Richard G. Christensen

Determination of Oltipraz in serum by high-performance liquid chromatography with optical absorbance and mass spectrometric detection

Three methods have been developed for the analysis of Oltipraz in serum. A method suitable for routine use employs spiking with a homologous internal standard, off-line solid-phase extraction, high-performance liquid chromatographic separation, and optical absorbance detection at 450 nm. Method detection limit is about 1 ng/ml. A second method, less susceptible to bias from co-eluting interferences, uses a stable isotope-labeled internal standard, similar extraction and separation, and detection by thermospray mass spectrometry. Method detection limit is about 0.2 ng/ml. A third method was developed which can be used without specially synthesized internal standards. It uses on-line solid-phase extraction, with quantification by comparison with external standards. Method detection limit is about 3 ng/ml. Good agreement was observed between these methods and with similar and different methods run in other laboratories. Calibration curves were linear over the entire range which was investigated, i.e., up to 500 ng/ml. Coefficients of variation were similar for all three methods, being about 5%.

Detectors for Liquid Chromatographic Analysis for Polynuclear Aromatic Hydrocarbons

Abstract A number of liquid chromatographic detectors of various types have been evaluated for both selectivity and sensitivity for the detection of polynuclear aromatic hydrocarbons (PAH). Detection limits for fixed and variable wavelength UV photometers, filter fluorimeters, and spectrofluorimeters have been determined. The utility of each of these types of detectors for use in the reversed-phase HPLC analysis of environmental extracts containing trace levels of PAHs is discussed.

Determination of dibenzothiophene in oils by liquid chromatography—tandem mass spectrometry

Abstract Quantitative trace analysis for organic compounds in complex matrices such as oils often requires time-consuming sample pretreatment. Two examples are shown of the use of a highly selective tandem mass spectrometer as a liquid chromatographic detector for the quantification of dibenzothiophene in a crude petroleum oil and in an alternate fuel oil. No sample preparation except an appropriate dilution was required. A preconcentrating liquid chromatography—mass spectrometry interface was used, allowing detection limits of ca . 20 ng to be attained.

Flow Immunoassay Using Solid-Phase Entrapment

A flow injection immunoassay was performed using a column packed with reversed-phase sorbents to effect separation of the immunoreacted species by entrapping free analyte and allowing antibody-conjugated analyte to pass unretained. Fluorescein-labeled analyte was measured in a competitive assay for the anticonvulsant drug phenytoin. The simplicity of the assay was the greatest advantage of the technique, which allowed for measurement of phenytoin in a 2-min assay time. The reliable detection limit for the assay was 5 nmol L(-)(1) of phenytoin in serum. The columns were regenerated with periodic injections of ethanol solutions to remove the entrapped analyte and prepare the column for subsequent analyses.

Determination of glycyrrhetinic acid in human plasma by high-performance liquid chromatography

A high-performance liquid chromatographic (HPLC) method has been developed for measuring 18 beta-glycyrrhetinic acid (GRA) in human plasma in the range of 0.1-3 micrograms/ml. The acetate ester of GRA is added to the plasma as an internal standard, plasma proteins are denatured with urea to release GRA, and the GRA and the internal standard are extracted in an ion-pairing solid-phase extraction process. An isocratic, reversed-phase HPLC separation is used, followed by ultraviolet absorbance detection at 248 nm. The results from the analysis of five GRA-fortified plasma pools show a mean relative standard deviation of 7% and are accurate to within 10%. With evaporative concentration of the extract, the limit of detection for GRA in plasma is approximately 10 ng/ml.

Characterization of a supercritical fluid chromatographic retention process with a large pressure drop by the temporal average density

The characterization of supercritical fluid chromatographic retention by different forms of the average density, viz., the temporal average density, the spatial average density and the arithmetic average density, is investigated in a system with appreciable pressure drop along the column. The logarithm of the capacity factor, when described in terms of the temporal average density, is independent of the pressure drop. Hence, supercritical fluid chromatographic retention processes can be characterized and represented by a hypothetical zero-pressure-drop system at a density equal to the temporal average density of the real system.

NIST reference materials to support accuracy in drug testing

Substance abuse is a major problem worldwide. There is considerable emphasis placed upon testing individuals for evidence of use of controlled substances. Because the consequences of a positive test can be quite severe, laboratories conducting such tests must rigorously follow a carefully designed quality assurance program. Such a QA program should include use of reference materials to assure that the methods used to detect and quantify drugs are providing accurate results. The National Institute of Standards and Technology (NIST) supports accuracy in drugs of abuse testing by providing Standard Reference Materials (SRMs) with certified concentrations of drugs of abuse in urine- and hair-based reference materials. NIST, working in collaboration with the College of American Pathologists (CAP), has developed urine-based SRMs for marijuana metabolite, cocaine metabolite, morphine and codeine, and morphine glucuronide and CAP Reference Materials for amphetamines and phencyclidine. Certification measurements performed at NIST involve two independent methods for each analyte, one of which always uses GC/MS with the other usually being an LC method with either MS or UV detection. Work has recently been completed on a seven component drug in urine SRM. In addition NIST conducts research in the analysis of hair for drugs of abuse. To assist laboratories testing hair for that purpose, NIST has developed two drugs in hair reference materials.

Certification of bilirubin SRM 916a

SummaryThe process used to certify Standard Reference Material (SRM) 916a Bilirubin is described. The certification involved the use of various analytical techniques to detect or quantitate impurities, as well as to characterize the SRM itself. Bilirubin (BR) is believed to exist, in human serum, as the IXα isomer. Samples prepared commercially, including this SRM, also contain the IIIα and XIIIα isomers which are believed to be formed during purification. For the SRM, the three isomers were measured by HPLC and TLC. 1H NMR was used to detect and quantitate chloroform in the BR. Biliverdine and mesobilirubin were not detected. Impurities insoluble in chloroform, the residue from the ashing of BR, and volatiles were measured, in addition to non-acidic impurities and impurities more acidic than BR. The absorptivity of BR in chloroform was measured. A pink fluorescent impurity was detected and measured by TLC. From these analyses, a best estimate of the total amount of impurities was determined, and the BR was issued as SRM 916a with a certified purity of 98.3±0.3%.