• Title/Summary/Keyword: HPLC analysis method

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Simultaneous Determination of Amitraz, Bromopropylate, Coumaphos, Cymiazole and 2,4-Dimethylaniline in Korean Honey Samples by High-Performance Liquid Chromatography

  • Ryoo, Jae-Jeong;Kim, Se-Hoon;Jeong, Young-Han;Do, Hwan-Suk;Ryu, Jeong-Eun;Kwon, Hae-Young;Jeong, Ji-Yoon;Park, Hye-Jin;Lee, Soon-Ho;Hong, Moo-Ki;Hong, Jong-Ki
    • Bulletin of the Korean Chemical Society
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    • v.29 no.5
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    • pp.1043-1047
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    • 2008
  • Simultaneous determination of amitraz, bromopropylate, coumaphos, cymiazole and 2,4-dimethylaniline in 200 honey samples purchased in Korea was performed by reversed-phase high-performance liquid chromatography with multiple UV detection. 2% Acetone in hexane was used for a liquid-liquid extraction and 20-40% water in acetonitrile solutions were used as mobile phases. The LOD for the analytes varied between 0.4 and 1.5 $\mu$g/L and the recoveries were yielded between 64 and 94%. Relative standard deviation of the repeatability of the method is less than 15%. Amitraz was not present in amount above 10 $\mu$ g/L and one for coumaphos and cymiazole and two for bromopropylate, and three for 2,4-dimethylanilne were detected in amount above 10 $\mu$ g/ L. Levels of the acaricide residues found were less than 50 $\mu$ g/L.

An Effective Method of Isolating Immunoglobulins from Bovine Plasma Proteins (도축혈액으로부터 면역단백질의 효과적인 분리법)

  • Choi, In-Wook;Lee, Hyun-Jung
    • Korean Journal of Food Science and Technology
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    • v.29 no.5
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    • pp.1067-1070
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    • 1997
  • Imunoglobulins from bovine plasma proteins were isolated by IMAC which $Cu^{2+}$ was chelated on a chelating sepharose fast flow gel. Most plasma proteins were eluted by 1st (0.01 M $Na_2HPO_4$, 0.5 M NaCl, pH 4.0) and 2nd elution buffers (0.01 M imidazol). According to the reverse phase HPLC analysis, it was found that proteins which were eluted by 1st elution buffer were mainly composed of serum albumin, while most IgG and transferrin were eluted by 2nd elution buffer. When protein fractions obtained by 2nd elution buffer was applied to ultra filtration system (molecular weight cut off: 100 kD), IgG was further purified. These results indicate that IMAC is an excellent tool for isolating imunoglobulins from plasma proteins.

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Preparation of Cyclosporin A-loaded Nanoparticles Containing Ethyl Myristate or Chitosan and Pharmacokinetics in Rats

  • Nam, Dae-Sik;;Lee, Woo-Kyoung
    • Journal of Pharmaceutical Investigation
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    • v.37 no.1
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    • pp.15-22
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    • 2007
  • An oil-in-water solvent evaporation method was used to prepare the cyclosporin A (CyA)-loaded nanoparticles varying in poly (D,L-lactide-co-glycolide) (PLGA) polymer (RG 502H, RG 503H) and the amount of additive ethyl myristate (EM) or chitosan (CS). The particles were characterized for drug loading and entrapment efficiency by HPLC, surface morphology by scanning electron microscopy, particle size by dynamic light scattering and surface charge by Zetapotential. The results showed drug loadings ranging from 10.9% to 15.8% with high encapsulation efficiency (82.0-97.8%). SEM and DLS studies showed discrete and spherical particles with smooth surfaces and mean size ranging 257.6-721.7 nm. The additive EM or CS did not change the mean sizes of the nanoparticles, whereas by the coating effect of CS, the Zetapotential values of the CS-added nanoparticles were moved to the more positive direction as the amount of CS was increased. From the pharmacokinetic analysis, the nanoparticles formulations showed the higher bioavailability and MRT than $Neoral^{\circledR}$ While little adding effect of EM or CS was detected in pharmacokinetic profile when RG 503H was used as polymer carrier, more noticeable different pharmacokinetic behaviors could be observed in case of RC 502H. EM incorporation was found to elevate the $K_{el}$, whereas CS coating resulted in the decrease of F and $K_{el}$, which seems to be due to the function of CS as a barrier and a mucoadhesive coating.

