• Analytical Science and Technology
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• v.19 no.6
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• pp.455-459
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• 2006

The extraction and separation of glabridin from licorice root by HPLC was performed in this work. First, by investigating the different extraction solvents, extraction methods and extraction times, a one-hour ultrasonic extraction procedure with ethyl acetate as the extraction solvent was optimized. Then the ethyl acetate extraction was applied to RP-HPLC for separation of glabridin. The column efficiencies and resolutions were experimentally investigated with different mobile phase compositions. Baseline separation of glabridin was obtained under the mobile phase composition of 50/50 vol.% (ACN/water). The retention time of glabridin was 20.3 min. The peak of glabridin was collected from the HPLC elution for several times and identified by LC/MS. Under the optimum extraction and HPLC separation methods, 1.26 g of glabridin per kg licorice root could be extracted.

• Korean Journal of Pharmacognosy
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• v.44 no.2
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• pp.125-130
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• 2013

Samchulgeonbi-tang (SC) is well-known traditional herbal medicine which is composed of fourteen medicine herbs. SC has been used for the treatment of the chronic gastritis, indigestion, gastric ulcers, gastroptosis and diarrhea disease. The variation in the amount of bioactive components of SC and its fermentation SC with ten Lactobacillus strains were investigated via high-performance liquid chromatography coupled with diode array detection (HPLC-DAD). Simultaneous qualitative and quantitative analysis of seven bioactive compounds; paeoniflorin, liquiritin, hesperidin, liquiritigenin, kaempferol, atractylenolide III, magnolol were achieved by comparing their retention times ($t_R$) and UV spectra with those of the standard compounds. In the results, the amount of paeoniflorin and hesperidin were 7.967 mg/g, 7.251 mg/g that were the main compounds in SC. The amounts of liquiritigenin was increased by all ten Lactobacillus strains, except strain 128. Especially, the liquiritigenin amount was highest in SC fermented with strain 145 (0.201 mg/g), which was increased by 158.39% compared to SC (0.081 mg/g). In the fermented SC using strains 344, almost components were increased than non-fermented SC, except paeoniflorin and kaempferol. Thus, these results considered that the strains 145 and 344 are most excellent fermentation strains among the 10 species of fermentation strains.

• Food Science of Animal Resources
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• v.37 no.3
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• pp.402-409
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• 2017

A novel peptide having free radical scavenging activity was separated, using an on-line high-performance liquid chromatography (HPLC) - ABTS screening method, from bovine skim milk fermented by Lactococcus lactis SL6 (KCTC 11865BP). It was further purified using reverse phase-HPLC (RP-HPLC) and sequenced by RP-HPLC-tandem mass spectrometry. The amino acid sequence of the identified peptide was determined to be Phe-Ser-Asp-Ile-Pro-Asn-Pro-Ile-Gly-Ser-Glu-Asn-Ser-Glu-Lys-Thr-Thr-Met-Pro-Leu-Trp (2,362 Da), which is corresponding to the C-terminal fragment of bovine ${\alpha}_{s1}$-casein (f179-199). The hydroxyl radicals scavenging activity ($IC_{50}$ $28.25{\pm}0.96{\mu}M$) of the peptide chemically synthesized based on the MS/MS data showed a slightly lower than that of the natural antioxidant Trolox ($IC_{50}$ $15.37{\pm}0.52{\mu}M$). Furthermore, derivatives of the antioxidant peptide were synthesized. The antioxidative activity of the derivatives whose all three proline residues replaced by alanine significantly decreased, whereas replacement of two proline residues in N-terminal region did not affect its antioxidative activity, indicating that $3^{rd}$ proline in C-terminal region is critical for the antioxidative activity of the peptide identified in this study. In addition, N-terminal region of the antioxidant peptide did not show its activity, whereas C-terminal region maintained antioxidative activity, suggesting that C-terminal region of the peptide is important for antioxidative activity.

• Journal of Dairy Science and Biotechnology
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• v.21 no.2
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• pp.114-119
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• 2003

Acidocin 4A produced by Lactobacillus acidophilus GP4A was purified to homogeneity by ammonium sulfate precipitation and sequential chromatographies containing Octyl sepharose CL-4B column, $C_{18}$ Sep-Pak Cartridge, $C_{18}$ RP HPLC and HPLC gel filtration. Tricine SDS-PACE resulted in a single band with estimated molecular mass of 4.1 kDa corresponding to the polypeptide weight marker. Electron microscopy of acidocin-treated indicator cells(L. delbrueckii subsp. lactis ATCC 4797) confirmed that acidocin 4A presented bacteriolytic effect, resulting in cell lysis. Curing trial using ethidium bromide (EtBr) was carried out to examine whether acidocin 4A determinant was encoded either by chromosome or on plasmid. The plasmid designated as pLA4A, being about 20 kb in size, was responsible for acidocin 4A production and immunity to host cells.

