• Title/Summary/Keyword: 2D GC-MS

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Development of Rapid and Simple Drug Identification and Semi Quantitative Analytical Program by Gas Chromatography-Mass Spectrometry (가스크로마토그래피/질량분석기를 이용한 약물의 확인 및 간이 정량분석 프로그램 개발)

  • Kim, Eun-Mi;Han, Eun-Young;Hong, Hyo-Jeong;Jeong, Su-Jin;Choe, Sang-Gil;Rhee, Jong-Sook;Jung, Jin-Mi;Yeom, Hye-Sun;Lee, Han-Sun;Lee, Sang-Ki
    • YAKHAK HOEJI
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    • v.55 no.2
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    • pp.106-115
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    • 2011
  • Systematic toxicological analysis (STA) means the process for general unknown screening of drugs and toxic compounds in biological fluids. In order to establish STA, in previous study we investigated pattern of drugs & poisons in autopsy cases during 2007~2009 in Korea, and finally selected 62 drugs as target drugs for STA. In this study, rapid and simple drug identification and quantitative analytical program by gas chromatography-mass spectrometry(GC-MS) was developed. The in-house program, "DrugMan", consisted of modified chemstation data analysis menu and newly developed macro modules. Total 55 drugs among 62 target drugs were applied to this program, they were 14 antidepressants, 8 anti-histamines, 5 sedatives/hypnotics, 5 narcotic analgesics, 3 antipsychotic drugs, and etc. For calibration curves, fifty five drugs were divided into four groups of range considering their therapeutic or toxic concentrations in blood specimen, i.e. 0.05~1 mg/l, 0.1~1 mg/l, 0.1~5 mg/l or 0.5~10 mg/l. Standards spiked bloods were extracted by solid-phase extraction (SPE) with trimipramine-D3 as internal standard. Parameters such as retention times, 3 mass fragment ions, and calibration curves for each drug were registered to DrugMan. A series of identification, semi quantitation of target drugs and reporting the results were performed automatically. Calibration curves for most drugs were linear with correlation coefficients exceeding 0.98. Sensitivity rate of DrugMan was 0.90 (90%) for 55 drugs at the level of 0.5 mg/l. For standard spiked bloods at the level of 0.5 mg/l for 29 drugs, semi quantitative concentrations were ranged 0.36~0.64 mg/l by DrugMan. If more drugs are registered to database in DrugMan in further study, it will be useful tools for STA in forensic toxicology.

Production of Phytol, an ACAT Inhibitor, from Callus Culture of Lettuce (Lactuca sativa L.) (상추 (Lactuca sativa L.) callus로부터 ACAT 억제 활성물질, phytol의 생산)

  • An, Kwang-Hee;Jang, Tae-O;Baek, Nam-In;Kim, Se-Young
    • Journal of Plant Biotechnology
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    • v.29 no.1
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    • pp.63-68
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    • 2002
  • The possibility for mass production of phytol, inhibitory diterpene against ACAT (Acyl-CoA: Cholesterol acyltransferase) was investigated by using callus culture of lettuce. The callus were induced from lettuce cotyledon explants on MS medium containing 0.5 mg.L$^{-1}$ NAA after 4 week's culture. Adventitious roots were formed from the explants on MS medium containing 0.5 mg.L$^{-1}$ IBA or 1.0 mg.L$^{-1}$ NAA. Adventitious shoots and roots were emerged from the callus when the callus was transferred to MS medium containing auxin alone, or with cytokinin. The plant growth regulators and their concentrations for the organogenesis were 1.0 mg.L$^{-1}$ NAA, 0.1 mg.L$^{-1}$ BA, 0.5 mg.L$^{-1}$ NAA with 0.1 mg.L$^{-1}$ kinetin, or 0.5 g.L$^{-1}$ 2.4-D with 1.0 mg.L$^{-1}$ kinetin. Analyses of chlorophyll contents showed that chlorophyll contents were higher in morphogenic calli than in non-morphogenic calli. However, the chemical analyses of gas chromatography indicated that phytol contents were not proportionate to the chlorophyll contents of callus. The content of phytol was higher in callus than in lettuce cotyledon.ledon.

