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Introduction: Pentobarbital (PB) is a euthanasia drug in doses of 2 to 10 grams, causing death within 15-30 minutes. We report a case of recovery from lethal pentobarbital deliberate self-poisoning with confirmatory serum drug concentrations., Case Report: A 45-year-old male purchased 20 grams of PB powder over the Internet. He ingested this powder and then alerted his mother 10 minutes later. She found him unresponsive and commenced cardiopulmonary resuscitation (CPR). Within 20 minutes of ingestion, emergency medical services arrived and initiated advanced life support. On arrival to the emergency department, heart rate was 116 bpm, BP 117/62 mmHg, on an epinephrine infusion. He was hypotonic and hypothermic, with absent brainstem reflexes. ECG and CT brain were normal. Activated charcoal was administered and he was admitted to ICU. He remained comatose with absent brainstem reflexes until day 5. Cerebral angiogram on day 3 was normal. Qualitative urine testing detected pentobarbital suggesting ongoing drug effects as the cause of coma. He was extubated on day 10, eventually making a full recovery. At 2.5 hours post-ingestion, PB concentration was 112 mg/L; PB peaked at 116 mg/L at 29 hours; PB was 2 mg/L at 190 hours and undetectable over 200 hours post-ingestion., Discussion: Average PB concentration in fatalities is reported around 30 mg/L. This patient survived higher serum concentrations with early CPR and prolonged cardiorespiratory support in the ICU. Assessment of brainstem death should be deferred until PB has been adequately eliminated.

Herb-drug interactions are an important safety concern and this study was conducted regarding the interaction between the natural top-selling antidepressant remedy Hypericum perforatum (Hypericaceae) and conventional drugs. This study examined the influence of acute pretreatment with different extracts of Hypericum perforatum from Serbia on pentobarbital-induced sleeping time, impairment of motor coordination caused by diazepam and paracetamol pharmacokinetics in mice. Ethanolic extract, aqueous extract, infusion, tablet and capsule of Hypericum perforatum were used in this experiment. The profile of Hypericum perforatum extracts as well as paracetamol plasma concentration was determined using RP-HPLC analysis. By quantitative HPLC analysis of active principles, it has been proven that Hypericum perforatum ethanolic extract has the largest content of naphtodianthrones: hypericin (57.77 µg/mL) and pseudohypericin (155.38 µg/mL). Pretreatment with ethanolic extract of Hypericum perforatum potentiated the hypnotic effect of pentobarbital and impairment of motor coordination caused by diazepam to the greatest extent and also increased paracetamol plasma concentration in comparison to the control group. These results were in correlation with naphtodianthrone concentrations. The obtained results have shown a considerable influence of Hypericum perforatum on pentobarbital and diazepam pharmacodynamics and paracetamol pharmacokinetics.

Objectives: To determine the pharmacokinetics of pentobarbital in neonates, infants, and young children with congenital heart disease after open-heart surgery., Study Design: Thirty-five subjects (3.0 days-4.4 years) after open-heart surgery who received pentobarbital as standard of care were enrolled. Serial pharmacokinetic blood samples were obtained. A population-based, nonlinear mixed-effects modeling approach was used to characterize pentobarbital pharmacokinetics., Results: A two-compartment model with weight as a co-variate allometrically expressed on clearance (CL), inter-compartmental clearance, central (V1) and peripheral volume of distributions, bypass grafting time as a co-variate on CL and V1, and age and ventricular physiology as co-variates on CL best described the pharmacokinetics. A typical infant (two-ventricle physiology, 6.9 kg, 5.2 months, and bypass grafting time of 60 minutes) had a CL of 0.12 L/hr/kg, V1 of 0.45 L/kg, and peripheral volume of distributions of 0.98 L/kg. The bypass grafting effect was poorly estimated. For subjects <12 months age, an age effect on CL remained after accounting for weight and was precisely estimated., Conclusions: Pentobarbital pharmacokinetics is influenced by age and weight. Subjects with single-ventricle physiology demonstrated a 15% decrease in clearance when compared with subjects with two-ventricle physiology., (Copyright © 2011 Mosby, Inc. All rights reserved.)

Two dogs, a 13-year-old spayed female and a 7-year-old neutered male, were diagnosed with pentobarbital poisoning. Follow-up investigation determined that the source of pentobarbital was the carcass of a horse that had been euthanized more than 2 years previously and that was also apparently responsible for the death of a least 1, and possibly 2, other dogs. The fact that the horse carcass remained lethally toxic more than 2 years after it was euthanized reemphasizes the necessity of proper disposal of euthanized animals.

Barbiturates are central nervous system depressants with sedative and hypnotic properties. Some barbiturates, with longer half-lives, are used as anticonvulsants. Their mechanism of action includes activation of gamma-aminobutyric acid (GABA) mediated neuronal transmission inhibition. Clinically used barbiturates include amobarbital, butalbital, pentobarbital, phenobarbital, secobarbital, and thiopental. Besides their therapeutic use, barbiturates are commonly abused. Their analysis is useful for both clinical and forensic proposes. Gas chromatography mass spectrometry is a commonly used method for the analysis of barbiturates. In the method described here, barbiturates from serum, plasma, or urine are extracted using an acidic phosphate buffer and methylene chloride. Barbital is used as an internal standard. The organic extract is dried and reconstituted with mixture of trimethylanilinium hydroxide (TMAH) and ethylacetate. The extract is injected into a gas chromatogram mass spectrometer where it undergoes "flash methylation" in the hot injection port. Selective ion monitoring and relative retention times are used for the identification and quantitation of barbiturates.

The Bexar County Medical Examiner's Office in San Antonio, Texas, has encountered 3 cases within a 15-month period involving decedents who were pronounced dead by brain death or cardiac death examination and who had elevated, if not toxic concentrations of pentobarbital present at the time of examination. The elevated levels of pentobarbital were discovered during an autopsy examination performed for medicolegal reasons. The diagnosis of brain death and the implications of pentobarbital intoxication during a brain death examination are discussed.

