A micro-extraction of volatile metabolites from body fluids for analysis by high resolution gas chromatography and GC-MS



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A solvent extraction method for sampling of volatile metabolites from human biological fluids developed by Zlatkis and Andrawes (45) was tested using plasma, urine, breast milk, and amniotic fluid collected from postpartum females and neonate on day 1 or soon after, depending on the fluid. The technique requires only 100 [mu]l plasma, 3 ml urine, 20 [mu]l breast milk, and 500 [mu]l amniotic fluid to be extracted with 500 [mu]l, 1 ml, 100 [mu]l, and 750 [mu]l ether, respectively, after saturation of the fluid with ammonium carbonate. The centrifuge tube containing the mixture is vortexed, centrifuged, revortexed, and centrifuged. The supernant fraction is distributed by a tubular syringe equipped with a 5" 22g needle onto glass wool contained in a tube that can be fitted directly into the injection port. After stripping the ether from the glass wool with helium, the tube is put into the inlet port. The volatile compounds are desorbed at 240°C into a stainless steel pre-column trap submerged into liquid nitrogen. After 15 minutes, gas chromatographic analysis is begun. The procedure is rapid, reproducible, requires only small volumes of biological fluids; thus, its use as a diagnostic test for pathological disorders is promising.