• Analytical Science and Technology
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• v.5 no.1
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• pp.63-71
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• 1992

Liquid chromatographic behavior of Pd(II), Ni(II) and Co(III) in N-Alkylisonitrosoacetylacetone imine(HIAA-NR) chelates was investigated by reversed phase high perfomance liquid chromatography. The optimum conditions for the separation of IAA-NR-metal chelates were examined respect to the flow rate and mobile phase strength. The metal-N-Alkylisonitrosoacetylacetone imine chelates in solution were successfully separated on Novapak $C_{18}$ column using acetonitrile/water mixture as mobile phase. The elution order of chelates is methyl>ethyl>propyl>butyl as N-alkyl group for ligand is varied. It was found that all IAA-NR-metal chelates were eluted in an acceptable range of capacity factor value($0{\leq}log\;k^{\prime}{\leq}1$). The dependence of log k' on the volume fraction of water in the binary mobile phase was examined. Also, the dependence of k' on the liquid-liquid extraction distribution ratio(Dc) in acetonitrile-water-alkane extraction system was investigated for IAA-NR-metal chelate. Both kinds of dependence are linear, which suggests that the retention of the electroneutral metal chelates on Novapak $C_{18}$ column is largely due to the hydrophobic effect.

• Asian-Australasian Journal of Animal Sciences
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• v.18 no.5
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• pp.741-746
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• 2005

To investigate the basic information and the possibility of ACE-inhibitory peptides for antihypertension materials, goat's caisin (CN) was hydrolyzed by various proteolytic enzymes and ACE-inhibitory peptides were separated and purified. ACE-inhibition ratios of enzymatic hydrolysates of goat's CN and various characteristics of ACE-inhibitory peptides were determined. ACE-inhibition ratios of goat's CN hydrolysates were shown the highest with 87.84% by pepsin for 48 h. By Sephadex G-25 gel chromatograms, Fraction 3 from goat's CN hydrolysates by pepsin for 48 h was confirmed the highest ACE-inhibition activity. Fraction 3 g and Fraction 3 gh from peptic hydrolysates by RP-HPLC to first and second purification were the highest in ACE-inhibition activity, respectively. The most abundant amino acid was leucine (18.83%) in Fraction 3 gh of ACE-inhibitory peptides after second purification. Amino acid sequence analysis of Fraction 3 gh of ACE-inhibitory peptides was shown that the Ala-Tyr-Phe-Tyr, Pro-Tyr-Tyr and Tyr-Leu. IC$_{50}$ calibrated in peptic hydrolysates at 48 h, Fraction 3, Fraction 3 g and Fraction 3 gh from goat's CN hydrolysates by pepsin for 48 h were 29.89, 3.07, 1.85 and 0.87 g/ml, respectively. Based on the results of this experiment, goat's CN hydrolysates by pepsin were shown to have ACE-inhibitory activity.

• KSBB Journal
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• v.31 no.2
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• pp.134-143
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• 2016

Lycii fructus has been traditionally used as a preventive and therapeutic medicine to treat enervation and diverse chronic diseases. In this study, we investigated whether fermentation of Lycii fructus extract (LE) increases the enzymatic activity of the alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH). The fermentation of LE by Bacillus subtilis subsp. subtilis and Saccharomyces cerevisiae IFO 2376 was shown to increase the enzymatic activity of ADH and ALDH. TLC analysis of LE and fermented LE (FLE) showed that ADH and ALDH activities increased in different spots. Fraction No. 66 of LE and fraction No. 68 of FLE by Silica gel chromatography showed increased ADH activity of 129.1% and 148.9%, respectively. Fractions No. 128 of LE and FLE by Silica gel chromatography showed increased ALDH activity of 134.1% and 148.1%, respectively. The fraction No. 68 of FLE obtained by HPLC showed new peaks at $R_t$ 11.938min, $R_t$ 22.072min and $R_t$ 28.842min, indicating that ADH activity was increased. The LE and FLE fractions with the greatest increases in ADH activity peaked at the same time ($R_t$ 13min),whereas the LE and FLE fractions with the greatest increases in ALDH activity peaked at different times ($R_t$ 16.307min and $R_t$ 36.640min, respectively).

