• Journal of Applied Biological Chemistry
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• 제66권
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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.

• 대한한의학방제학회지
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• 제18권2호
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• pp.251-257
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• 2010

Objectives : To develop and validate High-performance liquid chromatography-photodiode array methods for simultaneous determination of two constituents in Oryeong-san(ORS). Methods : Reverse-phase chromatography using a Gemini C18 column operating at $40^{\circ}C$, and photodiode array(PDA) detection at 280 nm, were used for quantification of the two marker components of ORS. The mobile phase using a gradient flow consisted of two solvent systems. Solvent A was $H_2O$ and solvent B was acetonitrile. Results : Calibration curves were acquired with correlation coefficient ($r^2$)>0.9999, and the relative standard deviation(RSD) values(%) for intra- and inter-day precision were not exceed 1.0%. The recovery rate of each compound was in the range of 93.01-104.16%, with an RSD less than 2.0%. The contents of two compounds in ORS were 1.10-3.72 mg/g. Conclusions : The established HPLC method will be helpful to improve quality control of ORS.

• Journal of Applied Biological Chemistry
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• 제66권
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• pp.320-327
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• 2023

Capsicum annuum belongs to the Solanaceae family, crops of which are extensively cultivated worldwide. It is a food source containing various nutrients and vitamins and also serves as a medicine for treating ailments. The burning feeling experienced while consuming Capsicum fruits is due to the presence of capsaicinoids, particularly capsaicin and dihydrocapsaicin. This study aimed to assess the content of these two compounds in 34 varieties of capsicum and paprika. High-performance liquid chromatography with a gradient elution system and a reverse-phase YMC Pack-Pro column with UV detection at 280 nm was employed. The results revealed that, among the 34 samples, only six samples (samples 1, 15, 20, 29, 32, and 34) contained capsaicin and dihydrocapsaicin, and their highest contents were found in sample 1 - variety name: Sungil-c (capsaicin: 3.42 mg/g extract, dihydrocapsaicin: 1.20 mg/g extract). These findings suggest that the content of these two compounds is attributed to the variety and is influenced by geographical location and environmental factors. Additionally, this study provides a basis for establishing a C. annuum variety with high capsaicin and dihydrocapsaicin contents.

• 분석과학
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• 제9권2호
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• pp.198-202
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• 1996

Histamine을 정확하고 신속하게 정량하기 위해 9-Fluorenylmethyl chloroformate를 형광유도체화제로 하여 역상 HPLC법으로 정량하였다. 히스타민을 형광유도체화할 때 반응액의 pH, 반응시간, 형광유도체화제의 농도 등 최적 반응 조건을 검토하였다. 이 방법으로 히스타민을 분석한 결과 $0.1{\sim}0.5{\mu}g/ml$의 농도범위에서 상관계수가 0.992인 양호한 직선성을 나타내였으며 검출한계는 $0.01{\mu}g/ml$였다.

• Journal of Ginseng Research
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• 제44권4호
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• pp.563-569
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• 2020

Background: White ginseng consists of the roots and rhizomes of the Panax species, and red ginseng is made by steaming and drying white ginseng. While red ginseng has both polar and nonpolar ginsenosides, previous studies showed white ginseng to have only polar ginsenosides. Because nonpolar ginsenosides are formed through the manufacture of red ginseng from white ginseng, researchers have generally thought that nonpolar ginsenosides do not exist in white ginseng. Methods: We developed a simultaneous quantitative method for six nonpolar ginsenosides in white ginseng using reverse-phase high-performance liquid chromatography coupled with integrated pulsed amperometric detection. The nonpolar ginsenosides of white ginseng were extracted for 4 h under reflux with 50% methanol. Results: Using the gradient elution system, all target components were completely separated within 50 min. Nonpolar ginsenosides were determined in the rhizome head (RH), main root (MR), lateral root, and hairy root (HR) of 6-year-old white ginseng samples obtained from several regions (Geumsan, Punggi, and Kanghwa). The total content in the HR of white ginseng was 37.8-56.8% of that in the HR of red ginseng. The total content in the MR of white ginseng was 5.9-24.3% of that in the MR of red ginseng. In addition, the total content in the RH of white ginseng was 28.5-35.8% of that in the HR of red ginseng Conclusion: It was confirmed that nonpolar ginsenosides known to be specific components of red ginseng were present at substantial concentrations in the HR or RH of white ginseng.

