• Journal of Applied Biological Chemistry
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• v.64 no.1
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• pp.57-61
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• 2021

Saikosaponin derivatives such as saikosaponins A, B1, B2, B3, B4, C, and D present in Bupleurum falcatum were analyzed by a high performance liquid chromatograph equipped with an evaporative light scattering detector, using different extraction solvents (water and 70% ethanol). The samples were injected into a YMC Pack Pro C18 column and separated using a gradient elution system with a mobile phase composed of acetonitrile and water at a flow rate of 1.1 mL/min. The content of saikosaponin derivatives was higher in 70% ethanol extract than in water extract. This study provides an efficient analytical method for determining the optimal conditions for extraction of saikosaponin derivatives, which can be used as a basis for development of functional foods and pharmaceutical products from B. falcatum.

• Journal of Applied Biological Chemistry
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• v.64 no.1
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• pp.13-17
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• 2021

Qantitative analysis of six compounds: (+)-catechin, benzoic acid, gallic acid methyl ester, paeonol, paeoniflorin, and albiflorin from Paeonia lactiflora extracts was performed using high-performance liquid chromatography and an ultraviolet (UV) detector, following different extraction methods. A reverse-phase column was used in a gradient elution system, and UV detection was performed at 280 nm. The results showed that the quantity of paeoniflorin was the highest in ethanol and water extracts (73.89 and 57.87 mg/g, respectively) among the six compounds. This study contributes a good analysis method for the contents of P. lactiflora and would be propitious for developing medicines and functional foods.

• Journal of Korean Society on Water Environment
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• v.25 no.5
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• pp.720-726
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• 2009

We investigated the most effective pre-treatment processes and LC/MS/MS condition for microcystins analysis. With a step-by-step pre-treatment, efficiencies of several established methods were compared. At the level of cell burst, sonication method was found to be the most efficient. As a mycrocystins first extraction solvent, 5% acetic acid showed the highest efficiency. An isolation and recovery rate of mycrocystins of ODS Sep-Pak $C_{18}$ cartridge was higher than HLB SPE cartridge. As a final elution solvent from cartridge, 100% MeOH had a better efficiency than others. Using a LC/MS/MS, effective analytical methods were established. C18 reverse column was used and gradient elution was performed with using acetonitrile, 0.1% formic acid as a mobile phase. We analysed to 0.8 mL/min flow rate fit to the $5{\mu}m$ particle size column and $55^{\circ}C$ housing temperature. The validity of established analytical method was evaluated that MDL as average $0.050{\pm}0.014{\mu}g/L$ and LOQ as average $0.160{\pm}0.045{\mu}g/L$ had a good sensitivity over 40 magnification rather than $2{\mu}g/L$ detection limit of HPLC.

• Analytical Science and Technology
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• v.19 no.6
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• pp.519-528
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• 2006

An analytical method for the determination of thyroid hormones in urine samples has been studied by using solid-phase extraction and high-performance liquid chromatography/diode array detector/electro-spray mass spectrometry. Seven thyroid hormones were successfully separated by gradient elution on the reverse phase Hypersil ODS column (4.6 mm I.D., 100 mm length, particle size $5{\mu}m$) with ammonium formate buffer and acetonitrile, and UV spectra and mass fragment could be confirmed. The extraction recoveries of thyroid hormones in the urine samples (pH 3) were in the range of 89.0-113.1% with solid-phase extraction by C18, followed by elution with 4 ml of methanol/ammonium hydroxide (9 : 1). The calibration curves showed good linearity with the correlation coefficients ($r^2$) varying from 0.992 to 0.998 and the detection limits of all analytes were obtained in the range of 2-4 ng/ml (3.8-13.0 pmol/ml).

• Korean Journal of Food Science and Technology
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• v.50 no.4
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• pp.377-382
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• 2018

An HPLC method was developed to quantitate a marker, oxyresveratrol (ORT), for the standardization of mulberry branch extracts (MBE) as a functional ingredient. HPLC was performed on a $C_{18}$ column with a gradient elution using 0.05% $H_3PO_4$ and acetonitrile at a flow rate of 0.8 mL/min, and detected at 320 nm. The HPLC method was validated according to Korea Food and Drug Administration (KFDA) guideline of analytical procedures with respect to specificity, linearity, accuracy and precision. Calibration curve of ORT showed high linearity ($R^2=1$), and limits of detection and quantification were 0.3 and $1.0{\mu}g/mL$, respectively. Relative standard deviation values from intra-and inter-day precision were less than 3.52 and 4.70%, respectively. Recovery rate ranged from 97.64% to 103.69%, and ORT content in MBE was approximately 3.78%. These results suggest that the HPLC method developed for the analysis of ORT in MBE is simple, efficient, and could contribute to the quality control of MBE.

