R. Duane Satzger

Evaluation of the renal effects of experimental feeding of melamine and cyanuric acid to fish and pigs.

OBJECTIVEnTo determine whether renal crystals can be experimentally induced in animals fed melamine or the related triazine compound cyanuric acid, separately or in combination, and to compare experimentally induced crystals with those from a cat with triazine-related renal failure.nnnANIMALSn75 fish (21 tilapia, 24 rainbow trout, 15 channel catfish, and 15 Atlantic salmon), 4 pigs, and 1 cat that was euthanatized because of renal failure.nnnPROCEDURESnFish and pigs were fed a target dosage of melamine (400 mg/kg), cyanuric acid (400 mg/kg), or melamine and cyanuric acid (400 mg of each compound/kg) daily for 3 days and were euthanatized 1, 3, 6, 10, or 14 days after administration ceased. Fresh, frozen, and formalin-fixed kidneys were examined for crystals. Edible tissues were collected for residue analysis. Crystals were examined for composition via Raman spectroscopy and hydrophilic-interaction liquid chromatography-tandem mass spectrometry.nnnRESULTSnAll animals fed the combination of melamine and cyanuric acid developed goldbrown renal crystals arranged in radial spheres (spherulites), similar to those detected in the cat. Spectral analyses of crystals from the cat, pigs, and fish were consistent with melamine-cyanurate complex crystals. Melamine and cyanuric acid residues were identified in edible tissues of fish.nnnCONCLUSIONS AND CLINICAL RELEVANCEnAlthough melamine and cyanuric acid appeared to have low toxicity when administered separately, they induced extensive renal crystal formation when administered together. The subsequent renal failure may be similar to acute uric acid nephropathy in humans, in which crystal spherulites obstruct renal tubules.

Determination of ephedrine compounds in nutritional supplements by cyclodextrin-modified capillary electrophoresis

Capillary electrophoresis was utilized for the separation, identification, and quantitation of ten stereoisomers in the ephedrine family. Chiral discrimination was accomplished through the use of hydroxypropyl-beta-cyclodextrin, and separation was enhanced at pH 2 in the presence of tetramethylammonium chloride and sodium dodecyl sulfate. Calibration plots of the ephedrines were linear over the range 4-100 micrograms/ml. This method was used in the analysis of nutritional supplements that contain Ma Huang, a Chinese herbal preparation that is made from plants in the genus Ephedra.

Electrothermal vaporisation interface for sample introduction in inductively coupled plasma mass spectrometry

An electrothermal vaporisation (ETV) device was developed for inductively coupled plasma mass spectrometry by modifying a commercial graphite furnace unit. A detailed description of the modifications are presented. The new ETV interface is designed to increase sample transport efficiency by converting analyte vapour into micro-particles in the furnace. The characterisation and optimisation of the device are also discussed. Ten consecutive firings of a 30-pg sample of Pb resulted in relative standard deviations (RSD) of 4% for peak-height and 1.6% for peak-area measurements. The absolute detection limit, determined by comparing the response obtained for a 3-µl sample of a 10 ng ml–1 Pb solution in 1% HNO3 with that obtained for blanks containing 1% HNO3 only, was 10 and 14 fg, for a dwell time of 10 µs, for peak area and peak height, respectively. The linear dynamic range for Pb extends over three orders of magnitude, from 300 fg to 300 pg.

DETERMINATION OF GAMMA-HYDROXYBUTYRATE (GHB) AND GAMMA-BUTYROLACTONE (GBL)BY HPLC/UV-VIS SPECTROPHOTOMETRY AND HPLC/THERMOSPRAY MASS SPECTROMETRY

This laboratory frequently receives samples that are suspected of being gamma-hydroxybutyrate (GHB), its lactone, gamma-butyrolactone (GBL), or a mixture of the two. We have developed an HPLC/UV-VIS spectrophotometric method for the separation and quantitation of GHB and GBL in illegal preparations that are available on the black market. The estimated detection limit is 50 ng injected onto the column. We are also reporting a simple and fast HPLC/thermospray mass spectrometric method for the confirmation of these compounds in illegal preparations. The characteristic mass spectrum for each compound could be obtained from as low as a 5 µg injection.

Detection of clenbuterol in bovine retinal tissue by high-performance liquid chromatography with electrochemical detection

A method for the detection of the beta-agonist drug clenbuterol in bovine retinal tissue has been developed. The extraction procedure involves sonication and centrifugation, followed by the addition of ethylenediaminetetraacetic acid (EDTA) to the supernatant. The pH of the supernatant is then brought to 12.2, which is then allowed to sit for 2 h. This is followed by a diethylether extraction. The diethylether extract is dried under nitrogen and the residue is dissolved in 1% formic acid. The quantitation of clenbuterol was accomplished by high-performance liquid chromatography with electrochemical detection. The electrochemical detector was an amperometric detection. The detector was set in the pulsed mode. The oxidizing potential of a carbon electrode was 1.3 V vs. a Ag/AgCl reference electrode and was pulsed to a reduction potential of 2.0 V vs. a Ag/AgCl reference electrode. The limit of detection for this method was 5 ng/ml of clenbuterol (S/N = 3). Typical spiked recoveries are 75%.