Isolation, Quality Evaluation, and Seasonal Changes of Bakkenolide B in Petasites japonicus by HPLC (머위로부터 Bakkenolide B의 순수분리, HPLC분석 방법 및 채취 시기별 함량 분석)

  • Kim, Tae Hoon;Kim, Do Youn;Jung, Won Jung;Nagaiya, Ravichandran;Son, Beung Gu;Park, Young Hoon;Kang, Jum Soon;Lee, Young Jae;Im, Dong-Soon;Lee, Young-Geun;Choi, Yung Hyun;Choi, Young-Whan
    • Journal of Life Science
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    • v.24 no.3
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    • pp.252-259
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    • 2014
  • The leaves of Peatasites japonicus are a traditional oriental medicine with diverse biological activities. A simple and specific analytical method for the quantitative determination of bakkenolide B constituents from methanolic extract of the leaves of P. japonicus was developed. Bakkenolide B was isolated from the leaves of P. japonicus, and its structure was elucidated based on 1D, 2D NMR, and GC-MS spectral data. A liquid chromatographic method was developed to evaluate the quality of P. japonicus through determination of major active compound, bakkenolide B. The wavelengths at 254 and 215 nm were chosen to determine bakkenolide B. The recovery of the method was in the range of 98.6 to 103.1%, and bakkenolide B showed good linearity ($r^2$=0.999) within test ranges. The developed method was applied to the determination of bakkenolide B in the plant part and seasonal changes. The results showed that the content of bakkenolide B in the leaf was higher than in the petiole and rhizome. In this study, a simple, rapid, and reliable high-performance liquid chromatography method was used to determine the percentage and composition of bakkenolide B in P. japonicus procured from different Petasites species plants in South Korea. The method can be employed in routine quantitative analysis and quality control of different products in the market.

Brief Introduction of Research Progresses in Control and Biocontrol of Clubroot Disease in China

