• Journal of Applied Biological Chemistry
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• v.56 no.3
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• pp.137-139
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• 2013

Free radical scavengers in the bisdemethoxycurcumin (BDMC), demethoxycurcumin (DMC) and curcumin of turmeric (Curcuma longa) were screened, identified, quantified and isolation using coupled off-line-2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) and on-line screening high-performance liquid chromatography (HPLC)-ABTS assay. There was a very small margin of error between the off-line-ABTS method and the on-line screening HPLC-ABTS method.

• Journal of Applied Biological Chemistry
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• v.57 no.4
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• pp.301-305
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• 2014

The Antioxidant activity screening identification of five kind compounds in Artemisiae annuae herba with the on-line screening high performance liquid chromatography (HPLC) $ABTS^+$ assay. The various experimental variables such as the extraction time (h) and extraction solvent composition (%) of dipping method were investigated efficiently extraction at the room temperature $25^{\circ}C$. The results, the highest yield of total extract amount (0.458 g, 15.250%) was obtained by dipping method with 100% water and extraction time to 3 h. And the on-line screening HPLC-$ABTS^+$ assay method was rapid and efficient to search for bioactivity from natural products.

• Archives of Pharmacal Research
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• v.17 no.5
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• pp.360-363
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• 1994

Column--swtiching technique with a reversed-phase high performance liquid chromatographic method has been developed for the routine analysis of radioiodinated insulin and its degadation products in biological fluids. The diluted biological samples were loaded onto a precolumn packed with LiChrosorb RP-8 $(25-40{\;}{\mu}m)$ using 0.1% trifuoroacetic acid (TFA) in water as a washing solvent. After valve switching, the concentrated insulins were eluted in the back-flush mode and separated by a W-Porex $C_{18}$ column with a gradient of 0.1% TFA in water and 0.1% TFA in acetonitrile as the mobile phase. The method showed good precision, accuracy and speed with the detection limit of 20 pg/ml. Total analysis time per sample was about 40 min and the coefficients of variation were less than 8, 2%.

• KSBB Journal
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• v.23 no.1
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• pp.96-100
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• 2008

In this work, we describes analysis of the antioxidant potential of Korean green tea phenolics using an high-performance liquid chromatography (HPLC) on-line $ABTS^{+}$ antioxidant screening method. In conjunction with the analysis of their 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid (ABTS+) radical scavenging ability, the extraction of catechine compounds from Korean green tea were performed by various temperature and time. The optimum operating conditions were experimentally determined to analyze the catechine compounds in the pretreatment extracts. From the results, the extraction temperature $60^{\circ}C$, time 3 min was selected as an optimal antioxidant activity condition. The analysis by $C_{18}$ column was performed, the flow rate of mobile phase and UV wavelength was fixed at 1.0 ml/min and 254 nm, respectively. the mobile phase was composed from acetonitrile and water, and the gradient elution mode were applied.

• KSBB Journal
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• v.24 no.2
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• pp.189-194
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• 2009

In this work, antioxidant substance homoorientin from Pseudosasa japonica and Phyllostachys bambusoides leaves wereextracted using ultrasonic system, and analyzed by reversed-phase high performance liquid chromatography (RP-HPLC) on-line ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) antioxidant screening method. Also, the various experimental variables were the frequency and time of ultrasonic system. From the results, homoorientin was high extracted at the experimental condition of low frequency 35 kHz and time 60 min. And the content of homoorientin in Phyllostachys bambusoides was remarkably higher than that in the Pseudosasa japonica extract.

• Biomolecules & Therapeutics
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• v.27 no.2
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• pp.193-200
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• 2019

Ceramide metabolism is known to be an essential etiology for various diseases, such as atopic dermatitis and Gaucher disease. Glucosylceramide synthase (GCS) is a key enzyme for the synthesis of glucosylceramide (GlcCer), which is a main ceramide metabolism pathway in mammalian cells. In this article, we developed a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to determine GCS activity using synthetic non-natural sphingolipid C8-ceramide as a substrate. The reaction products, C8-GlcCer for GCS, could be separated on a C18 column by reverse-phase high-performance liquid chromatography (HPLC). Quantification was conducted using the multiple reaction monitoring (MRM) mode to monitor the precursor-to-product ion transitions of m/z $588.6{\rightarrow}264.4$ for C8-GlcCer at positive ionization mode. The calibration curve was established over the range of 0.625-160 ng/mL, and the correlation coefficient was larger than 0.999. This method was successfully applied to detect GCS in the human hepatocellular carcinoma cell line (HepG2 cells) and mouse peripheral blood mononuclear cells. We also evaluated the inhibition degree of a known GCS inhibitor 1-phenyl-2-decanoylamino-3-morpholino-1-propanol (PDMP) on GCS enzymatic activity and proved that this method could be successfully applied to GCS inhibitor screening of preventive and therapeutic drugs for ceramide metabolism diseases, such as atopic dermatitis and Gaucher disease.

• KSBB Journal
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• v.23 no.4
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• pp.297-302
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• 2008

In this work, the amounts of Flavone C-glycosides homoorientin, orientin extracted from Black bamboo by various ultrasonic waves frequency (35, 72, 170 KHz, 300 Watt $\pm$1) time (15, 30, 60 min) and temperature ($25^{\circ}C$) were compared using 50% aqueous ethanol solution. And describesanalysis of the antioxidant potential of Black bamboo using an high-performance liquid chromatography (HPLC) on-line $ABTS^+$antioxidant screening method. In conjunction with the analysis of their 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid ($ABTS^+$) radical scavenging ability. The optimum operating conditions were experimentally determined to analyze the Flavone C-glycosides homoorientin, orientin in the pretreatment ultrasonic extracts. From the results, the effect on extraction yield of variations in frequency, solvent composition and extraction time was investigated. The highest yield of Black bamboo was obtained by ultrasonic waves with a frequency of 72 KHz and an extraction time of 60 min, The extraction frequency 35 KHz ($25^{\circ}C$), time 30 min was selected as an optimal antioxidant activity condition.

• 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.