• Title/Summary/Keyword: Reverse phase HPLC

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HPLC analytical method validation of Aralia elata extract as a functional ingredients (두릅 추출물의 기능성 원료 표준화를 위한 HPLC 분석법 검증)

  • Ahn, Eun-Mi;Choi, Song-Am;Choi, Ji-Young
    • Food Science and Preservation
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    • v.24 no.6
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    • pp.795-801
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    • 2017
  • Aralia elata Seemann (AE) has long been used as a folk medicine for the treatment of various diseases including diabetes mellitus, anti-arthritic, and anti-gastric ulcer agent in Korea, Japan, and China. This study was performed to establish a simple and reliable HPLC/UV analytical method for determination of most active anti-hypertensive compound, a 3-O-${\alpha}$-L-rhamnopyranosyl($1{\rightarrow}$2)-${\alpha}$-L-arabinopyranosyl hederagenin 28-O-${\beta}$-D-xylopyranosyl($1{\rightarrow}6$)-${\beta}$-D-glucopyranosylester (HE) for the standardization of the shoot extract of AE as a health functional food ingredient. The quantitative analytical method of HE was optimized by HPLC analysis using reverse-phase C18 column at $40^{\circ}C$ with $H_2O$ and acetonitrile (70:30, v/v) as an isocratic mobile phase at a flow rate of 1.0 mL/min and detection wavelength of UV 205 nm. This HPLC/UV analytical method showed good specificity and high linearity in the tested range of 0.03125-2.0mg/ml with excellent coefficient of determination ($R^2$) of 0.9999. The limit of detection and limit of quantification were $12.0{\mu}g/mL$ and $36.5{\mu}g/mL$, respectively. Relative standard deviation (RSD) values of data from intra- and inter-day precision were less than 0.2% and 0.1%, respectively. These results indicate that the established HPLC/UV analytical method is very simple, specific, precise, accurate, and reproducible and thus can be useful for the quantitative analysis of HE as a functional anti-hypertensive compound in AE extract.

Measurement of Carbonyl Compounds in Ambient Air using a DNPH Cartridge coupled with HPLC Method (DNPH 카트리지와 HPLC를 이용한 대기 중 카르보닐화합물의 농도측정)

  • 황윤정;박상곤;백성옥
    • Journal of Korean Society for Atmospheric Environment
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    • v.12 no.2
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    • pp.199-209
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    • 1996
  • The purpose of this study is to evaluate analytical method for the measurement of carbonyl compounds and to apply method to the measurement of indoor and outdoor concentrations of these compounds at public facilities. For sampling, 2.4-dinitrophenylhydrazine (DNPH) coated DNPH-Silica cartridges were uwed in this study. DNPH reacts with carbonyl compounds and forms carbonyl hydrazone, The carbonyl hydarzone was eluted from the cartridge with acetonitrile and analyzed by reverse-phase HPLC with UV detection. Possible interference caused by ozone during sampling was eliminated by using KI trap commected in series with the DNPH-Silica cartridge. A number of experimental studies were undertaken to evaluate and validate the analytical method, including collection efficiency, recovery, repeatability, lower limits of detection, and effect of ozone. Indoor and outdoor measurements of carbonyl compounds were simultaneously carried out at 14 public facilities located in Taegu city and Kyungsan city from June to July, 1995. Except for one or two sites, the indoor concentrations were found to be higher than the outdoor concentrations for carbonyl compounds. And the concentrations of carbonyl compounds measured in the morning and afternoon were showed higher than the concentrations measured in the evening.

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Analysis and Stability test of the Extract of Coptidis Rhizoma and Salviae Miltiorrhizae Radix for Toxicity Study (황련과 단삼의 독성평가를 위한 성분분석 및 안정성 시험)

  • Lee, Mi-Kyoung;Lee, Jun-Seok;Kwack, Seung-Jun;Kim, Ji-Myoung;Kang, Tae-Suk;Lee, Je-Hyun;Woo, Mi-Hee;Choi, Jae-Sue;Bae, Ki-Hwan;Min, Byung-Sun
    • Korean Journal of Pharmacognosy
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    • v.40 no.3
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    • pp.184-189
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    • 2009
  • A simple and reliable reverse phase HPLC method of Coptidis Rhizoma and Salviae Miltiorrhizae Radix were developed to determine pharmacologically active marker compounds. The water-extract of two natural medicines were evaluated by the HPLC methods. The stability test for two extract were examined for five months. However, no significant change in the content of the marker compounds of each extract observed during the time of investigation.