• KSBB Journal
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• v.19 no.3
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• pp.169-173
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• 2004

The Rhodiola Sachalinensis 5 g were mixed and extracted with methanol 150 $m\ell$ at the room temperature for 12 h. The effluents were collected and grouped into the two. Un this experimental condition, the mobile phase composition were linearly changed as follows; water/methanol : 90/10 - 30/70 (vol. %, for 5 min), 30/70 - 10/90 (vol. %, for 15 min) and an analytical column (3.9 ${\times}$ 25 em, 15 $\mu\textrm{m}$ particle size, and 300 ${\AA}$ pore size) was utilized. The performance of the extracted Rhodiola Sachalinensis as a whitening agent was not favorable, so it classifies the Rhodiola Sachalinensis extractions with two fractions and collects each fraction for whitening agent assay. For the in-vivo melanin production ratio assay that used melanin-a cell in 10 ppm concentration, it was 58.6%, the first fraction of the effluents collected between 1.0 and 4.0 min, while it was 60％ between 10.4 and 17.6 min for the second fraction, which were more efficient than that of arbutin, 45.6%.

• Natural Product Sciences
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• v.17 no.2
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• pp.147-159
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• 2011

A high-performance liquid chromatography-electrospray ionization-tandem mass spectrometric method was developed to determine simultaneously eight marker constituents of Forsythiae fructus, and subsequently applied it to classify its two botanical origins. The marker compounds of Forsythia suspensa were phillyrin, pinoresinol, phillygenin, lariciresinol and forsythiaside; those of F.viridissima were arctiin, arctigenin and matairesinol. Separation of the eight analytes was achieved on a phenyl-hexyl column (150${\times}$2.0 mm i.d., 3 ${\mu}M$) using gradient elution with the mobile phase: (A) 10% acetonitrile in 0.5% acetic acid, (B) 40% aqueous acetonitrile. A few fragment ions specific to the types of lignans, among the product ions generated by collisonally induced dissociation (CID) of molecular ion clusters, such as [M-H]$^-$ or [M+OAc]$^-$ were used not only for fingerprinting analysis but for the quantification of each epimer by using multiple-reaction monitoring mode. It was shown good linearity ($r^2{\geq}$ 0.9998) over the wide range of all analytes; intra- and inter-day precisions (RSD, %) were within 9.14% and the accuracy ranged from 84.3 to 115.1%. The analytical results of 40 drug samples, combined with multivariate statistical analyses - principal component analysis (PCA) and hierarchical cluster analysis (HCA) - clearly demonstrated the classification of the test samples according to their botanical origins. This method would provide a practical strategy for assessing the authenticity or quality of the herbal drug.

• Korean Journal of Pharmacognosy
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• v.47 no.3
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• pp.232-236
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• 2016

In traditional Korean medicine, Osmunda japonica Thunb has been used as hemostasis and antipyretic treatment. The main compound "cinnamtannin B-1" was obtained by column chromatographic separation, and its structure was determined by spectroscopic methods, including $^1H$, $^{13}C$ NMR, and IT-TOF-ESI MS. Ash, moisture and extract content and acidinsoluble ash were monitored as identification test to establish the analytical methods. The optimum reflux extraction condition was 100% methanol extracted 30 min for 2 times. A quantitative analysis using HPLC method exhibited that the main compound at 24.7 min and its content was 0.96% in methanol extraction.

• The Journal of Korean Medicine
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• v.18 no.1
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• pp.187-197
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• 1997

Major separation for the active ingredients and characterization of chemical properties in conjunction with screening test on animal were performed in order to analyze and standardize Ligustici Rhizoma or Angelicae Tenuissimae Radix as an important oriental herbal medicine for antiphlogistic or an important oriental herbal medicine for antiphlogistic or an anodyne. Furthermore the structure, composition and contents of ingredients for essential oil in Angelicae Tenuissimae Radix(Suckpo, Korea) were determined by means of Ge/MS followed by screening test on Z-ligustilide(82%) known as major ingredient as well as butylidenephthalide collected by HPLC with normal phase semiprep-column. The total active ingredient in Ligustici Rhizoma from China or Angelicae Tenuissimae harvested at Choonyang(Kyungnam, Korea), Jungsun(Kangwon, Korea), Suckpo(Kyungnam Korea), Youngchun(Kyungnam, Korea) have been determined showing higher abundant for three times on the product in Korea compared to that in China. In addition, the major component in Ahgelicae Tebyussunae Radux extract was found to be Z-ligustilide(70-80%) which is very different from that in Ligustici Rhizoma senkyunolide(39%) as major species. For screening test of Ligustici Rhizoma or Angelicae Tenuissimae Radix extracts toward the target animal, the efficiency has been shown the similarity on both extracts. Taking into account the level of ingredient, the total efficiency may be three times higher on Angelicae Tenuissimae Radix in Korea compared to Ligustici Rhizoma in China. As a result of present study, it is preferable to distinguish between Ligustici Rhizoma and Angelicae Tenuissimae Radix for better usage of oriental herbal medicine because of very different composition and abundant in spite of their similar screening effect.

