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Titolo:
DETERMINATION OF INTRACELLULAR AND EXTRACELLULAR NITRITE AND NITRATE BY ANION CHROMATOGRAPHY
Autore:
RADISAVLJEVIC Z; GEORGE M; DRIES DJ; GAMELLI RL;
Indirizzi:
LOYOLA UNIV,SCH MED,BURN SHOCK TRAUMA INST,BLDG 110 4TH,2160 S 1ST AVE MAYWOOD IL 60153
Titolo Testata:
Journal of liquid chromatography & related technologies
fascicolo: 7, volume: 19, anno: 1996,
pagine: 1061 - 1079
SICI:
1082-6076(1996)19:7<1061:DOIAEN>2.0.ZU;2-4
Fonte:
ISI
Lingua:
ENG
Soggetto:
OXIDE PRODUCTION; RELAXING FACTOR; L-ARGININE; OXIDATION; SERUM;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Science Citation Index Expanded
Citazioni:
20
Recensione:
Indirizzi per estratti:
Citazione:
Z. Radisavljevic et al., "DETERMINATION OF INTRACELLULAR AND EXTRACELLULAR NITRITE AND NITRATE BY ANION CHROMATOGRAPHY", Journal of liquid chromatography & related technologies, 19(7), 1996, pp. 1061-1079

Abstract

A highly sensitive method for the simultaneous direct detection as well as the ratio of intracellular and extracellular nitrite (NO2-) and nitrate (NO3-) in human macrophage (M phi), plasma and urine has' beendeveloped. Samples were deproteinized with the organic solvent acetonirile, lyophilized and reconstituted in buffer, determination and quantification of nitric oxide (NO) stable end products NO2- and No-3(-) was performed utilizing an isocratic high performance liquid chromatography (HPLC) with an anion exchange column. A mobile phase of 20 mM NaCl with 1 mM mono sodium phosphate-NaH2PO4 at pH = 7.0 was used. Analyte anions were detected by direct UV-wavelength at 210 nm. Sensitivity in intracellular and extracellular fluids were 0.01 mu mol/L for both anions with recovery rates of 99.6-99.4% for NO2- and NO3-. This method has successfully been applied to the determination of nitrite and nitrate in plasma and urine of normal human volunteers as well as in cell line U-937 macrophage cytosol/supernatant. The mean concentration ofNO2- in normal plasma (n=22) was 3.1 +/- 0.4 mu mol/L and NO3- 10.3 +/- 0.3 mu mol/L with ratio of NO2-:NO3- = 0.3; in normal urine (n=22) NO2- was 380 +/- 61 mu mol/L and NO3- 1345 +/- 270 mu mol/L with ratioof NO2-:NO3- = 0.3; in intracellular human macrophage fluid (n=21) NO2- was 0.66 +/- 0.08 mol/L/10(6) M phi, and NO3- 0.68 +/- 0.09 mol/L/10(6) M phi with ratio NO2-:NO3 = 1; and in extracellular human macrophage fluid (n=21) NO2- was 0.03 +/- 0.01 mol/L/10(6) M phi and NO3- 3.03 +/- 0.33 mol/L/10(6) M phi with. ratio NO2-:NO3- = 0.01.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 21/09/20 alle ore 11:35:28