• Bulletin of the Korean Chemical Society
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• v.35 no.7
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• pp.1956-1964
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• 2014

To provide the scientific corroboration of the traditional uses of Akebia quinata (Thunb.) Decne., a detailed analytical examination of A. quinata stems was carried out using a reversed-phase high performance liquid chromatography (RP-HPLC) method coupled to photodiode array detector (PDA) for the simultaneous determination of four phenolic substances; cuneataside D (1), 2-(3,4-dihydroxyphenyl)ethyl-O-${\beta}$-D-glucopyranoside (2), 3-caffeoylquinic acid (3) and calceolarioside B (4). Particular attention was focused on the main compound, 3-caffeoylquinic acid (3), which has a range of biological functions. In addition, 2-(3,4-dihydroxyphenyl)ethyl-O-${\beta}$-D-glucopyranoside (2) was considered as a discernible marker of A. quinata from its easy confuse plants. The contents of compounds 2 and 3 ranged from 0.72 to 2.68 mg/g and from 1.66 to 5.64 mg/g, respectively. The validation data indicated that this HPLC/PDA assay was used successfully to quantify the four phenolic compounds in A. quinata from different locations using relatively simple conditions and procedures. The pattern-recognition analysis data from 53 samples classified them into two groups, allowing discrimination between A. quinata and comparable herbs. The results suggest that the established HPLC/PDA method is suitable for quantitation and pattern-recognition analyses for a quality evaluation of this medicinal herb.

• YAKHAK HOEJI
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• v.45 no.2
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• pp.227-236
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• 2001

The complexity and variability of both the biologicals and the bioassays used to test them led to the use of the reference standard- a sample of the product of defined purity and potency, against which all preparations of that product must be calibrated. In order to prepare and establish KFDA reference standard for recombinant human growth hormone (somatropin), somatropin substance was filled in ampoules in National Institute for Biological Standards and Control (NIBSC). The candidate KFDA reference standard for somatropin (designated as 98/674) was evaluated to determine the suitability of serving as a KFDA reference standard for somatropin by the collaborative study, in which 10 laboratories participated. Physicochemical analysis and in vivo bioassay were performed by direct comparison with the international somatropin standard 88/624. 98/674 was identified as somatropin by SDS-PAGE, IEF, peptide mapping, and HPLC. Determination of somatropin content by SE-HPLC yielded a mean estimate of 2.01 mg somatropin per ampoule. Data from the study also yielded mean values of 0.39 $\pm$ 0.26% for high molecular weight impurities by SE-HPLC and mean values of 2.13 $\pm$ 1.29% for somatropin related proteins by RP-HPLC. Estimates of relative potency by weight gain bioassay in the hypophysectomised rats showed that relative potency of KS 98/674 was 1.07 aganist IS 88/624. Based on the results of the collaborative study, the candidate reference standard for somatropin is suitable to serve as a KFDA reference standard for somatropin.

• Korean Chemical Engineering Research
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• v.46 no.4
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• pp.660-664
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• 2008

In this work, analytical HPLC was utilized to obtain Tanshinone IIA (TIIA) from Salvia Miltiorrhiza Bunge (SMB). The optimum operating conditions were experimentally determined to analyze the TIIA in the pretreated extract. SMB was extracted with the various organic solvents of methanol, ethyl acetate, and ethanol, then the extract was analyzed to compare the amount of TIIA. From the results, the methanol showed the best extraction efficiency of TIIAd. The analysis by $C_{18}$ column was performed. The mobile phase was composed of methanol and water, and the isocratic elution mode was mainly applied. $2.154{\mu}g$ of TIIA/mg of SMB powder was extracted with methanol.

