• Title/Summary/Keyword: HPLC quantification

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Studies on the quantification of sobrerol by high-performance liquid chromatography

  • Lee, Dong-Mi;Won, Young-Jun;Yoon, Chang-Yong;Seo, Il-Won;Nam, Hye-Seon;Park, Dong-Hee;Lee, Hyang-Mi;Kim, Se-Sil;Jung, Eun-Mi
    • Proceedings of the PSK Conference
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    • 2003.04a
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    • pp.283.1-283.1
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    • 2003
  • This study was designed to develop and validate an isocratic reversed phase high performance liquid chromatographic(HPLC) method for the quantitation of sobrerol in drug preparations, and obtain the data pool that can be used in the revision of pharmacopoeia. The separation of sobrerol and the other compounds (S-carboxymethylcysteine, acetaminophen, methyl paraben, propyl paraben, and sobrerol degradants) was achieved in a C18 column with an acetonitrile-methanol-water(24.5:10.5:65.0) mobile phase. (omitted)

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Three Hydroxylated Ginsenosides from Heat Treatmented Ginseng (인삼의 열처리 과정 중 생성되는 3종의 수산화진세노사이드에 대한 연구)

  • Lee, Sang Myung
    • Korean Journal of Pharmacognosy
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    • v.51 no.4
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    • pp.255-263
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    • 2020
  • Ginsenosides are considered to be the most important ingredients in ginseng. They are chemically converted by endogenous organic acids contained in ginseng and the heat applied during red ginseng processing. During this procedure, various converted ginsenosides are produced through hydrolysis of substitute sugars of ginsenosides and forming double bonds through dehydration in the dammarane skeleton. In order to study the conversion mechanism of protopanaxadiol-type ginsenosides during the heat treatment process of ginseng, we purified the three final converted ginsenosides by heating fresh ginseng for a long time. The three isolated ginsenosides were identified as 25(OH)-ginsenoside Rg5, 25(OH)-ginsenoside Rz1 and 25(OH)-ginsenoside Rg3 through NMR spectrum analysis. As a result of quantification of ginseng heated at 100 ℃ for 0 to 6 days by HPLC/UV and TLC methods, the content of 25(OH)-ginsenosides tended to increase in proportion to the time exposed to heat. In particular, the content of 25(OH)-ginsenosid Rg5 was confirmed to be noticeably increased.

Development of Vitamin D Determination in Infant Formula by Column-Switching HPLC with UV Detector

  • Ko, Jin-Hyouk;Kwak, Byung-Man;Ahn, Jang-Hyuk;Shim, Sung-Lye;Kim, Kyong-Su;Yoon, Tae-Hyung;Leem, Dong-Gil;Jeong, Ja-Young
    • Food Science of Animal Resources
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    • v.32 no.5
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    • pp.571-577
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    • 2012
  • This study was carried out to develop an analytical method for the determination of vitamin D in infant formula. Vitamin D was determined by column-switching high-performance liquid chromatography (HPLC) equipped with a reversed phase column and UV detector after saponification and extraction of the formula with an organic solvent. A preseparation column ($C_8$), focusing column ($C_{18}$), analytical column ($C_{18}$) and UV-Vis detector (254 nm) were used. The limits of detection (LOD) and the limits of quantification (LOQ) for vitamin D were estimated to be $1.51{\mu}g/kg$ and $4.95{\mu}g/kg$, respectively. The linearity, recovery, precision and accuracy of the analytical method for vitamin D were evaluated through the application of a SRM (Standard Reference Material) 1846 (National Institute of Standard & Technology, USA). The linearity of this method was calculated with a value of the coefficient of determination ($r^2$) ${\geq}0.9999$. The recovery of vitamin D was $85.20{\pm}3.00%$. The intra-assay precision for vitamin D was between $1.68{\pm}0.03%$ and $5.75{\pm}0.33%$, and the inter-assay precision for vitamin D ranged from $1.73{\pm}0.03%$ to $2.96{\pm}0.09%$. The intra-assay accuracy for vitamin D was between $100.03{\pm}2.77%$ and $102.01{\pm}0.59%$, and the inter-assay accuracy for vitamin D ranged from $99.00{\pm}1.53%$ to $102.01{\pm}3.04%$. The proposed method is optimal for the separation and quantification of vitamin D from infant formula.

