대한 라니디엠®정 4 밀리그램의 생물학적동등성)

  • Lee, Yun-Young;Kim, Hye-Jin;La, Sookie;Cho, Kyung-Hee;Jang, Moon-Sun;Park, Young-Joon;Lee, Hee-Joo
    • Journal of Pharmaceutical Investigation
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    • v.40 no.2
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    • pp.125-131
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    • 2010
  • A bioequivalence study of LANIDIEM$^{(R)}$ tablet 4 mg (Samil. Co., Ltd.) to Vaxar$^{(R)}$ tablet 4 mg (GlaxoSmithKline Co., Ltd.) was conducted according to the guidelines of Korea Food and Drug Administration (KFDA). Forty healthy male Korean volunteers were enrolled in the study and thirty six volunteers completed the study according to the protocol. Thirty six volunteers received each medicine at the lacidipine dose of 4 mg in a $2{\times}2$ crossover study. There was one week wash-out period between the doses. Plasma concentrations of lacidipine were monitored by a high performance liquid chromatography - tandem mass spectrometry (LC-MS/MS) for over a period of 24 hours after drug administration. $AUC_t$ (the area under the plasma concentration-time curve from time zero to 24 hr) was calculated by the linear trapezoidal rule method. $C_{max}$ (maximum plasma drug concentration) and $T_{max}$ (time to reach $C_{max}$) were compiled from the plasma concentration-time data. Analysis of variance was carried out using logarithmically transformed $AUC_t$ and $C_{max}$. No significant sequence effect was found for all of the bioavailability parameters indicating that the crossover design was properly performed. The 90% confidence intervals of the $AUC_t$ ratio and the $C_{max}$ ratio for LANIDIEM$^{(R)}$/Vaxar$^{(R)}$ were log 0.8102~log 1.0417 and log 0.8493~log 1.1439, respectively. These values were within the acceptable bioequivalence intervals of log 0.80~log 1.25. Thus, our study demonstrated the bioequivalence of LANIDIEM$^{(R)}$ tablet 4 mg and Vaxar$^{(R)}$ tablet 4 mg with respect to the rate and extent of absorption.