• Mass Spectrometry Letters
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• v.4 no.3
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• pp.47-50
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• 2013

An ultra-fast generic LC-MS/MS method was developed for high-throughput quantification of discovery pharmacokinetic (PK) samples and its reliability was verified. The method involves a simple protein precipitation for sample preparation and the analysis by ultra-fast generic LC-MS/MS with the ballistic gradient program and selected reaction monitoring (SRM) mode. Approximately 290 new chemical entities (NCEs) (over 10,000 samples) from 5 therapeutic programs were analyzed. The calibration curves showed good linearity in the concentration range of 1, 2 or 5 to 2000 ng/mL. No significant ion suppression was observed in the elution region of all the NCEs. When approximately 300 plasma samples were continuously analyzed, the peak area of internal standard was constant and reproducible. In the repeated analysis of samples, the plasma concentrations and the area under the curve (AUC) were consistent with the results from the first analysis. These results showed that the present ultra-fast generic LC-MS/MS method is reliable in terms of selectivity, sensitivity, and reproducibility and could be useful for high-throughput quantification and other bioanalysis in drug discovery.

• Korean Journal of Microbiology
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• v.21 no.4
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• pp.221-228
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• 1983

An improved procedure for the rapid purification of glucose-6-phosphate dehydrogenase from extracts of Saccharomyces cerevisiae was developed by using affinity chromatography. Among six affinty media tested, $NADP^+ -agarose$ and Affi-gel Blue were more effective than others (i.e., Affi-gel Red, AMP-agarose, ATP-agarose, and $NAD^+ -agarose$). Conditions to desorb the enzyme bound to the affinity media were examined to increase the purity as well as yield. The best result was obtained when the column was developed with a linear gradient of KCl (0-1.0M). In case of Affi-gel Blue, introduction of $NAD^+$ (15mM) washing step prior to the salt gradient was most effective to remove $NAD^+ -binding$ proteins. For a large scale preparation of G-6-P dehydrogenase higher recovery was obtained by Affi-gel Blue than $NADP^+ -agarose$, however, the purity of the enzyme was decreased by 10 times if the former was used as the affinity medium. The capacity of Affi-gel Blue for G-6-P dehydrogenase was found to be 5 times higher than that of $NADP^+ -agarose$. Furthermore Affi-gel Blue could be reused repeatedly and its preparation is relatively easier and less expensive than $NADP^+ -agarose$.

• Journal of Microbiology and Biotechnology
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• v.7 no.5
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• pp.310-317
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• 1997

Enzymatic synthesis of glucosyl-sugar alcohols using various transglycosylating enzymes, such as cyclodextrin glucanotransferase (CGTase), ${\alpha}$-amylase, ${\alpha}$-glucosidase, and pullulanase was investigated using various sugar alcohols, such as sorbitol, xylitol, inositol, maltitol, and lactitol as glucosyl acceptors. CGTase showed the highest transglycosylating activity to sugar alcohols compared to other transglycosylating enzymes, and inositol and maltitol were the most suitable glucosyl acceptors. Soluble starch, extruded starch, cyclodextrins, and maltooligosaccharides were also identified to be adequate glucosyl donors for transglycosylation reaction of CGTase to sugar alcohols. The synthesis of glucosyl-maltitol in the reaction system using extruded starch as the glucosyl donor and maltitol as the glucosyl acceptor showed the best results showing the highest transglycosylation yield. The transglycosylation products were purified by activated carbon column chromatography with ethanol gradient elution. Chemical structures of above transglucosylated products were analyzed by nuclear magnetic resonance spectroscopy, and two products were identified to be maltotritol and maltotetraitol, in which one or two glucose molecules attached to the parent maltitol molecule by a ${\alpha}$-l,4-glucosidic bond, respectively.

• Food Science and Biotechnology
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• v.17 no.2
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• pp.430-433
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• 2008

A high performance liquid chromatography (HPLC) method has been successfully developed to identify and quantify major ginsenosides in Korean ginseng seeds and roots. Using gradient elution of acetonitrile and water without buffer, the 6 major ginsenosides ($Rb_1,\;Rb_2$, Rc, Rd, Re, and $Rg_1$) were identified. Compared with ginseng roots, the amount of ginsenoside Re and Rd in ginseng seeds were significantly higher than those in ginseng roots (p<0.05). In ginseng seeds, the content of protopanaxtriol (PPT) was higher than that of protopanaxdiol (PPD) and the ratio of PPT and PPD was approximately 2.2 : 1. However, the content of PPT was lower than that of PPD in ginseng roots. It should be mentioned that both content of PPT and PPD in ginseng seeds were much higher than those in ginseng roots.

• Journal of Haehwa Medicine
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• v.23 no.1
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• pp.93-104
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• 2014

Glycyrrhizae Radix et Rhizoma has been extensively used by human beings as a medicinal herb as well as natural sweetener. In this study, we performed quantification analysis of three major constituents including liquiritin, glycyrrhizin, and glycyrrhetinic acid in the 70% ethanol extracts of non-processed Glycyrrhizae Radix et Rhizoma and processed Glycyrrhizae Radix et Rhizoma using a high-performance liquid chromatography coupled with photodiode array detector. The analytical column for separation of the 3 constituents used a Gemini C18 column kept at $40^{\circ}C$ by the gradient elution of two mobile phase. The amounts of liquiritin, glycyrrhizin, and glycyrrhetinic acid in non-processed Glycyrrhizae Radix et Rhizoma were 2.57%, 3.52%, and not detected. After processing by roasting, the best roasting temperature and time of iquiritin, glycyrrhizin, and glycyrrhetinic acid were $160^{\circ}C$-15 min (2.46%), $160^{\circ}C$-15 min (3.67%), and $240^{\circ}C$-15 min (0.76%), respectively.