Background: Several drugs may affect the diagnosis of brain death by depressing the electroencephalographic signal. Serum levels of these drugs must be below their respective therapeutic ranges., Methods: A high performance liquid chromatography-based fast and simple method was developed for determination of thiopentone, pentobarbitone, phenobarbitone, methohexital and propofol in serum and validated according to international recommendations., Results: Separation of extracted analytes was performed on a reversed phase column [Agilent Zorbax SB C18, 5 microm, 4.6 x 250 mm; mobile phase 50% 50 mM NaH(2)PO(4) pH 4.6 mixed with 35% (v/v) acetonitrile and 15% (v/v) methanol]. Calibration curves were linear throughout the selected ranges (microg/mL, thiopentone 0.25-50, pentobarbitone 0.25-25, phenobarbitone 2.5-50, methohexital 0.125-2.50, propofol 0.25-5.0). The standard deviations for the regression line, recovery, imprecision and accuracy results were all highly satisfactory. The lower limits of quantification for propofol, thiopentone and pentobarbitone were set at 0.25 microg/mL, for phenobarbitone 2.5 microg/mL, and for methohexital 0.125 microg/mL, which are below the lowest pharmacologically relevant serum concentrations. Intra- and inter-day coefficients of variation were less than 10% throughout as determined with six replicates., Conclusions: The method presented is suitable for drug monitoring to help enhance the reliability of the diagnosis of brain death.

Toxicokinetic parameters were analyzed in 25 patients who were acutely intoxicated with pentobarbital. The serum pentobarbital concentrations were at hypnotic / therapeutic levels (1- 10 microg/mL) in 11 patients, at toxic levels (10-24 microg/mL) in 10 patients, and at lethal levels (>24 microg/mL) in 4 patients. One fatal case was encountered with a serum pentobarbital concentration of 37.27 microg/ mL. The relationships between serum pentobarbital concentrations and the drug, the induced toxic symptoms, such as respiratory / cardiac depression and hypotension, were in good correlation. The therapy of direct hemoperfusion carried out for many pentobarbital-intoxicated patients was highly effective.

Brain death diagnosis may be confounded by centrally acting drugs. The certainty of brain death diagnosis can be enhanced by demonstrating that the concentrations of such drugs are well below the therapeutic range. A combined high-performance liquid chromatography-based method was developed for the benzodiazepines midazolam, 1-hydroxymidazolam, 1-hydroxymidazolam glucuronide, diazepam, and nordiazepam and for the barbiturates thiopentone and pentobarbitone in serum or plasma of critically ill patients. The lower limits of detection of the assays for benzodiazepines and barbiturates were 2.5 ng/mL and 0.05 microg/mL. The lower limits of the working ranges of these assays were set at 25 ng/mL and 0.5 microg/mL, respectively, and are below the lowest pharmacologically active plasma concentrations of these drugs. Intra- and interday coefficients of variations were less than 2.5% and 11.0% throughout, as determined with six replicates (n = 6). These assays were accurate in that the relative difference between actually measured and expected concentration never exceeded 12%. Utilization of these assays will render the diagnosis of brain death more reliable.

We investigated a method for the simultaneous screening, identification, and quantitative determination of salicylic acid, acetaminophen, theophylline, barbiturates, and bromvalerylurea, drugs that frequently cause acute poisoning in Japan and therefore require rapid analysis for effective treatment in the clinical setting. The method employs liquid chromatography/electrospray mass spectrometry (LC/MS) of solid-phase extracted serum samples. For LC/MS ionization, the electrospray-ionization method was used, with acetaminophen in the positive-ion mode, and salicylic acid, theophylline, phenobarbital, bromvalerylurea, pentobarbital, amobarbital, and o-acetamidophenol (internal standard) in the negative-ion mode, the base ions were used in each case for quantitative analysis. Quantitation was possible for the following sample concentration ranges: salicylic acid and acetaminophen, 100 to 5 microg/ml; theophylline, 100 to 0.5 microg/ml; and phenobarbital, bromvalerylurea, pentobarbital, and amobarbital, 100 to 1 microg/ml. Using full-scan mass spectrometry, the lower detection limits of 1 microg/ml for salicylic acid and acetaminophen, 0.1 microg/ml for theophylline, and 0.5 microg/ml for phenobarbital, bromvalerylurea, pentobarbital, and amobarbital were adequate for identifying acute poisoning. When each compound was added to serum to a final concentration of 5 microg/ml and solid-phase extraction was performed using Oasis HLB 1-cc (30-mg), the mean recovery rate of each compound was 89.2 to 96.1% (n=5), and the coefficients of variation of the intraday and interday assays were 3.55 to 6.05% (n=5) and 3.68 to 6.38% (n=5), respectively, which are acceptable. When this method of analysis was applied in testing the sera of a female patient who had consumed a large amount of an unknown commercial drug, salicylic acid and bromvalerylurea were identified, and the treatment strategy could be determined in accordance with the serum concentration of those drugs.

Objective: The bispectral index (BIS), a processed variable derived from the raw electroencephalogram (EEG) used to guide sedation in the intensive care unit (ICU), has not been tested during barbiturate therapy for elevated intracranial pressure. We determined the BIS and suppression ratio (SR) values during traditional burst monitoring of the raw EEG during pentobarbital infusions., Design: Prospective, observational cohort study., Setting: A 42-bed multidisciplinary ICU in a tertiary care medical center., Patients: Twelve consecutive patients with elevated intracranial pressure treated with pentobarbital infusions., Intervention: All patients were monitored continuously with the Aspect Medical Systems A-1050 bedside EEG monitor using a bilateral referential montage. Pentobarbital doses were titrated based on the raw EEG to attain a burst-suppression pattern with a goal of 3-5 bursts/minute. Drug dosage, intracranial pressure, cerebral perfusion pressure values, EEG bursts/minute, BIS version 3.2, and SR were recorded daily., Measurements and Main Results: The 12 patients were monitored for 62 patient-days. Mean +/- SD age was 32 +/- 15 years, seven (58%) patients were male, mean Acute Physiology and Chronic Heath Evaluation II score was 17.0 +/- 5.0, and hospital mortality was 42%. The mean pentobarbital infusion rate was 124 +/- 49 mg/hour or 2.3 +/- 1.3 mg/kg/hour, and mean pentobarbital serum concentration was 29.7 +/- 13 microg/ml. The mean BIS value was 18 +/- 14, mean SR 56% +/- 36%; BIS correlated well with SR (r=-0.99, p<0.001). For patient-days with a burst-suppression pattern, BIS 3.2 (r=0.90, p<0.001) and SR (r=-0.89, p<0.001) strongly correlated with the number of bursts/minute. The mean BIS value corresponding to 3-5 bursts/minute was 15 (95% confidence interval [CI] 10-20); SR value was 71 (95% CI 61-80)., Conclusion: The Aspect A-1050 applied to patients and monitored by nurses and physicians works well as a bedside EEG monitor, providing a raw EEG signal to titrate barbiturate therapy. The continuous data trend and real-time digital output for the BIS and SR quantify the degree of EEG suppression well and may prove helpful in facilitating titration of barbiturate infusions.