• YAKHAK HOEJI
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• v.60 no.3
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• pp.112-117
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• 2016

A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the determination of caffeine in forensic aqueous sample. The centrifuged sample ($100{\mu}l$) was diluted 50-fold with distilled water. The diluted sample ($400{\mu}l$) was then diluted further with $200{\mu}l$ of 0.1% formic acid solution and $400{\mu}l$ of acetonitrile containing 500 ng of caffeine-(3-methyl-$^{13}C_3$) prior to LC-MS/MS analysis. The mobile phase was composed of 0.1% formic acid in distilled water (A) and acetonitrile (B). Chromatographic separation was performed by using a Zorbax SB-C18 ($100mm{\times}2.1mm$ i.d., $3.5{\mu}m$) column and caffeine was eluted within 1.1 min. Linear least-squares regression with a 1/x weighting factor was used to generate a calibration curve with the coefficients of determination ($r^2=0.9983$). The lower limit of quantification was $25ng/ml$ for the analyte. The process efficiency was 98.6~100.1%. Intra- and inter-day precisions were not more than 2.1% and 1.7%, while intra- and inter-day accuracies were ranged from -6.8 to 4.5%, respectively. The suitability of the method was examined by analyzing unknown forensic aqueous samples.

• Bulletin of the Korean Chemical Society
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• v.30 no.10
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• pp.2240-2244
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• 2009

A simple high performance liquid chromatographic method was developed to evaluate the quality of Moutan Cortex Radicis based on chromatographic fingerprints that characterize eight pharmacological compounds, namely, gallic acid, paeoniflorin, galloyl paeoniflorin, benzoic acid, quercetin, benzoylpaeoniflorin, paeoniflorigenone, and paeonol. These compounds were identified by their characteristic UV profiles and the mass spectroscopy data, and their contents were determined by HPLC. The chromatographic separation was performed on a $C_{18}$ column by gradient elution with 0.05% formic acid in water and acetonitrile. The methodological validation gave acceptable linearities (r = 0.9996) and recoveries (ranging from 99.4∼103.1%). The limits of detection (LOD) of these compounds ranged from 10 to 30 $\mu$g/mL. The representative chromatographic fingerprints of Moutan Cortex Radicis were obtained by analyzing 20 batches of samples collected from markets in Korea and China. For the efficient evaluation of quality for the commercial Moutan Cortex Radicis it is recommended that the total content of the six characteristic compounds should contain more than a minimum of 2% and that the content of total paeoniflorin and paeonol should exceed a minimum of 1.5% of dry weight of Moutan Cortex Radicis.

• Journal of Ginseng Research
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• v.40 no.4
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• pp.366-374
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• 2016

Background: In this study, we screened and identified an endophyte JG09 having strong biocatalytic activity for ginsenosides from Platycodon grandiflorum, converted ginseng total saponins and ginsenoside monomers, determined the source of minor ginsenosides and the transformation pathways, and calculated the maximum production of minor ginsenosides for the conversion of ginsenoside Rb1 to assess the transformation activity of endophyte JG09. Methods: The transformation of ginseng total saponins and ginsenoside monomers Rb1, Rb2, Rc, Rd, Rg1 into minor ginsenosides F2, C-K and Rh1 using endophyte JG09 isolated by an organizational separation method and Esculin-R2A agar assay, as well as the identification of transformed products via TLC and HPLC, were evaluated. Endophyte JG09 was identified through DNA sequencing and phylogenetic analysis. Results: A total of 32 ${\beta}$-glucosidase-producing endophytes were screened out among the isolated 69 endophytes from P. grandiflorum. An endophyte bacteria JG09 identified as Luteibacter sp. effectively converted protopanaxadiol-type ginsenosides Rb1, Rb2, Rc, Rd into minor ginsenosides F2 and C-K, and converted protopanaxatriol-type ginsenoside Rg1 into minor ginsenoside Rh1. The transformation pathways of major ginsenosides by endophyte JG09 were as follows: $Rb1{\rightarrow}Rd{\rightarrow}F2{\rightarrow}C-K$; $Rb2{\rightarrow}C-O{\rightarrow}C-Y{\rightarrow}C-K$; $Rc{\rightarrow}C-Mc1{\rightarrow}C-Mc{\rightarrow}C-K$; $Rg1{\rightarrow}Rh1$. The maximum production rate of ginsenosides F2 and C-K reached 94.53% and 66.34%, respectively. Conclusion: This is the first report about conversion of major ginsenosides into minor ginsenosides by fermentation with P. grandiflorum endophytes. The results of the study indicate endophyte JG09 would be a potential microbial source for obtaining minor ginsenosides.