Protein Methylase Inhibitor from Porcine Liver : Purification and Properties (돼지 간장 조직에서 얻은 단백질 메칠라제 저해제의 정제와 특성)

  • 박선미;박연호;백운기;이향우
    • YAKHAK HOEJI
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    • v.37 no.2
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    • pp.149-157
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    • 1993
  • Protein methylase inhibitor which is a modulator of biological methylation has been purified and characterized from porcine liver soluble fraction by cell fractionation, Sephadex G25 chromatography, reverse phase HPLC, size exclusion HPLC. The results are summarized as follows. 1) The purified inhibitor shows apparent homogeneity, as judged by HPLC. 2) A molecular weight of the purified inhibitor which is composed of 18 amino acid residues is about 1,400 daltons. 3) A single absorption peak of ultraviolet spectrum was observed at 260nm. 4) The inhibitor was not inactivated by heating at $100^{\circ}C$ until 60min. and its activity was not influenced by treatment with digestive enzymes, such as trypsin, pepsin, pronase, chymotrypin, lysozyme, DNase, and RNase. 5) The purified inhibitor inhibited protein rnethylase I, II, III and phospholipid methyltransferase activities. 6) The purified inhibitor inhibited noncompetitively protein methylase II from porcine liver, spleen, and testis. 7) The $K_{i}$ values for protein methylase II from porcine liver, spleen, and testis were 300nM, 250nM, 297nM, respectively.

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Determination of Luteolin in the Different Parts of Lithospermum erythrorhizon by HPLC (HPLC를 이용한 지치중의 부위별 Luteolin 정량)

  • Han, Saem;Park, Jun-Yeon;Lee, Sul-Lim;Mok, So-Youn;Kim, Hye-Min;Lee, Jeong-Min;Ahn, Young-Hee;Lee, Sang-Hyun
    • Korean Journal of Medicinal Crop Science
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    • v.18 no.5
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    • pp.311-314
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    • 2010
  • The content of luteolin in the different parts of Lithospermum erythrorhizon was determined by high performance liquid chromatography (HPLC). Luteolin was quantified by a reverse-phase column with gradient solvent program (water : ACN = 90 : 10 to 77 : 23 for 60 min). UV detection was conducted at 330 nm. The content of luteolin was measured in the seeds (0.28 mg/g), stems (0.30 mg/g), and roots (0.16 mg/g) of L. erythrorhizon. The presence of luteolin in Lithospermum species is important in agricultural crop production for increasing the amounts of clinically available medicine and health supplements.

Pharmacokinetic Characteristics of Oral Clonazepam in Korean (한국인에서의 경구용 클로나제팜의 약물동력학적 특성)

  • 정지훈;유대식;박준홍;이석용;손의동
    • YAKHAK HOEJI
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    • v.48 no.5
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    • pp.278-284
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    • 2004
  • In order to determine the bioavailability of c1onazepam, an anxiolytic drug, a simple, rapid and sensitive HPLC analysis was developed in healthy Korean volunteers. The analysis system was validated in specificity, accuracy, precision and linearity. The analysis condition we established was 2.58 min and 5 ng/$m\ell$ in retention time and limit of quantitation of c1onazepam, respectively, using reverse-phase C18 column connected to UV detector. Quantitation was performed at 235 nm wave length with p-hydroxybenzoic acid ethyl ester as internal standard. The method involved a simple extraction. In order to study blood level profiles as a function of time, eight volunteers were enrolled and orally took 6 mg clonazepam once. The blood samples were collected from 0 to 120 h after the drug administration. Mean AUC and Cmax value were 1028.17$\pm$568.165 (ng/$m\ell$$.$hr) and 41.25$\pm$10.82 (ng/$m\ell$), respectively. And mean Tmax and T$_{1}$2/ value were 1.08$\pm$0.42 (hr) and 30.78$\pm$3.26 (hr). From the results we determine the pharmacokinetic characteristics of clonazepam in Korean people using a newly developed and useful HPLC method.

Determination of Silymarin and Silybin Diastereomers in Korean Milk Thistle using HPLC/UV Analysis (HPLC 분석을 통한 한국산 밀크씨슬 중 실리마린과 실리빈의 정량)

  • Kim, Juree;Paje, Leo Adrianne;Choi, Jung Won;Lee, Hak-Dong;Shim, Jae Suk;Shim, Jeehyoung;Geraldino, Paul John L.;Lee, Sanghyun
    • Korean Journal of Pharmacognosy
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    • v.51 no.4
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    • pp.297-301
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    • 2020
  • Silymarin (SM) and silybin diastereomers (SD) in milk thistle (Silybum marianum) were determined using high-performance liquid chromatography and quantified using a reverse-phase column in a gradient elution system. UV detection was performed at 288 nm. The content of SM and SD in milk thistle was 3.236 and 0.553 mg/g DW, respectively. Determining the presence and quantifying the content of SM and SD in milk thistle are vital for the pharmaceutical industry to identify optimal sources for developing health supplements or therapeutics.