The effects of baclofen and pancuronium bromide on evoked electromyogram (EMG), cortical electroencephalogram (EEG) and auditory brainstem responses (ABR) were studied in pentobarbital anesthetized normal rabbits. Evoked EMG was measured in the gastrocnemius muscle by electrical stimulation of the sciatic nerve. Intravenous injection of baclofen decreased EEG and arterial blood pressure and light reflex, however, it had no significant influence on EMG or ABR at doses of 10 and 20 mg/kg/h. Pancuronium bromide immediately inhibited respiration, decreased EEG and EMG, however, it had no significant influence on arterial blood pressure, ABR, or light reflex, at doses of 0.4 and 1.0 mg/kg/h in anesthetized rabbits. ABR waves were observed until just before cardiac arrest with both of the muscle relaxants. It is suggested that ABR are not influenced by central or peripheral muscle relaxants, or by pentobarbital.

Background: Barbiturates enhance gamma-aminobutyric acid type A (GABA(A)) receptor function and also inhibit the alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) subtype of glutamate receptor. The relative contribution of these actions to the behavioral properties of barbiturates is not certain. Because AMPA receptor complexes that lack the GluR2 subunit are relatively insensitive to pentobarbital inhibition, GluR2 null mutant mice provide a novel tool to investigate the importance of AMPA receptor inhibition to the anesthetic effects of barbiturates., Methods: GluR2 null allele (-/-), heterozygous (+/-), and wild-type (+/+) mice were injected with pentobarbital (30 and 35 mg/kg intraperitoneally). Sensitivity to anesthetics was assessed by measuring the latency to loss of righting reflex, sleep time, and the loss of corneal, pineal, and toe-pinch withdrawal reflexes. In addition, patch-clamp recordings of acutely dissociated CA1 hippocampal pyramidal neurons from (-/-) and (+/+) mice were undertaken to investigate the effects of barbiturates on kainate-activated AMPA receptors and GABA-activated GABA(A) receptors., Results: Behavioral tests indicate that sensitivity to pentobarbital was increased in (-/-) mice. In contrast, AMPA receptors from (-/-) neurons were less sensitive to inhibition by pentobarbital (concentrations that produced 50% of the maximal inhibition [IC50], 301 vs. 51 microM), thiopental (IC50, 153 vs. 34 microM), and phenobarbital (IC50, 930 vs. 205 microM) compared with wild-type controls, respectively. In addition, the potency of kainate was greater in (-/-) neurons, whereas no differences were observed for the potentiation of GABA(A) receptors by pentobarbital., Conclusions: The GluR2 null mutant mice were more sensitive to pentobarbital anesthesia despite a reduced sensitivity of GluR2-deficient AMPA receptors to barbiturate blockade. Our results indicate that the inhibition of AMPA receptors does not correlate with the anesthetic effects of barbiturates in this animal model. We postulate that the increase in the sensitivity to anesthetics results from a global suppression of excitatory neurotransmission in GluR2-deficient mice.

The authors evaluated a new Cassette Serum Barbiturates fluorescence polarization immunoassay (FPIA) on the COBAS INTEGRA. The assay was calibrated with secobarbital standards at 0, 0.5, and 4.0 microg/mL. The assay range was 0.030 to 80 microg/mL using an automatic 1/20 postdilution feature. Precision was established for two COBAS INTEGRA instruments for ten days by assaying secobarbital target concentrations ranging from 0.125 to 2.2 microg/mL. The coefficients of variation (CV) for the above target concentrations for the first instrument ranged from 2.7% to 8.3%, and for a second instrument, 3.8 to 8.3%. Seven clinically elevated bilirubin samples were spiked with 0.46 and 1.77 microg/mL secobarbital. Bilirubin interference was less than 10.9 and less than 7.9%, respectively. The average recovery ranged from 85% to 94%. The mean difference in recovery in serum versus plasma was < or = 3%. Fifty-two clinical samples were analyzed for butalbital, pentobarbital, secobarbital, and phenobarbital by GC/MS, and the results were compared to the new Cassette Serum Barbiturates FPIA. The diagnostic sensitivity and specificity were 95% and 100%, respectively. Both FPIA and GC/MS assays are clinically efficacious for monitoring serum barbiturates.

A gas chromatographic-mass spectrometric (GC-MS) assay for the determination of thiopental and its main metabolite pentobarbital in human plasma is presented in this study. The sample preparation consists only in the addition of the internal standard barbital and an acidic extraction with ethyl acetate. Analytical separation is accomplished on a RTX-1 15 m x 0.25 mm capillary column with a film thickness of 0.5 micron. The effluent is observed by a mass selective detector operating in the single ion monitoring mode. The limits of detection are 5 ng/ml for pentobarbital and 10 ng/ml for thiopental, the intra-day variabilities are 2.2% and 4.0% and the inter-day variabilities are 3.3% and 7.1% at concentrations of 5 micrograms/ml, respectively. Applying this assay, the stability of thiopental and pentobarbital in human plasma was tested at concentrations of 5 micrograms/ml each. Thiopental is stable in human plasma at least over 41 days stored at -20 degrees C and 5 degrees C, respectively. A decay of about 2%/day is observed under storage at ambient temperature (19-20 degrees C). Pentobarbital is stable under all storage conditions. Methanolic solutions of thiopental are stable for 83 days under storage at 5 degrees C. Aqueous solutions of thiopental-sodium are stable for at least 23 days under storage at 5 degrees C or ambient temperature.

