• Title/Summary/Keyword: alcohol extraction

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Distillation and Quality Characteristics of Medicinal Herb Wines (약용주의 증류와 품질특성)

  • Jeong, Heon-Sang;Cho, Jung-Gun;Min, Young-Kyoo
    • Applied Biological Chemistry
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    • v.39 no.5
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    • pp.368-373
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    • 1996
  • Korean general medicinal herbs-sasam, gilkyung, jakyak, danggwi, hwangki, and chunkung-were added In the normal brewing procedure as a raw material or in the distilling procedure as a packing material. The distillates from the former procedure and those from the latter procedure were compared in quality and distillation properties. As distillation proceeded, pH of the medicinal herb wine distillate and the control(not added herbs) distillates were decreased, whereas that of the herb packing distillate was increased slowely of $0.05{\sim}0.97$ during $1{\sim}4$ fractions and decreased remarkably of $0.92{\sim}0.98$ afterward. Average pH was the highest of 5.70 in jakyak and lowest of 4.37 In gilkyung. Absorbances of the herb Packing distillate were decreased rapidly of $0.60{\sim}1.59$ in the $1{\sim}4$ fractions but slowely of $0.19{\sim}0.54$ in the next fractions. During distillation both fractional alcohol concentration of the distillates and distillation rate were decreased. Their values were decreased more slowly than the control. Distillation rates of medicinal herb wine distillate were varied by medicinal herb varieties and alcohol concentration of fermented wine. Danggwi and control showed the highest average distillation rate as $0.12\;m{\ell}/sec$ and gilkyung the lowest value as $0.073\;m{\ell}/sec$. Maximum concentration of index component, paeoniflorin of jakyak was observed as 293 mg% in the 5th fraction of herb packing distillate and decrusin of danggwi as 3514 mg% In the 1st fraction of herb packing distillate. The extraction rate was 41.3% for paeoniflorin and 20.5% for decrusin. From sensory evaluation, the highest overall Qualify was observed in the medicinal herb wine distillate of hwangki added wine, the next in those of danggwi and jakyak added wine.

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Composition and Cell Cytotoxicity of Essential Oil from Caryopteris incana Miq. in Korea (층꽃나무(Caryopteris incana Miq) 정유의 성분 분석과 세포 독성 평가)

  • Kim, Song-Mun
    • Applied Biological Chemistry
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    • v.51 no.3
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    • pp.238-244
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    • 2008
  • The essential oil was obtained from the aerial part of Caryopteris incana Miq. by steam distillation, samples were collected by headspace (HS) and solid-phase microextraction (SPME) methods, and the compositions of the essential oil were analyzed by gas chromatography-mass spectrometry (GCMS). The fragrance of the essential oil was fougere and woody. There were sixty-nine constituents in the essential oil: 28 carbohydrates, 22 alcohols, 7 acetates, 7 ketones, 3 aldehydes, and 2 others. Major constituents were 4,6,6-trimethyl [1S-($1{\alpha},2{\beta},5{\alpha}$)]-bicyclo[3.1.1]hept-3-en-2-ol (11.8%), taucadinol (9.4%), myrtenyl acetate (9.2%), pinocarvone (7.0%), 1-hydroxy-1,7-dimethyl-4-isopropyl-2,7-cyclodecadiene (6.3%), ${\delta}$-3-carene (6.2%). By SPME extraction, forty-nine constituents were identified: 22 hydrocarbons, 16 alcohols, 6 acetates, 3 ketones, and 2 ethers. Major constituents of the SPME-extracted sample were ${\delta}$-3-carene (12.6%), (-)-myrtenyl acetate (11.2%), 6,6-dimethyl-2-methylene-bicycol [3.1.1] heptan-3-o1 (10.9%), pinocarvone (9.3%). By HS extraction, ten constituents were identified: 5 hydrocarbons, 2 amines, 1 alcohol, and 2 others. Major constituents of the HS-extracted sample were (Z)-2-fluoro-2-butene (34.9%), ${\delta}$-3-carene (6.9%), 6-(4-chlorophenul)tetrahydro-2-methyl-2H-1,2-oxazine (5.9%). The $IC_{50}$ value (0.011 ${\mu}g/mg$) in MTT assay using HaCaT keratinocyte cell line was lower than those of commercially-selling rosemary and tea tree, suggesting more toxicological studies are needed for commercial use of the essential oil of Caryopteris incana Miq.

