Comparison of HPLC method and commercial ELISA assay for asymmetric dimethylarginine (ADMA) determination in human serum.
The performance of a new ELISA assay kit (DLD Diagnostika GmbH, Hamburg, Germany) for the determination of asymmetric dimethylarginine (ADMA) was evaluated against a reversed phase HPLC method. ADMA concentrations of 55 serum samples were measured with both methods. The intra-assay CV for ADMA-ELISA was 19% (n=10). Inter-assay CVs for ADMA-ELISA were 9% for kit control 1 (0.410+/-0.037 microM) and 14% for kit control 2 (1.174+/-0.165 microM). The intra- and inter-assay CVs for HPLC assay for ADMA were 2.5% (0.586+/-0.015 microM) and 4.2% (0.664+/-0.028 microM), respectively. There was no correlation between these two methods (R(2)=0.0972). The effect of storage conditions of the samples on ADMA concentrations was investigated by HPLC. ADMA concentration was stable after four freezing and thawing cycles. Overall, the HPLC method offered better sensitivity, selectivity and, very importantly, simultaneous determination of ADMA, SDMA, l-homoarginine and l-arginine.
Valtonen
Pirjo
P
Laboratory Centre, Kuopio University Hospital, P.O. Box 1777, FIN-70211 Kuopio, Finland. pirjo.valtonen@kuh.fi
Karppi
Jouni
J
Nyyssönen
Kristiina
K
Valkonen
Veli-Pekka
VP
Halonen
Toivo
T
Punnonen
Kari
K
D001120:Arginine / Q000031:analogs & derivatives* / Q000097:blood / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004355:Drug Stability; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D011786:Quality Control; D015203:Reproducibility of Results
C487735:dimethylarginine; D001120:Arginine