• KOREAN JOURNAL OF CROP SCIENCE
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• v.50 no.spc1
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• pp.181-186
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• 2005

This experiment was conducted to know the appropriate methods for extraction and determination of rutin contained buckwheat plants. The efficient HPLC analytical condition of rutin contained buckwheat plants was developed. The gradient elution employed a $250mm\times4.6mm$ i.d. Tosoh ODS 120T column. The gradient system was used two mobile phases. A gradient elution was performed with mobile phase A, consisting of $2\%$ Acetic $acid-45\%$ Acetonitrile, and mobile phase B, comprising $2\%$ aqueous acetic acid, and delivered at a flow rate of 1mL/min as follows: 0-18 min, $50-100\%$ A; 18-20 min, $100-50\%$ A; 20-22 min, $50\%$ A. The UV detection wavelength was set at 355 nm. The limit of detection (LOD) for rutin standard compound was 20 ng/mL. And, the higher content of rutin in the extracts was obtained by $80^{\circ}C$ reflex extraction for 120 min. from plants of buckwheat using ethanol.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.53 no.2
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• pp.224-232
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• 2008

The health benefits associated with tea consumption have resulted in the wide inclusion of green tea extracts in botanical dietary supplements, which are widely consumed as adjuvants for complementary and alternative medicines. Tea contains polyphenols such as catechins or flavan-3-ols including (-)-epicatechin (EC), (-)-epigallocatechin (EGC), (-)-epicatechin gallate (ECG), and (-)-epigallocatechin gallate (EGCG), as well as the alkaloid, caffeine. The contents of catechins and caffeine in green tea are considered as a standard of quality evaluation of green tea. Therefor, the purpose of this study was to investigate the most suitable HPLC condition for simultaneous determination of catechins and caffeine in green tea extracts. The efficient HPLC analytical condition of catechins and caffeine contained green tea extracts was developed. The gradient elution employed a $250\;mm\;{\times}\;4.6\;mm$ i.d. YMC-pak ODS-AM 303 column. The gradient system was used two mobile phases. A gradient elution was performed with mobile phase A, consisting of 0.1% aqueous phosphoric acid, and mobile phase B, comprising 100% MeOH, and delivered at a flow rate of 1 mL/min as follows: $0{\sim}25\;min$, 80% A; $26{\sim}50\;min$, $80{\sim}70%$ A; 51 min, 80% A. $51{\sim}55\;min$, 80% A. The UV detection wavelength was set at 280 nm. The limit of detection (LOD) for catechins and caffeine standards were under 50 ng/mL.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.53 no.4
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• pp.359-368
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• 2008

Black soybean has been widely utilized as foods and oriental medicinal materials. The pigmentation in the seed coat of black soybean is due to accumulate anthocyanins in the epidermis palisade layer. The anthocyanins of black soybean seed coat are considered as a parameter of quality evaluation of black soybean. Therefore, the purpose of this study was to investigate the most suitable HPLC condition for simultaneous determination of anthocyanins in black soybean seed coats extracts. The efficient HPLC analytical condition of D3G, C3G, and Pt3G contained extracts of black soybean seed coats was developed. The gradient elution employed a $250\;mm\;{\times}\;4.6\;mm$ i.d. YMC-pak ODS-AM 303 column. The gradient system was used two mobile phases. A gradient elution was performed with mobile phase A, consisting of 5% aqueous formic acid, and mobile phase B, comprising 5% formic acid - acetonitrile, and delivered at a flow rate of 0.7 mL/min as follows: $0{\sim}35\;min$, $90%\;A{\sim}60%\;A$; 36 min, 90% A; 46 min, 90% A. The UV-VIS. detection wavelength was set at 520 nm. The limit of detection (LOD) for D3G, C3G, and Pt3G were under 10 ng/mL.

• Archives of Pharmacal Research
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• v.24 no.5
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• pp.407-411
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• 2001

Reversed-phase high performance liquid chromatographic method was applied for the determination of eleutheroside B and E in the various Acanthopanax species collected In Korea. The stationary phase used was Zorbax 300 SB $C_{18}$ and a mobile phase program was used, which started at 6% acetonitrile for 2 min, and then a linear gradient was operated for the next 18 min to 17% acetonitrile at a flow rate of 1.0 ml/min. The column effluent was monitored at UV 210 nm. Identification was carried out by comparing the retention time and the LC/MS spectrum of each peak corresponding to eleutheroside B and E from sample with those of standards. In general, the contents of eleutheroside B and 1 in stems were higher than those In roots. Acanthopanax species could be classified into two groups based upon the contents of eleutheroside B and E: one group contains no or very little eleutheroside B and another contains both eleutheroside B and E.

