Peter Kissinger

Purdue University | Purdue · Department of Chemistry

This is a story of the 100-year path for voltammetric electroanalytical chemistry from the serendipitous discovery of polarography by Jaroslav Heyrovsky in 1922 to the miniature biosensors of today. In spite of issues with the dropping mercury electrode (DME), polarography was dominant for almost 50 years due to the good quantitative results it pro...

The advent of electrochemical affinity assays and sensors evolved from pioneering efforts in the 1970s to broaden the field of analytes accessible to the selective and sensitive performance of electrochemical detection. The foundation of electrochemical affinity assays/sensors is the specific capture of an analyte by an affinity element and the sub...

Scope: In this glossary, bioanalysis is defined as the quantitative or qualitative measurement of endogenous or exogenous analytes in biological matrices, including tissue, blood, serum, urine, or other bodily fluidsThe broader use of the term used by biochemists is excluded. This glossary lists terms and definitions: * directly associated with bio...

We examined the kinetic and transport processes involved in Mg production from MgO via electrolysis at ca 1250 K with in a eutectic mixture of MgF2 – CaF2, using a Mo cathode, and carbon anode. Exchange current densities, transfer coefficients, and diffusion coefficients of the electroactive species were established using a combination of cyclic an...

Good data can only come from good samples. Technology is now available to collect, transport, store and document samples like never before. It is critical that these new tools be given as much attention as the bioanalytical instruments to which the samples will be fed.

Ambient ionization methods for MS enable direct, high-throughput measurements of samples in the open air. Here, we report on one such method, desorption electrospray ionization (DESI), which is coupled to a linear ion trap mass spectrometer and used to record the spatial intensity distribution of a drug directly from histological sections of brain,...

A thin-layer electrochemical cell constructed from engineering plastics is described. The sandwich design can accommodate carbon or metal electrodes embedded in the wall of a rectangular channel. Miniature grid electrodes can be placed across the channel. The new transducer has been used in both static and hydrodynamic experiments including amperom...

Electroanalytical chemistry has been a primary activity in Ph.D.-granting institutions since the 1940s. It has proved to be an excellent tool for research, enabling students to think in time and space and grasp thermodynamics, kinetics, and instrumentation principles as applied to accessible measurements. This personal perspective is an informal re...

Microdialysis has progressed from an inspired academic curiosity in the early 1970s to a widely used biomedical research tool 35 years later. This chapter reviews the evolution of the instrumentation from laboratory built manual apparatus to fully automated systems today that are used in both preclinical and clinical research. A systems view is pre...

A reverse phase LC method with amperometric electrochemical detection is described for determination of several common analgesics in blood serum and urine. The retention properties as a function of organic mobile phase modifier and pH are graphically illustrated and discussed. Electrochemical properties of the analgesics and possible internal stand...

In order to understand the origin of numerous metabolites found in human urine following ingestion of bananas, a study of aromatic metabolism in the banana has been initiated. Thin-layer chromatography and reverse-phase ion-pair liquid chromatography with electrochemical detection have been combined to yield specific information about aromatic amin...

A liquid chromatography method with multi-channel electrochemical detection is reported for the identification and determination of polyphenols in green tea, black tea, and extracted green tea powder. Following a simple aqueous extraction, (−)-epigallocatechin (EGC), (+)-catechin (C), (−)-epicatechin (EC), (−)-epigallocatechin gallate (EGCG), (−)-g...

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...

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 mi...

The preclinical bioanalytical process with animal models begins with dosing and then sampling biological fluids and tissue. How an animal is housed, dosed and then sampled is equally important to how the samples are analyzed. Animal science and bioanalytical chemistry are very different professions, but integrating them is essential to the process...

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-chan...

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 anim...

Aim Methods A method for determination of naringenin (NG-E) in rat plasma by using liquid chromatography/electrochemistry(LCEC) has been developed and validated. NG-E was extracted from 100 µL of rat plasma with ethyl acetate. The analyte was separated on a microbore C 18 column 150 x 1 mm with a C 18 guard column 14 x 1 mm. The mobile phase was 20...

