title: HPLC determination of plasma dimethylarginines: method validation and preliminary clinical application.
abstract: BACKGROUND
  Asymmetric dimethylarginine (ADMA) has been suggested as a possible marker of endothelial dysfunction, and interest in its use in clinical practice is increasing. However, the potential role of symmetric dimethylarginine (SDMA) as an endogenous marker of renal function, has been less widely investigated. The aims of the present study were therefore to determine reference values for dimethylarginines in plasma after method validation, and to ascertain ADMA plasma concentrations in patients with disorders characterized by endothelial dysfunction; a further end-point was to investigate the relationship between SDMA plasma concentrations and estimated GFR (eGFR) as well as plasmatic creatinine in patients with chronic kidney disease (CKD).
  
  
  METHODS
  HPLC with fluorescence detection was used for the determination of plasma dimethylarginines. To verify the clinical usefulness of ADMA and SDMA, values from 4 groups of patients at a high risk of cardiovascular complications as well renal dysfunction (chronic heart failure n=126; type II diabetes n=43; pulmonary arterial hypertension n=17; chronic kidney disease n=42) were evaluated, and compared with the reference values, obtained from 225 blood donors.
  
  
  RESULTS
  The intra- and inter-assay CVs (<5.2%), the absolute and relative recoveries (96-106%) were highly satisfactory. ADMA levels were significantly elevated in all groups of patients compared with controls (p<0.001) with the exception of samples from patients with type II diabetes. SDMA levels were significantly elevated both in the patients with chronic kidney disease and in the patients with type II diabetes complicated by renal insufficiency, the values being closely correlated with both eGFR (R=0.740) and plasmatic creatinine (R=0.700).
  
  
  CONCLUSIONS
  The findings made in the present study shows that ADMA levels are significantly increased in patients with diseases associated with endothelial dysfunction This molecule might, therefore, be used as a biochemical marker for the evaluation of endothelial function. Furthermore, the preliminary results reported suggest that SDMA might be a reliable marker of renal function, especially in peadiatric populations, for which the use of eGFR is not recommended.
authors.affiliation: Department of Laboratory Medicine, University-Hospital, Padova, Italy.
authors.forename: Mariela
authors.initials: M
authors.lastname: Ivanova
authors.forename: Carlo
authors.initials: C
authors.lastname: Artusi
authors.forename: Giovanni Maria
authors.initials: GM
authors.lastname: Boffa
authors.forename: Martina
authors.initials: M
authors.lastname: Zaninotto
authors.forename: Mario
authors.initials: M
authors.lastname: Plebani
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D001120:Arginine / Q000031:analogs & derivatives* / Q000097:blood; D002318:Cardiovascular Diseases / Q000175:diagnosis; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003401:Creatine / Q000097:blood; D004730:Endothelium, Vascular / Q000503:physiopathology; D005260:Female; D005919:Glomerular Filtration Rate; D006801:Humans; D007677:Kidney Function Tests; D008297:Male; D008875:Middle Aged; D051436:Renal Insufficiency, Chronic / Q000150:complications / Q000175:diagnosis*; D055815:Young Adult
chemical_list: C487735:dimethylarginine; C024917:symmetric dimethylarginine; C018524:N,N-dimethylarginine; D001120:Arginine; D003401:Creatine