Drug-analysis technology: overview and state of the art.

Analytical techniques are available to reliably detect and, if necessary, quantify drugs of abuse and their metabolites in urine. A variety of different immunoassays are likely to remain at the heart of initial screening methods but with an increasing emphasis on confirmation of positive findings by GC-MS. The quality of work in laboratories (which may make considerable financial gain through increased volume of samples) must be guarded by both internal quality assurance and external proficiency testing, but most of all by ensuring that staff are truly forensic toxicologists with adequate credentials and experience. Laboratories must be credible to the courts and to their clients, which is simply not the case today. For the past 20 years, the toxicologist has played an increasingly important role as an ombudsman of public-health issues; this new dimension of professional responsibility has almost frightening proportions. The alarm that is rightly felt in the profession can only be assuaged and brought into perspective by increasing the knowledge base from which toxicological opinions are rendered, and by recognizing that, however critical, the analytical laboratory report is only a part, and never the whole case. Just a few areas of immediate needs have been mentioned, but they are critical, they cannot be avoided, and they must receive appropriate attention very soon.

The influence of recombinant DNA-derived human and murine gamma interferons on mouse hepatic drug metabolism.

Human gamma interferon given for up to 5 days by subcutaneous infusion or intraperitoneal injection did not significantly alter mouse hepatic microsomal oxidative drug-metabolizing enzyme activities. In contrast, murine gamma interferon and human alpha interferon given for 5 days at the same dose (10(7) units/kg) caused 25 and 50% decreases, respectively, in hepatic microsomal cytochrome P-450 concentrations. The human alpha interferon-induced decline in cytochrome P-450 was accompanied by a significant drop in p-nitroanisole demethylase activity and significant elevations in serum alanine aminotransferase and cytosolic glutathione S-transferase activities. An elevation in glutathione-S-transferase was the only significant change found following human gamma interferon administration. Microsomal UDP-glucuronosyltransferase activity was unaffected by any interferon.

Concentrations of lidocaine and monoethylglycylxylidide (MEGX) in lidocaine associated deaths.

Concentrations of lidocaine and MEGX were determined in a variety of tissues and other samples collected at autopsy. In 13 of the cases examined in which lidocaine was associated with death, tissue concentrations were greater than 15 mg/kg. Tissue concentrations in other patients treated with lidocaine were significantly lower.

Activation of human polymorphonuclear neutrophil functions by interferon-gamma and tumor necrosis factors.

Recombinant human interferon-gamma (rHuIFN-gamma) and natural human tumor necrosis factor beta (nHuTNF-beta) (previously called lymphotoxin), purified to homogeneity, were used to assess their effects on certain functions of human polymorphonuclear neutrophils (PMN) in vitro. The treatment of PMN with 100 U of either rHuIFN-gamma or nHuTNF-beta for 20 min significantly increased their ability to phagocytize 1.5-microns latex beads as detected by flow cytometry. Preparations of recombinant human TNF-beta (rHuTNF-beta) showed activities similar to those of its natural counterpart in activating phagocytosis. In addition, a significant enhancement in PMN-mediated antibody-dependent cellular cytotoxicity was observed after treatment for 2 hr with IFN gamma and both TNF-alpha and TNF-beta. The enhancement by treatment with a combination of rHuIFN-gamma and nHuTNF-beta exceeded the enhancement caused by either agent alone. We also show that although lipopolysaccharide (LPS) is a potent stimulator of PMN function, polymyxin B can block LPS-induced but not lymphokine-induced activation. These data demonstrate new activities for both TNF-alpha and TNF-beta in augmenting the phagocytic and cytotoxic activities of PMN.

In vivo augmentation of natural killer activity by combined treatment with recombinant gamma interferon and interleukin-2.

