• Analytical Science and Technology
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• v.34 no.3
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• pp.115-121
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• 2021

Ethyl glucuronide (EtG) and ethyl sulfate (EtS), which are ethanol metabolites, are direct indicators of ethanol intake; they have been studied in a variety of biological samples in forensic science. It is necessary to analyze ethanol metabolites to determine whether the ethanol detected in autopsy cases was due to alcohol consumption before death or due to the ethanol produced from post-mortem decay. In general, EtG and EtS are detected in the blood together with ethanol; however, it may be difficult to secure blood depending on the extent of decay. Therefore, the aforementioned method should be replaced by detecting the ethanol metabolites using tissue biological samples. In this study, we determined the optimal experimental conditions for detecting EtG and EtS from spleen samples using Liquid Chromatography - Tandem Mass Spectrometry (LC-MS/MS). Herein, the test method was validated, and an analysis method was applied to the actual autopsy cases.

• Bulletin of the Korean Chemical Society
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• v.34 no.8
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• pp.2413-2418
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• 2013

Forensic and legal medicine require reliable data to indicate excessive alcohol consumption. Ethanol is oxidatively metabolized to acetate by alcohol dehydrogenase and non-oxidatively metabolized to ethyl glucuronide (EtG), ethyl sulfate (EtS), phosphatidylethanol, or fatty acid ethyl esters (FAEE). Oxidative metabolism is too rapid to provide biomarkers for the detection of ethanol ingestion. However, the non-oxidative metabolite EtG is a useful biomarker because it is stable, non-volatile, water soluble, highly sensitive, and is detected in body fluid, hair, and tissues. EtG analysis methods such as mass spectroscopy, chromatography, or enzyme-linked immunosorbent assay techniques are currently in use. We suggest that nuclear magnetic resonance (NMR) spectroscopy could be used to monitor ethanol intake. As with current conventional methods, NMR spectroscopy doesn't require complicated pretreatments or sample separation. This method has the advantages of short acquisition time, simple sample preparation, reproducibility, and accuracy. In addition, all proton-containing compounds can be detected. In this study, we performed $^1H$ NMR analyses of urine to monitor the ethanol and EtG. Urinary samples were collected over time from 5 male volunteers. We confirmed that ethanol and EtG signals could be detected with NMR spectroscopy. Ethanol signals increased immediately upon alcohol intake, but decreased sharply over time. In contrast, EtG signal increased and reached a maximum about 9 h later, after which the EtG signal decreased gradually and remained detectable after 20-25 h. Based on these results, we suggest that $^1H$ NMR spectroscopy may be used to identify ethanol non-oxidative metabolites without the need for sample pretreatment.

• Archives of Pharmacal Research
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• v.27 no.2
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• pp.173-176
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• 2004

Three phenolic compounds, rosmarinic acid (1), methyl rosmarinate (2), ethyl rosmarinate (3), and two flavonoids, luteolin (4), luteolin-7-O-$\beta$-D-glucuronide methyl ester (5) were isolated from the aerial part of Lycopus lucidus (Labiatae). Their structures were determined by chemical and spectral analysis. Compounds 1-5 exhibited potent antioxidative activity on the NBT superoxide scavenging assay. The $IC_{50}$ values for compounds 1-5 were 2.59, 1.42, 0.78, 2.83, and 3.05 $\mu\textrm{g}$/mL respectively. In addition, five compounds were isolated from this plant for the first time.

• Analytical Science and Technology
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• v.33 no.3
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• pp.151-158
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• 2020

Alcohol, which can easily be obtained in the same way as ordinary beverages, is harmful enough to cause death due to excessive drinking and chronic alcohol intake, so it is important to maintain a proper amount of drinking and healthy drinking habits. In addition, the incidence of behavioral disturbances and impaired judgments that can be caused by chronic alcohol drinking of more than adequate amounts of alcohol is also significant. Accordingly it is very useful for forensic science to check whether the person involved is drunken or is alcoholism state in various accidents. Currently, in Korea, alcohol consumption is determined by detecting the level of alcohol or alcohol metabolism 'ethyl glucuronide (EtG)' in blood or urine samples. However, analysis of alcohol or EtG in blood or urine can only provide information about the current state of alcohol consumption because of a narrow window of detection time. Therefore, it is important to analyze the EtG as a long-term direct alcohol metabolite bio-marker in human hair and to investigate relationship between alcohol consumption and EtG concentration for the evaluation of chronic ethanol consumption. In this study, we established an analytical method for the detection of EtG in Korean hair efficiently and validated selectivity, linearity, limits of detection (LOD), limits of quantification (LOQ), matrix effect, recovery, process efficiency, accuracy and precision using liquid chromatography tandem mass spectrometry (LC-MS/MS). In addition, the assay performance was evaluated in Korean social drinker's hair and the postmortem hair of a chronic alcoholism. The results of this study can be useful in monitoring the alcohol abuse of Korean in clinical cases and legal procedures related to custody and provide a useful tool to evaluate postmortem diagnosis of alcoholic ketoacidosis in forensics.