Nutrient Analyses of Sustainable Ready-to-Use-Supplemental Food (RUSF) developed with East African Ethnic Plant Resources (동아프리카 자원식물을 활용하여 개발된 지속가능한 영양보충식품의 영양성분 분석)

  • Lee, Kyoung-Ae;Kim, Ye-Jung;Koh, Kwangoh;Kim, Hee-Seon
    • Journal of the East Asian Society of Dietary Life
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    • v.26 no.5
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    • pp.466-472
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    • 2016
  • The objectives of the study were to analyze the various nutrient contents of ready-to-use-supplemental-food (RUSF) developed with Eastern African ethnic plants and compare them with commercially available supplemental food (CASF) for infants. RUSF was made with three African plants, including Ficus sur, Amaranthus caudatus, and Prosopis juliflorafa and locally available food ingredients in DR Congo and Ethiopia and was cooked at $180^{\circ}C$ for 20 min in a conventional oven. Macronutrients were analyzed by the AOAC method. Contents of 17 amino acids were analysed with amino acid analyzer, and vitamin $B_1%$ niacin, vitamin $B_6$, and vitamin C were analyzed with HPLC. Isomers of vitamin E were analyzed by GC-FID. The price of RUSF was 0.133 USD per 100g. Contents of macronutrients were not significantly different between developed RUSF and CASF. Results of amino acid analysis showed that most amino acid contents were similar or higher in RUSF compared with CASF. However, glutamic acid and lysine contents were less in RUSF than in CASF. Vitamin E and vitamin $B_1$ contents were more than five times higher in RUSF (22.40, 6.26 mg/100 g, respectively) than in CASF (4.00, 0.80 mg/100 g, respectively). Niacin was about three times higher while vitamin $B_6$ was almost 20 times higher in RUSF than in CASF. However, ascorbic acid was significantly lower in RUSF (3.43 mg/100 g) than in CASF (42.08 mg/100 g). Our study found that the developed localized RUSF showed higher nutrient contents in most micronutrients analyzed except for ascorbic acid. Out of 17 amino acids, glutamic acid and lysine contents in RUSF were less than in CASF. Therefore, further study is needed in order to improve these nutrients contents.

Therapeutic Anti-inflammatory Effect of Ginkgo Terpene on Arthritis due to Candida albicans (Ginkgo Terpene의 Candidate albicans로 인한 관절염에 대한 치료효과)

  • Lee, Soon-Hyun;Lee, Jue-Hee;Han, Yong-Moon
    • YAKHAK HOEJI
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    • v.49 no.2
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    • pp.140-146
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    • 2005
  • Candida albicans, a polymorphic fungus, causes systemic and local infections. Recent reports show that the fungus is a main etiological agent for the arthritis. For trea tment, antifungal drugs and/or rheumatoid drugs are used, but resistance and side effects limit application of the drugs. In search of new sources for treatment of the fungal arthritis, we choose Egb 761 (extract of Ginkgo leaves 761), one of the most popular over-the-counter herbal medicines. The Egb 761 contains two major ingredients such as terpene and flavonoid. In the present study, we examined if the terpene portion of Egb 761 had anti-inflammatory activity against C.albicans-caused arthritis. The terpene was extracted with combination of methanol and water from the Egb 761, followed by gel-permeation chromatography. Presence of terpene was determined by the Salkowski colorimetric method and HPLC analysis. For an animal model of inflammation induction, mice were given an emulsion form of C.albicans cell wall mixed with Complete Freund's Adjuvant (CFA) by footpad-injection. Results showed that intraperitoneal administration of the water-soluble portion that contained terpene and flavonoid reduced the inflammation. Whereas the terpene had anti-inflammatory activity, flavonoid portion had no such activity, For determination of possible mechanism of the activity, the terpene seemed to be suppression of nitric oxide (NO) production from LPS-treated macrophages. Taken together the Ginkgo terpene may have anti-inflammatory effect against C.albicans-caused arthritis, possibly by blocking NO production.