Aim Recently resveratrol has come to scientific attention for its cardiovascular and anti-cancer effects. Laboratory tests have clearly demonstrated that resveratrol may help prevent cardiovascular disease and cancer. However, a little is known about the absorption and clearance of resveratrol, the identities of its metabolic products, or its effec...

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...

Multi-channel electrochemical detector has been developed as a new generation of electrochemical product, which has several features like easy to be used, fast and accuracy for the assay, eliminating the interference and suitable for the analysis of components in a complex system. A novel scheme of liquid chromatography with four-channel electroche...

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 Cule...

A simple gradient reverse-phase liquid chromatographic method has been developed for the separation of daidzin, genistin, daidzein, and genistein in various natural sources and dietary supplements. Peak assignment at various retention times was confirmed by comparison of the simultaneous response of sample peaks to standards, at glassy carbon elect...

A liquid chromatography/tandem mass spectrometry (LC/MS/MS) method has been developed for the determination of brain basal nucleosides (inosine, guanosine and adenosine) in microdialysates from the striatum and cortex of freely moving rats. A microdialysis probe was surgically implanted into the striatum or cortex of individual rats and Ringer's so...

A simple gradient reverse-phase liquid chromatographic method has been developed for the separation of daidzin, genistin, daidzein, and genistein in various natural sources and dietary supplements. Peak assignment at various retention times was confirmed by comparison of the simultaneous response of sample peaks to standards, at glassy carbon elect...

A liquid chromatography method with multi-channel electrochemical detection was developed for the determination of epigallocatechin gallate (EGCG) in rat plasma. After administration of EGCG, blood samples were periodically collected by Culex (an automated blood sampling robot). EGCG was extracted from 50 μl of diluted blood (blood and saline at a...

A scheme using liquid chromatography with four-channel electrochemical detection for the identification and quantification of phenolic acids is reported. After a preliminary identity assignment based on spiking samples with standard compounds, an electrochemical detector was employed to obtain a voltammetric characterization of eluting compounds. P...

The biomedical accelerator mass spectrometry (AMS) initiative at PRIME Lab including the status of equipment and sample preparation is described. Several biomedical projects are underway involving one or more of the nuclides: 14C, 26Al and 41Ca. Routine production of CaF2 and graphite is taking place. Finally, the future direction and plans for imp...

A method using liquid chromatography/tandem mass spectrometry (LC/MS/MS) has been developed for the determination of basal acetylcholine (ACh) in microdialysate from the striatum of freely moving rats. A microdialysis probe was surgically implanted into the striatum of the rats and Ringer's solution was used as the perfusion medium at a flow rate o...

A sensitive and selective liquid chromatography/ electrochemistry method with multi-channel detection was developed for the determination of the natural product trans-resveratrol in wines, grape juice, and grape seed capsules. Samples were prepared with an automated solid phase extraction workstation. Chromatographic separation was achieved on a C1...

A sensitive and selective multichannel liquid chromatography–electrochemistry method was developed for the determination of the natural product trans-resveratrol (resveratrol) in rat blood. After administration of resveratrol, blood samples were periodically collected by an automated blood sampling device. Resveratrol was extracted from 150 μl of d...

Biosensors have been under development for over 35 years and research in this field has become very popular for 15 years. Nevertheless, for electrochemical biosensors, sensors for only one analyte (glucose) have achieved widespread commercial success. This review provides some arguments related to the unrealistic expectations for electrochemical (a...

A review of Thin-Layer Chromatography, Fourth Edition.

The career of Prof. Ralph N. Adams spans the evolution of electroanalytical chemistry from an academic curiosity based on polarography to a widely used tool with important applications across the chemical disciplines, from environmental science to neuropharmacology, and from heterogeneous catalysis to battery electrodes for cellular telephones. Ada...

Liquid chromatography/tandem mass spectrometry (LC/MS/MS) has been coupled to in vivo microdialysis for on-line monitoring of melatonin in a freely moving rat for a period of 15 hours. A microdialysis probe was surgically implanted into the jugular vein of the rat, and deionized water was used as the perfusion medium at a flow rate of 1.0 microL/mi...