• Journal of Soil and Groundwater Environment
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• v.14 no.1
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• pp.51-60
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• 2009

A series of experiments was performed to develop an optimized analytical procedure for the analysis of explosives in soil by HPLC with soil samples collected at two live-fire military shooting ranges. The minimum amount of soil to be collected, Wmin, for the analysis of explosive compounds was 125g, based on the segregation and homogeneity constants that account for soil heterogeneity and non-homogeneous distribution of target explosive compounds. The optimization of extraction and HPLC analytical conditions were also studied based on analytes CV values. The most effective soil/ extractant ratio was estimated to be 10g-pretreated soil/20 mL acetonitrile as extractant. The optimized HPLC elution conditions for the separation of US EPA designated 14 explosive compounds, were column temperature 30${\circ}C$, eluents ratio of isopropanol: acetonitrile: water = 18 : 12: 70, and flow rate of 0.8 mUmin at 230 nm. However, UV wavelength 254 nm was better for the analysis of NB, 2,4-DNT, 2NT, 4NT, and 3NT.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.35 no.1
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• pp.11-17
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• 2009

Betamethasone propionate, an anti-inflammatory glucocorticosteroid, was detected in cosmetics with no indication on the label of this compound as an ingredient. The product was formulated as a topical spray or shampoo and labeled to contain zinc pyrithione as the active ingredient. A thin-layer chromatographic analysis was carried out on silica gel plates to provide a first indication about the presence of a compound with steroid structure and reactivity; then high-performance liquid chromatography (HPLC) separation allowed the identification of the corticosteroid agent and its quantification. To identify the corticosteroid agent from these commercial samples we collected the fractions suspected to have ketol steroids by prep HPLC and identified the compound as betamethasone propionate by NMR and MS spectrometry. Then we synthesized the standard for the betamethasone 17-propionate and 21-propionate and quantitate the corticosteroids from the sample by HPLC with that standards. By this method we identified the corticosteroid compounds from some commercial cosmetics such as zinc pyrithione sprays. The finding of betamethasone propionate in the products was shown by comparison to an authenticated standard of betamethasone propionate by retention time on reverse-phase HPLC. Two of the tested products contained betamethasone propionate at the levels of 0.005 ${\sim}$ 0.02% and the others were free of betamethasone propionate.

• Journal of Life Science
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• v.25 no.2
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• pp.180-188
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• 2015

The bacterial strain isolated from Kimchi showed antibacterial activity against Micrococcus luteus IAM 1056. The selected strain was identified as Lactococcus lactis by 16S rRNA nucleotide sequence analysis and named as Lactococcus sp. KD 28. The treatment of culture supernatant with proteinase K removed antibacterial activity, indicating its proteinaceous nature, a bacteriocin. This bacteriocin was sensitive to hydrolytic enzymes such as ${\alpha}$-chymotrypsion, trypsin, proteinase K, lipase, ${\alpha}$-amylase and subtilisin A. The bacteriocin was highly thermostable and resistant to heating at $80^{\circ}C$ for up to an hour but 50 % of the total activity was remained at $100^{\circ}C$ for 30 min. The pH range from 2.0 to 8.0 had no effect on bacteriocin activity and it was not affected by solvents such as acetonitrile, isopropanol, methanol, chloroform and acetone up to 50% concentration. The bacteriocin showed antibacterial activity against M. luteus IAM 1056, Lactobacillus delbrueckii subsp. lactis KCTC 1058, Enterococcus faecium KCTC 3095, Bacillus cereus KCTC 1013, B. subtilis KCTC 1023, Listeria ivanovii subsp. ivanovii KCTC 3444, Staphylococcus aureus subsp. aureus KCTC 1916, B. megaterium KCTC 1098 and B. sphaericus KCTC 1184. The bacteriocin was purified through ammonium sulfate concentration, SP-Sepharose chromatography and RP-HPLC. The molecular weight was estimated to be about 3.4 kDa by tricine-SDS-PAGE analysis.

