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액체크로마토그래프-탠덤질량분석기(LC-MS/MS)를 이용한 소변 내 D-, L- Lactate 분리 및 정량
문철진,양송현,Moon, Chul Jin,Yang, Song Hyun 대한유전성대사질환학회 2015 대한유전성대사질환학회지 Vol.15 No.2
사람의 신체내에서 주로 존재하는 lactate는 L-lactate로서 몇몇 선천성대사이상과 관련하여 증가된다. 최근 2형 당뇨병(type 2 diabetes)과 만성적인 염증성 위장질환(inflammatory bowel disease)과 관련하여 증가되는 D-lactate를 L-lactate와 분리해야 하는 요구도가 높아졌다. 이에 액체크로마토그래프-탠덤질량 분석기를 사용하여 소변 내 D-, L-lactate를 분리하는 방법을 확립하였으며, 분석시간과 정밀성, 정확성, 특이성 등에서 신뢰성 있는 방법임을 확인하였다. Purpose: Lactate has two optical isomers, L-lactate and D-lactate. In human L-lactate is the most abundant enantiomer of lactate. As plasma and urinary levels of L-lactate is associated with inherited metabolic disorders in general, D-lactate have been linked to the presence of diabetes and inflammatory bowel disease. Previously developed techniques have shown several limitations to further evaluate D-lactate as a biomarker for this condition. In this paper, we describe a highly sensitive, specific and fast liquid chromatography tandem mass spectrometry (LC-MS/MS) method for the analysis of D-, L-lactate in urine. Methods: D- and L-lactate were quantified using high performance liquid chromatography tandem mass spectrometry (LC-MS/MS) with labelled internal standard. Samples were derivatized with (+)-O,O'-diacety-L-tartaric anhydride (DATAN) and seperated on a Poroshell 120 EC-C18 column. Results: Quantitative analysis of D-, and L-lactate was achieved successfully. Calibration curves were linear (r>0.999) over $0.5-100{\mu}g/mL$. Stabilities for samples were within the 10% varation. Inter- and Intra-day assay variations were below 10%. Conclusion: The presented method proved to be suitable for the quantitation of D- and L-lactate and opens the possibility to explore the use of D-lactate as a biomarker.
알루미늄을 투여한 흰쥐의 해마와 대뇌피질에서 Reactive Oxygen Species 생성으로 인한 생체거대분자의 산화적 손상
문철진(Chul-Jin Moon),고현철(Hyun-Chul Koh),신인철(In-Chul Shin),이은희(Eun-Hee Lee),문해란(Hae-Ran Moon) 한국독성학회 2004 Toxicological Research Vol.20 No.3
This work aimed to study the effectiveness of cellular oxidative parameter (malondialdehyde,<br/> protein carbonyl, and 8-hydroxy-2'deoxyguanosine). The experimental groups were aluminum<br/> treated rats and control rats. Aluminum treatd rats were given intraperitoneally aluminum nitrate<br/> nonahydrate (Al3+, 0.2 mmol/kg) daily for 30 days except Sunday. Control rats were injected 1 ml of<br/> saline. After the dose, rats were decapitated and hippocampus and cerebral cortex were removed.<br/> The measured parameters were tissue malondialdehyde (MDA, index of lipid peroxidation), protein<br/> carbonyl (index of protein oxidation), 8-hydroxy-2'-deoxy-guanosine (8-OHdG, index of DNA oxidation),<br/> reduced glutathione (GSH) levels as well as glutathione reductase (GR) and catalase. Al concentrations<br/> in the tissues were also measured. All results were corrected by tissue protein levels. The<br/> results were as followed; 1. The concentrations of Al in the cortex and hippocampus were significantly<br/> higher in the Al-treated rats than in the control rats. 2. Antioxidative enzyme's activity, catalase<br/> and GR, were significantly higher in the Al-treated rats than the control rats. GSH levels were also<br/> higher in the Al-treated rats. 3. MDA, protein carbonyl, and 8-OHdG concentration of Al-treated rats<br/> were significantly higher than those of control rats. 4. The concentrations of antioxidants, and oxidative<br/> stress parameter were correlated with the concentrations of Al in hippocampus and cerebral cortex.<br/> Catalase and GR activity were also correlated with the concentration of Al. Based on these<br/> results, it can be suggested that intraperitoneally injected Al was accumulated in the brain and<br/> induced the increase of antioxidant levels and antioxidatve enzyme activity. Also, the oxidative products<br/> of cellular macromolecules are significantly related to tissue Al concentration. Therefore MDA,<br/> protein carbonyl, and 8-OHdG are useful markers for oxidative stress on cellular macromolecules.