Amperometric bienzyme electrodes based on coupled L-glutamate oxidase (GlOx) and horseradish peroxidase (HRP) were constructed for the direct monitoring of L-glutamate in a flow injection (FI)-system. The bienzyme electrodes were constructed by coating solid graphite rods with a premixed solution containing GlOx and HRP crosslinked with a redox pol...

The utility of microdialysis as a quantitative sampling technique for in vitro drug metabolism studies was demonstrated by investigating the stereoselective metabolism of D-, L- and DL-amphetamine by the cytochrome P-450 enzymes. Microdialysates containing the isomers of amphetamine and its metabolite were derivatized with the fluorescent chiral de...

A new post-column photolysis technology has been developed based on the use of a low pressure, low temperature UV lamp and TiO2 coated knitted reaction coil. As a test case the developed technique was used for the determination of 3-nitro-L-tyrosine by liquid chromatography with electrochemical detection. Different photolysis lamps and reactor tubi...

A liquid chromatographic method has been developed for the determination of 3-nitro-L-tyrosine. Different detection methods, including UV, oxidative and redox electrochemistry, and postcolumn photolysis followed by electrochemical detection, have been optimized and compared in terms of analysis time, detection limit and dynamic range. It was demons...

Membrane probes are implantable devices, which can be used to sample from the interstitial fluid of the tissues in which they are implanted. Two types of membrane probes, one based on microdialysis and the other on ultrafiltration, were developed and tested in vitro for the following analytes: sodium, potassium, chloride, glucose and lactate. These...

An amperometric three-enzyme based biosensor for determination of acetylcholine has been developed with possible use for monitoring of brain microdialysates by co-immobilising acetylcholinesterase (AchE), choline oxidase (ChOx) and horseradish peroxidase (HRP) in an Os-based redox polymer on solid graphite electrodes. The redox hydrogel was formed...

Simple and reliable methods for liquid chromatography/electrochemistry (LCEC) and LC/MS/MS with atmospheric pressure chemical ionization (APCI) have been developed for the determination of melatonin in commercially available products. The dissolution of melatonin from the melatonin-containing formulations was simply obtained by dissolving the table...

Determinations of neurotransmitters in rat brain microdialysates using microbore liquid chromatography with electrochemical detection are described. Catecholamines, serotonin and their metabolites in microdialysates were sensitively measured with detection limits of 0.1 to 0.5 pg per injection. Acetylcholine and choline were measured by using a pos...

Amperometry is very attractive for analytical chemistry work on a small scale. There are now many examples whereby it has been demonstrated in volumes of a few microliters or even less for selected analytes. The advantages of a direct conversion of chemical events into a measurable current are substantial. The full promise of this feature is often...

Titanium dioxide was used as a photocatalyst to improve the yield and rate of product formation in a post-column derivatization reactor for microbore liquid chromatography. The photo-catalytic reactor demonstrated an increase in sensitivity of greater than one order of magnitude for the electrochemical detection of phenylalanine compared to the mor...

Current methods for studying in vitro drug metabolism involve add-incubate-separate-measure approach. Separation of the desired analytes requires removal of protein which is typically accomplished by precipitation and centrifugation and extraction of the analytes into an organic phase. The analysis scheme then becomes more complex resulting in a de...

A flow-through electrode with a high conversion efficiency has been developed for chromatographic detection as well as other on-line electrochemical applications. A small length of reticulated vitreous carbon material of high pore density permits excellent conversion of so ne organic compounds. This can easily be employed in dual electrode applicat...

An amperometric flow cell was developed for continuous monitoring of lactate in the subcutaneous microdialysate from a rat. The lactate measuring flow cell is based on a platinum- electrode covered with a lactate oxidase containing sandwich membrane. The functional stability of the lactate probe was more than one-week and the results from the enzym...