Evaluation of a New Handheld Instrument for the Detection of Counterfeit Artesunate by Visual Fluorescence Comparison

There is an urgent need for accurate and inexpensive handheld instruments for the evaluation of medicine quality in the field. A blinded evaluation of the diagnostic accuracy of the Counterfeit Detection Device 3 (CD-3), developed by the US Food and Drug Administration Forensic Chemistry Center, was conducted in the Lao Peoples Democratic Republic. Two hundred three samples of the oral antimalarial artesunate were compared with authentic products using the CD-3 by a trainer and two trainees. The specificity (95% confidence interval [95% CI]), sensitivity (95% CI), positive predictive value (95% CI), and negative predictive value (95% CI) of the CD-3 for detecting counterfeit (falsified) artesunate were 100% (93.8–100%), 98.4% (93.8–99.7%), 100% (96.2–100%), and 97.4% (90.2–99.6%), respectively. Interobserver agreement for 203 samples of artesunate was 100%. The CD-3 holds promise as a relatively inexpensive and easy to use instrument for field evaluation of medicines, potentially empowering drug inspectors, customs agents, and pharmacists.

Preliminary investigation of a tangential flow torch with coupling probe injector for helium microwave induced plasma mass spectrometry

A stable, low gas-flow torch has been developed for use with a helium microwave induced plasma (MIP). A toroidal plasma with central analyte introduction is obtained by the addition of a tantalum coupling probe injector tube. This injector penetrates through 100% of the total cavity depth and aids in the efficiency of power transfer to the cavity, in plasma initiation, and in circumventing the effects of a lack of homogeneity in the microwave field on analyte distribution in the plasma. The tangential helium flow was 41/min and the microwave power was 60 W.

Death and serious illness following influenza vaccination: a multidisciplinary investigation.

To evaluate a possible association between influenza vaccination and four deaths and four serious illnesses among 114 recent influenza vaccinees in a long‐term care facility (LTCF) and two deaths from a nearby physicians office. All had received vaccine from the same lot (Lot A).

Assessment of the effectiveness of the CD3+ tool to detect counterfeit and substandard anti-malarials.

BackgroundThe US FDA recently developed CD3+, a counterfeit detection tool that is based on sample illumination at specific wavelengths of light and visual comparison of suspect sample and packaging materials to an authentic sample. To test performance of the CD3+ in field conditions, a study was conducted in Ghana which compared the CD3+ side-by-side with two existing medicine quality screening technologies—TruScan™ Portable Raman spectrometer and GPHF Minilab®.MethodsA total of 84 anti-malarial test samples comprising artemether–lumefantrine tablets and artesunate–amodiaquine tablets were used. The technologies were evaluated for sensitivity in determining counterfeit/substandard (The term counterfeit or falsified is used in this article to refer to medicines that carry a false representation of identity or source or both. The term substandard is used to refer to medicines that do not meet the quality specifications given in the accepted pharmacopeia.) medicines, specificity in determining authentic products, and reliability of the results. Authentic samples obtained from manufacturers were used as reference standards. HPLC analysis data was used as the “gold standard” for decisions regarding a sample being authentic or substandard/counterfeit.ResultsCD3+ had a sensitivity of 1.00 in detecting counterfeit/substandard products compared to Minilab (0.79) and TruScan (0.79). CD3+ had a lower specificity (0.53) in determining authentic products compared to the specificities reached by Minilab (0.99) and TruScan (1.00). High sensitivity in this context means that the technology is effective in identifying substandard/counterfeit products whereas the low specificity means that the technique can sometimes mischaracterize good products as substandard/counterfeit. Examination of dosage units only (and not packaging) using CD3+ yielded improved specificity 0.64. When only assessment of sample identification was done, the TruScan provided sensitivity (1.00) and specificity (0.99); and the Minilab provided sensitivity (1.00) and specificity (1.00). All three technologies demonstrated 100xa0% reliability when used to analyse the same set of samples over 3xa0days by a single analyst and also when used to determine the same set of samples by three different analysts. Eight of the field samples were confirmed to be counterfeits with no active pharmaceutical ingredient content. All three technologies identified these samples as counterfeits.ConclusionsThe study revealed the relative effectiveness of the technologies as quality control tools. Using a combination of CD3+, with either the Minilab or TruScan, to screen for medicine quality will allow for complete examination of both the dosage units and the packaging to decide whether it is authentic or counterfeit.

Guidelines for the Identification of Unknown Samples for Laboratories Performing Forensic Analyses for Chemical Terrorism

Abstract:u2002 Since the early 1990s, the FBI Laboratory has sponsored Scientific Working Groups to improve discipline practices and build consensus among the forensic community. The Scientific Working Group on the Forensic Analysis of Chemical, Biological, Radiological and Nuclear Terrorism developed guidance, contained in this document, on issues forensic laboratories encounter when accepting and analyzing unknown samples associated with chemical terrorism, including laboratory capabilities and analytical testing plans. In the context of forensic analysis of chemical terrorism, this guidance defines an unknown sample and addresses what constitutes definitive and tentative identification. Laboratory safety, reporting issues, and postreporting considerations are also discussed. Utilization of these guidelines, as part of planning for forensic analysis related to a chemical terrorism incident, may help avoid unfortunate consequences not only to the public but also to the laboratory personnel.