• Microbiology and Biotechnology Letters
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• v.38 no.3
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• pp.278-282
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• 2010

In this study, the antibacterial activity and component analysis of Persicaria perfoliata extracts were investigated. MIC (Minimum Inhibitory Concentration) values of ethyl acetate fraction from P. perfoliata on Escherichia coli, Propionibacterium acnes, and Staphylococcus aureus were 0.25%, equally. The results showed that the antibacterial activity of the ethyl acetate fraction was the highest in the S. aureus and P. acnes. Ethyl acetate fraction from P. perfoliata extracts showed 5 bands in TLC and 8 peaks in HPLC (320nm). In HPLC chromatogram of ethyl acetate fraction, first main peak 2 (24.99%) and peak 3 (22.65%) were identified as quercetin-3-o-glucuronide and isoquercitrin in the order of elution time. Also, in HPLC chromatogram of deglycosylated fraction, main peak 2 (76.93%) was identified as quercetin in the order of elution time. And these components are very effective as antioxidants. Thus, these results indicate that extract/fractions of P. perfoliata can be applicable to new functional cosmetics for antioxidant, antibacterial activity.

• Journal of Microbiology and Biotechnology
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• v.31 no.11
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• pp.1576-1582
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• 2021

Bacterial β-glucuronidase in the intestine is involved in the conversion of 7-ethyl-10-hydroxycamptochecin glucuronide (derived from irinotecan) to 7-ethyl-10-hydroxycamptothecin, which causes intestinal bleeding and diarrhea (side effects of anti-cancer drugs). Twelve compounds (1-12) from Polygala tenuifolia were evaluated in terms of β-glucuronidase inhibition in vitro. 4-O-Benzoyl-3'-O-(O-methylsinapoyl) sucrose (C3) was highly inhibitory at low concentrations. C3 (an uncompetitive inhibitor) exhibited a k_i value of 13.4 μM; inhibitory activity increased as the substrate concentration rose. Molecular simulation revealed that C3 bound principally to the Gln158-Tyr160 enzyme loop. Thus, C3 will serve as a lead compound for development of new β-glucuronidase inhibitors.

• Proceedings of the PSK Conference
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• 2003.10b
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• pp.200.1-200.1
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• 2003

Cephalotaxus koreana Nakai is an endemic species in Korea. The EtOH extract of leaf and branch from the plant showed potent antitumor activity in Teruhiro's method. The tumor volume inhibition ratio value is 25.2% with 20mg/kg in the BDF1 mouse injected LLC cell. We isolated one flavone, sciadopitysin (1), two flavone O-glycosides, quercetin 3-O-${\beta}$-D-glucuronide 6"-ethyl ester (2), apigenin 7-neohesperidoside (3) in comparison with literatures data. Compounds 1-3 showed stronger antitumor activity than Taxol used as positive control. The inhibition ratio values of compounds 1-3 is 34.9, 31.6, 34.0%, respectively, and Taxol is 27.0 % compared with control group.

• Clinical and Experimental Reproductive Medicine
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• v.9 no.1_2
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• pp.43-53
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• 1982

여성의 난소기능은 뇨중 Oestrone-3-glucuronide를 간편한 solid-phase 의 화학발광성 면역학적 측정법 (Chemiluminescence Imrnunoassay(CIA) 에 의하여 그 기능이 탐지될 수 있다. Oestrone-3-g1ucuronyl-6-bovine serum albumine에 대한 antiserum의 IgG fraction은 polystyrene 실험관벽에 흡착시켰으며, 항원으로서는 est r one-3- gl ucuronyI-6-aminoethyl-ethyl-isoluminol 을 항원 (antigen) 에 labeI 시킨 것이다. 시험 대상물인 뇨는 매일아침뇨(early morning urine) 을 희석 (1:1000 V/V)한 후 100mcl 를 취하여 이를 각기 이중분석액으로 택하였다. 시험관 내에서 결합반응 (1 hour at $4^{\circ}C)이 일어난 후에는 시험관내의 액체를 전부 흡입 폐기시켰으며, 항체반응이 일어난 후 ( antibody-bound fraction )에는 완충액 (400mcl)으로 한번 세척시켰다. 그후 염화수산화물(2N , 200mcl)을 가지고 $22^{\circ}C$에 60 분간 방치 혼합케 한 후 효소(microperoxidase) 와 과산화수소를 가하면서 산화작용에서 발생되는 발광양을 10초동안 측정하여 그 결과를 분석하였다. 위에 기술한 분석방법을 평가하면 다음과 같은 결론을 얻었다. Calibration curve sensitivity$3.12{\pm}0.75$ PG/tube ($mean{\pm}SD$)였고, lntra-assay precision(CV%) 9.52 (20 replicates;$38.4{\pm}3.66$nmol/1) 와 8.81 (15 replicates; $102.4{\pm}8.82$nmol/1)였다. Inter-assay precision(CV%) 은 11.9 (mean of 4 pools-7.03, 23.16, 52.11 과 117.53 nmol/1)로 2개월 동안에 걸쳐 시행되었고, 평균 비이어스(mean bias)는 -0.78 로 28에서 448 nmol 범위로서 매일아침 "뇨"의 차이분(different aliquots)은 좋은 결과를 얻었다. 건강한 여성으로부터 채취된 뇨중 Oestrone-3-glucuronide 의 농도(nmol/1)를 보면 월경주기의 여포기와 배난기 및 황체기에 있어서 각기 $40.2{\pm}9.9$ , $102.3{\pm}39.4$와 $84.3{\pm}13.3$nmol/1였다. 이와같은 결과는 동일한 검사뇨를 방사면역학적 방법(RIA)으로 측정 (6 menstrual cycle)한 결과와 유사한 측정치를 얻으므로서 간편하고 진보된 좋은 방법중의 하나라고 사료되는바이다.