Erythromycin is determined in both urine and plasma samples using microbore reversed-phase liquid chromatography with tris(2,2'-bipyridyl)ruthenium(II) [Ru(bpy)3(2+)] electrogenerated chemiluminescence (ECL) detection. Ru(bpy)3(2+) is included in the mobile phase thus eliminating band broadening caused by post-column reagent addition. Extra column...

Tetrahydrobiopterin and seven of its analogues (neopterin, xanthopterin, biopterin, pterin, tetrahydropterin, 6-methyltetrahydropterin, and 6-methylpterin) were separated on a 1 × 150 mm C18 microbore column. These analytes were detected by dual-electrode amperometry and UV absorption. Both a conventional glassy carbon electrode and an interdigitat...

Ultrafiltrate probes were implanted subcutaneously and intramuscularly in five adult horses. After collecting baseline samples, 28 gm i.v. CaCl2 was infused. Plasma, ultrafiltrate of plasma, ultrafiltrate of subcutaneous interstitial fluid, and ultrafiltrate of intramuscular interstitial fluid were analyzed for Na+, K+, and Ca2+. Post infusion conc...

An eight-sector array (split disk) electrode was designed for a low flow rate (<100 μL/min) amperometric detector. This electrode was fabricated photolithographically for dimensional accuracy and reproducibility. This array of a pie-shaped electrode was combined with a thin-layer radial flow cell, and a conversion efficiency of 94% was achieved at...

A mercury thread electrode modified with hydrophilic dialysis membrane was first introduced in 1994. Incorporation of this electrode in a flow stream is now introduced. The flow of solution and mercury are conveniently achieved using two syringe pumps. The cell can be used with a sample loop of a desired volume; hence, it allows flow injection appl...

Thin-layer cross-flow amperometric detectors with enzyme modified working electrodes were developed for the determination of oxidase substrates in flowing streams. The enzyme was covalently immobilized in an osmium redox polymer film on the electrode surface. Electrodes modified with glucose oxidase or lactate oxidase were prepared for the continuo...

This review article presents an overview of current research on the use of amperometric electrochemical detectors in bioanalytical chemistry. Topics covered include microdialysis and ultrafiltration membranes for in-vivo sampling; microbore liquid chromatography; capillary electrophoresis; enzyme, photochemical, and chemical post-column reactions;...

The electrochemical properties of carbon film based ring-disk and split-disk electrodes in small active volume (0.24–2μL) radial flow cells were studied in order to employ them in conjunction with microbore liquid chromatography (LC) for determining catecholamines. The high conversion of dopamine (DA) at the disk of a ring-disk electrode makes the...

This chapter observes that the Microdialysis and ultrafiltration are complementary sampling techniques, developed for studying the interstitial space in vivo. Microdialysis and ultrafiltration are separation techniques that involve moving a chemical across a semipermeable membrane. They have many characteristics in common. Both methods sample the e...

A method for the highly sensitive determination of acetylcholine (ACh) and choline (Ch) was developed using platinum (Pt) black microarray electrodes as detectors in a microbore liquid chromatography (LC) with a post column enzyme reactor. The electrodes were prepared by plating a gold (Au) film electrode with sub-μm Pt-black particles. Since hydro...

Aim: To develop a sensitive method for determination of serotonin in biological samples. Methods: A combination of microdialysis sampling and microbore liquid chromatography with electrochemical detection (LCEC) was established. Results: Changes of serotonin in fg or pg in microdialysates from brain striatum of the free moving rat were easily...

The detection limit of catecholamines can be lowered by using a carbon-based interdigitated array (IDA) microelectrode as a detector for liquid chromatography (LC). The IDA electrode is more sensitive than conventional glassy carbon electrodes due to the high current density caused by radial diffusion at each microband, and redox cycling between tw...

A liquid chromatography-electrochemistry (LC-EC) method is described for the determination of basal acetylcholine (ACh) in microdialysate from the striatum of freely moving rats. This method is based on the separation of ACh and choline (Ch) by microbore liquid chromatography followed by passage of the effluent through a post-column immobilized enz...