• Journal of the Korean Applied Science and Technology
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• v.29 no.4
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• pp.585-593
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• 2012

The EO separation and quantitative determination of polysorbate 20, polysorbate 40, polysorbate 60 and polysorbate 80 was carried out by reversed phase HPLC. The water/acetonitrile was used for the mobile phase of gradient conditions. An YMC Pack Ph ($250mm{\times}4.6mm$ i.d., $5{\mu}m$) and Phenomenex C4 ($250mm{\times}4.6mm$ i.d., $5{\mu}m$) and the selected ELSD detector was applied. The analysis results of HPLC showed good linearity with correlation coefficient of $r^2$=0.997 in the rage of $180.2{\sim}980.5{\mu}g/mL$ and detection limit.

• Korean Journal of Pharmacognosy
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• v.43 no.3
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• pp.250-256
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• 2012

In order to extract the curcuminoid such as curcumin, demethoxycurcumin (DMC), and bisdemethoxycurcumin (BDMC) in turmeric (Curcuma longa), solvent extraction methods (dipping and ultrasonic extraction method) and solid-phase extraction (SPE) were used. RP-HPLC (reverse-phase high-performance liquid chromatography) and TLC (thin-layer chromatography) were used for identification and analysis the three curcuminoid. From the experimental results, it is evident that the percentage of curcuminoid extracted from turmeric by ultrasonic extraction method was higher than dipping method. The percentage of curcumin extracted from turmeric by pure methanol was higher than any aqueous methanolic composition. Moreover, the total peak area of three curcuminoid was above 92% in RP-HPLC using solid-phase extraction. These results will form a database for investigating the constituents of natural products and the resources of pharmaceutical, nutrition, and cosmetic products.

• Preventive Nutrition and Food Science
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• v.7 no.1
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• pp.12-17
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• 2002

Simultaneous determination of nine water-soluble vitamins contained in multi-nutrient tablets was carried out by reversed phase high-performance liquid chromatography (RP-HPLC) equipped with analytical $C_{18}$ column and UV (270 nm) detector. Those standard vitamins were successfully separated within 23 minutes by gradient elution with solvent A (0.5 M potassium phosphate monobasic) and solvent B (0.25 M potassium phosphate monobasic-methanol, 1:1). Calibration curves showed good linealities with correlation coefficients (> 0.92) in tested ranged respectively. The detection limits were considered to be 2.1 ng for ascorbic acids 60 ng for Vit B$_{6}$ 3 ng for p-aminobenzoic acid, 9 ng for niacinamide, 9 ng for thiamin, 5.0 ng for folic acid and 1.5 ng for riboflavin at 0.05 a.u.f.s. Solid phase extraction through Sep-Pak (C$_{18}$ ) cartridge was successfully applied for purification of water soluble vitamins in commercial multi-nutrient tablets.ts.

• Analytical Science and Technology
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• v.26 no.3
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• pp.182-189
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• 2013

The purpose of this research is to separate and quantitate selenium species in some food samples with HPLC-ICP-MS. Cation exchange chromatography showed efficient separation only for inorganic Se species while reversed phase ion pair chromatography showed good separation for both inorganic and organic Se species. $C_8$ column ($Symmetryshield^{TM}\;RP_8$, 3.5 ${\mu}m$, $4.6{\times}150$ mm) was used with optimum condition of 5% methanol mobile phase, 0.05% of nonafluorovaleric acid ion pairing reagent. Five standard Se species of Se(IV), Se(VI), SeCys(selenocystein), SeMet(selenomethionine) and Se-M-C(seleno methyl cystein) were separated successfully under the optimum condition (mobile phase; 5% methanol, ion-pairing reagent; 0.05% nonafluorovaleric acid, flow rate; 0.9 mL $min^{-1}$). To extract Se species, microwave assisted and enzyme-assisted extraction methods were studied. In enzyme-assisted extraction method, protease I for garlic, protease I plus trypsin for pork and mackerel, and protease XIV for tuna showed the best extraction efficiency. With the optimum condition for each sample, it was found that mostly inorganic Se, SeCys and SeMet are present in the sample studied ranging from few ${\mu}g$ $g^{-1}$ to few tens of ${\mu}g$ $g^{-1}$.

• Analytical Science and Technology
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• v.26 no.6
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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.

• Journal of Ginseng Research
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• v.14 no.3
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• pp.442-446
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• 1990

As a part of studies on the quality control of crude drug-drink preparations, ginger components were identified by TLC and 6-gingerol was determined quantitatively by HPLC. Ginger components were identified by TLC with benzene/acetone (4:1, v/v, on silica gel plate by spraying a vanillinsulfuric acid reagent. 5-Gingerol contents were determined at 280 nm by HPLC on Lichro CART RP-18 column with acetonitrile/wate(38:62, v/v). Its transfer rate in the 3 types of crude drug extract drinks was 65.4-85.1% compared to the content in the ginger extract.