Quantification of Entacapone in Human Plasma by HPLC Coupled to ESI-MS/MS Detection: Application to Bioequavalence Study (체외에서 ESI-MS/MS 탐지에 연결된 HPLC에 의한 Entacapon의 수량화: 생물학적 동등성 연구에 적용)

  • Balasekhara Reddy., Ch.;Baburao., Ch.;Chandrasekhar., K.B.;Kanchanamala., K.;RihanaParveen., S.K.;Ravikumar., Konda
    • Journal of the Korean Chemical Society
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    • v.54 no.5
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    • pp.523-532
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    • 2010
  • The proposed method is simple, sensitive and specific Liquid chromatography-tandem mass spectrometry (LCESI-MS/MS) method for the quantification of Entacapone (EA) in human plasma using Entacapone-d10 (EAD10) as an internal standard (IS). Chromatographic separation was performed on Zorbax SB-C18, $2.1{\times}50\;mm$, $5\;{\mu}m$ column, mobile phase composed of 10 mM Ammonium formate (pH 3.0): Acetonitrile (60:40 v/v), with a flow-rate of 0.7 mL/min, followed by Liquid-liquid extraction. EA and EAD10 were detected with proton adducts at m/z $306.1{\rightarrow}233.1$ and $316.3{\rightarrow}233.0$ in multiple reaction monitoring (MRM) positive mode respectively. The method was validated over a linear concentration range of 1.00 - 2000.00 ng/mL with correlation coefficient ($r^2$) $\geq$ 0.9993. Intra and inter-day Precision within 3.60 to 7.30 and 4.20 to 5.50% and Accuracy within 97.30 to 104.20 and 98.30 to 105.80% proved for EA. This method is successfully applied in the bioequivalence study of healthy Indian human volunteers.

Biological Treatment of TNT-containing Wastewater (pink water) by Stenotrophomonas maltophilia OK-5, and RT-PCR Quantification of the Nitroreductase (pnrB) Gene (Stenotrophomonas maltophilia OK-5에 의한 TNT 함유폐수 (pink water)의 생물학적 처리 와 Nitroreductase (pnrB) 유전자의 RT-PCR 정량화)

  • Cho, Su-Hee;Cho, Yun-Seok;Oh, Kye-Heon
    • KSBB Journal
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    • v.24 no.6
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    • pp.556-562
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    • 2009
  • The biological treatment of TNT-containing wastewater, known commonly as pink water, was investigated using a stirred tank reactor with Stenotrophomonas maltophilia OK-5 bacterial culture. S. maltophilia OK-5 exhibited effective degradation of TNT contained in pink water, completely degrading TNT (100 mg/L) within 6 days of incubation. The dark-red brown color derived from Hydride-Meisenheimer complex became more pronounced during the incubation period, which was determined quantitatively. High-pressure liquid chromatography was used to measure residual TNT, which also resolved the metabolic intermediates (i.e., 2,4-dinitrotoluene, 2,6-dinitrotoluene and 2,4-dinitro-6-hydroxytoluene). Gas chromatography-mass spectrometry was used to verify these intermediates. Quantification of the nitroreductase (pnrB) gene isolated from S. maltophilia OK-5 growing in pink water was performed with real-time PCR. The amount of pnrB gene copies increased to $10^3$-fold after 5 days of incubation time.

Antioxidant Properties and Quantification of Phenolic Compounds from Safflower (Carthamus tinctorius L.) Seeds

  • Kim, Eun-Ok;Oh, Ji-Hae;Lee, Sung-Kwon;Lee, Jun-Young;Choi, Sang-Won
    • Food Science and Biotechnology
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    • v.16 no.1
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    • pp.71-77
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    • 2007
  • The antioxidant properties of twelve phenolic compounds, including matairesinol 4'-O-$\beta$-D-glucoside, 8'-hydroxyarctigenin 4'-O-$\beta$-D-glucoside, matairesinol, 8'-hydroxyarctigenin, N-feruloylserotonin 5-O-$\beta$-D-glucoside, N-(p-coumaroyl)-serotonin-5-O-$\beta$-D-glucoside, N-feruloylserotonin, N-(p-coumaroyl)serotonin, luteolin 7-O-$\beta$-D-glucoside, luteolin, acacetin 7-O-$\beta$-glucuronide, and acacetin, isolated from defatted safflower (Carthamus tinctorius L.) seeds were evaluated with regard to the DPPH, superoxide and hydroxyl radicals. Additionally, levels of phenolic compounds were determined by HPLC in two cultivars of safflower seeds. Among them, four serotonin derivatives showed potent DPPH ($IC_{50}=10.83-21.75\;{\mu}M$) and hydroxyl ($IC_{50}=75.93-374.63\;{\mu}M$) radical scavenging activities, and their activities were significantly stronger than that of ${\alpha}-tocopherol$. Four flavonoids ($IC_{50}=170.65-275.83\;{\mu}M$) and four lignans ($IC_{50}=114.22-406.10\;{\mu}M$) exhibited significant superoxide and hydroxyl radical scavenging activities, respectively, whereas these compounds contained less activity toward the DPPH and hydroxyl radicals than serotonin derivatives. The levels of serotonin derivatives, lignans and flavonoids in safflower seeds of two cultivars ranged from 49.30 to 260.40, 3.72 to 158.90, and 11.72 to 214.97 mg% (dry base), respectively. Of the two cultivars, 'Cheongsu' had somewthat higher concentrations of phenolic compounds than 'Uisan'. These results suggest that phenolic compounds in safflower seeds may playa role as protective phytochemical antioxidants against reactive oxygen-mediated pathological diseases.