Using capillary zone electrophoresis (CZE) with a 75 mM phosphate buffer at pH 8.5 containing 5 mM hydroxypropyl-gamma-cyclodextrin (OHP-gamma-CD) as chiral selector, the separation of the enantiomers of thiopental and its oxybarbiturate metabolite, pentobarbital, is reported. Enantiomer assignment was performed via preparation of enantiomerically enriched fractions using chiral recycling isotachophoresis (rITP) processing of racemic barbiturates and analysis of rITP fractions by chiral CZE and circular dichroism spectroscopy. Thiopental and pentobarbital enantiomers in plasma were extracted at low pH using dichloromethane and extracts were reconstituted in acetonitrile or 10-fold diluted, achiral running buffer. The stereoselectivity of the thiopental and pentobarbital metabolism was assessed via analysis of 12 plasma samples that stemmed from patients undergoing prolonged or having completed long-term racemic thiopental infusion. The data obtained revealed a modest stereoselectivity with R-(+)-thiopental/S-(-)-thiopental and R-(+)-pentobarbital/S-(-)-pentobarbital plasma ratios being < 1 (P < 0.05 compared to data obtained with racemic controls) and > 1 (P < 0.001), respectively. The total S-(-)-thiopental plasma concentration was found to be on average about 24% higher compared to the concentration of R-(+)-thiopental, whereas the total R-(+)-pentobarbital plasma level was observed to be on average 29% higher compared to the S-(-)-pentobarbital concentration.

The Swiss German chapter of the Exit Association provides conditional assistance to individuals wishing to end their own lives. The Exit Association advocates death with dignity and fights for the right to freely choose the timing of one's own death. According to the Swiss criminal code (articles 114 and 115), altruistic assistance to suicide is not punishable. Active euthanasia is punished by imprisonment. An individual commits active euthanasia if he or she is driven by honorable motives (e.g., pity) and causes the death of another person wishing to die who seriously and insistently requests such action. Based on our information, the preparation for suicide and its completion relies on a well-defined protocol. First, the candidate's eligibility for Exit Association assistance is verified. The candidate then writes a farewell declaration that explicitly confirms the will to terminate his or her own life. A written report describes the events during the suicide procedure. Depending on the circumstances, the investigative judge requests a forensic autopsy and toxicologic analyses. The results of the forensic investigations conducted in the cases presented here are in agreement with the scenario described in the reports of the Exit Association, namely, suicide by massive ingestion of pentobarbital.

Four commonly used barbiturates (phenobarbital, butalbital, pentobarbital and thiopental) were analyzed in human serum using supercritical fluid extraction (SFE) and negative ionization LC/ESI-MS/MS. Barbital was used as the internal standard. Carbon dioxide SFE was performed at 40 degrees C and 500 atm, with a total extraction time of 35 min. The analytes were collected off-line in a liquid trap containing absolute methanol. Samples were then concentrated by vacuum centrifugation. The high performance liquid chromatography separation utilized gradient elution with a total analysis time of 21 min. The precursor and major product ions for the four barbiturates were monitored on a triple quadrupole mass spectrometer with negative ion electrospray ionization (ESI) in the multiple reaction monitoring mode as follows: (1) thiopental (m/z 241.20-->58.00), (2) phenobarbital (m/z 231.10-->188.0), (3) pentobarbital (m/z 225.10-->181.90) and (4) butalbital (m/z 222.80-->179.90). In the case of phenobarbital, pentobarbital and butalbital, the most abundant product ion arises from the loss of 43 u (HCNO loss). However, in the case of thiopental, the most abundant product ion was observed at m/z 58.0 (the [M-183]-ion, or NCS-). Mechanisms for the formation of the collision induced dissociation reaction products of these barbiturates are proposed.

Pentobarbital is a sedative hypnotic which is used in the treatment of people suffering from anxiety related conditions and to manage elevated intracranial pressures and cerebral ischemia due to neurosurgical procedures. This paper develops and validates a rapid and sensitive method for the determination of S-(-)- and R-(+)-enantiomers in serum using capillary electrophoresis (CE) and UV detection. Stereoselective resolution was accomplished using 40 mM hydroxypropyl-gamma-cyclodextrin contained in 50 mM phosphate buffer pH 9.0. The method involves a solid-phase extraction of both the enantiomers and the internal standard, aprobarbital, from serum using C18 Bond-Elut cartridges. The CE system consists of a 52 cmX75 microm I.D. fused-silica capillary maintained at a run voltage of 15 kV with sample detection performed at 254 nm. The detection and quantitation limits for S-(-)- and R-(+)-pentobarbital are 1 microg/ml from serum. Linear calibration curves from 1 to 60 microg of both S-(-)- and R-(+)-enantiomers show a coefficient of determination of more than 0.999. The precision and accuracy of the method calculated as R.S.D. and error are 0.20-2.20% and 0.00-4.40%, respectively for R-(+)-pentobarbital and 0.30-2.19% and 0.30-6.40%, respectively for S-(-)-pentobarbital.

Purpose: To characterize the pharmacodynamic relationships between plasma pentobarbitone and thiopentone concentrations and nocifensive reflexes during emergence from anaesthesia., Methods: Forty-nine rats were studied. Plasma barbiturate concentrations were measured with high performance liquid chromatography. Nocifensive reflexes were assessed with the hindlimb withdrawal latency (WL) to heat and the somatic motor response threshold (SMRT) to tail pressure. In Protocol I, SMRT, WL, sedation, and the presence of paw-licking and the righting reflex were assessed in unrestrained rats before and every 10 min for two hours after an intraperitoneal injection of pentobarbitone (30 mg.kg-1). Plasma pentobarbitone kinetics were determined in a separate group of rats. In Protocol II, SMRT and drug concentrations were measured concurrently in partially restrained animals before and for 35 min after a computer-controlled i.v. bolus of thiopentone. In Protocol III the SMRT-plasma thiopentone relationship was determined during increasing and decreasing plasma thiopentone concentrations., Results: Enhancement of both nocifensive reflexes was observed in the unrestrained animals. Enhancement of SMRT was maximal [175% (153-197) of control values] at a mean plasma thiopentone concentration of 11 (9-13) micrograms.ml-1. The SMRT-plasma thiopentone curve showed a mean efflux-influx difference in plasma thiopentone concentration of 4(2.3-5.7) micrograms.ml-1., Conclusions: Barbiturate-associated nocifensive reflex enhancement occurs in unrestrained animals with both thermal and pressure stimuli. The SMRT-plasma thiopentone concentration relationship during emergence from anaesthesia was similar to that observed previously during induction. The thiopentone plasma concentration-SMRT plot showed an equilibrium delay similar to that previously described by others for thiopentone at an electroencephalographic effect site.

