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Analysis of Free Amino Acids and Flavors in Fermented Jujube Wine by HPLC and GC/MS (GC-MS 및 HPLC를 이용한 대추발효주의 유리 아미노산 및 향기성분 분석)

  • Chun, Myoung Sook;Kim, Soon Jin;Noh, Bong Soo
    • Korean Journal of Food Science and Technology
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    • v.44 no.6
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    • pp.779-784
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    • 2012
  • Characteristic chemical compositions of jujube wine using different preparation methods including fermentation were investigated. Fermentation for jujube wine started using whole fruit (JW1), seed-removed fruit (JW2) and whole fruit heated at $100^{\circ}C$ for 2 h and then extracted (JW3). The free amino acids and flavors were analyzed quantitatively by HPLC and GC-MS. A total of 18 amino acids were identified in all samples. The amount of total free amino acids was detected from 141-210 ppm (JW1), 147-342 ppm (JW2), and 336-362 ppm (JW3). Large amounts of proline, aspartate, glutamate, arginine and alanine were detected in jujube wine. Thirteen kinds of volatile compounds including six alcoholic compounds (ethyl alcohol, iso-butyl alcohol, n-butyl alcohol, iso-amyl alcohol, n-amyl alcohol, and phenethyl alcohol), four ester (ethyl acetate, hexyl acetate, ethyl caprylate, and phenethyl acetate) and three aldehydes (diethylacetal, furfural, and benzaldehyde) were detected. Ethyl alcohol (30.50-33.95% peak area), benzaldehyde (2.55-15.97% ratio), furfural (0.07-15.28% ratio), iso-amyl alcohol (1.04-14.73% ratio), and phenethyl acetal (0.78-9.28% ratio) were abundant in jujube wine.

Electrospray ionization tandem mass fragmentation pattern of camostat and its degradation product, 4-(4-guanidinobenzoyloxy)phenylacetic acid (Camostat 및 분해산물 4-(4-guanidinobenzoyloxy)phenylacetic acid의 전자분무 이온화 텐덤 질량 fragmentation 패턴)

  • Kwon, Soon-Ho;Shin, Hye-Jin;Park, Ji-Myeong;Lee, Kyoung-Ryul;Kim, Young-Jin;Lee, Sang-Hoo
    • Analytical Science and Technology
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    • v.24 no.2
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    • pp.78-84
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    • 2011
  • The fragmentation patterns of a serine protease inhibitor, camostat, and its degradation product, 4-(4-guanidinobenzoyloxy)phenylacetic acid (GBPA), were for the first time investigated by a triple quadrupole tandem mass spectrometry equipped with an electrospray source (ESI-MS/MS) in positive and/or negative ion mode under collision-induced dissociation (CID). The positive CID spectrum of camostat showed distinctly that the single bond (C-O) cleavage between carbonyl group and oxygen atom of the ester bonds of the compound favorably occurred and then the loss of N,N-dimethylcarbamoylmethyl group was more susceptible than that of guanidine moiety. In the positive ion CID spectrum of GBPA, the initial cleavage between the carbonyl group and oxygen atom of 4-guanidinobenzoyloxy group also occurred, yielding the most abundant fragment ion at m/z 145. On the other hand, the negative CID spectrum of GBPA characteristically showed the occurrence of the most abundant peak at m/z 226 resulting from the sequential neutral losses of $CO_2$ and HN=C=NH from the parent ion at m/z 312.

Secondary Metabolites from the Mycelial Culture Broth of Phellinus linteus (상황(Phellinus linteus) 배양 균사체의 2차 대사산물에 대한 화학적 연구)