The in vivo regulation of natural killer (NK) cell function by recombinant murine gamma interferon (rMuIFN-gamma) and recombinant human interleukin-2 (rHuIL-2) was investigated. Peritoneal exudate-derived natural killer (PE-NK) cells of mice treated with rMuIFN-gamma or rHuIL-2 exhibited significantly enhanced cytolytic activities against YAC-1 target cells. Compared with animals receiving treatment with either rMuIFN-gamma or rHuIL-2 alone, the sequential treatment of mice with both agents resulted in a significantly greater enhancement of PE-NK cell function, especially when rMuIFN-gamma was administered 24 h before treatment with rHuIL-2. The cytolytic activities were significantly diminished after pretreatment of effector cells with anti-Qa-5 antiserum and complement but not with anti-Lyt-3.2 antiserum. These data constitute the first evidence demonstrating the association between IFN-gamma and IL-2 in the regulation of NK cell function in vivo, reinforce the potential benefit of combined treatment with biological response modifiers, and show that the schedule of administration may be a critical requirement for optimal benefits of combined lymphokine treatment in vivo.

Effect of murine gamma-interferon on the mouse liver and its drug-metabolizing enzymes: comparison with human hybrid alpha-interferon.

Murine IFN(gamma) and human IFN(alpha)-AD:Bgl were compared over a limited dose range and after single and multiple dosing for their effect on male mouse liver oxidative and conjugative drug metabolizing enzymes. Both IFNs depressed the microsomal cytochrome P-450 concentration but did not alter cytosolic glutathione S-transferase nor microsomal UDP-glucuronosyltransferase activity. Both IFNs showed some slight hepatotoxicity (elevated serum ALT), alpha AD:Bgl more than gamma, especially after multiple dosing. While the IFNs did not produce significant increases in liver weight, they did increase the yield of microsomal protein. The increased endoplasmic reticulum may compensate for the decreased cytochrome P-450 concentration and so account for the lack of observed effect of the IFNs on hexobarbital sleep times in vivo. Overall, the minimal effects of murine gamma-IFN on the mouse liver were no different than those of human alpha AD:Bgl.

Drugs in fatally injured young male drivers.

One or more drugs were detected in 81 percent of 440 male drivers, aged 15-34, killed in motor vehicle crashes in California; two or more drugs were detected in 43 percent. Alcohol, the most frequently found drug, was detected in 70 percent of the drivers, marijuana in 37 percent, and cocaine in 11 percent. Each of 24 other drugs was detected in fewer than 5 percent. Except for alcohol, drugs were infrequently found alone; typically, they were found in combination with high blood alcohol concentrations. The causal role of drugs in crashes was assessed by comparing drivers with and without drugs in terms of their responsibility for the crash. Alcohol was associated with increased crash responsibility; the role of other drugs could not be adequately determined.

Absorption of ipecac alkaloids in emergency patients.

Syrup of ipecac contains the nauseant alkaloids emetine and cephaeline. Although thousands of doses are given yearly, no data exist on the absorption of these alkaloids in man. We gave 30 mL of USP Syrup to ten adult patients. Blood and urine samples were obtained at approximately one-half and two hours after administration, and the entire volume of vomitus was saved. The samples were then analyzed for cephaeline and emetine by a high-performance liquid chromatographic (HPLC) assay developed in our laboratory. All patients vomited within 30 minutes, but the amounts of alkaloid regurgitated varied from 22 +/- 14% in six patients to 80 +/- 16% in the remaining four. Only six patients had emetine or cephaeline in their blood by two hours (range, 5 to 73 ng/mL), although ten patients had detectable concentrations of the alkaloids in their urine. Measured over two hours, no patient eliminated more than 0.5% of the dose by the urinary route. In our study ipecac was absorbed by all who received it; the extent of absorption varied widely, and elimination by the renal route was small.

Self-poisoning with dextropropoxyphene and dextropropoxyphene compounds: the USA experience.