The effect of empenthrin, a synthetic pyrethroid, on pentobarbital (PTB)-induced sleeping time was examined in mice and rats. In mice, pretreatment with empenthrin prolonged PTB-induced sleeping time in a dose-dependent manner. The maximum effect on PTB-sleeping time was noted when mice were pretreated orally with empenthrin 2-4 h before PTB injection. However, empenthrin did not change the sleeping time induced by diethyl ether which is hardly metabolized in liver. Empenthrin inhibited the clearance of serum PTB in mic, but did not change the PTB concentration in serum at which animals recovered from sleeping. To examine the effect of PTB on metabolic enzymes in mouse liver, PTB was incubated aerobically with a hepatic microsomal fraction in the presence of NADPH at 37 degrees C. Empenthrin inhibited the vitro metabolism of PTB dose-dependently. In rats, empenthrin neither changed the PTB sleeping time, nor inhibited the clearance of serum PTB. No inhibitory effect of empenthrin was observed on the in vitro metabolism of PTB using rat hepatic microsomal fraction. These findings indicate that empenthrin prolongs PTB-sleeping time in mice through an inhibition of the PTB-metabolizing enzyme(s) in the liver , an effect that does not occur in rats. Also, there is a clear species-specificity in the inhibitory effect of empenthrin on the PTB-metabolizing enzyme(s).

An HPLC method was developed to determine the plasma concentrations of R(+)- and S(-)-thiopentone for pharmacokinetic studies in sheep. The method required separation of the thiopentone enantiomers from the corresponding pentobarbitone enantiomers which are usually present as metabolites of thiopentone. Phenylbutazone was used as an internal standard. After acidification, the plasma sample were extracted with a mixture of ether and hexane (2:8). The solvent was evaporated to dryness and the residues were reconstituted with sodium hydroxide solution (pH 10). The samples were chromatographed on a 100 mm x 4 mm I.D. Chiral AGP-CSP column. The mobile phase was 4.5% 2-propanol in 0.1 M phosphate buffer (pH 6.2) with a flow-rate of 0.9 ml/min. This gave k' values of 1.92, 2.92, 5.71, 9.30 and 11.98 for R(+)-pentobarbitone, S(-)-pentobarbitone, R(+)-thiopentone, S(-)-thiopentone, and phenylbutazone, respectively. At detection wavelength of 287 nm, the limit of quantitation was 5 ng/ml for R(+)-thiopentone and 6 ng/ml for S(-)-thiopentone. The inter-day coefficients of variation at concentrations of 0.02, 0.1 and 8 micrograms/ml were, respectively, 4.8, 4.4 and 3.5% for R(+)-thiopentone and, respectively, 5.0, 4.3 and 3.9% for S(-)-thiopentone (n = 6 each enantiomer). At the same concentrations, the intra-day coefficients of variation from six sets of replicates (measured over six days) were, respectively, 8.0, 8.0 and 8.8% for R(+)-thiopentone and 8.8, 7.4 and 9.6% for S(-)-thiopentone. Linearity over the standard range, 0.01-40 micrograms/ml, was shown by correlation coefficients > 0.998. This method has proven suitable for pharmacokinetic studies of thiopentone enantiomers after administration of rac-thiopentone in human plasma also and would be suitable for pharmacokinetic studies of the pentobarbitone enantiomers.

A method for the determination of ethynyloestradiol in samples of rabbit plasma containing pentobarbital and heparin, the former used as an anaesthetic and the latter as an anticoagulant, has been developed. Quantification was carried out using a reversed-phase high-performance liquid chromatographic (HPLC) method in isocratic mode at room temperature, with electrochemical detection at an applied potential of +1 V vs. Ag/AgCl. Under these conditions, the retention time for ethynyloestradiol was ca. 2.9 min, the average recovery from plasma was 74.5%, and the limit of detection was 10 pg, corresponding to a plasma concentration of 50 pg/ml using 1 ml of plasma. Natural oestrogens, oestriol, oestradiol and oestrone showed peaks that did not interfere with ethynyloestradiol, and retention times of ca. 0.8, 2.4 and 3.4 min, respectively.

The lot-to-lot reproducibilities of Bond Elut Certify and Clean Screen DAU columns are described. The recoveries of five test drugs obtained from twelve lots of Bond Elut Certify columns ranged from 84 to 104% with standard deviations of less than 9%. The recoveries of five test drugs obtained from six lots of Clean Screen DAU columns ranged from 81 to 103% with standard deviations of less than 7%. The 95% confidence intervals of the means as obtained by ANOVA demonstrate that there are no significant differences between the tested lots of Bond Elut Certify and Clean Screen DAU columns. Comparison of the two brands shows that both Bond Elut Certify and Clean Screen DAU columns are well acceptable for routine drug screening in systematic toxicological analysis, with a slightly higher overall recovery for the former.

The development of cross-tolerance to various alcohols and pentobarbital was examined in ethanol (EtOH)-treated mice. Chronic EtOH treatment (dosage rising in steps from 3.5-4.5 g/kg i.p. daily during a 23-day period) produced tolerance to its hypnotic effect. Such tolerance was seen as a reduction in the duration of loss of righting reflex (LRR), as well as higher blood EtOH levels at the offset of LRR, in EtOH-treated mice as compared to saline-treated controls. Cross-tolerance was shown by shifts in dose-response curves for the LRR induced by n-propanol and t-butanol. Such treatment, however, did not confer functional cross-tolerance to n-butanol and pentobarbital. Because n-butanol and pentobarbital are more lipid-soluble, whereas EtOH, n-propanol and t-butanol have low degrees of lipid solubility, the development of cross-tolerance among these sedative-hypnotic drugs might be related to their relative degrees of lipid solubility.

Although the pentobarbital-anesthetized dog is often used as a model in studies of respiratory muscle activity during spontaneous breathing, there is no information regarding the stability of the pattern of breathing of this model over time. The electromyograms of several inspiratory and expiratory muscle groups were measured in six dogs over a 4-h period by use of chronically implanted electrodes. Anesthesia was induced with pentobarbital sodium (25 mg/kg iv), with supplemental doses to maintain constant plasma pentobarbital concentrations. Phasic electrical activity increased over time in the triangularis sterni, transversus abdominis, and external oblique muscles (expiratory muscles). The electrical activity of the costal diaphragm, crural diaphragm, and parasternal intercostal muscles (inspiratory muscles) was unchanged. These changes in electrical activity occurred despite stable plasma levels of pentobarbital and arterial PCO2. They were associated with changes in chest wall motion and an increased tidal volume with unchanged breathing frequency. We conclude that expiratory muscle groups are selectively activated with time in pentobarbital-anesthetized dogs lying supine. Therefore the duration of anesthesia is an important variable in studies using this model.