Surfactant Enhanced In-Situ Soil Flushing Pilot Test for the Soil and Groundwater Remediation in an Oil Contaminated Site (계면활성제 원위치 토양 세정법을 이용한 유류 오염 지역 토양.지하수 정화 실증 시험)

  • 이민희;정상용;최상일;강동환;김민철
    • Journal of Soil and Groundwater Environment
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    • v.7 no.4
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    • pp.77-86
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    • 2002
  • Surfactant enhanced in-situ soil flushing was performed to remediate the soil and groundwater at an oil contaminated site, where had been used as a military vehicle repair area for 40 years. A section from the contaminated site (4.5 m $\times$ 4.5 m $\times$ 6.0 m) was selected for the research, which was composed of heterogeneous sandy and silt-sandy soils with average $K_d$ of 2.0$\times$$10^{-4}$cm/sec. Two percent of sorbitan monooleate (POE 20) and 0.07% of iso-prophyl alcohol were mixed for the surfactant solution and 3 pore volumes of surfactant solution were injected to remove oil from the contaminated section. Four injection wells and two extraction wells were built in the section to flush surfactant solution. Water samples taken from extraction wells and the storage tank were analyzed on a gas-chromatography (GC) for TPH concentration in the effluent with different time. Five pore volumes of solution were extracted while TPH concentration in soil and groundwater at the section were below the Waste Water Discharge Limit (WWDL). The effluent TPH concentration from wells with only water flushing was below 10 ppm. However, the effluent concentration using surfactant solution flushing increased to 1751 ppm, which was more than 170 times compared with the concentration with only water flushing. Total 18.5 kg of oil (TPH) was removed from the soil and groundwater at the section. The concentration of heavy metals in the effluent solution also increased with the increase of TPH concentration, suggesting that the surfactant enhanced in-situ flushing be available to remove not only oil but heavy metals from contaminated sites. The removal efficiency of surfactant enhanced in-situ flushing was investigated at the real contaminated site in Korea. Results suggest that in-situ soil flushing could be a successful process to remediate contaminated sites distributed in Korea.

Evaluation of in-vitro Antithrombosis Activity of Lees of Korean Traditional Wine (전통주 주박의 항혈전 활성 평가)

  • Kim, Mi-Sun;Lee, Ye-Seul;Kim, Jong Sik;Shin, Woo-Chang;Sohn, Ho-Yong
    • Journal of Life Science
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    • v.24 no.8
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    • pp.865-872
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    • 2014
  • In this study, ethanol and hot water extracts of lees from Korean traditional wine (J-B, J-S, J-Y, J-H, and J-W) were prepared, and their effects on blood coagulation, platelet aggregation, and hemolysis of human red blood cells (hRBCs) were investigated to develop functional food ingredients from lees. The pH and brix of the lees ranged from 3.90 to 4.29 and 5.0 to $27.0^{\circ}$, respectively, and there was a huge difference in the water and ethanol content among the lees. The nuruk and additives used affected the color and physicochemical properties of lees. The J-W takju made from only rice and traditional nuruk, which has $13^{\circ}$ brix and 1.8% of alcohol, has potential as functional food ingredient. With regard to the extraction yields of lees, higher yields were obtained from J-H, which contains different medicinal plants, in ethanol, followed by J-W, J-B, J-S, and J-Y. Higher extraction yields of lees were obtained from J-S in hot water, followed by J-B, J-W, J-H, and J-Y, respectively. The ethanol extract of J-H and the hot water extract of J-Y had the highest contents of total polyphenol and total flavonoids among the lees extracts. The 10 lees extracts did not show hemolysis activity against hRBCs up to 5 mg/ml. In an anticoagulation activity assay, the ethanol extracts of three yakju lees (J-B, J-S, and J-Y) and the hot water extract of J-W inhibited thrombin activity, whereas the hot water extract of J-B, J-S, and J-H inhibited blood coagulation factors. In an antiplatelet aggregation activity assay, only the J-W takju lees showed significant inhibition activity. Our results suggest that lees from traditional wine had high potential as a novel antithrombosis agent.

