• Natural Product Sciences
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• 제16권4호
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• pp.233-238
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• 2010

In order to facilitate the quality control of Artemisia capillaris, a simple, accurate and reliable HPLC method was developed for the simultaneous determination of the six bioactive compounds: scopolin (1), chlorogenic acid (2), 2,4-dihydroxy-6-methoxyacetophenone 4-glycoside (3), hyperoside (4), isorhamnetin 3-Orobinobioside (5), and scoparone (6), which were selected as the chemical markers of A. capillaris. Separation was achieved on an Agilent Eclipse XDB-C18 column with a gradient solvent system of 0.1% trifluoroacetic acid aqueous-acetonitrile at a flow-rate of 1.0 mL/min and detected at 254 nm. All six calibration curves showed good linearity ($R^2$ > 0.998). A simple reversed phase HPLC method was developed for extracting pharmacologically active compounds scopolin, chlorogenic acid, 2,4-dihydroxy-6-methoxyacetophenone 4-glycoside, hyperoside, isorhamnetin 3-O-robinobioside, and scoparone from A. capillaris using a binary gradient of acetonitrile : 0.1% trifluoroacetic acid with UV detection at 254 nm. The scopolin (1), chlorogenic acid (2), 2,4-dihydroxy-6-methoxyacetophenone 4-glycoside (3), hyperoside (4), isorhamnetin 3-O-robinobioside (5), and scoparone (6) contents of the herb of A. capillaris collected from fifteen district markets in Korea were 0.00~0.90 mg/g, 0.06~7.29 mg/g, 0.06~0.91 mg/g, 0.07~5.05 mg/g, 0.42~13.11 mg/g, and 1.11~29.82 mg/g, respectively. The results demonstrated that this method is simple and reliable for the quality control of A. capillaris.

• Parasites, Hosts and Diseases
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• 제48권2호
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• pp.127-132
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• 2010

Biomphalaria alexandrina snails play an indispensable role in transmission of schistosomiasis. Infection rates in field populations of snails are routinely determined by cercarial shedding neglecting prepatent snail infections, because of lack of a suitable method for diagnosis. The present study aimed at separation and quantification of oxalic, malic, acetic, pyruvic, and fumaric acids using ion-suppression reversed-phase high performance liquid chromatography (HPLC) to test the potentiality of these acids to be used as diagnostic and therapeutic biomarkers. The assay was done in both hemolymph and digestive gland-gonad complex (DGG) samples in a total of 300 B. alexandrina snails. All of the studied acids in both the hemolymph and tissue samples except for the fumaric acid in hemolymph appeared to be good diagnostic biomarkers as they provide not only a good discrimination between the infected snails from the control but also between the studied stages of infection from each other. The most sensitive discriminating acid was malic acid in hemolymph samples as it showed the highest F-ratio. Using the Z-score, malic acid was found to be a good potential therapeutic biomarker in the prepatency stage, oxalic acid and acetic acid in the stage of patency, and malic acid and acetic acid at 2 weeks after patency. Quantification of carboxylic acids, using HPLC strategy, was fast, easy, and accurate in prediction of infected and uninfected snails and possibly to detect the stage of infection. It seems also useful for detection of the most suitable acids to be used as drug targets.

• BMB Reports
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• 제39권3호
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• pp.270-277
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• 2006

Helices 4 and 5 of the Bacillus thuringiensis Cry4Ba $\delta$-endotoxin have been shown to be important determinants for mosquito-larvicidal activity, likely being involved in membrane-pore formation. In this study, the Cry4Ba mutant protein containing an additional engineered tryptic cleavage site was used to produce the $\alpha4$-$\alpha5$ hairpin peptide by an efficient alternative strategy. Upon solubilization of toxin inclusions expressed in Escherichia coli and subsequent digestion with trypsin, the 130-kDa mutant protoxin was processed to protease-resistant fragments of ca. 47, 10 and 7 kDa. The 7-kDa fragment was identified as the $\alpha4$-loop-$\alpha5$ hairpin via N-terminal sequencing and mass spectrometry, and was successfully purified by size-exclusion FPLC and reversed-phase HPLC. Using circular dichroism spectroscopy, the 7-kDa peptide was found to exist predominantly as an $\alpha$-helical structure. Membrane perturbation studies by using fluorimetric calcein-release assays revealed that the 7-kDa helical hairpin is highly active against unilamellar liposomes compared with the 65-kDa activated full-length toxin. These results directly support the role of the $\alpha4$-loop-$\alpha5$ hairpin in membrane perturbation and pore formation of the full-length Cry4Ba toxin.