Development of an analytical method for the quantification of oleanonic acid from mastic gum using HPLC/PDA

  • Hak-Dong Lee;Chang-Dae Lee;So Yeon Choi;Sanghyun Lee
    • Journal of Applied Biological Chemistry
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    • v.66
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    • pp.67-72
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    • 2023
  • A simple and accurate method was developed for the quantitative analysis of oleanonic acid (OA) from mastic gum. The analysis was carried out using reverse-phase high-performance liquid chromatography combined with a photodiode array detector (HPLC/PDA). Our optimized method was validated by measuring various parameters, using an INNO C18 column fitted with a gradient elution system. The results revealed limits of detection and quantification of 0.34 and 1.042 ㎍/mL, respectively. The OA calibration curve exhibited excellent linearity over the concentration range of 0.0625 to 2.0 mg/mL, with r2 =0.9996. Accuracy tests revealed a high recovery rate of 99.44-103.66%, with precision values below 0.15%. These results suggest that the present analytical method can identify and quantify OA in mastic gum with high precision. The HPLC approach developed in this study might be applied to routine analyses and large-scale extraction procedures for OA content quantification.

Fractionation of Angiotensin Converting Enzyme(ACE) Inhibitory Peptides from Soybean Paste (된장으로부터 Angiotensin Converting Enzyme(ACE) 저해 Peptide의 분획)

  • Shin, Zae-Ik;Ahn, Chang-Won;Nam, Hee-Sop;Lee, Hyung-Jae;Lee, Hyung-Joo;Moon, Tae-Hwa
    • Korean Journal of Food Science and Technology
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    • v.27 no.2
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    • pp.230-234
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    • 1995
  • Angiotensin converting enzyme(ACE) inhibitory peptides lowering blood pressure were fractionated from a commercial soybean paste(Doenjang). When the freeze-dried sample of soybean paste was extracted with cold water, the recovery yield of total nitrogen(TN) was shown to be 73.3% in 30 minutes. The cold water extract was filtered through PM-10 membrane(Amicon) for 3 hours in order to remove high molecular weight polypeptides. The TN and salt of ultrafiltrate were recovered to 80.8% and 99.2%, respectively, and its ACE $IC_{50}$ was $41.8{\mu}g/ml$. When the ultrafiltrate was divided into 7 fractions by reverse phase prep-HPLC, F5 fraction showed the highest ACE inhibitory activity ($IC_{50}=6.8{\mu}g/ml$) and salt could be collected into F1 fraction. Subsequently, the F5 fraction was divided into another five fractions by ion exchange prep-HPLC, all of which appeared to be high ACE inhibitory activity($IC_{50}=2.5{\sim}8.3{\mu}g/ml$). Among them, F53 fraction had the highest ACE inhibitory activity, and its main amino acid component was found to be histidine.

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Isolation of Bacillus velezensis SSH100-10 with Antifungal Activity from Korean Traditional Soysauce and Characterization of Its Antifungal Compounds (전통재래 간장으로부터 항진균 활성 B. velezensis SSH100-10의 분리와 그 항진균 물질의 특성 구명)

  • Chang, Mi;Moon, Song Hee;Chang, Hae Choon
    • Food Science and Preservation
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    • v.19 no.5
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    • pp.757-766
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    • 2012
  • The SSH100-10 bacterial strain, which exhibits strong antifungal (anti-mold and anti-yeast) activity, was isolated from traditional korean soysauce aged 100 years. The strain was identified as Bacillus velezensis based on Gram-staining, the biochemical properties and 16S rRNA gene sequence determination. B. velezensis SSH100-10 showed strong proteinase activity and NaCl tolerance, but did not produce enterotoxin. Two-antifungal compounds from B. velezensis SSH100-10 were purified using SPE, preparative HPLC, and reverse phase-HPLC. The purified antifungal compounds were identified as $C_{14}$ and $C_{15}$ iturin through MALDI-TOF-MS and amino acid composition analysis. The stability characteristics of the antifungal compounds after temperature, pH, and enzyme treatments suggested that B. velezensis SSH100-10 produced more than two antifungal compounds; pH-stable $C_{14}$ iturin A and $C_{15}$ iturin A, and unidentified pH-unstable compounds. The results suggested that B. velezensis SSH100-10 can be used in soybean fermentation as a starter. Moreover it has potential as a biopreservative in the food and feed industry and as a biocontrol agent in the field of agriculture.