Quality Properties of White Lotus Leaf Fermented by Mycelial Paecilomyces japonica (동충하초 균사체로 발효시킨 백련잎차의 품질특성)

  • Kim, Jong-Suk;Wang, Su-Bin;Kang, Seong-Koo;Cho, Young-Sook;Park, Seok-Kyu
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.5
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    • pp.594-600
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    • 2009
  • Quality characteristics of white lotus leaf tea (LLT) fermented with or without mycelial Paecilomyces japonica were investigated. Extraction yield and browning index of hot water extract from non fermented and fermented LLTs were higher than those of ethanol extract (p<0.05). In all LLTs, nutritional components such as total free sugar, free amino acids and minerals of hot water extracts were higher than those of ethanol extracts except for total organic acids (p<0.05). Contents of total free sugar and organic acids were markedly increased through fermentation process of mycelial Paecilomyces japonica. in the same solvent extracts (p<0.05). Contents of most taste components of fermented LLT were increased by mycelial solid fermentation (p<0.05), but total free amino acids of two extracts were decreased in the range of $37.1{\sim}67.2%$ as compared to non-fermented LLT. Fifty-nine volatile compounds were identified by GC and GC-MS, including 11 aldehydes, 14 alcohols, 11 ketones, 11 hydrocarbones and 12 acids. Aldehyde and ketone compounds were more identified in fermented LLT than in non-fermented LLT being abundant alcohol compounds by simultaneous steam distillation and extraction. The most abundant compounds of LLT identified in this study were curcumene followed by 2,6-bis(1,1-dimethylethyl)-4-methyl-phenol and cyclohexen. Main compounds of fermented LLT were 2,6-bis(1,1-dimethylethyl)-4-methyl-phenol, butanoic acid, furfural, benzaldehyde, hexanoic acid and 2(3H)-furanone.

