An HPLC method for determination of inosine and hypoxanthine in human plasma from healthy volunteers and patients presenting with potential acute cardiac ischemia.
A simple and sensitive high-performance liquid chromatography (HPLC) method utilizing ultraviolet (UV) detection was developed for the determination of inosine and hypoxanthine in human plasma. For component separation, a monolithic C(18) column at a flow rate of 1.0 mL/min with an aqueous mobile phase of trifluoroacetic acid (0.1% TFA in deionized water pH 2.2, v/v) and methanol gradient was used. The method employed a one-step sample preparation utilizing centrifugal filtration with high component recoveries (approximately 98%) from plasma, which eliminated the need of an internal standard. The method demonstrated excellent linearity (0.25-5 microg/mL, R>0.9990) for both inosine and hypoxanthine with detection limits of 100 ng/mL. This simple and cost effective method was utilized to evaluate potential endogenous plasma biomarker(s), which may aid hospital emergency personnel in the early detection of acute cardiac ischemia in patients presenting with non-traumatic chest pain.
Farthing
Don
D
Department of Internal Medicine, Division of Nephrology, Clinical Pharmacology and Hypertension, VCU Medical Center, Virginia Commonwealth University, Richmond, VA 23298, USA.
Sica
Domenic
D
Gehr
Todd
T
Wilson
Bill
B
Fakhry
Itaf
I
Larus
Terri
T
Farthing
Christine
C
Karnes
H Thomas
HT
D016022:Case-Control Studies; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005260:Female; D006801:Humans; D019271:Hypoxanthine / Q000097:blood*; D007288:Inosine / Q000097:blood*; D017202:Myocardial Ischemia / Q000097:blood*; D011683:Purine-Nucleoside Phosphorylase / Q000097:blood; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D019271:Hypoxanthine; D007288:Inosine; D011683:Purine-Nucleoside Phosphorylase