Chemical Standardization of Poria cocos (복령의 품질표준화를 위한 지표성분 탐색 및 정량법 개발)

  • Hoang, Lam;Kwon, Soon-Ho;Kim, Kyung-Ah;Hur, Jong-Moon;Kang, Young-Hwa;Song, Kyung-Sik
    • Korean Journal of Pharmacognosy
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    • v.36 no.3 s.142
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    • pp.177-185
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    • 2005
  • For the purpose of the quality control of Poria cocos, three major compounds were isolated and identified as pachymic acid (1), $3{\beta}-hydroxylanosta-7,9(11),24-trien-21-oic$ acid (2), and dehydroeburicoic acid (3). The optimal extraction conditions for the quantification of pachymic acid its analogues were the 3 hours of reflux with 15g of P. cocos in 100ml 95% ethanol. HPLC conditions were as follows: Column; ZORBAX Eclipse XDB C18 $(4.6{\times}250\;mm,\;Agillent)$, mobile phase; 1% HOAc in 70% $MeOH{\rightarrow}1%$ HOAc in 100% MeOH for 25 min, then 1% HOAc in 100% MeOH for 15 min, detector, ELSD, flow rate; 1ml/min. The mean contents of 1,2, and 3 in Poria cocos cultivated at 18 different site were $0.65{\pm}0.19,\;0.88{\pm}0.72,\;and\;0.84{\pm}0.54\;mg/g$, respectively, and values might be the guide line for the quality control of P. cocos.

Establishment and validation of an analytical method for quality control of health functional foods derived from Agastache rugosa

  • Park, Keunbae;Jung, Dasom;Jin, Yan;Kim, Jin Hak;Geum, Jeong Ho;Lee, Jeongmi
    • Analytical Science and Technology
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    • v.32 no.3
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    • pp.96-104
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    • 2019
  • Agastache rugosa, known as Korean mint, is a medicinal plant with many beneficial health effects. In this study, a simple and reliable HPLC-UV method was proposed for the quantification of rosmarinic acid (RA) in the aqueous extracts of A. rugosa. RA was selected as a quantification marker due to its easiness in procurement and analysis. The developed method involved chromatographic separation on a $C_{18}$ column ($250{\times}4.6mm$, $5{\mu}m$) at room temperature. The mobile phase consisted of water and acetonitrile both containing 2 % acetic acid and was run at a flow rate of $1mL\;min^{-1}$. The method was validated for specificity, linearity, precision, and accuracy. It was specific to RA and linear in the range of $50-300{\mu}g\;mL^{-1}$ ($r^2=0.9994$). Intra-day, inter-day, and inter-analyst precisions were ${\leq}0.91%\;RSD$, ${\leq}1.40%\;RSD$, and 1.94 % RSD, respectively. Accuracy was 93.3-95.9 % (${\leq}1.21%\;RSD$). The method could be applied to three batches of bulk samples and three batches of lab scale samples, which were found to be $0.64({\pm}0.04)mg\;g^{-1}$ and $0.48({\pm}0.02)mg\;g^{-1}$ for the dried raw materials of A. rugosa. The results show that the proposed method can be used as a readily applicable method for QC of health functional foods containing the aqueous extracts of A. rugosa.

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.

Establishment of an Analytical Method for Azorubine, an Undesignated Food Colorant in Korea

  • Kim, Min-ji;Park, Ju-hee;Suh, Hee-Jae;Lee, Chan
    • Journal of Food Hygiene and Safety
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    • v.31 no.5
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    • pp.311-318
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    • 2016
  • Azorubine is a synthetic tar color containing azo-bond in the molecular structure. This food colorant has been allowed to be used for beverages, cheese and dried fruits in the European Union and for some food in Australia. Even though it is applicable as a food color in many countries, this compound has not been permitted in Korea so far as a food additive. Thus, this study was performed to establish an analysis method for azorubine in Korea by comparison of three HPLC analysis methods for azorubine and other azo-compounds which are officially used in the European Food Safety Authority (EFSA, EU), the Food Standard Agency (FSA, England) and the National Institute of Food and Drug Safety Evaluation (NIFDS, Korea). The analysis method of the FSA for azorubine showed the best linearity ($r^2=0.999$), limit of detection (LOD, $0.07{\mu}g/mL$), limit of quantification (LOQ, $0.20{\mu}g/mL$), precision (0~0.5%) and accuracy (98.6~100.7%) among tested HPLC methods using a C-18 column and diode array detector (DAD) with ammonium acetate solution and acetonitrile as an eluent solution. Finally selected method of FSA was further verified by inter-day and intra-day experiments with linearity, LOD, LOQ, precision and accuracy. Recovery test showed the recover ratios of 97~103%, 95~101%, and 93~102% in beverages, breads/snacks and other foods, respectively. Inter-laboratory test represented the absolute value of z-score of less than 2 which means satisfactory levels in this test. Selected method of FSA showed reliable analytical results in application test using food samples collected in commercial markets in Europe.