• BMB Reports
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• v.29 no.5
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• pp.436-441
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• 1996

The palmitoyl acyl carrier protein (ACP) specific thioesterase (EC 3.1.2.14) from Iris pseudoacorus was purified and characterized. The thioesterase which was very unstable in relatively high salt concentrations was eluted using a co-gradient of Triton X-100 and low concentration of KCl or Na-phosphate from Q-Sepharose, DEAE-Sepharose, and hydroxyapatite chromatography. SDS-PAGE analysis showed a single band with a molecular weight of 35,000. The native molecular weight of approximately 37,000 was estimated by Sephacryl S-200 chromatography, indicating that the enzyme is a monomer. The thioesterase activity was inhibited about 75% and 50% by N-ethylmaleimide (2 mM) and phenylmethylsulfonyl fluoride (2 mM). respectively. The N-ethylmaleimide-inactivation was protected by sodium palmitate but the inactivation with phenylmethylsulfonyl fluoride was not protected. Oxidation of thiols by 2 mM 5.5'-dithio-bis-(2-nitrobenzoic acid) resulted in 65% inactivation of the enzyme. These results suggest that a cysteinyl residue is essential to the catalytic reaction of the enzyme. The enzyme activity was increased by sodium citrate and also by $Cu^{2+}$

• Journal of the Korean Society of Tobacco Science
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• v.31 no.1
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• pp.45-50
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• 2009

In this study, a simple gradient RRLC method for rapid determination of carbonyl compounds of cigarette smoke was developed. Within 10 min, 8 carbonyl compounds have been separated and identified on ZORBAX Eclipse XDB-C18 column ($4.6{\times}50\;mm$, $1.8\;{\mu}m$) with gradient elution using water and acetonitrile as a mobile phase. RRLC was used for the quantification of carbonyl compounds in mainstream smoke of 3R4F reference cigarette, and evaluated those efficiencies in the recovery, repeatability and reproducibility. The correlation coefficients ($r^2$) for calibration curves of carbonyl compounds were over 0.9998. The developed RRLC method was successfully applied to the analysis of smoke samples and the recoveries of carbonyl compounds were in the range of 97.5~102.1% with RSD<3.1%.

• KSBB Journal
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• v.15 no.6
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• pp.605-608
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• 2000

Oligodeoxynucleotides (ODNs) were separated by high-performance membrane chromatography (HPMC), a combined system of chromatography and membrane. The separation mechanism involved anion-exchange, and the stationary phase was cation CIM (Convective Interaction Media) DEAE disk (16${\times}$3 mm). Two types of mobile phase were used, buffer A (20mM Tris-HCl, pH 7.4) and buffer B (buffer A + 1M NaCl). As the amount of NaCl dissolved in buffer linearly increased, the retention time shortened, which enabled a gradient elution mode. Based on the number of theoretical plates and resolution observed, the optimum mobile phase and operating condition (Buffer A/Buffer B=50/50 - 20/80 vol%, gradient time 2 min) were experimentally determined. In this experimental condition, ODNs were separated within 2 min at a mobile phase flow rate of 6 ml/min.

• Natural Product Sciences
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• v.16 no.1
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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.

• Applied Chemistry for Engineering
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• v.17 no.3
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• pp.317-320
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• 2006

The optimum mobile phase condition for analysis of the six purine derivatives (caffeine, guanine, hypoxanthine, purine, theobromine, and theophylline) were determined by a HCI program. Reversed-phase HPLC system was used with the binary mobile phase, water and methanol. Three retention models (Snyder, Langmuir, and Binary polynomial) were considered to predict the retention factors. The elution profiles were calculated by the plate theory based on the binary polynomial retention model. From the final calculated results, the binary polynomial retention model showed the best agreements between the calculated and experimental data. In the isocratic mode, the optimum mobile phase composition of water/methanol is 93/7(v/v). However, we used step-gradient mode to decrease the run-time ($1^{st}$ mobile phase : water/methanol = 93/7 (v/v), gradient time : 5 min, $2^{nd}$ mobile phase : water/methanol = 75/25 (v/v)). The experimental and simulated profiles of above the two conditions show a good agreement.

• Journal of Applied Biological Chemistry
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• v.62 no.1
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• pp.31-38
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• 2019

Scutellaria baicalensis Georgi (Scutellariae Radix) has been widely used as a dietary ingredient and traditional herbal medicine such as diuretic, hyperlipidemia, antibacterial, anti-allergy, anti-inflammatory and anticancer properties. In this study, the isolation of biomarkers or bioactive compounds from complex S. baicalensis extracts represents an essential step for de novo identification and bioactivity assessment. The bioactive fraction consisted of eight compounds which was chromatographed on an analytical high performance liquid chromatography column using two different gradient runs. A simulative replacement of the analytical column with a medium pressure liquid chromatography and open column allowed the determination of gradient profile to allow sufficient separation in the preparative scale. From the optimized method, eight standard compounds have been identified in the fractions. In addition, MS, UV, HRMS detection was provided by ultraperformance liquid chromatographyequadrupole time-of-flight mass spectrometry (UPLC-QTof-MS) of all fractions. Therefore, this scale up procedure was successfully applied to a S. baicalensis extract.