This survey continues and expands earlier studies (Finkle et al., 1976a, b). A total of 27 medical examiner or coroner offices across the USA and Canada were visited. The combined jurisdictional population of these 27 sites is 64.8 million people: 56.5 million people in the USA (27% of the US population) and 8.3 million in Canada (Ontario). Since 1969 through mid-1983, a total of 4412 cases provided information sufficient for inclusion in this study. In each of the cases the presence of propoxyphene, and often its major metabolite, in the blood or tissues of the deceased was confirmed by toxicological analysis at the survey site. The following is a summary of the survey findings: The incidence of propoxyphene-associated deaths reached a peak in 1977, then declined by 22.2% from 1977 to 1978 and by 33.3% from 1978 to 1979, and by a further 10-18% since then. This continuing decline has occurred despite increased interest in propoxyphene misuse and the existence of improved analytical methods for the detection of propoxyphene and its metabolites. The decline is greater than can be accounted for by the decline in propoxyphene prescribing. The most common manner of death was suicide, accounting for about 45% of the cases; it can be safely assumed that the suicides were under-reported. A large majority of the suicides involving propoxyphene were multiple-drug intoxications, including alcohol. Propoxyphene alone was noted in about one-sixth of the suicides. These findings confirm those of the earlier studies, i.e., that a high proportion of the deaths associated with propoxyphene are suicides, and, in most cases, the deceased were victims of multiple-drug toxicity. More than 90% of cases involved persons between the ages of 20 and 40 years. There were very few instances of paediatric, adolescent or older adult deaths associated with propoxyphene. There is no evidence to support the view that the deceased were part of the street drug-abuse population. A history of heroin abuse appeared in less than 5% of the cases. Even fewer people had been known to have abused propoxyphene before their deaths, but 18% of the population had been known to 'self-medicate', using multiple drugs without appropriate medical supervision. The distribution of males to females approximated that of the US population.(ABSTRACT TRUNCATED AT 400 WORDS)

Quantitative analysis of emetine and cephaeline by reversed-phase high performance liquid chromatography with fluorescence detection.

A versatile method for the quantitation of emetine and cephaeline in biological samples is described. Two milliliters of samples containing N-propylprocainamide as the internal standard are buffered to pH 9 and extracted with n-butyl chloride. After subsequent back extraction into 0.01 M hydrochloric acid, a portion of the acid layer is analyzed by reversed-phase high performance liquid chromatography with fluorescence detection. Routinely, the minimum level of detection for both drugs is 5 ng/mL and linearity is demonstrated from 5 to 2500 ng/mL.

Drugs and driving: a systematic analytical approach.

To collect useful epidemiological data about drug involvement in highway safety, it is essential that sensitive and specific analytical procedures be used to establish the presence of and to determine the concentrations of drugs and metabolites in samples collected from drivers. This paper describes a comprehensive and systematic screening procedure requiring 6 mL of blood, which has been used for the analysis of samples collected from injured and fatally injured drivers. The procedure uses radioimmunoassay, gas chromatography with selective detectors, and high performance liquid chromatography. Drugs and metabolites presumptively identified are then confirmed primarily using gas chromatography--chemical ionization mass spectrometry.

Capillary and packed column GC determination of propoxyphene and norpropoxyphene in biological specimens: analytical problems and improvements.

Several problems associated with the gas chromatographic determination of DPX and NDPX are discussed and an improved method for their extraction and quantification is described. The method involves two separate extractions, one for DPX with SKF 525A as the internal standard and one for NPDX with dinor-LAAM as the internal standard. The use of two internal standards improved quantitative reproducibility by almost 50%. It was also found that routine DPX and NDPX assays were better determined on packed columns than on capillary columns because the quality of the samples and of the columns was critical. The use of two IS and the double extraction procedure is recommended for general toxicological analyses.

Proficiency testing in forensic toxicology: a feasibility study.

This study has shown that a national proficiency testing program in forensic toxicology is feasible. Samples that resemble typical case specimens were prepared and shipped to approximately 100 laboratories. Participation varied between 61 and 73%. Tissue samples obtained from laboratory animals can be used to simulate those encountered by forensic toxicologists. This has been demonstrated using liver homogenates from animals administered pentobarbital and methaqualone and propoxyphene and acetaminophen. There was a large coefficient of variation however, for the quantitation of acetaminophen in liver. The qualitative data obtained during the course of this study showed a very low incidence of false positives. However, there was a disappointingly low percentage of positive responses for (a) low concentrations of secobarbital and (b) the opiate narcotics (morphine and codeine) in blood, despite the fact that sensitive immunoassay procedures are available for detecting these particular compounds in blood samples. The quantitative determination of drugs and metabolites, other than ethanol, shows wide interlaboratory variation. This variation is presumably not a result of the use of different analytical techniques, since gas liquid chromatography was used by the majority of participants to quantitate drugs and metabolites. Forensic toxicologists are willing to participate in a voluntary proficiency testing program conducted by an independent agency. The performance data developed in this study can serve as a baseline for current forensic toxicology laboratory functional capability in the assessment of future changes and improvements in analytical forensic toxicology.