An Assessment of Korean Housewives Exposed to Polycyclic Aromatic Hydrocabons(PAHs) in Indoor Air (일부 주부의 실내공기 중 PAHs 노출에 관한 연구)

  • Lee, Tae-Hyung;Kim, Yun-Sin;Son, Bu-Soon
    • Journal of Environmental Science International
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    • v.16 no.3
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    • pp.323-331
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    • 2007
  • Polycyclic aromatic hydrocarbons (PAHs) are well known for strong carcinogen. However, the human exposure analysis of PAHs is quite difficult and unreliable because of hard for estimation of actual expose dose. Then urinary 1-hydroxypyrene (1-OHP) has been a biological marker of exposure to PAHs. The purpose of this study was to investigate total amount from exposure to PAHs soused by indoor occupational exposure, and residence at Seoul metropolitan area and Kyeonggi province in Korea. Thirty-five housewives were included in this study from April 2003 through February 2004. Dietary habit and general characteristics such as age, type of building, existence of passive smoking, period of residence, fuel type for heating and ventilation type were obtained by self administered questionnaire. Urine samples were collected at morning and freeze quickly. Urinary creatinine was measured for converting into 24 hr urine. Concentration of the indoor PAHs was examined by NIOSH method number 5506. Urinary 1-OHP and PAHs were analysed by HPLC. Correlation coefficient between urinary 1-OHP levels and pyrene concentration of indoor air was 0.66 and statistically significant(P<0.01). The difference of urinary 1-OHP level due to dietary habits were not significant. Urinary 1-OHP level of Spring, Summer, Autumn, and Winter were $0.21{\pm}0.12,\;0.10{\pm}0.17,\;0.16{\pm}0.12,\;0.17{\pm}0.14{\mu}g/g$ cr, respectively. The arithmetic means of urinary 1-OHP for four season tee $0.16{\pm}0.14 {\mu}g/g$ cr. There was a trend that urinary 1-OHP level of residents who dwelling in apartment were higher compared with detached home, Comparison of 1-OHP level between heating by kerosene and LPG, Much higher gas heating type than kerosene type (P<0.05). This result implies that the urinary 1-OHP can be applied as the PAHs exposure indices.

Bioequivalence of GomcillinTM Capsule to FamoxinTM Capsule (Amoxicillin 500 mg) (파목신 캅셀(아목시실린 500 mg)에 대한 곰실린 캅셀의 생물학적동등성)

  • Lee, Yun-Young;Choi, Mee-Hee;Lee, Kyung-Ryul;Lee, Hee-Joo
    • Journal of Pharmaceutical Investigation
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    • v.34 no.4
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    • pp.311-317
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    • 2004
  • A bioequivalence study of $Gomcillin^{TM}$ capsules (DAEWOONG Pharmaceutical Co., Korea) to $Famoxin^{TM}$ capsules (Dong Wha Pharm. Ind. Co., Korea) was conducted according to the guideline of Korea Food and Drug Administration (KFDA). Twenty four healthy male Korean volunteers received each medicine at the amoxicillin dose of 500 mg in a $2{\times}2$ crossover study. There was a one-week wash out period between the doses. Plasma concentrations of amoxicillin were monitored by a high-performance liquid chromatography for over a period of 8 hours after the administration. $AUC_t$ (the area under the plasma concentration-time curve from time zero to 8 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 $Gomcillin^{TM}/Famoxin^{TM}$ were $log0.91\;{\sim}\;log1.03$ and $;log0.93\;{\sim}\;log1.10$, respectively. These values were within the acceptable bioequivalence intervals of $log0.80\;{\sim}\;log1.25$. Thus, our study demonstrated the bioequivalence of $Gomcillin^{TM}$ and $Famoxin^{TM}$ with respect to the rate and extent of absorption.

Development and validation of LC-MS/MS for bioanalysis of hydroxychloroquine in human whole blood

  • Park, Jung Youl;Song, Hyun Ho;Kwon, Young Ee;Kim, Seo Jin;Jang, Sukil;Joo, Seong Soo
    • Journal of Biomedical and Translational Research
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    • v.19 no.4
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    • pp.130-139
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    • 2018
  • This study aimed to analyze a high-performance liquid chromatography (HPLC) separation using a pentafluorophenyl column of parent drug hydroxychloroquine (HCQ) and its active metabolite, desethylhydroxchloroquine (DHCQ) applying to determine bioequivalence of two different formulations administered to patients. A rapid, simple, sensitive and specific liquid chromatography-tandem mass spectrometry (LC-MS/MS) method has been developed and validated for bioanalysis of HCQ and its metabolite DHCQ in human whole blood using deuterium derivative $hydroxychloroquine-D_4$ as an internal standard (IS). A triple-quadrupole mass spectrometer was operated using electrospray ionization in multiple reaction monitoring (MRM) mode. Sample preparation involves a two-step precipitation of protein techniques. The removed protein blood samples were chromatographed on a pentafluorophenyl (PFP) column ($50mm{\times}4.6mm$, $2.6{\mu}m$) with a mobile phase (ammonium formate solution containing dilute formic acid) in an isocratic mode at a flow rate of 0.45 mL/min. The standard curves were found to be linear in the range of 2 - 500 ng/mL for HCQ; 2 - 2,000 ng/mL for DHCQ in spite of lacking a highly sensitive MS spectrometry system. Results of intra- and inter-day precision and accuracy were within acceptable limits. A run time of 2.2 min for HCQ and 2.03 min for DHCQ in blood sample facilitated the analysis of more than 300 human whole blood samples per day. Taken together, we concluded that the assay developed herein represents a highly qualified technology for the quantification of HCQ in human whole blood for a parallel design bioequivalence study in a healthy male.