• Advances in Traditional Medicine
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• 제8권3호
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• pp.222-227
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• 2008

Citrus aurantium var. amara L., commonly known as 'bitter orange' or 'sour orange', of the family Rutaceae, has traditionally been used in the treatment of various ailments, and it possesses different types of pharmacological properties. As a part of our on-going studies on the plantsfrom the Iranian flora, the extract of C. aurantium var. amara has been studied for its weight lossproperties using the mice model. While the Sep-Pak fraction, 20% methanol (MeOH) in water, of the hydro-methanolic extract of the peels of C. aurantium var. amara fruits, when injectedintraperitoneal (i.p.) at a dose of 10 mg/kg, significantly decreased the level of weight gain of the mice in comparison with control the group (P < 0.01), the Sep-Pak fraction 80% MeOH in water decreased the initial weight of mice by 0.44% in six weeks. The administration of the total extract(10 and 20 mg/kg, i.p.), and the Sep-Pak fractions, 40% and 60% MeOH in water (10 mg/kg, i.p.)did not show any significant change of weight of the test mice. Of the two active fractions, the80% MeOH in water fraction did not show any noticeable adverse effects on mice, and was therefore analysed by reversed-phase preparative high performance liquid chromatography resulting in the isolation and identification of four major components, two coumarins, meranzin hydrate (1) and bergamottin (2), and two flavonoids, xanthomicrol 5,4'-di-methyl ether (tangeritin, 3) and hymenoxin 5,7-di-methyl ether (nobiletin, 4).

• Journal of Korean Neurosurgical Society
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• 제46권5호
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• pp.459-467
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• 2009

Objective : The purpose of this study was to assess memory dysfunction in patients with mild and moderate traumatic brain injury (TBI) with and without frontal lobe injury (FLI). Methods : The subjects were 110 TBI patients, who had recovered from the acute clinical phase, and comprised 20 (18.2%) mild TBI (MTBI) patients with FLI, 16 (14.5%) MTBI patients without FLI, 51 (46.4%) moderate TBI (MOTBI) patients with FLI and 23 (20.9%) MTBI patients without FLI. All patients were administrated the Korean version of the Memory Assessment Scale (K-MAS). Results : Almost all the Summary Scale scores on the K-MAS failed to show any differences between TBI patients with and without FLI, but differences did emerge by types at severities. TBI patients with FLI showed higher Global Memory ability than TBI patients without FLI if their TBI was only mild, but when their TBI was more severe, this finding was reversed, and TBI patients with FLI showed lower Verbal and Global Memory abilities than TBI patients without FLI. Conclusion : Different kinds of assessment tools are needed for the measurement of memory abilities in TBI patients with FLI, and that the selection of the appropriate tool depends on the severity of the TBI.

• Journal of Crop Science and Biotechnology
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• 제11권1호
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• pp.45-50
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• 2008

Lignans and neolignans are optically active plant secondary metabolites. Research on biosynthesis of lignans has already been advanced especially for the formation of (+) pinoresinol but information on the biosynthesis of 8-O-4'- neolignans is still limited. Moreover, the chemical structure(position of substituents on aromatic rings) and stereochemistry of 8-O-4' neolignans is not clear. Katayama and Kado discovered that incubation of cell-free extracts from E. ulmoides with coniferyl alcohol in the presence of hydrogen peroxide gave (+)-erythro- and (-)-threo- guaiacylglycerol 8-O-4'-(coniferyl alcohol) ether (GGCE)(diastereomeric ratio, 3:2) which is the first report on enzymatic formation of optically active -8-O-4' neolignans from an achiral monolignol. In this aspect, enzymatic formation of guaiacyl 8-O-4' neolignan is noteworthy to clarify its stereochemistry from incubation of coniferyl alcohol with enzyme prepared from Eucommia ulmoides. In this experiment, soluble and insoluble enzymes prepared from E. ulmoides were incubated with 30 mM coniferyl alcohol(CA) for 60 min. The enzyme catalyzed GGCE, dehydrodiconiferyl alcohol(DHCA), and pinoresinol identified by reversed phase HPLC. Consequently, diastereomeric compositions of GGCE were determined as erythro and threo isomer. Enantiomeric composition was determined by the chiral column HPLC. Both enzyme preparations enantioselectively formed (-)-erythro, (+)-erythro and (+)-threo, (-)-threo-GGCEs respectively.

• 한국미생물·생명공학회지
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• 제36권4호
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• pp.326-335
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• 2008

Bacillus subtilis AH18균주는 auxin, siderophore 그리고 cellulase를 동시에 생산하는 PGPR 균주이자 생물방제균주로 항진균성 siderophore의 특성을 확인한 결과 catechol type의 siderophore로 확인하였다. $Siderophore_{AH18}$ 의 정제는 Amberlite XAD-2, sephadex LH-20 column chromatography 그리고 HPLC를 통해 정제 및 정제여부를 확인하였으며, GC-MS, $^1H$-NMR, 그리고 $^{13}C$-NMR을 통해 구조 및 분자량을 확인하였다. 그 결과 B. subtilis AH18이 생산하는 siderophore은 분자량 883의 bacillibactin임을 확인하였으며, 포자발아억제활성을 나타냄을 확인하였다. B. subtilis AH18은 P. capsici에 의한 고추역병을 효과적으로 방제하였으며(방제력 55%), 이는 bacillibactin에 의한 효과가 포함되리라 추측된다.

