✦ LIBER ✦

HPLC determination of glutathione and other thiols in human mononuclear blood cells

✍ Scribed by M. A. Raggi; R. Mandrioli; G. Casamenti; D. Musiani; M. Marini

Publisher: John Wiley and Sons
Year: 1998
Tongue: English
Weight: 94 KB
Volume: 12
Category: Article
ISSN: 0269-3879
DOI: 10.1002/(sici)1099-0801(199809/10)12:5<262::aid-bmc744>3.0.co;2-j

No coin nor oath required. For personal study only.

✦ Synopsis

A simple and sensitive HPLC method is proposed for the determination of glutathione (GSH) in human mononuclear cells, based on the derivatization of the tripeptide with Ellman's reagent. The mobile phase was composed of a mixture of methanol and ammonium formate (10:90 v/v, with a flow rate of 1 mL/min). The stationary phase was a C18 (4.6 microns, 250 x 4 mm) reversed phase column. The detection of GSH was performed at 280 nm, resulting in a neat chromatographic peak at 5.8 min. A calibration curve showed good linearity over the concentration range 3 x 10(-6) - 6 x 10(-5) M, with a satisfactory precision. The method was found to yield a quantitative recovery of glutathione (96%), to be sensitive (down to 30 pmol of glutathione per injection) and to have a high precision (R.S.D.% approximately equal to 2). The proposed HPLC method allows for the separation and quantitation of cysteine and N-acetylcysteine, if present in biological samples. Furthermore, the method allows for the determination of total thiol present in human mononuclear cells.

📜 SIMILAR VOLUMES

Endosulfan-induced apoptosis and glutath

Endosulfan-induced apoptosis and glutathione depletion in human peripheral blood mononuclear cells: Attenuation by N-acetylcysteine

✍ Tanzeel Ahmed; Ashok K. Tripathi; Rafat S. Ahmed; Shukla Das; Sanvidhan G. Suke; 📂 Article 📅 2008 🏛 John Wiley and Sons 🌐 English ⚖ 131 KB 👁 1 views

## Abstract Present study investigated whether endosulfan, an organochlorine pesticide is able to deplete glutathione (GSH) and induce apoptosis in human peripheral blood mononuclear cells (PBMC) in vitro. The role of oxidative stress in the induction of apoptosis was also evaluated by the measurem

Detection of human respiratory syncytial

Detection of human respiratory syncytial virus sequences in peripheral blood mononuclear cells

✍ Ikuko Yui; Akiyoshi Hoshi; Yukiko Shigeta; Takeshi Takami; Tetsuo Nakayama 📂 Article 📅 2003 🏛 John Wiley and Sons 🌐 English ⚖ 146 KB

## Abstract Peripheral blood mononuclear cells (PBMC) obtained from patients with lower respiratory infections were examined for the detection of human respiratory syncytial virus (RSV) sequences in the N region using the reverse transcription polymerase chain reaction (RT‐PCR). RSV infection was c

Quantitative PCR determination of human

Quantitative PCR determination of human cytomegalovirus in blood cells

✍ J. Satou; T. Funato; N. Satoh; Y. Abe; K.K. Ishii; T. Sasaki; M. Kaku 📂 Article 📅 2001 🏛 John Wiley and Sons 🌐 English ⚖ 71 KB

## Abstract We evaluated a rapid and sensitive method to determine human cytomegalovirus (CMV) DNA levels in blood cells using a quantitative polymerase chain reaction (PCR) technique. This method is based on real‐time detection of PCR using a dual fluorescence‐labeled probe and a sequence detector

Catabolism of Deoxycytidine in Human Per

Catabolism of Deoxycytidine in Human Peripheral Blood Mononuclear Cells and Its Interference with the Determination of in Situ Thymidylate Synthase Activity

✍ E.S.J. Arner; S. Eriksson 📂 Article 📅 1993 🏛 Elsevier Science 🌐 English ⚖ 301 KB

Resting and stimulated human peripheral blood lymphocytes and monocyte-derived macrophages were incubated with tritiated deoxycytidine labeled at the 5position. Release of tritiated water into the medium was thereupon detected utilizing its lack of binding to active charcoal, which is an established

Direct determination of glutathione in h

Direct determination of glutathione in human blood by micellar electrokinetic chromatography: Simultaneous determination of lipoamide and lipoic acid

✍ Karina C. Pañak; Oscar A. Ruiz; Sergio A. Giorgieri; Dr. Luis E. Díaz 📂 Article 📅 1996 🏛 John Wiley and Sons 🌐 English ⚖ 323 KB 👁 2 views

A reproducible, rapid procedure for the determination of glutathione in human blood by micellar electrokinetic chromatography has been developed. Whole blood samples were deproteinized with 5% w/v sulfosalicylic acid (final concentration). After centrifugation, the supernatant was directly injected

Acellular components of Chlamydia pneumo

Acellular components of Chlamydia pneumoniae stimulate cytokine production in human blood mononuclear cells

✍ Mihai G. Netea; Craig H. Selzman; Bart Jan Kullberg; Jochem M. D. Galama; Adrian 📂 Article 📅 2000 🏛 John Wiley and Sons 🌐 English ⚖ 124 KB 👁 1 views

Accumulating evidence suggest that infection with Chlamydia pneumoniae is associated with atherosclerosis, but the mechanisms involved remain unclear. Inflammation is important in the initial phase of atherogenesis, and cytokines are important in the initiation and progression of inflammation. The a