Development and validation of a fast quantitative method for plasma dimethylarginines analysis using liquid chromatography-tandem mass spectrometry.

OBJECTIVES: The aim of this work was to implement a fast, accurate and simple method to quantify plasma ADMA and SDMA, in a run time suitable for routine analysis. DESIGN AND METHODS: We developed and validated a hydrophilic interaction chromatographic method coupled to tandem mass spectrometry (HILIC-MS/MS) for separation and simultaneous quantification of Arginine (Arg) and its dimethylarginines, ADMA and SDMA, with a short run time (less than 5 min) using a small volume of human plasma (0.02 mL). RESULTS: Correlation coefficients (r) of the calibration curves ranged from 0.9926 to 0.9984. Within-day and between-day imprecision (CV%) and inaccuracy (%), carry-over and recovery were also evaluated for validation. Preliminary data of Arg, ADMA and SDMA from 30 apparently healthy subjects and type 2 diabetic patients (n=33) with and without kidney dysfunction were calculated and some statistical differences occurred among them (p<0.05). CONCLUSIONS: Data from calibration curves and quality controls reveal that the method is accurate and precise. Healthy subjects and diabetic patients' values are in agreement with those reported in other studies.

Cigarette smoking is associated with low glomerular filtration rate in male patients with type 2 diabetes.

OBJECTIVE: The relationship between cigarette smoking and renal dysfunction in diabetes has predominantly been documented in patients with type 1 diabetes. The aim of the present study was to explore the relationship between cigarette smoking and glomerular filtration rate (GFR) in a large cross-sectional study carried out in male subjects with type 2 diabetes. The role of metabolic syndrome in modulating this relationship was also investigated. RESEARCH DESIGN AND METHODS: One hundred fifty-eight current smokers and 158 never smokers with type 2 diabetes were consecutively recruited. Low GFR was defined as GFR <60 ml/min per 1.73 m(2). RESULTS: The proportion of patients affected by low GFR was significantly higher in current smokers (20.9 vs. 12.0%, P = 0.03). The adjusted risk (odds ratio [OR]) of low GFR in current smokers was 2.20 (95% CI 1.14-4.26, P = 0.02) and markedly higher in patients from the first tertile of disease duration (4.27 [1.26-14.40], P = 0.02). When metabolic syndrome was added to the statistical model exploring the relationship between smoking and low GFR, the risk of low GFR showed a small change, although it did not become any more significant (1.84 [0.98-3.45], P = 0.06). Current smokers showed even higher free oxygen radical test unit values (560.0 +/- 91.5 vs. 442.7 +/- 87.2, P < 0.0001). CONCLUSIONS: In a large population of male patients with type 2 diabetes, the risk of low GFR is markedly enhanced by smoking and is at least partially mediated by metabolic syndrome.