  • He, Yueqiu;Wu, Yixin;He, Pengfei;Li, Xinyu
    • 한국균학회소식:학술대회논문집
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    • 2015.05a
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    • pp.45-46
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    • 2015
  • Clubroot disease of crucifers has occurred since 1957. It has spread to the whole China, especially in the southwest and nourtheast where it causes 30-80% loss in some fields. The disease has being expanded in the recent years as seeds are imported and the floating seedling system practices. For its effective control, the Ministry of Agriculture of China set up a program in 2010 and a research team led by Dr. Yueqiu HE, Yunnan Agricultural University. The team includes 20 main reseachers of 11 universities and 5 institutions. After 5 years, the team has made a lot of progresses in disease occurrence regulation, resources collection, resistance identification and breeding, biological agent exploration, formulation, chemicals evaluation, and control strategy. About 1200 collections of local and commercial crucifers were identified in the field and by artificiall inoculation in the laboratories, 10 resistant cultivars were breeded including 7 Chinese cabbages and 3 cabbages. More than 800 antagostic strains were isolated including bacteria, stretomyces and fungi. Around 100 chemicals were evaluated in the field and greenhouse based on its control effect, among them, 6 showed high control effect, especially fluazinam and cyazofamid could control about 80% the disease. However, fluzinam has negative effect on soil microbes. Clubroot disease could not be controlled by bioagents and chemicals once when the pathogen Plasmodiophora brassicae infected its hosts and set up the parasitic relationship. We found the earlier the pathogent infected its host, the severer the disease was. Therefore, early control was the most effective. For Chinese cabbage, all controlling measures should be taken in the early 30 days because the new infection could not cause severe symptom after 30 days of seeding. For example, a biocontrol agent, Bacillus subtilis Strain XF-1 could control the disease 70%-85% averagely when it mixed with seedling substrate and was drenching 3 times after transplanting, i.e. immediately, 7 days, 14 days. XF-1 has been deeply researched in control mechanisms, its genome, and development and application of biocontrol formulate. It could produce antagonistic protein, enzyme, antibiotics and IAA, which promoted rhizogenesis and growth. Its The genome was sequenced by Illumina/Solexa Genome Analyzer to assembled into 20 scaffolds then the gaps between scaffolds were filled by long fragment PCR amplification to obtain complet genmone with 4,061,186 bp in size. The whole genome was found to have 43.8% GC, 108 tandem repeats with an average of 2.65 copies and 84 transposons. The CDSs were predicted as 3,853 in which 112 CDSs were predicted to secondary metabolite biosynthesis, transport and catabolism. Among those, five NRPS/PKS giant gene clusters being responsible for the biosynthesis of polyketide (pksABCDEFHJLMNRS in size 72.9 kb), surfactin(srfABCD, 26.148 kb, bacilysin(bacABCDE 5.903 kb), bacillibactin(dhbABCEF, 11.774 kb) and fengycin(ppsABCDE, 37.799 kb) have high homolgous to fuction confirmed biosynthesis gene in other strain. Moreover, there are many of key regulatory genes for secondary metabolites from XF-1, such as comABPQKX Z, degQ, sfp, yczE, degU, ycxABCD and ywfG. were also predicted. Therefore, XF-1 has potential of biosynthesis for secondary metabolites surfactin, fengycin, bacillibactin, bacilysin and Bacillaene. Thirty two compounds were detected from cell extracts of XF-1 by MALDI-TOF-MS, including one Macrolactin (m/z 441.06), two fusaricidin (m/z 850.493 and 968.515), one circulocin (m/z 852.509), nine surfactin (m/z 1044.656~1102.652), five iturin (m/z 1096.631~1150.57) and forty fengycin (m/z 1449.79~1543.805). The top three compositions types (contening 56.67% of total extract) are surfactin, iturin and fengycin, in which the most abundant is the surfactin type composition 30.37% of total extract and in second place is the fengycin with 23.28% content with rich diversity of chemical structure, and the smallest one is the iturin with 3.02% content. Moreover, the same main compositions were detected in Bacillus sp.355 which is also a good effects biocontol bacterial for controlling the clubroot of crucifer. Wherefore those compounds surfactin, iturin and fengycin maybe the main active compositions of XF-1 against P. brassicae. Twenty one fengycin type compounds were evaluate by LC-ESI-MS/MS with antifungal activities, including fengycin A $C_{16{\sim}C19}$, fengycin B $C_{14{\sim}C17}$, fengycin C $C_{15{\sim}C18}$, fengycin D $C_{15{\sim}C18}$ and fengycin S $C_{15{\sim}C18}$. Furthermore, one novel compound was identified as Dehydroxyfengycin $C_{17}$ according its MS, 1D and 2D NMR spectral data, which molecular weight is 1488.8480 Da and formula $C_{75}H_{116}N_{12}O_{19}$. The fengycin type compounds (FTCPs $250{\mu}g/mL$) were used to treat the resting spores of P. brassicae ($10^7/mL$) by detecting leakage of the cytoplasm components and cell destruction. After 12 h treatment, the absorbencies at 260 nm (A260) and at 280 nm (A280) increased gradually to approaching the maximum of absorbance, accompanying the collapse of P. brassicae resting spores, and nearly no complete cells were observed at 24 h treatment. The results suggested that the cells could be lyzed by the FTCPs of XF-1, and the diversity of FTCPs was mainly attributed to a mechanism of clubroot disease biocontrol. In the five selected medium MOLP, PSA, LB, Landy and LD, the most suitable for growth of strain medium is MOLP, and the least for strains longevity is the Landy sucrose medium. However, the lipopeptide highest yield is in Landy sucrose medium. The lipopeptides in five medium were analyzed with HPLC, and the results showed that lipopeptides component were same, while their contents from B. subtilis XF-1 fermented in five medium were different. We found that it is the lipopeptides content but ingredients of XF-1 could be impacted by medium and lacking of nutrition seems promoting lipopeptides secretion from XF-1. The volatile components with inhibition fungal Cylindrocarpon spp. activity which were collect in sealed vesel were detected with metheds of HS-SPME-GC-MS in eight biocontrol Bacillus species and four positive mutant strains of XF-1 mutagenized with chemical mutagens, respectively. They have same main volatile components including pyrazine, aldehydes, oxazolidinone and sulfide which are composed of 91.62% in XF-1, in which, the most abundant is the pyrazine type composition with 47.03%, and in second place is the aldehydes with 23.84%, and the third place is oxazolidinone with 15.68%, and the smallest ones is the sulfide with 5.07%.

