Direct automated EMIT d.a.u. analysis of N,N-dimethylformamide-modified serum, plasma, and postmortem blood for amphetamines, barbiturates, methadone, methaqualone, phencyclidine, and propoxyphene.

The addition of two volumes of N,N-dimethylformamide (DMF) to serum, plasma, and postmortem blood with subsequent centrifugation resulted in supernatant that could be directly analyzed by EMIT d.a.u. urine reagents on the Syva autocarousel. Application of this method to the drugs below gave cutoff concentrations in milligrams of immunochemically cross-reactive analyte equivalents/L as follows: 0.05 for amphetamine, 0.05 for secobarbital, 0.075 for methadone, 0.05 for methaqualone, 0.025 for phencyclidine, and 0.05 for propoxyphene. Quantitative "false" negative/positive noncongruence between total EMIT cross-reactants and free-drug analyses by gas chromatography/mass spectrometry were 3/4 (n = 50) for amphetamines, 2/0 (n = 60) for barbiturates, 0/0 (n = 47) for methadone, 0/0 (n = 48) for methaqualone, 1/0 (n = 44) for phencyclidine, and 1/2 for propoxyphene (n = 53). Within-day precision, as indicated by the coefficient of variation, of quantitative estimates using low and high controls ranged from 3.7 to 11% and 1.8 to 10.3%, respectively. Using the same control levels, between-day precision of quantitative estimates varied from 5.8 to 30.3% and 3.0 to 11.8%, respectively.

Determination of lysergic acid diethylamide (LSD) in urine by instrumental high-performance thin-layer chromatography.

An instrumental high-performance thin-layer chromatographic (HPTLC) technique for the determination of lysergic acid diethylamide (LSD) in urine was developed. Before chromatographic separation, a single-step extraction with alkaline wash is performed. The procedure can detect less than 1 microgram LSD/L urine. Results of HPTLC determination are compared with those from a radioimmunoassay (RIA) procedure.

Direct automated EMIT d.a.u. analysis of N,N-dimethylformamide-modified serum, plasma, and postmortem blood for benzodiazepines, benzoylecgonine, cannabinoids, and opiates.

Serum, plasma, and postmortem blood treated with two volumes of N,N-dimethylformamide (DMF) and centrifuged, were directly analyzable by means of the EMIT d.a.u. reagents on the Syva Autocarousel in the same manner as urine. Cutoff values in milligrams (immunochemically cross-reactive analyte equivalents)/L are 0.05 for morphine (MOR), 0.15 for benzoylecgonine (BZE), 0.20 for oxazepam (OX), and 0.02 for 11-nor-delta 9-tetrahydrocannabinol carboxylic acid (THCC). The relationship of concentrations (micrograms/mL serum) to absorbance changes (delta A) were S-shaped up to greater than 3.0 for MOR, 4.0 for BZE, greater than 5.0 for OX, and 0.2 for THCC. Beyond these maximal concentrations, delta A values declined. Thus, negatives should be repeated on substantially diluted aliquots to avoid missing extraordinarily high positives. "False" quantitative negative/positive noncongruence between total EMIT cross-reactives and free-drug analyses by gas chromatography/mass spectrometry (GC/MS) were 0/17 (N = 75) for opiates, 8/0 (N = 119) for cocaine products, and 19/5 (N = 103) for cannabinoids. For benzodiazepines (N = 58) the "false" negative/"false" positive ratio of EMIT (total)/high performance liquid chromatography (HPLC) (free) was 4/4. Within-day precision as coefficient of variation (CV) of quantitative estimates was 8-18%. For between-day precision, quantitative estimates varied by 8% for MOR, 15% for BZE, 18% for OX, and 34% for THCC.

Oleandrin distribution in a fatality from rectal and oral Nerium oleander extract administration.

In a fatal (cardiotoxic) case of oleander extract poisoning of a young female, ethanol extracts of blood and tissue homogenates were purified by lead acetate. After removal of excess lead by ammonium sulfate, oleandrin was extracted into chloroform. Oleandrin in the extract concentrates was detected by thin-layer chromatography, with location by fluorescence and chromogenically by means of p-anisaldehyde. Quantitation was performed on dried extracts reconstituted in water/methanol, reacted with hydrogen peroxide, ascorbic acid, and hydrochloric acid, and analyzed by fluorescence spectrophotometry. Excitation was at 355 nm, and fluorescence scanning from 340 to 580 nm. The fluorescence peak at 460 nm was used for the quantitative measurement. The concentrations of oleandrin measured in blood, stomach wall, colon tissue, liver, heart, lung, brain, spleen, and kidney ranged from 10 to 39 micrograms/g, with 200 micrograms/mL in the total gastric content residue submitted for analysis.