Analytical Sciences

Abstract − Analytical Sciences, 29(2), 221 (2013).

Gas Chromatographic Analysis of Guanidino Compounds in Sera and Urine of Uremic Patients Using Glyoxal and Ethyl Chloroformate as Derivatizing Reagents
Subhan Ali MAJIDANO and Muhammad Yar KHUHAWAR
Institute of Advanced Research Studies in Chemical Sciences, University of Sindh, Jamshoro, Pakistan
Gas chromatographic (GC) method has been developed for the determination of the guanidino compounds: guanidine (G), methylguanidine (MG), guanidinoacetic acid (GAA), guanidinopropionic acid (GPA), guanidinobutyric acid (GBA) and guanidinosuccinic acid (GSA) was carried out after precolumn derivatization with glyoxal and ethyl chloroformate from the column HP-5 (30 m × 0.32 mm i.d.) at 90°C for 3 min, followed by a heating rate 25°C/min up to 260°C with a nitrogen flow rate of 2 ml/min. Detection was by FID. The linear calibrations were obtained within 0.1 – 20.0 μmol/L, with limits of detection (LODs) within 0.014 – 0.024 μmol/L. The separation and derivatization was repeatable (n = 6) with relative standard deviations (RSD) within 0.8 – 1.9% in retention time and 0.5 – 1.8% in peak height/peak area. A number of additives and amino acids did not affect the determination. The method was applied for the determination of guanidino compounds from the serum and urine of 9 healthy volunteers and 8 uremic patients and the amounts found were in the range 0.08 – 0.48 and below the limit of detection (LOD) – 345 μmol/L and 1.82 – 13.88 and 0.77 – 432.0 μmol/L with RSDs within 4.2%, respectively.