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Volatile Flavor Components in Mash of Takju prepared by using Aspergillus oryzae Nuruks. (Aspergillus oryzae 누룩으로 담금한 탁주 술덧의 발효 과정 중 휘발성 향기성분)

  • Lee, Taik-Soo;Han, Eun-Hey
    • Korean Journal of Food Science and Technology
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    • v.33 no.3
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    • pp.366-372
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    • 2001
  • Volatile flavor components in the mash of Takjus prepared by using Aspergillus oryzae nuruk were identified by using Gas Chromatography and Gas Chromatography-Mass Spectrometry. Twenty-four esters, 21 alcohols, 10 acids, 9 aldehydes and 4 others were found in the mash of Takju. Thirty six components including 13 esters and 12 alcohols were detected in the beginning of fermentation. Twenty nine components were more detected after second day of fermentation and 68 components were detected after 12 days of fermentation. Thirty five flavor components including 12 alcohols such as ethanol, 2-methyl-1-propanol, 3-methyl-1-butanol and benzeneethanol, 13 esters such as ethyl acetate, ethyl caprylate, ethyl butyrate and isoamyl acetate, 4 aldehydes and 6 acids were usually detected in the fermentation process. Ethanol was predominantly found in the range of $79.86{\sim}89.54%$ as a major component by using relative peak area. 3-Methyl-1-butanol, ethyl caprylate and benzeneethanol were some of the major volatile components through the fermentation respectively. Peak area of 2-methyl-1-propanol, 1-hexanol, 1-dodecanol, ethyl acetate, monoethyl butanoate, acetic acid and isobutylaldehyde among the same group were higher than other components depending upon fermentation time.

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The Pyrolytic Behaviors of Tobacco Constituents and Additives by Double-Shot Pyrolyzer (열분해 조건에 의한 담배 성분과 첨가제의 열분해 특성)

  • Lee Jae Gon;Jang Hee Jin;Kwag Jae Jin;Lee Dong Wook;Lee Chang Gook
    • Journal of the Korean Society of Tobacco Science
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    • v.26 no.2 s.52
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    • pp.141-151
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    • 2004
  • This study was conducted to evaluate the characterization of the pyrolysis products of tobacco constituents such as cellulose, lignin and tobacco additives. The pyrolysis condition was designed to simulate the pyrolysis/distillation zone$(200\~600^{\circ}C)$ and combustion zone$(700\~950^{\circ}C)$of burning com in the smoking cigarette. The pyrolysis products were determined by GC/MS after pyrolysis using Double-Shot pyrolyzer. In the case of cellulose and lignin, the number of pyrolysis product in the condition that simulate the pyrolysis/distillation zone was much more than the combustion zone simulating one. The major products of cellulose were levoglucosan, furfural, and 1, 6-anhydro-$\beta$-D-glucofuranose and that of lignin were phenol, 2-methoxy phenol, and 1, 2-dimethoxy benzene. In the case of tobacco additives such as 2, 6-dimethyl pyrazine, maltol, and piperonal, the pyrolysis products of these additives were evaporated from the pyrolyszer at least $96\%$ intactly. These results indicate that tobacco constituents such as cellulose and lignin were thermally degraded at the pyrolysis/distillation zone and thoroughly broke down at the combustion zone, but tobacco additives were intactly evaporated from burning com of smoking cigarette.