Sesquiterpenoids Bioconversion Analysis by Wood Rot Fungi

  • Lee, Su-Yeon;Ryu, Sun-Hwa;Choi, In-Gyu;Kim, Myungkil
    • 한국균학회소식:학술대회논문집
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    • 2016.05a
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    • pp.19-20
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    • 2016
  • Sesquiterpenoids are defined as $C_{15}$ compounds derived from farnesyl pyrophosphate (FPP), and their complex structures are found in the tissue of many diverse plants (Degenhardt et al. 2009). FPP's long chain length and additional double bond enables its conversion to a huge range of mono-, di-, and tri-cyclic structures. A number of cyclic sesquiterpenes with alcohol, aldehyde, and ketone derivatives have key biological and medicinal properties (Fraga 1999). Fungi, such as the wood-rotting Polyporus brumalis, are excellent sources of pharmaceutically interesting natural products such as sesquiterpenoids. In this study, we investigated the biosynthesis of P. brumalis sesquiterpenoids on modified medium. Fungal suspensions of 11 white rot species were inoculated in modified medium containing $C_6H_{12}O_6$, $C_4H_{12}N_2O_6$, $KH_2PO_4$, $MgSO_4$, and $CaCl_2$ for 20 days. Cultivation was stopped by solvent extraction via separation of the mycelium. The metabolites were identified as follows: propionic acid (1), mevalonic acid lactone (2), ${\beta}$-eudesmane (3), and ${\beta}$-eudesmol (4), respectively (Figure 1). The main peaks of ${\beta}$-eudesmane and ${\beta}$-eudesmol, which were indicative of sesquiterpene structures, were consistently detected for 5, 7, 12, and 15 days These results demonstrated the existence of terpene metabolism in the mycelium of P. brumalis. Polyporus spp. are known to generate flavor components such as methyl 2,4-dihydroxy-3,6-dimethyl benzoate; 2-hydroxy-4-methoxy-6-methyl benzoic acid; 3-hydroxy-5-methyl phenol; and 3-methoxy-2,5-dimethyl phenol in submerged cultures (Hoffmann and Esser 1978). Drimanes of sesquiterpenes were reported as metabolites from P. arcularius and shown to exhibit antimicrobial activity against Gram-positive bacteria such as Staphylococcus aureus (Fleck et al. 1996). The main metabolites of P. brumalis, ${\beta}$-Eudesmol and ${\beta}$-eudesmane, were categorized as eudesmane-type sesquiterpene structures. The eudesmane skeleton could be biosynthesized from FPP-derived IPP, and approximately 1,000 structures have been identified in plants as essential oils. The biosynthesis of eudesmol from P. brumalis may thus be an important tool for the production of useful natural compounds as presumed from its identified potent bioactivity in plants. Essential oils comprising eudesmane-type sesquiterpenoids have been previously and extensively researched (Wu et al. 2006). ${\beta}$-Eudesmol is a well-known and important eudesmane alcohol with an anticholinergic effect in the vascular endothelium (Tsuneki et al. 2005). Additionally, recent studies demonstrated that ${\beta}$-eudesmol acts as a channel blocker for nicotinic acetylcholine receptors at the neuromuscular junction, and it can inhibit angiogenesis in vitro and in vivo by blocking the mitogen-activated protein kinase (MAPK) signaling pathway (Seo et al. 2011). Variation of nutrients was conducted to determine an optimum condition for the biosynthesis of sesquiterpenes by P. brumalis. Genes encoding terpene synthases, which are crucial to the terpene synthesis pathway, generally respond to environmental factors such as pH, temperature, and available nutrients (Hoffmeister and Keller 2007, Yu and Keller 2005). Calvo et al. described the effect of major nutrients, carbon and nitrogen, on the synthesis of secondary metabolites (Calvo et al. 2002). P. brumalis did not prefer to synthesize sesquiterpenes under all growth conditions. Results of differences in metabolites observed in P. brumalis grown in PDB and modified medium highlighted the potential effect inorganic sources such as $C_4H_{12}N_2O_6$, $KH_2PO_4$, $MgSO_4$, and $CaCl_2$ on sesquiterpene synthesis. ${\beta}$-eudesmol was apparent during cultivation except for when P. brumalis was grown on $MgSO_4$-free medium. These results demonstrated that $MgSO_4$ can specifically control the biosynthesis of ${\beta}$-eudesmol. Magnesium has been reported as a cofactor that binds to sesquiterpene synthase (Agger et al. 2008). Specifically, the $Mg^{2+}$ ions bind to two conserved metal-binding motifs. These metal ions complex to the substrate pyrophosphate, thereby promoting the ionization of the leaving groups of FPP and resulting in the generation of a highly reactive allylic cation. Effect of magnesium source on the sesquiterpene biosynthesis was also identified via analysis of the concentration of total carbohydrates. Our current study offered further insight that fungal sesquiterpene biosynthesis can be controlled by nutrients. To profile the metabolites of P. brumalis, the cultures were extracted based on the growth curve. Despite metabolites produced during mycelia growth, there was difficulty in detecting significant changes in metabolite production, especially those at low concentrations. These compounds may be of interest in understanding their synthetic mechanisms in P. brumalis. The synthesis of terpene compounds began during the growth phase at day 9. Sesquiterpene synthesis occurred after growth was complete. At day 9, drimenol, farnesol, and mevalonic lactone (or mevalonic acid lactone) were identified. Mevalonic acid lactone is the precursor of the mevalonic pathway, and particularly, it is a precursor for a number of biologically important lipids, including cholesterol hormones (Buckley et al. 2002). Farnesol is the precursor of sesquiterpenoids. Drimenol compounds, bi-cyclic-sesquiterpene alcohols, can be synthesized from trans-trans farnesol via cyclization and rearrangement (Polovinka et al. 1994). They have also been identified in the basidiomycota Lentinus lepideus as secondary metabolites. After 12 days in the growth phase, ${\beta}$-elemene caryophyllene, ${\delta}$-cadiene, and eudesmane were detected with ${\beta}$-eudesmol. The data showed the synthesis of sesquiterpene hydrocarbons with bi-cyclic structures. These compounds can be synthesized from FPP by cyclization. Cyclic terpenoids are synthesized through the formation of a carbon skeleton from linear precursors by terpene cyclase, which is followed by chemical modification by oxidation, reduction, methylation, etc. Sesquiterpene cyclase is a key branch-point enzyme that catalyzes the complex intermolecular cyclization of the linear prenyl diphosphate into cyclic hydrocarbons (Toyomasu et al. 2007). After 20 days in stationary phase, the oxygenated structures eudesmol, elemol, and caryophyllene oxide were detected. Thus, after growth, sesquiterpenes were identified. Per these results, we showed that terpene metabolism in wood-rotting fungi occurs in the stationary phase. We also showed that such metabolism can be controlled by magnesium supplementation in the growth medium. In conclusion, we identified P. brumalis as a wood-rotting fungus that can produce sesquiterpenes. To mechanistically understand eudesmane-type sesquiterpene biosynthesis in P. brumalis, further research into the genes regulating the dynamics of such biosynthesis is warranted.