  • Song, K.S.;Cho, S.M.;Ko, K.S.;Han, M.W.;Yoo, I.D.
    • Applied Biological Chemistry
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    • v.37 no.2
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    • pp.100-104
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    • 1994
  • From the 48 hour-cultured mycelial broth of Phellinus linteus, six compounds were isolated by means of ethyl acetate extraction, silica gel column chromatography and preparative thin layer chromatography, consecutively. Compound 1 was identified as a succinic acid by the comparison of its spectral data with authentic sample. Compounds 2 and 3 were identified as p-hydroxyphenyl acetic acid methyl ester and p-hydroxybenzaldehyde by spectroscopic studies, respectively. NMR and MS studies of compound 6 revealed that it was 2,5-dihydroxymethyl furan. Compound 4, which showed similar NMR absorptions and MS fragmentation pattern with those of compound 6 was identified as 2-hydroxymethyl-5-methoxymethylfuran. These structures were verified by the spectral data of the acetate derivatives of the compounds. Compound 5 was supposed to be a N-acetyltyramine from its $^1H-NMR$ and EI-MS data, and its structure was confirmed by a synthesis starting from tyramine.

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Establishment of Genetic Characteristics and Individual Identification System Using Microsatellite loci in Domestic Beef Cattle (초위성체 DNA표지인자를 이용한 국내 육우집단의 품종특성 및 개체식별 체계설정)

  • Kim, Sang-Wook;Jang, Hee-Kyung;Kim, Kwan-Suk;Kim, Jong-Joo;Jeon, Jin-Tae;Yoon, Du-Hak;Kang, Seong-Ho;Jung, Hyo-Il;Cheong, Il-Cheong
    • Journal of Animal Science and Technology
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    • v.51 no.4
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    • pp.273-282
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    • 2009
  • DNA marker information is used to identify or distinguish cattle breeds or individual animal. The purpose of this study was to apply Bovine Genotypes Kit Version 1.1/2.1 to bovine DNA samples (National Institute of Animal Science) taken from Australian / American beef (n=148), Holstein beef (n=170) and Hanwoo cattle (n=177) bred in Jeongeub, Jeonbuk, Korea, so that it could distinguish Hanwoo breed. The Bovine Genotype Kits consist of 16 ISAG MS markers, which were used to build a database of genotypes in each group. Genotyping results were analyzed using MS Tool kit and Phylip program to create phylogenetic tree. The GeneClass 2.0 was used to estimate breed identification. These analyses found that this kit had 100% capacity to distinguish Hanwoo beef, 95.3% capacity to differentiate Australian / American beef and 90% capacity to identify Korean Holstein steer beef. Hence, it is expected that 16 commercial microsatellite markers is useful to categorizegenetic characteristics of Hanwoo breed and also identify Hanwoo individuals and the origin of beef. In particular, it is expected that these markers will be advantageous in discriminating domestic Holstein beef from Australian / Americanbeef.

Quantitative Analysis of Brain Metabolite Spectrum Depending on the Concentration of the Contrast Media in Phantom (팬텀 내 조영제 농도에 따른 뇌 대사물질 Spectrum의 정량분석)

  • Shin, WoonJae;Gang, EunBo;Chun, SongI
    • Journal of the Korean Society of Radiology
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    • v.9 no.1
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    • pp.47-53
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    • 2015
  • Quantitative analysis of MR spectrum depending on mole concentration of the contrast media in cereberal metabolite phantom was performed. PRESS pulse sequence was used to obtain MR spectrum at 3.0T MRI system (Archieva, Philips Healthcare, Best, Netherland), and the phantom contains brain metabolites such as N-Acetyl Asparatate (NAA), Choline (Cho), Creatine (Cr) and Lactate (Lac). In this study, optimization of MRS PRESS pulse sequency depending on the concentration of contrast media (0, 0.1 and $0.3mmol/{\ell}$) was evaluated for various repetition time(TR; 1500, 1700 and 2000 ms). In control (cotrast-media-free) group, NAA and Cho signals were the highest at TR 2000 ms than at 1700 and 1500 ms. Cr had the highest peak signal at TR 1500 ms. When concentration of contrast media was $0.1mmol/{\ell}$, the metabolites were increased NAA 73%, Cho 249%, Cr 37% at TR 1700 ms compared with other TR, and also signal increased at $0.3mmol/{\ell}$, In $0.5mmol/{\ell}$ of contrast agent, cerebral metabolite peaks reduced, especially when TR 1500 ms and 2000 ms they decreased below those of control group. The ratio of metabolite peaks such as NAA/Cr and Cho/Cr decreased as the concentration of the contrast agent increased from 0.1 to $0.5mmol/{\ell}$. Authors found that the optimization of PRESS sequence for 0.3T MRS was as follows: low density of contrast agent ($0.1mmol/{\ell}$ and $0.3mmol/{\ell}$) made the highest signal intensity, while high density of contrast agent reveals the least reduction of signal intensity at 1700 ms. In conclusion, authors believe that it is helpful to reduce TR for acquiring maximum signal intensity.