Antioxidant Activities of Essential Oils from Citrus × natsudaidai (Yu. Tanaka) Hayata Peels at Different Ripening Stage

  • YANG, Jiyoon;CHOI, Won-Sil;LEE, Su-Yeon;KIM, Minju;PARK, Mi-Jin
    • Journal of the Korean Wood Science and Technology
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    • v.50 no.4
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    • pp.272-282
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    • 2022
  • The essential oil extracted from Citrus × natsudaidai (Yu. Tanaka) Hayata peels is known to have various biological properties. However, the chemical composition of essential oil is influenced by the ripening stages of fruits, which then affects related biological activities. This study investigates the antioxidant activities of essential oils extracted from Citrus × natsudaidai peels at different ripening stages (immature, mature, and overripe). The essential oils were extracted using the hydro-distillation method. As a result of gas chromatography-mass spectrometry (GC-MS) analysis, d-limonene was dominant and was increased as matured. However, 𝛄-terpinene was decreased. The antioxidant properties and their total phenolic content (TPC) were influenced by the ripening stages. The TPC was highest in the immature stage of essential oil (1,011.25 ± 57.15 mg GAE/100 g). 2,2-Diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity was excellent in the immature stage (EC50 = 15.91 ± 0.38 mg/mL). 2,2'-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging activity was superior in overripe stage (EC50 = 20.43 ± 0.37 mg/mL). The antioxidant activity measured using ferric reducing antioxidant power (FRAP) assay showed higher values for the essential oils in immaturity (1,342.37 ± 71.07 mg Fe2+/100 g). Comprehensively, the essential oil in the immature stage showed the best antioxidant activity. Finally, knowing the chemical composition and antioxidant activity at different ripening stages will provide data for selecting the right fruit.

Determination of Trichlorfon Pesticide Residues in Milk via Gas Chromatography with μ-Electron Capture Detection and GC-MS

  • Hem, Lina;Khay, Sathya;Choi, Jeong-Heui;Morgan, E.D.;El-Aty, A.M. Abd;Shim, Jae-Han
    • Toxicological Research
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    • v.26 no.2
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    • pp.149-155
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    • 2010
  • The pesticide trichlorfon is readily degraded under experimental conditions to dichlorvos. A method has therefore been developed by which residues of trichlorfon in milk are determined as dichlorvos, using gas chromatography with ${\mu}$-electron capture detection. The identification of dichlorvos was confirmed by mass spectrometry. Milk was extracted with acetonitrile followed by centrifugation, freezing lipid filtration, and partitioning into dichloromethane. The residue after partitioning of dichloromethane was dissolved in ethyl acetate for gas chromatography. Recovery concentration was determined at 0.5, 1.0, and 2.0 of times the maximum permitted residue limits (MRLs) for trichlorfon in milk. The average recoveries (n = 6) ranged from 92.4 to 103.6%. The repeatability of the measurements was expressed as relative standard deviations (RSDs) ranging from 3.6%, to 6.7%. Limit of detection (LOD) and limit of quantification (LOQ) were 3.7 and $11.1{\mu}g/l$, respectively. The accuracy and precision (expressed as RSD) were estimated at concentrations from 25 to $250{\mu}g/l$. The intra- and inter-day accuracy (n = 6) ranged from 89.2% to 91% and 91.3% to 96.3%, respectively. The intra- and inter-day precisions were lower than 8%. The developed method was applied to determine trichlorfon in real samples collected from the seven major cities in the Republic of Korea. No residual trichlorfon was detected in any samples.