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Effects of Drying Conditions on the Profile of Volatile Terpenoid and Colour of Schizandra Fruit(Schizandra Chinensis fructus) (건조 조건이 오미자의 휘발성 terpene류 및 색도에 미치는 영향)

  • Kim, Yun-Je;Lee, Young-Guen;Choi, Young-Whan;Kim, Yong-Chul
    • Journal of Life Science
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    • v.18 no.8
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    • pp.1066-1071
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    • 2008
  • Schizandra fruit (Schizandra chinensis fructus) were dried by three processes, $50^{\circ}C$ (50HAD), $70^{\circ}C$ hot air (70HAD) and freeze drying process (FRD). Terpenoid were collected by SDE(Simultaneous Steam Distillation-Extraction) and followed by GC-MSD analysis. Also colour profile of each dried samples were measured by Hunter colorimeter. From fresh schizandra fruit, were detected 15 kinds of monoterpene, 28 kinds of sesquiterpene and 7 kinds of terpene alcohol. Myrcene(56.97 ${\mu}g/g$) and ${\gamma}$-terpinene(58.49 ${\mu}g/g$) were the major monoterpenes, ${\beta}$-elemene(120.16 ${\mu}g/g$), ${\alpha}$-bergamotene (103.45 ${\mu}g/g$), ${\gamma}$-selinene (75.97 ${\mu}g/g$), ${\beta}$-cubebene(66.69 ${\mu}g/g$), aristolene (51.25 ${\mu}g/g$) and ${\alpha}$-ylangene(28.06 ${\mu}g/g$) were the sesquiterpenes, and T-muurolol (96.45 ${\mu}g/g$) and terpinen-4-ol(46.02 ${\mu}g/g$) were the terpene alcohols. The dried samples lost more than half of terpenoid content of fresh schizandra fruit during early stage of drying process, and then the level of terpenoid content was not significantly changed. The content of sesquiterpenes appeared to increase until 6 day of FRD. The amount of residual terpene alcohols contained in schizandra fruit dried by FRD was more than those remained after drying by other processes, and schizandra fruit dried by 70HAD exhibited the least residual terpene alcohols. Brightness parameter $L^{\ast}$ decreased with the rise in the level of drying temperature, to which redness parameter $a^{\ast}$ and yellowness $b^{\ast}$ appeared to be similar.