• Applied Chemistry for Engineering
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• v.28 no.4
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• pp.479-484
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• 2017

In this study, antioxidative activities and cellular protective effects of 70% ethanol extracts and fractions from lavender were evaluated. The scavenging activity ($FSC_{50}$) of free radical (1,1-phenyl-2-picrylhydrazyl, DPPH) was 46.6, 45.5 and $477.5{\mu}g/mL$ in the 70% ethanol extract, ethyl acetate fraction and aglycone fraction, respectively. The reactive oxygen species scavenging activities (${OSC_{50}$) of 70% ethanol extract, ethyl acetate fraction and aglycone fraction were 8.1, 3.3 and $17.6{\mu}g/mL$, respectively, and they showed lower antioxidative activity than that of using L-ascorbic acid ($1.5{\mu}g/mL$). However, the aglycone fraction showed higher photohemolysis protective effect than that of using the 70% ethanol extract and ethyl acetate fraction. At $50{\mu}M$ concentration, the cellular protective effect (${\tau}_{50}$) of 70% ethanol extract, ethyl acetate fraction and aglycone fraction from lavender was 70.6, 87.2 and 165.2 min, respectively. In particular, the lavender aglycone fraction showed 3.8 times higher cellular protective effect than that of (+)-${\alpha}$-tocopherol. The lavender fractional components including luteolin 7-O-glucuronide, vitextin, rosmarinic acid, luteolin, and apigenin were identified using TLC and LC-MS. However, the lavender aglycone fraction did not show any significant increase in flavonoids (luteolin and apigenin) compared to that of the ethyl acetate fraction. In conclusion, it is suggested that lavender may be applied as an antioxidant material in cosmetic industries.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.43 no.2
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• pp.175-187
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• 2017

In this study, the antioxidant effect and component analysis for extract and fractions of Cardiospermum halicacabum leaf were investigated. All experiments were performed with 50% ethanol extract, ethyl acetate fraction and aglycone fraction obtained from dried C. halicacabum leaf. The yields of extract and fractions were 16.4, 0.9 and 0.3% per dried powder, respectively. DPPH (1,1-phenyl-2-picrylhydrazyl) radical scavenging activity ($FSC_{50}$) of ethyl acetate fraction ($92.5{\mu}g/mL$) was the greatest radical scavenging activity, but lower than (+)-${\alpha}$-tocopherol ($8.9{\mu}g/mL$). In reactive oxygen species (ROS) scavenging activity (total antioxidant capacity, $OSC_{50}$) on ROS generated in $Fe^{3+}-EDTA/H_2O_2$ system, aglycone fraction ($4.2{\mu}g/mL$) was the highest total antioxidant capacity and similar to L-ascorbic acid ($1.5{\mu}g/mL$). The cellular protective effects of C. halicacabum leaf extract and fractions on the $^1O_2$-induced cellular damage of human erythrocytes were exhibited at all concentration-dependent ($5.0-25.0{\mu}g/mL$). Especially, aglycone fraction (${\tau}_{50}$, 76.4 min) in $25.0{\mu}g/mL$ showed the most protective effect among extracts. Components of the ethyl acetate fraction obtained from C. halicacabum extracts were analyzed by TLC, HPLC chromatogram and LC/ESI-MS. Results showed that the ethyl acetate fraction contained some flavonoids, such as apigenin-7-O-glucuronide, apigenin-7-glucosdie and quercitrin hydrate. These results suggest that the extracts and fractions of C. halicacabum leaf may be applied as antioxidant functional cosmetic raw materials.