Endonuclease Restriction Patterns of Chloroplast DNA in Somatic Hybrids Obtained by Protoplast Fusion of Nicotiana tabacum and N. glutinosa (Nicotiana tabacum과 N. glutinosa간 원형질체융합 식물체에 있어서 엽록체 DNA의 제한효소단편의 유형)

  • 김준철
    • Journal of Plant Biology
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    • v.34 no.1
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    • pp.37-43
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    • 1991
  • Mesophyll protoplasts of Nicoliana labacum ($NR^{-}/SR^{+}$) and N glulinosa were electrofused with AC field of 0.5 MHz and 1 kV DC pulse for 2 ms. Fused protoplasts were selected and cultured to the green cell clusters in $MSNO_3$ medium containing 1.2 mg!ml streptomycin sulfate. Four plant lines regenerated from selected colonies showed both parental morphological characteristics of leaf and flower and these plant lines were confirmed as somatic hybrids based on electrophoretic patterns of leaf peroxidase. In XhoI restriction patterns of chloroplast DNA, these hybrid plant lines expressed both parent common restriction sites and parent specific sites. One of these hybrid lines exhibited interspecific pattern of both parental chloroplast genomes. indicating nine both parent common sites, one N labacum specific site and two N glutinosa specific sites. sites.

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Study on the Synthesis of N,N'-Dicyclohexylcarbodiimide from N,N'-Dicyclohexylurea (디사이클로헥실우레아로부터 디사이클로헥실카르보디이미드의 합성에 관한 연구)

  • Kim, Jae Young;Chung, Dae-Won
    • Applied Chemistry for Engineering
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    • v.22 no.3
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    • pp.319-322
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    • 2011
  • N,N'-Dicyclohexylcarbodiimide (DCC) known as powerful dehydrating reagent in amide or ester synthesis is converted into N,N'-dicyclohexylurea (DCU) during the reaction. In the paper, DCU was recovered from the reaction for the synthesis of the hydrophilic derivative of ${\beta}$-sitosterol, and the purification of the recovered DCU and the dehydration of DCU into DCC were investigated. In the presence of tosyl chloride, (TsCl) and triethylamine (TEA), DCU was converted into DCC, and the optimum molar ratio of [DCU] : [TsCl] : [TEA] was found to be 1.0 : 1.5 : 3.0. Pure DCC was obtained with a 46% yield by the sublimation after the purification process, and characterized by GC/MS, FT-IR and $^{13}C-NMR$.

Development of Analytical Method for Fipronil Residues in Agricultural Commodities Using GC-ECD/MS (GC-ECD/MS를 이용한 농산물 중 Fipronil의 잔류 분석법 개발)

  • Ahn, Kyung-Geun;Kim, Gyeong-Ha;Kim, Gi-Ppeum;Hwang, Young-Sun;Kang, In-Kyu;Lee, Young Deuk;Choung, Myoung-Gun
    • Korean Journal of Environmental Agriculture
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    • v.34 no.4
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    • pp.309-317
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    • 2015
  • BACKGROUND: An analytical method was developed using GC-ECD/MS to precisely determine the residue of fipronil, a phenylpyrazole insecticide used to control a wide range of foliar and soil-borne pests.METHOD AND RESULTS: Fipronil residue was extracted with acetone from representative samples of five raw products which comprised hulled rice, soybean, Kimchi cabbage, green pepper, and apple. The extract was diluted with saline water, and fipronil was partitioned into n-hexane/dichloromethane (20/80, v/v) to remove polar co-extractives in the aqueous phase. Florisil column chromatography was additionally employed for final purification of the extract. Fipronil was separated and quantitated by GC-ECD using a DB-17 capillary column. Accuracy of the proposed method was validated by the recovery from crop samples fortified with fipronil at 3 levels per crop in each triplication.CONCLUSION: Mean recoveries ranged from 86.6% to 106.0% in five representative agricultural commodities. The coefficients of variation were less than 10%. Limit of quantitation of fipronil was 0.004 mg/kg as verified by the recovery experiment. A confirmatory technique using GC/MS with selected-ion monitoring was also provided to clearly identify the suspected residue. Therefore, this analytical method was reproducible and sensitive enough to determine the residue of fipronil in agricultural commodities.