A Study on The Content of Liver Protein, Nucleic Acids, and Guanine Deaminase Activity of Mouse During Acute Starvation (급성(急性) 기아(饑餓)마우스의 간단백질(肝蛋白質), 핵산(核酸) 및 Guanine Deaminase 활성(活性)에 관(關)한 연구(硏究))

  • Park, Seung-Hee;Kim, Seung-Won
    • Journal of Nutrition and Health
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    • v.1 no.2
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    • pp.107-115
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    • 1968
  • Number of aspects, not only nutritional but social as well as political involved in human starvation pose nowadays global problems. In order to help establish the minimum nutritional requirements in the daily life of a man and to free people as well from either undernourishment, malnutrition or even starvation many workers have devoted themselves so far on the research programs to know what and how number of metabolic events take place in animals in vivo. It is the purpose of the present paper to examine in effect to what extent both of the protein and nucleic acids (DNA & RNA) together with an enzyme, guanine deaminase, which converts guanine into xanthine and in turn ends up to uric acid as an end product, undergo changes, quantitatively during acute starvation, using the mouse as an experimental animal. The mouse was strictly inhibited from taking foods except drinking water ad libitum and was sacriflced 24, 48, and 72 hours following starvation thus acutely induced. The animals consisted of two experimental groups, one control and another starvation groups, each being consisted of 6-24 mice of whose body weights ranged in the vicinity of 10 g. The animals were sacriflced by a blow on the head, followed by immediate excision of their livers into ice-cold distilled water, washing adherent blood and other contaminant tissues. The liver was minced foramin, by an all-glass homogenizer immersing it in an ice-bath, followed by subsequent fractionatin of the homogenate (10% W/V in 0.25M sucrose solution made up with 0.05M phosphate buffer of pH 7.4). For the liver protein and guanine deaminase assay, the 10% homogenate was centrifuged at 600 x g for 10 minutes to eliminate the nuclear fraction; and for the estimation of DNA and RNA, the homogenate was prepared by the addition of 10% trichloroacetic acid in order to free the homogenate from the acid-soluble fraction, the remaining residue being delipidate by the addition of alcohol and dried in vacuo for later KOH (IN) hydrolysis. The changes in body and liver wegihts during acute starvation were checked gravimetrically. Protein contents in the liver were monitored by the method of Lowry et al; and guanine deaminase activities were followed by the assay of liberated ammonia from the substrate utilizing the Caraway's colorimetry. The extraction of both DNA and RNA was performed by the Schmidt-Thannhauser's method, which was followed by Marmur's method of purification for DNA and by Chargaff's method of purification for RNA. The determinations of both DNA and RNA were carried out by the diphenylamine reaction for the former and by the orcinol reaction for the latter. The following resume was the results of the present work. 1. It was observed that the body as well as liver weights fall abruptly during starvation, and that the loss of body weight showed no statistical correlation with the decreases in the content of liver protein. 2. The content of liver protein and activity of liver guanine deaminase activity as well decline dramatically, and the specific activities of the enzyme (activity/protein), however, decreased gradually as starvation proceeded. 3. Both of the nucleic acids, DNA and RNA, showed decrements in the liver of mouse during acute starvation; the latter, however, being more striking in the decline as compared to the former. 4. The decreases in the liver protein content as resulted from the acute starvation had no statistically significant correlation with the decrements of DNA in the same tissue, but had regressed with a significant statistical correlation with the fall of RNA in the tissue. 5. The decrease in the activity of guanine deaminase in the liver of mouse during acute starvation was functionally more proportional to the decrease in RNA than DNA, and moreover correlated with the changes in the content of the liver protein. 6. The possible mechanisms involved during in this acute starvation as bring the decreases in the contents of DNA, protein, and guanine deaminase were discussed briefly.