This study addressed the commonly held, but seldom tested, notion that faster rates of increase of drug effects are associated with more positive subjective effects. Sodium pentobarbital was administered to normal healthy volunteers in either a single oral dose or in a series of divided, cumulating doses, and subjective responses were monitored. Twelve subjects participated in three weekly sessions, during which they received capsules containing placebo, 150 mg pentobarbital in a single dose (SIN) or 180 mg pentobarbital administered in six divided doses (DIV) of 30 mg every 30 min. Doses of pentobarbital in the SIN and DIV were selected to produce similar peak plasma levels. Blood samples were obtained at regular intervals for plasma drug level determinations, and throughout the session subjects completed self-report mood questionnaires (e.g., Profile of Mood States, visual analog ratings of drug liking and drug "high") and psychomotor performance tests (e.g., Digit Symbol Substitution Test). As expected, the SIN and DIV conditions yielded similar peak levels of pentobarbital, but the peak was attained more rapidly in the SIN condition. Despite the similarity in peak plasma levels, subjects reached greater peaks in ratings of "high" and wanted more of the drug when they were in the SIN condition. On an end-of-session liking questionnaire they also reported significantly greater liking of the drug in the SIN condition. On other measures of drug effects (e.g., sedation and psychomotor impairment) no significant differences were observed between the conditions. Thus, the rate of increase of the drug's effects specifically influenced subjects' ratings on subjective measures (e.g., "high" and liking) that may be associated with risk for abuse.(ABSTRACT TRUNCATED AT 250 WORDS)

Controversy exists regarding the optimal means for monitoring the patient receiving pentobarbital therapy during medical coma. Serum pentobarbital levels have been used traditionally to gauge cerebral penetration and efficacy of the drug. These peripheral levels have been assumed to reflect pentobarbital concentrations in the cerebrospinal fluid (CSF) and, therefore, the physiological effect on the central nervous system. To determine the relative accuracy of serum versus CSF pentobarbital levels, continuous electroencephalographic (EEG) monitoring in 10 consecutive patients was studied prospectively. Each patient received pentobarbital therapy for cerebral protection in the face of a traumatic injury. Simultaneous serum and CSF pentobarbital levels were obtained 1) before and after the initial barbiturate bolus, 2) every 12 hours during constant infusion therapy, and 3) before and after subsequent boluses necessary because of elevated intracranial pressure (ICP) (ICP greater than 15 mm Hg) or loss of burst suppression by continuous EEG monitoring (defined as greater than five bursts per minute). ICP and relevant clinical events were recorded hourly. Serum and CSF levels ranged from 33 to 74 mg/L and 4 to 54 mg/L, respectively. There was poor correlation between serum and CSF pentobarbital levels at any given time, although patients remained in burst suppression 73% of the time during their therapy. The EEG monitoring not only provided dynamic physiological monitoring, but it also permitted the lowest pentobarbital dose to maintain burst suppression for a specific patient's metabolism, reducing the likelihood of toxicity. In conclusion, CSF pentobarbital levels are of no greater accuracy than serum pentobarbital levels in predicting physiological effect.(ABSTRACT TRUNCATED AT 250 WORDS)

We developed a method for measuring pentobarbital in samples that also contain phenobarbital. The phenobarbital is destroyed by adding sodium hydroxide and then heating at 95 degrees C for 60 min. Pentobarbital is not affected by this pretreatment and can then be measured with an Abbott TDx barbiturates kit. Using this method, we obtained an average analytical recovery of 98% of added pentobarbital and a correlation of y = 0.953x + 3.4 vs HPLC. The intra-assay CV was 3.7% at 25.8 mg/L; the interassay CV was 6.2% at 16.1 mg/L. Other long-acting barbiturates, e.g., hexobarbital, are also effectively destroyed by this alkali pretreatment. Other short-acting barbiturates, e.g., secobarbital, are not removed and would produce an interference.

Pentobarbital was administered to 4 groups of rats: 1) intermittently before testing on the rotarod (RR) (experienced, EXP), 2) chronically (CHR) before testing on the RR (EXP), 3) intermittently (INT) after being tested on the RR (NONEXP), and 4) chronically (CHR) after being tested on the RR (NONEXP). On postchronic testing, Group 1 (INT/EXP) failed to show tolerance to the RR decrement, related to prechronic scores, while Group 3 (INT/NONEXP) actually showed an enhanced RR decrement. Group 2 (CHR/EXP) and Group 4 (CHR/NONEXP) both exhibited prominent tolerance to RR impairment at the postchronic test, with a nonsignificant trend for greater tolerance in Group 2. The lack of an expressed behavioral tolerance in INT/EXP rats and the enhanced RR decrement in INT/NONEXP subjects at the postchronic test was attributed to repeated use of a towel wrap restraint during the chronic treatment period. When the prechronic tests for INT/EXP animals were separated into the first 3 and last 3 days, pentobarbital impairment of RR during days 4-6 was significantly less than during days 1-3. This tolerance in INT/EXP rats was lost at the postchronic testing, while INT/NONEXP subjects had by then developed an enhanced RR impairment to pentobarbital. Following postchronic testing, chronic pentobarbital (CHR/EXP and CHR/NONEXP groups) and chronic vehicle (INT/EXP and INT/NONEXP groups) were discontinued for 9 days (withdrawal), after which an intermediate dose of the drug was tested on RR performance. Next, 9 days of extinction training involved vehicle injection daily before testing RR performance, after which the intermediate drug dose was again tested. INT/EXP and INT/NONEXP groups showed no change in RR impairment at the postwithdrawal and postextinction tests. However, in CHR/EXP rats pentobarbital tolerance was partly lost at the postwithdrawal test, with a significantly greater loss at the postextinction test.(ABSTRACT TRUNCATED AT 250 WORDS)