Chemical Derivatization of Catecholamines for Gas Chromatography-Mass Spectrometry

  • Park, Sun-Young;Kang, Bo-Xin;Li, Quing;Kim, Hoon-Sik;Lee, Jun-Gae;Hong, Jong-Ki
    • Bulletin of the Korean Chemical Society
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    • v.30 no.7
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    • pp.1497-1504
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    • 2009
  • GC/MS analysis of catecholamines (CAs) in biological sample may produce poor reproducible quantitaion when chemical derivatization is used as the technique to form a volatile derivative. Significant quantities of the side products can be formed from CAs with primary amine during the derivatization reaction under un-optimized conditions. We have tested various chemical derivatization techniques in an attempt to find an optimum derivatization method that will reduce side product formation, enable to separate several catecholamine derivatives in GC chromatogram, and obtain significant improvement of detection sensitivity in GC/MS analysis. Whereas several derivatization techniques such as trimethylsilylation (TMS), trifluoroacylation (TFA), and two step derivatization methods were active, selective derivatization to form O-TMS, N-heptafluorobutylacyl (HFBA) derivative using N-methyl-N-(trimethylsilyl)-trifluoroacetamide (MSTFA) and N-methyl-bis(heptafluorobutyramide) (MBHFBA) reagents was found to be the most effective method. Moreover, this derivative formed by selective derivatization could provide sufficient sensitivity and peak separation as well as produce higher mass ion as base peak to use selected ion in SIM mode. Calibration curves based on the use of an isotopically labeled internal standard show good linearity over the range assayed, 1 ~ 5000 ng/mL, with correlation coefficients of > 0.996. The detection limits of the method ranged from 0.2 to 5.0 ppb for the different CAs studied. The developed method will be applied to the analysis of various CAs in biological sample, combined with appropriate sample pretreatment.

Somatic embryogenesis and in vitro plant regeneration in moth bean [Vigna aconitifolia (Jacq.) Marechal]: a recalcitrant grain legume

  • Choudhary, Kailash;Singh, M.;Rathore, M.S.;Shekhawat, N.S.
    • Plant Biotechnology Reports
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    • v.3 no.3
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    • pp.205-211
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    • 2009
  • An efficient in vitro regeneration protocol for moth bean [Vigna aconitifolia (Jacq.) Marechal] via somatic embryogenesis has been developed. Embryogenic callus cultures were established from the cotyledonary node as explant on semi-solid Murashige and Skoog (MS) medium supplemented with $0.75mg\;1^{-1}$ 2,4-dichlorophenoxyacetic acid (2,4-D) and $1.5mg\;1^{-1}$ 6-benzylaminopurine (BA) and with various additives ($50mg\;1^{-1}$ ascorbic acid and $25mg\;1^{-1}$ each of adenine sulphate, citric acid and $_L-arginine$). Numerous somatic embryos differentiated on MS basal nutrient medium supplemented with $0.25mg\;1^{-1}$ 2,4-D and $0.5mg\;1^{-1}$ of kinetin (Kin). Sustained cell division resulted in the formation of cell aggregates, which progressed to the globular- and heart-shaped somatic embryos and then, if they differentiated properly, to the torpedo shape and cotyledonary stages. The transfer of embryos onto fresh MS basal medium containing $0.2mg\;1^{-1}$ BA and $2.0mg\;1^{-1}$ gibberellic acid enabled the embryos to achieve complete maturation and germination. More than 80% of somatic embryos were converted into true-to-type fertile plants. In vitro-regenerated plantlets with well-developed roots were successfully hardened in a greenhouse and established in soil.