Rapid, sensitive micro blood lead analysis: a mass screening technique for lead poisoning.

A rapid micro blood lead method is described. Analyses were performed on 20-microL blood samples spotted on filter paper, collected in graduated heparinized capillary glass tubes following finger pricks. The samples were air dried on filter paper and mailed to the laboratory in glassine envelopes. These samples stored on filter paper are stable for at least six months. The blood spots were punched out with a 1/4-in. diameter hole punch and placed in Delves cups for insertion into the flame atomic absorption spectrometer. The innovation of this method is that an ashing step precedes sample introduction into the flame. In phase 1, the Delves cup with the blood sample is pushed 1 cm from the flame. The heat is sufficient for the filter paper to ignite and burn to completion in seconds. After the smoke dissipates, the samples are introduced into the flame for lead analysis, reading the signal at 283.3 nm. The entire analysis time is 15 s per sample. The limit of quantitation is 4 micrograms/dL of lead. Standard curves were linear from 4-42 micrograms/dL. The average CV for this range is 8.2%. The comparative study between the MIBK extraction method and this method yielded a correlation coefficient r = .99 (n = 55). The method is fast, practical, economical, and easily adaptable to screen large numbers of micro lead samples.

Capillary gas-liquid chromatography separation of phenethylamines in amphetamine-positive urine samples.

Good gas chromatography (GC) separation of molecules is essential for clean gas chromatography/mass spectrometry (GC/MS) confirmation of compounds. The trifluoro derivatives of ephedrine (E) and methamphetamine (MA) coelute on dimethyl silicone capillary columns, such as DB-1, which are most commonly used by chromatographers. Methods are described to separate E and MA to aid GC/MS confirmations of methamphetamine, ephedrine, or both E and MA together, whichever may be present in Enzyme Immunoassay (EIA)-analyzed amphetamine-positive urine samples. The use of the heptafluoro derivatives of E and MA on a DB-1 column, or the trifluoro derivatives of E and MA on a DB-17 column, is suggested for good gas chromatographic separation.

Rapid confirmation of enzyme multiplied immunoassay technique (EMIT) cocaine positive urine samples by capillary gas-liquid chromatography/nitrogen phosphorus detection (GLC/NPD).

A rapid gas-liquid chromatographic (GLC) method was developed for the confirmation of benzoylecgonine (BE) positive urine samples screened by the enzyme multiplied immunoassay technique (EMIT) assay. The procedure is performed by solvent extraction of BE from 0.1 or 0.2 mL of urine, followed by an aqueous wash of the solvent and evaporation. The dried residue was derivatized with 50 microL of pentafluoropropionic anhydride and 25 microL of pentafluoropropropanol at 90 degrees C for 15 min. The derivatizing reagents were evaporated to dryness, and the derivatized BE, and cocaine if present, were reconstituted and injected into the gas chromatograph. The column was a 15-m by 0.2-mm fused silica capillary column, coated with 0.25 micron of DB-1, terminating in a nitrogen phosphorus detector (NPD). Cocaine and the pentafluoro BE derivatives retention times were 3.2 and 2.6 min, respectively. Nalorphine was used as reference or internal standard with a retention time of 4.78 min. The complete procedure can be performed in approximately 1.5 h. The EMIT cutoff between positive and negative urine samples is 300 ng/mL of BE. The lower limit of sensitivity of this method is 25 ng of BE extracted from urine. Validation studies resulted in confirmation of 101 out of 121 EMIT cocaine positive urine samples that could not be confirmed by thin-layer chromatography (TLC). This represents 84% confirmation efficiency.

Loss of the cortisol response to naltrexone in Alzheimer's disease.

The administration of a single dose of the opiate antagonist naltrexone (NT) was accompanied by significant elevations in plasma cortisol in normal elderly subjects; in contrast, the cortisol response to NT was absent in individuals of comparable age with Alzheimer's disease (AD). The differential effect of AD on the cortisol response was not accompanied by a significant group difference in plasma prolactin in response to NT administration. Furthermore, this differential cortisol response to NT was not associated with any evident differences in age, sex ratio, plasma levels of naltrexone or its major metabolite beta-naltrexol, or with differences in measures of nonspecific stress, such as plasma free MHPG, pulse, or blood pressure, between the two groups. The absence of the well-characterized cortisol response to NT in AD, together with other reports of abnormal responses to other pharmacological challenges, suggests that neuroendocrine abnormalities might be an important concomitant and possibly a central contributor to the pathophysiology of Alzheimer's disease.

Quantitation of l-alpha-acetylmethadol and its metabolites in human serum by capillary gas-liquid chromatography and nitrogen detection.

A procedure is described for the simultaneous measurement of l-alpha-acetylmethadol and its two pharmacologically active metabolites: noracetylmethadol and dinoracetylmethadol. In the method an intramolecular conversion reaction of the two metabolites to their amide configuration is utilized. The reaction is performed while the metabolites are still in the serum. Following solvent extraction the samples are analyzed by capillary gas-liquid chromatography coupled with nitrogen detection. Quantitation is achieved by internal standardization. The lower limit of sensitivity is 5 ng/ml in serum. Absolute sensitivity is 0.1 ng for all three compounds. The advantages over other procedures are: speed due to the single extraction step; increased recovery of noracetylmethadol and dinoracetylmethadol due to decreased polarity of the amides; greater stability of the metabolites in the amide configuration; better chromatographic quantitation and separation because detector response for the amides is greater than it is for the original configuration of the metabolites and the area of the chromatographic tracing is free of interfering substances.

