Faculty Collaboration Database

Simultaneous determination of adenosine, inosine, hypoxanthine, xanthine, and uric acid in microdialysis samples using microbore column high-performance liquid chromatography with a diode array detector. Anal Biochem 1996 Jun 15;238(1):34-9

Date

06/15/1996

Pubmed ID

8660582

DOI

10.1006/abio.1996.0246

Scopus ID

2-s2.0-0029948194 (requires institutional sign-in at Scopus site)   62 Citations

Abstract

In the myocardial interstitial space, adenosine and its metabolites are important markers of ischemia, regulators of blood flow, and may produce cardioprotection against ischemia. A fast and sensitive method to assess the concentrations of adenosine and its metabolites is necessary to determine their involvement in mediating these effects. A method for the simultaneous determination of adenosine, inosine, hypoxanthine, xanthine, and uric acid in the interstitial fluid of the canine myocardium was developed using microdialysis, microbore column high-performance liquid chromatography, and a photo diode array detector (DAD). The microdialysis samples were injected directly onto a microbore C18 reverse-phase column without any prior sample preparation. Use of a DAD in this method provided many advantages. First, a DAD allowed the simultaneous detection of UV absorbance at multiple wavelengths, allowing the detection of each compound at their maximal UV absorbance. Further, the full UV absorption spectrum was recorded for each detected peak, confirming peak purity and identity. Using a microbore HPLC column and detection of UV absorbance at the maximal absorbance for each compound improve the sensitivity for all compounds. The detection limit of these compounds is 50 fmol (signal-to-noise ratio, S/N = 3). This method is useful in analyzing the temporal effect of a prolonged period of myocardial ischemia and reperfusion upon interstitial adenosine, inosine, hypoxanthine, xanthine, and uric acid concentrations in an in vivo canine model.

Author List

Mei DA, Gross GJ, Nithipatikom K

MESH terms used to index this publication - Major topics in bold

Adenosine
Animals
Calibration
Chromatography, High Pressure Liquid
Dogs
Extracellular Space
Hypoxanthine
Hypoxanthines
Inosine
Microdialysis
Myocardial Reperfusion Injury
Sensitivity and Specificity
Time Factors
Uric Acid
Xanthine
Xanthines