The development of behavioral tolerance to pentobarbital-induced hypothermia, as separable from cellular and metabolic tolerance, was established. Pentobarbital (PB) was administered to 4 groups of rats, 2 groups of which received intermittent (INT) IP PB treatment. One of these groups, INT/EXP, experienced the hypothermic (measured as rectal body temperature) drug effect after PB injection. The other group, INT/NONEXP, was monitored for body temperature functions (room temperature) before receiving PB (vehicle administration) and then prevented from experiencing PB-induced hypothermia by maintenance of body temperature with a towel wrap restraint and a heating lamp. The INT/EXP group also received equivalent exposure to this towel wrap after vehicle administration. Two other groups received chronic PB treatment (IP and in ground chow), one with experience for hypothermia after injections (CHR/EXP) and one prevented from experiencing the hypothermia (CHR/NONEXP). These groups also received equivalent exposure to the body temperature (at room temperature) testing and towel wrap restraint, EXP rats after vehicle injections and NONEXP after drug injections. A postchronic test of all groups compared the extent of PB hypothermia to prechronic test effects to assess the degree of tolerance. The INT/EXP group demonstrated behavioral tolerance for PB-induced hypothermia, as contrasted with the INT/NONEXP group which demonstrated little or no tolerance. Prominent tolerance was noted in both chronic groups for PB hypothermia, without a significant difference between them. After the postchronic test, chronic treatment was discontinued for 9 days (withdrawal) followed by 9 days of extinction training (vehicle behavioral testing). The two intermittent groups demonstrated no change in the hypothermic drug response during the postwithdrawal and postextinction drug tests.(ABSTRACT TRUNCATED AT 250 WORDS)

Hypothermia and motor impairment (tilt-plane test) were used to assess the phenomenon of rapid cross-tolerance between ethanol and pentobarbital in rats. The hypothermic and motor-impairment responses were significantly reduced on day 2 in animals receiving ethanol on day 1, compared to the control group pretreated with saline. Ethanol pretreatment, however, did not result in rapid cross-tolerance to pentobarbital on either test. Pentobarbital pretreatment on day 1 resulted in rapid tolerance to pentobarbital on day 2. However, in contrast to the lack of rapid cross-tolerance to pentobarbital after pretreatment with ethanol, pentobarbital pretreatment clearly conferred rapid cross-tolerance to ethanol. Determination of ethanol and pentobarbital blood levels suggested that pharmacokinetic alterations did not contribute significantly to the observed rapid tolerance and cross-tolerance. The asymmetry of rapid cross-tolerance seen in these studies mimics the results obtained by us in chronic tolerance and cross-tolerance studies reported recently. These results suggest that rapid tolerance and cross-tolerance can be used as predictors of chronic tolerance and cross-tolerance.

Pentobarbital and alpha-chloralose are widely used for experimental anesthesia, but their direct electrophysiological actions at anesthetic concentrations are unknown. Trough and peak concentrations measured by high-performance liquid chromatography averaged 27 +/- 3 and 45 +/- 13 mg/l (means +/- SD) for pentobarbital and 41 +/- 15 and 103 +/- 13 mg/l for alpha-chloralose in dogs receiving them for general anesthesia. The direct effects of each agent on papillary muscle action potentials obtained from guinea pigs killed by decapitation were studied in vitro. Pentobarbital increased action potential duration to 95% by 24 +/- 6 and 33 +/- 4% at 25 and 50 mg/l (P less than 0.001 for each), respectively, and caused corresponding increases in effective refractory period. Furthermore, pentobarbital reduced maximum rate of voltage change (Vmax) of phase 0 in a voltage-, rate-, and concentration-dependent fashion, suggesting use-dependent sodium channel blocking actions. The voltage dependence of Vmax was shifted by 3.7 +/- 1.7 (P less than 0.01) and 6.5 +/- 1.8 mV (P less than 0.001) by 25 and 50 mg/l pentobarbital, respectively. In canine ventricular muscle, pentobarbital caused rate- and concentration-dependent decreases in Vmax and increases in action potential duration and refractory period over a concentration range of 5-100 mg/l. alpha-Chloralose was devoid of direct electrophysiological effects in both species. We conclude that pentobarbital has potentially important electrophysiological actions on ventricular tissues at concentrations required for general anesthesia and may confound the results of in vivo studies.

Saline or ascorbic acid (AA), 1000 mg/kg and 1400 mg/kg was administered intraperitoneally (IP) to 3 groups of female Swiss-Webster mice after they had been anesthetized with sodium pentobarbital (SP). Both doses of AA given IP after SP anesthesia significantly prolonged sleeping time. An additional 50 mice were injected IP with 0.3 ml SP containing 6 mg/ml unlabeled SP and 0.6 ng C14 labeled SP. Immediately 25 mice received 40 mg AA in 0.4 ml saline, IP, and the remainder received saline. The serum half life of the radioactively labeled SP was more than 3 times longer in AA treated mice than in the controls.

A single i.v. dose (0.1 mmol Be2+/kg) of beryllium chloride prolonged the duration of pentobarbital-induced sleep and zoxazolamine-induced paralysis, in rats. The effects are correlated with changes of the pharmacokinetic parameters and with the in vitro inhibition of both aliphatic and aromatic hydroxylation of pentobarbital and zoxazolamine. In vitro N-demethylation of meperidine and aminopyrine was partially inhibited while O-demethylation of quinidine was unaffected by liver microsomes of rats pretreated with beryllium salt. The findings give clues that beryllium chloride inhibits some forms of cytochrome P-450, especially those responsible for hydroxylation of substrates, like pentobarbital and zoxazolamine.

The commercially available fluorescence polarization immunoasay (FPIA) method for phenobarbital is demonstrated to be useful in the quantification of pentobarbital in serum. The effective range for the method is 1.0 to 80.0 mg/L. The method is simple, precise, and fast. The application of the method is limited to patients receiving pentobarbital as the only barbiturate. The FPIA is recommended for use in clinical laboratories when a rapid estimation of pentobarbital is required in monitoring patients undergoing high-dose pentobarbital therapy.

Studies in rats were performed to evaluate the effect of sorbitol on the antidotal efficacy of activated charcoal against four test drugs and to investigate the influence of storage upon the antidotal effect of activated charcoal-sorbitol suspension. The antidotal potency of activated charcoal was not diminished by sorbitol solution 70% w/v. In fact, it was enhanced by the sorbitol solution, as indicated by greater decrease in peak tissue drug concentration, compared to the effect produced by activated charcoal in aqueous suspension. Furthermore, storage of activated charcoal in sorbitol for as long as 1 year did not reduce the antidotal-efficiency of the absorbent.