Quantitative determination of haloperidol in human plasma by high-performance liquid chromatography.

We present a method for the quantitative determination of haloperidol in human plasma. The high-performance liquid chromatographic method of analysis is a significant advancement in terms of ease and speed of haloperidol determination. The drug and the internal standard, chlorohaloperidol, are extracted from 2.0 ml of serum or plasma, back-extracted into the aqueous mobile phase, separated on a C-18 reversed-phase column, and monitored at 254 nm. The potential interference by other drugs was evaluated and was found to be negative. The method is sensitive to at least 5 ng/ml of extracted material and is suitable for drug measurement in the therapeutic range (5-20 ng/ml) and in the toxic range (greater than 50 ng/ml).

A rapid, quantitative GLC method for the simultaneous determination of nicotine and cotinine.

Published gas-liquid chromatographic (GLC) methods for the determination of nicotine and cotinine have proved impractical for the analysis of a large number of clinical samples. Significant improvements over other methods have been achieved, being low sample volume (0.5 mL plasma), rapid two-step extraction from plasma, no evaporation step, and good separation. The lower limits of sensitivity for nicotine and cotinine were 1 and 5 ng/mL, respectively. The method was validated by the analysis of plasma samples from cigarette-smoking volunteers. The method described permits the quick, routine determination of nicotine and cotinine in a large number of samples.

Confirmation of EMIT benzodiazepine assay with GLC/NPD.

A simple, rapid, and reliable gas-liquid chromatographic (GLC) confirmation procedure was developed for urine samples found positive by the EMIT-dau benzodiazepine metabolite assay. The procedure involves acid hydrolysis, organic extraction, and identification using GLC-nitrogen-phosphorus detection (NPD). The method was validated in three subjects who took 10 mg diazepam daily for five days. Urinary excretion of the diazepam related substances was monitored quantitatively for 12 days. Considerable differences in diazepam metabolism was observed, despite the small number of subjects used. The data further indicated that both the physical characteristics and the metabolic profile of each individual may determine whether or not their occasional benzodiazepine use would be detected by the EMIT procedure. In some individuals taking daily diazepam, 48 to 72 hours may be required before sufficient metabolites accumulate in the urine to give a positive EMIT reaction.

Propoxyphene and norpropoxyphene kinetics after single and repeated doses of propoxyphene.

Plasma concentrations of propoxyphene (P) and its pharmacologically active metabolite norpropoxyphene (NP) were determined in normal subjects after single 130-mg oral doses and during and after 13 consecutive oral doses of 130 mg P, and in former heroin addicts who were maintained on 900 to 1200 mg of P per day. The data were analyzed using a first-pass elimination pharmacokinetic model. Both P and NP cumulated during repeated dosing to levels 5 to 7 times those after the first dose. In contrast, "maintenance" patients exhibited steady-state trough plasma NP cumulation that exceeded that of P by a factor of 13. Several changes in P and NP kinetics occurred during repeated dosing with P to the normal subjects: P clearance decreased from 994 to 508 ml/min, NP clearance decreased from 454 to 2210 ml/min, P half-life (t 1/2) increased from 3.3 to 11.8 hr, NP t 1/2 increased from 6.1 to 39.2 hr, and area under the concentration time curves for P and NP were doubled. These changes in kinetics during repeated dosing resulted in more extensive cumulation of P and NP than would be predicted from the single-dose kinetic profile. Changes in the extent of first-pass elimination of P result in variability in plasma P and NP that may contribute to P-induced toxicity.

Estimation of phencyclidine in leafy mixtures and in vapor phase after smoking.

Five street samples of leafy material coated with phencyclidine (PCP) were analyzed by a gas chromatographic nitrogen detection assay. The samples contained 15.6+1.8% PCP by weight or 32.2+ 13.8 mg PCP per "joint". An aliquot of a joint was smoked with a laboratory apparatus and the vaporized PCP was collected on a filter. Only 22.6+8.0% of the PCP or 6.7+2.1 mg PCP, reached the filter. This amount is in an approximation of the dose of PCP which becomes available to the oral and pulmonary mucosa following the smoking of a single PCP coated joint.

Simultaneous determination of nicotine and cotinine in human plasma by nitrogen detection gas-liquid chromatography.

Human plasma levels of nicotine and its principal metabolite, cotinine, were simultaneously quantitated by gas-liquid chromatography combined with nitrogen selective detection. Nicotine, cotinine, and the added internal standard ketamine are extracted from plasma at basic pH into methylene chloride, back-extracted into acid, and then re-extracted into methylene chloride. Analysis is carried out on a packed glass column of 3% SE-30 while column temperature is programmed from 150 to 200 degrees C. Detector response is linea over the range of 2 to 50 ng/mL nicotine and 50 to 500 ng/mL cotinine. The method was validated on 150 plasma samples obtained from habitual smokers. Mean levels of 19.5 and 219 ng/mL were found for nicotine and cotinine, respectively. Both the mean and the range of the levels were in agreement with previously reported plasma levels for nicotine and cotinine.