Optimization of short-column gas chromatography/electron ionization mass spectrometry conditions for the determination of underivatized anabolic steroids

A gas chromatographic/mass spectrometric method based on the use of short capillary gas chromatograph columns (3‐5 m) and electron ionization mass spectrometry has been optimized and evaluated for the determination of underivatized anabolic steroids. The short‐column gas chromatographic/mass spectrometric method was shown to result in short analysis times and to require minimal sample preparation, but suffered from some loss in sensitivity and chromatographic resolution compared with conventional gas chromatographic/mass spectrometric techniques for derivatized steroids. Therefore, short‐column gas chromatographic conditions were optimized to maximize the sample transfer efficiency (sensitivity) from the gas chromatograph into the ion source of the mass spectrometer, while maintaining chromatographic integrity and minimizing thermal decomposition. Mass spectrometric conditions were optimized to maximize ionization efficiency with respect to the intensity of the molcular ion and degree of fragmentation such that positive identication of each steroid could be made based on the resulting mass spectra. Under optimized conditons, we have shown that underivatized anabolic steroids spiked into urine samples can be determined at low‐nanogram levels using short‐column chromatography/full‐scan electron ionization mass spectrometry.