Electrospary Tandem Mass를 이용한 혈중 글리퀴돈의 정량법 개발 및 생체이용률 시험
문철진(Chul-Jin Moon),이은희(Eun-Hee Lee),양송현(Song-Hyun Yang),문해란(Hae-Ran Moon) 대한약학회 2005 약학회지 Vol.49 No.3
A rapid, sensitive and selective electrospray tandem mass spectrometric (ESI-LC/MS/MS) method for the quantitation of gliquidone in human plasma was developed. A bioavailability study of gliquidone tablet (30 mg gliquidone, Boehringer Ingelheim Korea Co.) was performed using the validated ESI-LC/MS/MS method. The dose of 30 mg of gliquidone (1 tablet) was orally administered to 9 healthy Korean subjects. After administration, blood was taken at 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 7, 9, 12, 24, and 33 hour. The validation data were as follows; the standard curve was linear (γ2=0.999) over the concentration range of 10~1000 ng/ml. The coefficient of variation for intra- and inter-day assay were 8.30~18.86, and 2.19~12.92%, respectively. The lower limit of quantification for gliquidone was 10 ng/ml. The pharmacokinetic parameters obtained were as follows; AUCt was 3861.17±1328.61 ng.hr/ml, Cmax was 831.02±227.99 ng/ml, tmax was 2.94±0.77 hr, Ke was 0.19±0.06 1/hr, and t1/2 was 4.47±3.52 hr. Based on the validated analytical method and pharmacokinetic parameters, a standard guideline of the bioavailability test of gliquidone dosage forms was prepared successfully and could be used for the bioequivalence test of gliquidone preparation.
Tandem Mass를 이용한 혈중 니코란딜의 정량법 개발 및 생체이용률시험
문철진(Chul-Jin Moon),이은희(Eun-Hee Lee),양송현(Song-Hyun Yang),문해란(Hae-Ran Moon) 대한약학회 2005 약학회지 Vol.49 No.3
A rapid, sensitive and selective tandem mass spectrometric method (LC-MS/MS) for the quantitation of nicorandil in human plasma was developed. A bioavailability study of Sigmat tablet(5 mg nicorandil, Choongwae Co.) was performed using the validated LC-MS/MS method. The dose of 5 mg of nicorandil (1 tablet) was orally administered to 9 healthy Korean subjects. After administration, blood was taken at 0.25, 0.5, 1, 2, 3, 4, 5, 6, 9, 12, and 24 hour. The validation data were as follows; the standard curve was linear (γ2=0.999) over the concentration range of 0.5~200.0 ng/ml. The coefficient of variation for intra- and inter-day assay were 3.55~7.44, and 2.17~9.10%, respectively. The lower limit of quantification for nicorandil was 0.5 ng/ml. The pharmacokinetic parameters obtained were as follows; AUCt was 145.9±83.0 ng.hr/ml, Cmax was 83.8±32.2 ng/ml, tmax was 0.42±0.13 hr, Ke was 0.56±0.23 1/hr, and t1/2 was 1.42±0.52 hr. Based on the validated analytical method and pharmacokinetic parameters, a standard guideline of the bioavailability test of nicorandil dosage forms was prepared successfully and could be used for the bioequivalence test of nicorandil preparation.
외부 수압을 받는 필라멘트 와인딩 후판 복합재 원통의 좌굴 연구
문철진(Chul-jin Moon),허성화(Seong-Hwa Hur),안정희(Jung-Hee Ahn),권진회(Jin-Hwe Kweon),최진호(Jin-Ho Choi),조종래(Jong-Rae Cho),조상래(Sang-Rae Cho) 한국항공우주학회 2009 韓國航空宇宙學會誌 Vol.37 No.2
외부 수압을 받는 필라멘트 와인딩 후판 복합재 원통의 좌굴 및 파손을 유한요소법과 시험으로 연구하였다. 자체 개발 프로그램인 ACOS와 상업용 프로그램인 M5C.NASTRAN(선형)과 MSC.MARC(비선형)를 이용한 유한요소해석을 수행하였다. 복합재 원통 시편은 T700 카본-에폭시로 필라멘르 와인딩 기법에 의해 [±30/90]<SUB>FW</SUB>, [±45/90]<SUB>FW</SUB>, [±60]<SUB>FW</SUB>, [±60/90]<SUB>FW</SUB>의 각도로 제작하여 수압시험을 수행하였다. 사용한 세 가지 프로그램 중 ACOS가 시험값과 1.7~14.3%의 차이를 보이며 좌굴 압력을 가장 잘 예측하였다. 모든 경우에 좌굴 후 지지하중이 초기 좌굴하중보다 커지지 못하고 최종파손으로 연결되었다. The buckling and failure of filament wound thick composite cylinders under external hydrostatic pressure were investigated by the finite element analysis and test. ACOS, MSC.NASTRAN, and MSC.MARC were used for finite element analysis. T700 carbon-epoxy filament wound composite cylinders were fabricated to have winding angles of [±30/90]<SUB>FW</SUB>, [±45/90]<SUB>FW</SUB>, [±60]<SUB>FW</SUB>, and [±60/90]<SUB>FW</SUB>, and tested to verify the finite element analysis. Among the softwares, ACOS predicted buckling load the best with about 1.7~14.3% deviation from test. Analysis and test shows cylinders do not recover the initial buckling pressure after buckling and directly lead to final failure.