Routine identification of drugs of abuse in human urine : II. Development and application of the XAD-2 resin column method

Abstract

The methods described in this report were developed and applied to the detection of drugs of abuse in human urine in a laboratory analyzing in excess of one thousand urines per day. Routinely 2 ml of urine were analyzed fluorimetrically for morphine and quinine. The positive morphine and/or quinine samples were confirmed by subjecting 15-ml aliquots of urine to acid hydrolysis, extracting and applying the extract to thin-layer chromatograms followed by specific chromogenic spray reagents. 25 ml of urine were applied to polypropylene columns containing 1.1 g of Amberlite XAD-2 resin. The flow rates were controlled with the hydraulic flow control apparatus and the resin eluted with 15 ml chloroform—isopropanol (3:1). The organic solvent extracts were shaken with 1 ml of saturated sodium bicarbonate and the separated organic phases evaporated to dryness in a water bath under a stream of air. The residues were dissolved in methanol and applied equally to silica gel plates and sheets. The plates were developed in ethyl acetate—methanol—water—ammonia (85:10:3:1) and the sheets in chloroform—methanol—ammonia (90:10:1). The plates were sprayed with 0.3% ninhydrin—acid, 5% H 2SO 4, iodoplatinate and p-nitroaniline followed by NaOH. The Polygram sheets were sprayed with 0.4% ninhydrin—acetone, HgSO 4, diphenylcarbazone, NaI and iodoplatinate. The opiates, opioids, barbiturates, amphetamines, phenothiazines, antihistamines and minor tranquilizers were detected by specific color reactions and R F values. The plates and sheets were cross-compared for final interpretation of the results. These methods were unusually simple, extremely rapid, inexpensive and very effective in detecting psychoactive drugs at concentrations ranging from 0.1 to 3 μg/ml of urine.