Quantification of the Bioactive Components of the Rhizomes of Curcuma wenyujin and Assessment of Its Anti-inflammatory Effect in Benign Prostatic Hyperplasia-1 Cells

  • Seo, Chang-Seob;Park, Eunsook;Lee, Mee-Young;Shin, Hyeun-Kyoo
    • Natural Product Sciences
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    • v.25 no.3
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    • pp.238-243
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    • 2019
  • In this study, the marker compounds of Curcumae Rhizoma (CR) were simultaneously quantified by high-performance liquid chromatography equipped with a photodiode array detector and the anti-inflammatory effects of CR extract and marker compounds in human benign prostatic hyperplasia epithelial-1 (BPH-1) cell lines were investigated. The marker components (4S,5S)-(+)-germacrone-4,5-epoxide, furanodienone, and germacrone, were separated on Gemini $C_{18}$ columns ($250mm{\times}4.6mm$, $5{\mu}m$) at $40^{\circ}C$ by using a gradient of two mobile phases eluting at 1.0 mL/min. Prostaglandin $E_2$ ($PGE_2$) levels in Human BPH-1 cells were determined with an ELISA kit. The coefficients of determination in a calibration curve of each analyte were all 0.9997. The limits of detection and quantification of the three compounds were $0.10-0.32{\mu}g/mL$ and $0.30-0.98{\mu}g/mL$, respectively. The content of three compounds, (4S,5S)-(+)-germacrone-4,5-epoxide, furanodienone, and germacrone, in the CR sample were found to be 5.79 - 5.92 mg/g, 4.72 - 4.86 mg/g, and 1.06 - 1.09 mg/g, respectively. Regarding pharmacological activity against benign prostatic hyperplasia, CR and its components significantly suppressed $PGE_2$ levels of BPH-1 cells. The established analysis method will help to improve quality assessment of CR samples and related products. In addition, CR and its components exhibit antiinflammatory activity in BPH-1 cells, suggesting the inhibitory efficacy of these compounds against the pathogenesis of BPH.

Water-soluble Vitamin Content in Dishes Containing Meat and Seafood (육류 및 수산물을 이용한 조리식품에 함유된 수용성 비타민 함량)

  • Jin, Minguen;Kim, Byung Hee;Kim, Min-Hee;Yoon, Sung Won;Kim, Younghwa
    • Journal of the Korean Society of Food Culture
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    • v.36 no.5
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    • pp.502-511
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    • 2021
  • In this study, the content of water-soluble vitamins such as vitamin B1 (thiamin), B2 (riboflavin), B3 (niacin), B5 (pantothenic acid), B6 (pyridoxine), B7 (biotin), B12 (cyanocobalamin), and C (ascorbic acid) in dishes containing meat and seafood consumed in Korea were determined by high-performance liquid chromatography (HPLC) with ultraviolet and fluorescence detection. All analyses were performed under strict quality control of vitamins B1, B2, B3, B5, B6, B7, B12, and C. The highest content of vitamin B1 was observed in Bugeo-gangjeong (1.373 mg/100 g) and the highest level of vitamin B2 (5.162 mg/100 g) was found in pig liver. Bugeo-gangjeong showed the highest content of vitamin B3 (21.676 mg/100 g), and kkomak-muchim contained considerable amounts (43.310 mg/100 g) of vitamin B5. Vitamin B6 was not detected in most seafood dishes except for yangnyeom myeongran-jeot (0.274 mg/100 g) and was present at low levels or not present at all in meat dishes. The highest content of vitamin B7 was 6.506 ㎍/100 g in saeu-jeon and kkomak-muchim showed the highest content (21.132 ㎍/100 g) of vitamin B12. The highest content of vitamin C was in yangnyeom myeongran-jeot (84.508 mg/100 g). In addition, the analysis methods of each water-soluble vitamin were verified. These results showed that seafood-based ingredients in several dishes could be a good source of water-soluble vitamins.