• 한국수산과학회지
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• 제39권2호
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• pp.59-65
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• 2006

Fluoroquinolones are the most common group of antibacterial agents currently used in the Korean aquaculture industry, and use of these agents has been increasing steadily. High performance liquid chromatography (HPLC) with fluorescence detection was used for the simultaneous determination of five fluoroquinolones in fish and shellfish: ofloxacin (OFL), pefloxacin (PEF), norfloxacin (NOR), ciprofloxacin (CIP), and enrofloxacin (ENRO). Fish and shellfish muscle was homogenized, and protein, lipid, and low molecular weight pigments were then excluded from the homogenate. The final eluates were analyzed by HPLC equipped with a Shiseido UG-120 type C18 reverse-phase column ($4.6{\times}250 mm$, $5{\mu}m$) and a fluorescence detector (excitation at 280 nm, emission at 450 nm). The mobile phase was 0.1 M phosphoric acid and acetonitrile solution (91:9, v/v) and tetrahydrofuran (THF) was added to it at a rate of 5 mL per a liter of the mobile phase. Adequate chromatography separation was obtained using the above method. Average recoveries of fortified samples at levels from 0.05 to 0.5 mg/kg were $72.3{\pm}2.5-84.5{\pm}1.2%$ for OFL, $82.7{\pm}3.3- 109.3{\pm}7.5%$ for NOR, $85.3{\pm}6.6-116.0{\pm}7.9%$ for PEF, $76.0{\pm}4.3-109.3{\pm}12.4%$ for CIP, and $78.7{\pm}5.9-100.0{\pm}9.8%$ for ENRO. The limit of detection of OFL was $5{\mu}g/L$, the others were $1{\mu}g/L$. We concluded that the new analytical method was suitable for the determination of fluoroquinolones in fish and shellfish.

• 분석과학
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• 제26권6호
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• pp.357-363
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• 2013

Selenium-containing proteins were separated from selenium-enriched yeast (SEY) using Trizol$^{(R)}$ reagent followed by anion exchange (AE) chromatography. This method is simpler and less time consuming than electrophoresis. Five selenium containing proteins were identified by on-line AE HPLC-ICP/MS (high performance liquid chromatography-inductively coupled plasma/mass spectrometry). Each protein was enzymatically hydrolyzed to seleno-amino acids and separated with RP (reverse phase) HPLC for the identification of selenoproteins.

• Archives of Pharmacal Research
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• 제21권3호
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• pp.291-297
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• 1998

Cop protein has been overexpressed in Escherichia coli using a T7 RNA polymerase system. Purification to apparent homogeneity was achieved by the sequential chromatography on ion exchange, affinity chromatography, and reverse phase high performance liquid chromatography system. The molecular weight of the purified Cop was estimated as 6.1 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). But the molecular mass of the native state Cop was shown to be 19 kDa by an analytical high performance size exclusion chromatography, suggesting a trimer-like structure in 50 mM Tris-HCI buffer (pH 7.5) containing 100 mM NaCl. Cop protein Was calculated to contain $39.1% {\alpha}-helix, 16.8% {\beta}-sheet$, 17.4% turn, and 26.8% random structure. The DNA binding property of Cop protein expressed in E. coli Was preserved during the expression and purification process. The isoelectric point of Cop was determined to be 9.0. The results of amino acid composition analysis and N-terminal amino acid sequencing of Cop showed that it has the same amino acid composition and N-terminal amino acid sequence as those deduced from its DNA sequence analysis, except for the partial removal of N-terminal methionine residue by methionyl-aminopeptidase in E. coli.

• Natural Product Sciences
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• 제16권1호
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• pp.39-42
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• 2010

The content of hyperin in Acanthopanax species was determined by high performance liquid chromatography (HPLC). Hyperin was quantified by a reverse-phase column with elution program [initially gradient solvent (acetonitrile : water = 85 : 15 to 80 : 20 for 20 min), then isocratic solvent (acetonitrile : water = 80 : 20 for 20 min), and finally gradient solvent (acetonitrile : water = 80 : 20 to 65 : 35 for 20 min)]. UV detection was conducted at 210 nm. The content of hyperin in the fruits of Acanthopanax was measured in the species A. chiisanensis (2.04 mg/g), A. sessiliflorus (1.13 mg/g), A. divaricatus (0.98 mg/g), A. koreanum (0.75 mg/g) and A. senticosus (0.05 mg/g). The content of hyperin in A. chiisanensis was higher than that of other Acanthopanax species.

• Journal of Applied Biological Chemistry
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• 제66권
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• pp.153-158
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• 2023

Pinus species are native to the Northern Hemisphere and some parts of the tropics to temperate regions in the Southern Hemisphere. They were used as food and medicine in prehistoric times. Massonianoside B is a compound found in pine trees and possesses antioxidant activity. In order to determine the presence and content of this compound in Pinus species, three different parts (needles, branches, and bark) of three Pinus species were extracted and investigated. High-performance liquid chromatography with a gradient elution system along with a reverse-phase INNO column with photodiode array detector was employed. Results showed that the branches of the three Pinus species had higher massonianoside B content (5.502 to 9.751 mg/g DW) than either the needles or bark. Furthermore, among the three species, P. rigida × P. taeda had the highest concentration of total massonianoside B (11.557 mg/g DW). These findings thus provide evidence of biological activity in Pinus species and establish a foundation for further research.