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Amount and Chemical Characteristics of the Epicuticular Waxes on Leaves at Active Tillering and Heading Stages of Rice Varieties (벼 품종들의 분얼성기 및 출수기의 엽표면Wax의 양 및 화학적 조성)

  • Yong Woong, Kwon;Bong Jin, Chung
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.37 no.2
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    • pp.185-197
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    • 1992
  • Differences in the amount and chemical characteristics of the epicuticular waxes on rice leaves were studied for the active tillering and heading stages of rice varieties differing widely in gross leaf-surface property and genetics. The amount of waxes on surfaces of rice leaf-blades was determined by extraction with chloroform and chemical composition of the waxes was characterized by thin layer chromatography, gas liquid chromatography and infrared spectrophotometry. The amount of waxes varied by variety and significantly with growth stage. The amount at the heading stage was 1.7 to 3.6 mg/g fresh weight of leaves, which was two to three times as much as that at the tillering stage of 0.8 to 1.8 mg/g fresh weight. The waxes consisted of seven chemical classes, namely diols, fatty acids, fatty alcohols, fatty aldehydes, fatty esters, saturated and unsaturated hydrocarbons. Diols and unsaturated hydrocarbons were identified as new chemical classes of the rice epicuticular waxes. The polar constituents such as dials, fatty acids and fatty alcohols and the non-polars such as fatty aldehydes, fatty esters, and saturated and unsaturated hydrocarbons were identified at the heading stage, but at the tillering stage only the non-polar compounds were identified. In the carbon numbers (C) of the chemical classes, diols were composed entirely of C30 and acids were mainly of C30 and C31. In alcohols, primary alcohols were composed of C13 and C32, and the secondary alcohols were of C14, C16 and / or C30 regardless of the rice varieties. The acid portion of fatty esters, mainly composed of C22 and C23, showed low cabon numbers compared with the aldehydes. The alcohol portion of them showed a wide distribution in carbon numbers from C13 to C26 depending on the rice varieties. Hydrocarbons had odd carbon numbers, consisting mainly of C29 and C31.

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Development and Validation of the Determination of Sorafenib in Human Plasma using Tandem Mass Spectrometry Coupled with Liquid Chromatography (고속액체크로마토그래피 텐덤질량분석기법을 이용한 사람 혈장 내 소라페닙 농도분석법의 개발 및 검정)

  • Park, Daejin;Lee, Sunggon;Kim, Woomi
    • Journal of Life Science
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    • v.22 no.11
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    • pp.1456-1462
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    • 2012
  • Sorafenib is a multikinase inhibitor and an oral anticancer drug approved for the treatment of patients with advanced renal cell carcinoma and those with unresectable hepatocellular carcinoma. The purpose of this study was to develop an efficient method of the determination of sorafenib in human plasma using tandem mass spectrometry coupled with liquid chromatography (LC/MS/MS) and validate the method by the guidelines of the Korean Food and Drug Administration (KFDA). Plasma samples ($100{\mu}l$) were added with chlorantraniliprole as an internal standard and then mixed with the 0.1% formic acid-containing extraction solution composed of isopropyl alcohol and ethyl acetate (1:4, v/v). After centrifugation, the supernatant was concentrated at $45^{\circ}C$ under negative pressure and centrifugal force. The residue was reconstituted with a mobile phase and injected into the HPLC instrument using a reverse phase Waters XTerra$^{TM}$ C18 column (particle size $3.5{\mu}m$). Liquid chromatography was carried out within the run time of 5 min using a mobile phase composed of buffer (0.1% formic acid and 10 mM ammonium formate), methanol, and acetonitrile (1:6:3, v/v/v). The analytes were monitored by tandem mass spectrometry in the multiple reaction monitoring method programmed to detect sorafenib at 'm/z 465.2 ${\rightarrow}$ 252.5' and chlorantraniliprole at 'm/z 484.4 ${\rightarrow}$ 286.2' with positive electrospray ionization mode ($ES^+$). The result showed the proper linearity ($r^2$ > 0.99) over the range of 2,000-5,000 ng/ml with good accuracy (90.7-103.9%) and precision (less than 10%). The newly developed method using LC/MS/MS was validated by the guideline of KFDA and identified as more sensitive compared to the previous methods.