• Nutritional Sciences
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• v.9 no.3
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• pp.152-158
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• 2006

Helicobacter pylori (H. pylori) infection rapidly stimulated either COX-2 or 5-LOX and released arachidonic acid metabolites that have been considered as pivotal mediators in H. pylori-induced inflammatory responses. To determine whether red ginseng extract (RGE) can suppress the biosynthesis of 5(S)-hydroxyeicosatetraenoic acids (HETE), a precursor metabolite of leukotrienes B4 (LTB4) in H. pylori-provoked inflammatory responses in gastric epithelial cells, the biosynthesis of monohydroxy fatty acids was measured using radioactive arachidonic acid and validated by RP-HPLC using non-radioactive AA as substrate in AGS cells cocultured with H. pylori (ATCC 43504) with or without pretreatment of RGE. Among three known major HETEs, H. pylori infection specifically induced the biosynthesis of $^{14}C-5(S)-HETE$ rather than the complex of $^{14}C-15S-/^{14}C-12(S)-HETE$ from $^{14}C-AA$, concomitantly obtained by HPLC(p<0.01). RGE, 1 to $100{\mu}g/ml$, selectively suppressed H. pylori-stimulated $^{14}C-5(S)-HETE$ production implying the attenuation of 5-lipoxygenase activity, of which was similar to known LOX inhibitor NDGA $(10{\mu}M)$ (p<0.01). However, the amount of 5(S)-HETE was significantly reduced by higher dose of RGE $(100{\mu}g/ml)$ (p<0.05). These results indicated that LOX pathway might be one of principle pathogenic mechanisms of H. pylori and red ginseng could be a nutraceutical against H. pylori infection through inhibiting action of LOX activity.

• Korean Journal of Pharmacognosy
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• v.30 no.3
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• pp.335-339
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• 1999

Separation and quantitative determination of 20-hydroxyecdysone from Achyranthis Radix has been conducted by using HPLC method. 20-hydroxyecdysone in a methanol extract from the raw drug was separated on a reverse phase column using a $CH_3CN-H_2O\;(18:82)$ solvent system and the average content is $0.0931{\pm}0.0048%$. For the preparation of authentic standard, we isolated 20-hydroxyecdysone from the roots of Achyranthes fauriei by $SiO_2$ and RP-18 column chromatography.

• Food Science and Preservation
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• v.24 no.4
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• pp.542-549
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• 2017

In this study, a peptide exhibiting antioxidant activity was isolated from sandfish (Arctoscopus japonicus) roe hydrolysate (SRH) in order to evaluate their practical uses as materials for manufacturing functional foods. The A. japonicus roe protein was hydrolyzed using Collupulin MG, and isolation of antioxidant peptide was performed using ultrafiltration (UF), prep-HPLC, and RP-HPLC. The SRH with a molecular weight below 3 kDa constituted about 38% of the whole hydrolysate, and the fraction with a molecular weight below 3 kDa showed significantly greater antioxidant activity compared to the original SRH and other fractions. The isolation fold of the antioxidant peptide isolated from SRH throughout the four-step procedure was 7.11-fold, and protein yield was 14.8%. The DPPH radical scavenging activity of isolated antioxidant peptide was above 90% at a concentration of 1.0 mg/mL, which was similar to that of the Trolox at a concentration of 0.1 mg/mL. These results suggested that the antioxidant peptide derived from A. japonicus roe could be a useful additive for producing functional foods and protein supplements. However, it is necessary to perform further study the structural characteristics of this antioxidant peptide isolated from A. japonicus roe.

• Archives of Pharmacal Research
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• v.25 no.5
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• pp.613-616
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• 2002

A high-performance liquid chromatographic method has been developed for the separation and quantification of chrysin and synthetic chrysin derivatives (12 chrysin alkyl and 7 chrysin acyl derivatives). The chromatography was performed using a $Nova-Pak^{\circledR}C_{18}$ column. A RP-HPLC was performed by using a binary mixture (MeOH-10 mM H$_3$PO$_4$) as a mobile phase, and the column temperature was maintained at room temperature. A flow rate was 1.0 ml/min, and the effluent was monitored at a wavelenth of 280 nm. The retention times for chrysin acyl and alkyl derivatives were within 10 minutes and 20 minutes, respectively. The absolute recovery of samples were all over 96%. The detection limits were 0.1~18 ng at S/N = 3 ratio.