• Journal of Microbiology and Biotechnology
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• 제27권1호
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• pp.26-35
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• 2017

The hydrolysis of proteins constitutes an invaluable tool, granting access to a variety of peptide fragments with potentially interesting biological properties. Therefore, a hemoglobin (Hb) hydrolysate of Crocodylus siamensis was generated by digestion under acidic conditions. The antibacterial and antioxidant activities of the Hb hydrolysate were assessed in comparison with intact Hb. A disc diffusion assay revealed that the Hb hydrolysate exhibited antibacterial activity against eight strains of gram-positive bacteria and showed a higher efficacy than intact Hb. Moreover, the antioxidant activity of intact Hb and its hydrolysate was evaluated using ABTS and DPPH radical scavenging assays. The Hb hydrolysate exhibited free radical scavenging rates of 6-32%, whereas intact Hb showed a slightly higher activity. In addition, non-toxicity to human erythrocytes was observed after treatment with quantities of Hb hydrolysate up to $10{\mu}g$. Moreover, active fragmented Hb (P3) was obtained after purifying the Hb hydrolysate by reversed-phase HPLC. Scanning electron microscopy demonstrated the induction of bacterial cell membrane abnormalities after exposure to P3. Antibacterial and antioxidant activities play crucial roles for supporting the wound healing activity. Consequently, an in vivo mice excisional skin wound healing assay was carried out to investigate the effects of intact Hb treatment on wound healing in more detail. The results clearly demonstrate that intact Hb is capable of promoting 75% wound closure within 6 days. These findings imply that intact Hb of C. siamensis and its acid hydrolysate may serve as valuable precursors for food supplementary products benefitting human health.

• Journal of Pharmaceutical Investigation
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• 제27권4호
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• pp.331-335
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• 1997

Xerostomia is caused by organic or functional changes affecting the salivary system at different levels. Patients suffering from xerostomia may also complain of an oral burning sensation, ulceration or soreness, difficulty in swallowing, and poor denture retention. And pilocarpine is administered orally to induce salivary secretion. In Seoul National University Hospital(SNUH) pharmacy, the pilocarpine chewing tablets are prepared and supplied to patients of xerostomia in request of the dental hospital in SNUH. And we tested the salivary flow induction and the dissolution patterns of these products in saliva by a double-blind, sequential cross-over trials to eight healthy human volunteers with placebo. The pilocarpine chewing tablet contained 5 mg of pilocarpine, and placebo consisted of same materials as test drug, but didn't contain pilocarpine. In vivo experiment, all subjects were instructed to chew as 60-80 times/min. Mixed saliva was collected in the ranges of intervals such as 0-2, 2-5, 5-10, 10-15, 15-20, 20-30, 30-45 and 45-60 min after pilocarpine chewing tablet or placebo administration. Saliva volume was measured in each collecting time interval, and saliva pilocarpine concentrations were determined by reversed phase HPLC. The 82.5 percent $(4.13{\pm}0.69\;mg)$ of pilocarpine was extracted from chewing tablets during mastication of 60-80 times per minute for 60 minutes. Among these dissolved amounts, 90 percent was extracted within 20 minutes. The salivary flow rates were more increased in a group who administered pilocarpine chewing tablet at the interval of 5-10, 10-15, 20-30 and 45-60 min rather than a placebo-group, but only extracted amount of pilocarpine at 45-60 min interval is significanly different between two groups (p<0.05). But total amounts of saliva secreted for 1 hour in two group-pilocarpine and placebo treated- were $46.36{\pm}9.72\;ml\;and\;39.09{\pm}7.81\;ml$, respectively, and were not significantly different between two groups.

• 분석과학
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• 제30권2호
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• pp.102-111
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• 2017

Here we reported a methodology for identification of triacylglycerols (TAGs) and diacylglycerols (DAGs) in coix seed by preparative thin layer chromatography (prep-TLC) and non-aqueous reversed-phase liquid chromatography (NARP LC)-atmospheric pressure chemical ionization (APCI) tandem mass spectrometry (MS/MS). Lipid components were extracted from coix seed by reflux extraction using n-hexane for 3 hr. TAGs and DAGs in coix seed extract were effectively purified and isolated from matrix interferences by prep-TLC and then analyzed by LC-APCI-MS and MS/MS for identification. TAGs were effectively identified taking into consideration of their LC retention behavior, APCI-MS spectra patterns, and MS/MS spectra of $[DAG]^+$ ions. In MS/MS spectra of TAGs, diacylglycerol-like fragment $[DAG]^+$ ions were useful to identify TAGs with isobaric fragment ions. Based on an established method, 27 TAGs and 8 DAGs were identified in coix seed extract. Among them, 15 TAGs and 8 DAGs were for the first time observed in coix seed. Interestingly, some of TAGs isolated by prep-TLC were partly converted into DAGs through probably photolysis process during storing in room temperature. Thus, degradation phenomenon of TAGs should be considered in the quality evaluation and nutritional property of coix seed. LC-APCI-MS/MS combined with prep-TLC will be practical method for precise TAG and DAG analysis of other herbal plants.