This study describes the tolerance characteristics of barbital compared to pentobarbital, the standard drug, during "chronically equivalent" treatment. Barbiturate tolerance was assessed as the increase in dose from the beginning to the end of treatment required to achieve equieffective peak effect. Dispositional tolerance was assessed as a reduction in the elimination half-life of barbiturate from blood. Functional tolerance was assessed as the increase in blood concentration of barbiturate at the time of peak effect. Overall, greater tolerance was developed to pentobarbital than to barbital. For pentobarbital, tolerance was both dispositional and functional; the dispositional tolerance developed rapidly and was almost complete at 1 week. For barbital, tolerance was exclusively functional. A most interesting finding was that functional tolerance to barbital and pentobarbital developed at the same slow rate for chronically equivalent treatment. This finding suggests that functional tolerance development is independent of the particular barbiturate reflecting the adaptability of the central nervous system to chronic depression.

In order to test the possible differences in the clinical effects between benzodiazepines and barbiturates, 5 mg of diazepam and 60 mg of pentobarbital were used in a double-blind study as rectal premedication in 75 small children before major surgery. There were no significant differences between the two groups in the six parameters tested (demeanour in the anaesthetic room, preoperative emetic effect, response to venepuncture, degree of salivary suppression, postoperative state in the recovery room, postoperative emetic effect). The concentrations of pentobarbital and diazepam (and its 3 active metabolites) in the plasma were determined by gas chromatography and correlated with the clinical effects. No significant correlation was found between the total active benzodiazepine concentrations and the clinical effects of rectal diazepam. In contrast to diazepam, a weak but significantly positive correlation between the plasma concentrations of pentobarbital and the total score was found. Similarly, the plasma level of pentobarbital and the response to venepuncture and the degree of salivary secretion were in correlation with each other. In the plasma concentrations of both pentobarbital and total benzodiazepines, a wide interindividual variation was observed, indicating irregular rectal drug absorption.

The pharmacokinetics of pentobarbital were examined in 11 children with Reye syndrome, hypoxic encephalopathy, or acute head injury. Nine of these patients were hypothermic (less than 32 degrees C). The total systemic clearance and volume of distribution at steady state of pentobarbital were significantly reduced in these patients when compared to previous data in normothermic adult volunteers following intravenous doses of pentobarbital. Pentobarbital elimination half-life was not significantly different from control values. The diminished systemic clearance of pentobarbital may result from decreases in intrinsic enzyme activity that accompany hypothermia, as well as hepatic dysfunction in patients with Reye syndrome. Less extensive distribution of pentobarbital is likely the result of either differences in body fat composition or hypothermia-induced decreases in regional blood flow. The reduced clearance and distribution of pentobarbital may partially explain the enhanced reduction in cerebral metabolism that occurs on addition of hypothermia to barbiturate therapy in patients with elevated intracranial pressure.

Thiopental and pentobarbital have been used in high doses to protect the brain from injury following hypoxia or to reduce intracranial pressure. This study was performed to determine whether these barbiturates differ in cardiovascular effects when present in plasma concentrations that produce equivalent CNS effects. The effects of thiopental and pentobarbital on heart rate, stroke volume/kg, cardiac output/kg, systemic vascular resistance, mean arterial pressure, and central venous pressure were statistically indistinguishable at plasma concentrations of each barbiturate ranging from 50% to 100% of their concentration producing EEG silence. Three of the seven dogs given thiopental developed ventricular bigeminy at plasma concentrations ranging from 45% to 65% of their concentration producing EEG silence. Lidocaine (1.4-2.0 mg/kg intravenously) reversed the bigeminy to sinus rhythm. When given more than the amount needed to produce a flat EEG, five of the seven dogs given thiopental died, but all dogs given pentobarbital survived. Pentobarbital may be a better choice than thiopental when large doses are indicated.

Pharmacokinetics and duration of anesthesia of methohexital, pentobarbital, thiamylal, and thiopental in Greyhound and non-Greyhound, mixed-breed dogs were compared. In all dogs evaluated, pentobarbital induced the longest duration of anesthesia and methohexital induced the shortest duration. Pharmacokinetics of pentobarbital and methohexital were similar in both groups of dogs. Thiobarbiturates induced longer anesthetic effects in Greyhound dogs than in mixed-breed dogs. Plasma thiobarbiturate concentrations remained above normal longer in Greyhound dogs than in mixed-breed dogs. Disposition of thiobarbiturates in Greyhound dogs was characterized by nonlinearity from 45 minutes to 8 hours after dosing.

The effect of electrolytic lesions in the median and dorsal raphe nuclei was tested on acute tolerance development to ethanol and pentobarbital in the rat, as measured by motor impairment on the moving belt test. Acute tolerance to ethanol (1.7 g/kg, IP) or pentobarbital (17.5 mg/kg, IP) was monitored at 12.5, 25, or 50 min in separate subgroups tested only once each. One week of recovery was allowed between ethanol and pentobarbital tests. Median raphe lesions delayed the development of acute tolerance, whereas dorsal raphe lesions produced a negligible effect. These results were seen with both ethanol and pentobarbital. The mesolimbic 5-HT pathway from the median raphe nucleus is important in the development of acute tolerance to ethanol and pentobarbital, as was shown to be the case previously for chronic tolerance.

The Corvac integrated serum separator tube (TM Monoject Scientific) has been examined for use in processing specimens for drug analysis. This study was undertaken because the SST serum separator tube (Becton-Dickinson) was shown to cause decreases in drug levels of lidocaine, pentobarbital, and phenytoin, which were dependent upon blood volume and time of contact with the gel. Pools of donor blood were spiked with lidocaine or phenytoin and processed in Corvac tubes and red top tubes. Lidocaine levels were depressed by 5.2% when Corvac was used. Phenytoin levels were unchanged. The lidocaine depression was independent of blood volume, time of gel contact, or tube lot number. Paired patient specimens for quinidine, lidocaine, or theophylline analysis were obtained in Corvac and red top tubes. Lidocaine levels were depressed by 6.0% (p less than 0.01). Quinidine and theophylline levels were unchanged. All drug analyses were carried out using the Syva enzyme multiplied immunoassay technique. The Corvac tube has been shown to cause decreased levels of lidocaine but not of the other drugs tested. Corvac (silicone gel) results differ from SST (polyester gel) results because of the different construction of these tubes.