Diffusible and Volatile Antifungal Compounds Produced by an Antagonistic Bacillus velezensis G341 against Various Phytopathogenic Fungi

  • Lim, Seong Mi;Yoon, Mi-Young;Choi, Gyung Ja;Choi, Yong Ho;Jang, Kyoung Soo;Shin, Teak Soo;Park, Hae Woong;Yu, Nan Hee;Kim, Young Ho;Kim, Jin-Cheol
    • The Plant Pathology Journal
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    • v.33 no.5
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    • pp.488-498
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    • 2017
  • The aim of this study was to identify volatile and agardiffusible antifungal metabolites produced by Bacillus sp. G341 with strong antifungal activity against various phytopathogenic fungi. Strain G341 isolated from four-year-old roots of Korean ginseng with rot symptoms was identified as Bacillus velezensis based on 16S rDNA and gyrA sequences. Strain G341 inhibited mycelial growth of all phytopathogenic fungi tested. In vivo experiment results revealed that n-butanol extract of fermentation broth effectively controlled the development of rice sheath blight, tomato gray mold, tomato late blight, wheat leaf rust, barley powdery mildew, and red pepper anthracnose. Two antifungal compounds were isolated from strain G341 and identified as bacillomycin L and fengycin A by MS/MS analysis. Moreover, volatile compounds emitted from strain G341 were found to be able to inhibit mycelial growth of various phytopathogenic fungi. Based on volatile compound profiles of strain G341 obtained through headspace collection and analysis on GC-MS, dimethylsulfoxide, 1-butanol, and 3-hydroxy-2-butanone (acetoin) were identified. Taken together, these results suggest that B. valezensis G341 can be used as a biocontrol agent for various plant diseases caused by phytopathogenic fungi.

Metabolites profiling and hypolipidemic/hypocholesterolemic effects of persimmon (Diosyros kaki Thumb.) by different processing procedures: in vitro and in vivo studies (제조방법에 따른 떫은감 (Diosyros kaki Thumb.)의 대사체 프로파일링과 중성지질/콜레스테롤 대사 관련 유전자발현 연구 : in vitro 및 in vivo 연구)

  • Park, Soo-Yeon;Oh, Eun-Kyung;Lim, Yeni;Shin, Ji-Yoon;Jung, Hee-Ah;Park, Song-Yi;Lee, Jin Hee;Choe, Jeong-Sook;Kwon, Oran
    • Journal of Nutrition and Health
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    • v.51 no.4
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    • pp.275-286
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    • 2018
  • Purpose: Our previous study demonstrated that persimmon (Diospyros kaki Thumb.) at different stages of ripening provided different protective effects against high-fat/cholesterol diet (HFD)-induced dyslipidemia in rats. In this study, we compared the metabolites profile and gene expressions related to triglyceride (TG)/cholesterol metabolism in vitro and in vivo after treating with persimmon water extracts (PWE) or tannin-enriched persimmon concentrate (TEP). Methods: Primary and secondary metabolites in test materials were determined by GC-TOF/MS, UHPLC-LTQ-ESI-IT-MS/MS, and UPLC-Q-TOF-MS. The expression of genes related to TG and cholesterol metabolism were determined by RT-PCR both in HepG2 cells stimulated by oleic acid/palmitic acid and in liver tissues obtained from Wistar rats fed with HFD and PWE at 0, 150, 300, and 600 mg/d (experiment I) or TEP at 0, 7, 14, and 28 mg/d (experiment II) by oral gavage for 9 weeks. Results: PLS-DA analysis and heatmap analysis demonstrated significantly differential profiling of metabolites of PWE and TEP according to processing of persimmon powder. In vitro, TEP showed similar hypolipidemic effects as PWE, but significantly enhanced hypocholesterolemic effects compared to PWE in sterol regulatory element-binding protein 2 (SREBP2), HMG-CoA reductase (HMGCR), proprotein convertase subtilisin/kexin type 9 (PCSK9), cholesterol $7{\alpha}-hydroxylase$ (CYP7A1), and low density lipoprotein receptor (LDLR) gene expression. Consistently, TEP and PWE showed similar hypolipidemic capacity in vivo, but significantly enhanced hypocholesterolemic capacity in terms of SREBP2, HMGCR, and bile salt export pump (BSEP) gene expression. Conclusion: These results suggest that column extraction after hot water extraction may be a good strategy to enhance tannins and long-chain fatty acid amides, which might cause stimulation of hypocholesterolemic actions through downregulation of cholesterol biosynthesis gene expression and upregulation of LDL receptor gene expression.