Lab evaluation of chemical and biological terrorism.
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Bioterrorism↗
Biomedical subjects
Peter T Kissinger
Publications and source records attributed to Peter T Kissinger.
Biosensors-a perspective.
Biosensors have been under development for over 35 years and research in this field has become very popular for 15 years. Electrochemical biosensors are the oldest of the breed, yet sensors for only one analyte (glucose) have achieved widespread commercial success at the retail level. This perspective provides some cautions related to expectations for biosensors, the funding of science, and the wide gap between academic and commercial achievements for sensor research. The goal of this commentary is not to arrive at any particular truth, but rather to stimulate lively discussion.
Biosensing Techniques↗
Liquid chromatography/tandem mass spectrometry for the determination of carbamazepine and its main metabolite in rat plasma utilizing an automated blood sampling system.
A liquid chromatography/tandem mass spectrometry (LC/MS/MS) method for the simultaneous determination of carbamazepine and its main metabolite carbamazepine 10,11-epoxide in rat plasma is described. The method consists of a liquid-liquid extraction procedure and electrospray LC/MS/MS analysis. The chromatographic separation was achieved within 5 min using a C(8) (150 mm x 2.1mm) 5 microm column with a mobile phase composed of water/acetonitrile/acetic acid (69.5:30:0.5, v/v/v) at a flow rate of 0.4 ml/min. D(10)-carbamazepine is used as the internal standard for all compounds. Analytes were determined by electrospray ionization tandem mass spectrometry in the positive ion mode using selected reaction monitoring (SRM). Carbamazepine was monitored by scanning m/z 237-->194, carbamazepine 10,11-epoxide by m/z 253-->210 and d(10)-carbamazepine by m/z 247-->204. The lower limit of quantitation (LLOQ) is 5 ng/ml for each analyte, based on 0.1 ml aliquots of rat plasma. The extraction recovery of analytes from rat plasma was over 87%. Intra-day and inter-day assay coefficients of variations were in the range of 2.6-9.5 and 4.0-9.6%, respectively. Linearity is observed over the range of 5-2000 ng/ml. This method was used for pharmacokinetic studies of carbamazepine and carbamazepine 10,11-epoxide in response to two different blood sampling techniques (i.e., manual sampling versus automated sampling) in the rat. Several differences between the two sampling techniques suggest that the method of blood collection needs to be considered in the evaluation of pharmacokinetic data.
Animals↗
Good preclinical bioanalytical chemistry requires proper sampling from laboratory animals: automation of blood and microdialysis sampling improves the productivity of LC/MSMS.
The preclinical bioanalytical process with animal models begins with sampling biological fluids and tissue. The goal is to understand oral absorption kinetics, distribution, metabolism, excretion, blood brain barrier penetration, drug-drug interactions, and the influences on biomarkers, hematology, electrophysiology, cardiology, blood pressure and behavior. An overview is obtained by periodic blood sampling of 8-12 samples over a total time span of 10-24 h. Urine, feces, bile and microdialysates can augment the information available from whole blood. In today's preclinical environment, the majority of samples are processed by LC/MSMS augmented by robotic sample preparation tools. These tools save labor and improve precision for smaller volume/lower concentration samples. Our laboratories have been engaged in a project that is focused on improving both the quality and throughput for laboratory animal studies, while providing for reduced numbers of animals and enhanced animal comfort. We have implemented a robotic system that can accomplish most of the above goals for laboratory rats, dogs and primates. Studies with mice are at an earlier stage, but feasibility has been demonstrated. This presentation is a progress report on this evolving research program in cooperation with multiple pharmaceutical and drug development companies. We will illustrate results and discuss future directions.
Animals↗
Liquid chromatography coupled with multi-channel electrochemical detection for the determination of daidzin in rat blood sampled by an automated blood sampling system.
Daidzin, a soy-derived biologically active natural product, has been reported to inhibit mitochondrial aldehyde dehydrogenase and suppress ethanol intake. This paper describes a method for the determination of daidzin in rat blood. After administration of daidzin, blood samples were periodically collected from awake, freely moving animals by a Culex automated blood sampler. Daidzin was extracted from 50 microl of diluted blood (blood and saline at a ratio of 1:1) with ethyl acetate. Chromatographic separation was achieved within 12 min using a microbore C(18) (100 x 1.0 mm) 3 microm column with a mobile phase containing 20 mM sodium acetate, 0.25 mM EDTA, pH 4.3, 4% methanol and 11% acetonitrile at a flow-rate of 90 microl/min. Detection was attained using a four-channel electrochemical detector with glassy carbon electrodes using oxidation potentials of +1100, 950, 850, 750 mV vs. Ag/AgCl. The limit of detection for daidzin in rat plasma was 5 ng/ml at a signal-to-noise ratio of 3:1. The extraction recovery of daidzin from rat plasma was over 74%. Linearity was obtained for the range of 25-1000 ng/ml. The intra- and inter-assay precisions were in the ranges of 2.7-6.6 and 1.9-3.7%, respectively. This method is suitable to routine in vivo monitoring of daidzin in rat plasma.
Animals↗
The academic career as a platform for launching a company.
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Career Choice↗
Identification of the major vanilloid component in Capsicum extract by HPLC-EC and HPLC-MS.
A sensitive multi-channel HPLC-electrochemical (EC) method has been developed to determine the vanilloid content in the complex Capsicum annuum extract Capsibiol. Chromatographic separation was achieved within 10 min using a YMC Basic S5 column with a mobile phase containing chloroacetic acid, heptane sulphonic acid and acetonitrile. The multi-channel detector simultaneously applied four potentials between +500 and +800 mV (referenced to a silver/silver chloride electrode) to four glassy carbon working electrodes. The most abundant (0.94 mg/g) vanilloid analogue in the Capsibiol sample demonstrated an electrochemical reactivity and retention time similar to that of vanillic acid in HPLC-EC analysis. Its identity was confirmed by HPLC-MS using a Zorbax SB-CN column with a mobile phase containing formic acid and methanol.
Capsicum↗
An automated blood sampler for simultaneous sampling of systemic blood and brain microdialysates for drug absorption, distribution, metabolism, and elimination studies.
INTRODUCTION: A major problem in preclinical drug development where blood sampling from small animals is a routine practice is the time and labor involved in the serial sampling of small blood volumes from small animals such as rats for the duration of pharmacokinetic/pharmacodynamic (PK/PD) studies. The traditional method of manually drawing blood from the animal requires the animal to be anesthetized or restrained with some device, both of which cause stress to the animal. METHODS: An automated blood sampler (ABS) was developed to simultaneously collect blood and brain microdialysate samples at preprogrammed time points from awake and freely moving animals. The samples are delivered to fraction collectors and stored at 4 degrees C until use. The lost blood volume during collection is replaced with sterile saline to prevent fluid loss from the animal. In addition, the system is capable of collecting urine and feces for metabolism studies and monitoring the animal activity for behavioral studies. In the present study, blood samples were collected for 24 h after dosing rats orally with a 5 mg/kg dose of olanzapine (OLAN). Brain dialysates were collected for the same duration from a microdialysis probe implanted in the striatum. RESULTS: The pharmacokinetic parameters, obtained after an oral dose, are in good agreement with reported values in literature. The pharmacodynamic information obtained from brain dialysates data show that OLAN elevates the concentration of dopamine (DA) in the brain and remains in the brain even after it is cleared from the plasma. DISCUSSION: The ABS described here is a very useful tool in drug development to accelerate the pace of preclinical in vivo studies and to simultaneously provide pharmacodynamic and physiological information.
Administration, Oral↗