Enzymatic determination of free glutamic acid in dried soups and in minced sausages: NMKL collaborative study.

An enzymatic method for the determination of free glutamic acid in meat products and dried soups was collaboratively studied in 11 laboratories. In the presence of the enzyme glutamate dehydrogenase, L-glutamic acid is oxidatively deaminated by nicotinamide adenine dinucleotide (NAD) to 2-oxoglutarate. In a reaction catalyzed by diaphorase, the NADH thus formed converts 2-(p-iodophenyl)-3-(p-nitrophenyl)-5-phenyltetrazolium chloride to a formazan, which is measured in the visible range at 492 nm. Fourteen samples (7 samples of minced sausage and 7 samples of dried cauliflower soup) with glutamate contents varying between 0.4 and 16 g/kg were included in the study. Materials were distributed to participants as blind duplicates and as split level pairs. The mean relative standard deviation (RSDR) for reproducibility for the dried soup material containing glutamate between 7 and 16 g/kg was 4.6%. RSDR values for samples of minced sausage containing glutamate at lower levels (0.4-1.3 g/kg) were between 12 and 16%.     

Determination of benzene in polypropylene food-packaging materials and food-contact paraffin waxes

An analytical procedure was developed for determination of benzene in polypropylene food packaging and was adapted for determination of benzene in commercial paraffin waxes intended for food-contact use. The polymer was dissolved in hexadecane at 150 degrees C. The wax was melted in an 80 degrees C oven. A simple helium-sparging apparatus was used to remove the volatile chemical from the polymer or wax. The contaminant was collected in methanol, distilled water was added, and the resulting solution was analyzed by headspace gas chromatography. The instrument was equipped with a 30 m fused silica open tubular capillary column and a photoionization detector. Average recoveries of benzene from polymer and paraffin wax at low parts-per-billion concentrations were 63 and 70%, respectively. Limits of detection and quantitation for analysis of polypropylene were 8 and 17 ppb, respectively; the limit of quantitation for analysis of paraffin wax was 2 ppb. In several commercial polypropylene products examined, benzene levels ranged from none detected to 426 ppb. In 3 commercial waxes examined, concentrations of 16-73 ppb benzene were determined. The presence of benzene was confirmed by gas chromatography/mass spectrometry.     

Comparison of microwave digestion with conventional wet ashing and dry ashing digestion for analysis of lead, cadmium, chromium, copper, and zinc in shellfish by flame atomic absorption spectroscopy

A closed vessel microwave digestion procedure was developed for shellfish samples. This procedure was compared with wet and dry ash procedures for levels of lead, cadmium, chromium, copper, and zinc. Results obtained for microwave and conventional wet ash digestion were comparable. The dry ashing procedure produced results consistently lower than either of the other methods. Recoveries ranged from 80-92% for microwave and wet ashing procedures and 54-72% for the dry ashing procedure. Accuracy was also determined by analyzing lobster hepatopancreas marine reference material. Values for Pb, Cd, and Cr fell within the range specified for the reference material for all 3 digestion procedures; however, values were lower for Cu and Zn. Results of this study show that microwave digestion is comparable to wet ashing.     

DETERMINATION OF 14C IN SOIL BY A GEL SUSPENSION METHOD

Radioactivity in soils labelled with ¹⁴C was determined by suspending the finely ground soil in a thixotropic gel, dispersing it by ultrasonics and counting the dispersion in a liquid scintillation spectrometer. The relationship between the weight of sample and counting efficiency was found to be almost linear. Using 30 mg soil, the counting efficiency is 50 per cent and ¹⁴C may be determined with an average coefficient of variation of 1.5 per cent to within 5 per cent of the value determined by oxidative procedures.     

Fluxes,residence times,and sources of some elements to Lake Michigan

Fluxes of 28 elements to Lake Michigan are calculated from literature data on sedimentation rates and concentrations in sediments and water. Lake Michigan residence times are roughly 10⁴ lower than oceanic residence times. A mass balance has been formulated for 24 elements, with soil and aerosol as the only sources. These sources provide fourfold excesses of Cu, Hg, Sb, Se, and V, and order of magnitude deficits for Ca, Mg, Na, and Cl, but account (to within 50%) for the observed inputs of Ag, Al, As, Br, Co, Cr, Fe, K, La, Mn, S, Si, Sc, Th, and Zn. Except for Al, Co, La, Si, and Th, deposition from aerosol accounts for at least one-fifth of the total input.     

Determination of phosphatase activities of soils and animal wastes

A method is described to determine acid and alkaline phosphatase activities from the rate of decomposition of p-nitrophenylphosphate in the presence of large amounts of organic matter, such as occur in the surface layers of soils or in animal wastes. The p-nitrophenol formed is separated by high pressure liquid chromatography on a cellulose column from p-nitrophenylphosphate and other organic compounds present in the soil or waste extract. After separation, p-nitrophenol is measured on-line in a spectrophotometric flow cell at a wavelength of 405 nm. In this way p-nitrophenol concentrations down to 0.1 μm can be measured, making it possible to work with substrate concentrations of 1 μm.

The necessity of correcting the phosphatase activity measured in this way for adsorption of enzyme, substrate (p-nitrophenylphosphate) and product (p-nitrophenol) is discussed.

Acid and alkaline phosphatases are inhibited strongly at phosphate concentrations greater than 0.1 mm, consequently substrate concentrations in the range of 0.01 to 0.1 mm were used.

The method was applied to a number of sandy soils and to pig slurry. Air drying or freeze drying of soils was found to decrease the phosphatase activity. Freeze drying did not affect the phosphatase activity of pig slurry. Michaelis-Menten kinetics were found to apply reasonably well. The resulting kinetic parameters are compared with values from the literature. Phosphatase activities are correlated with organic P and organic matter contents of soils and pig slurry.     

Microcomputer controlled titration for determination of protein nitrogen in feeds and wheat.

The determination of protein nitrogen in feeds and wheat by microcomputer controlled titration is described. The method involves direct titration of ammonia with standard hypochlorite titrant in the presence of bromide. The titrant is delivered by an automatic buret, and the microcomputer controlled, automatically computed potentiometric end points are precise to 0.1% over a 5-fold concentration range of nitrogen. Digestions performed with both mercury and copper catalysts show comparable results. Samples are weighed before digestion by an electronic balance interfaced to the computer which records sample number and weight. An automatic pipet aliquots, dilutes, and buffers samples directly from the digestion tubes; the samples can be immediately titrated with the automatic titrator. The results for protein in NBS standards and check feed samples from an offical testing program compare closely with average values reported for these standards. Results show that feed and wheat samples contained 10-100% protein. Precision for successive aliquots of the same digests is 0.1-0.4%relative standard deviation; precision for multiple digestions of the same sample is 0.1-0.8%.     

Chemical comparison of goldenseal (Hydrastis canadensis L.) root powder from three commercial suppliers

The characterization of herbal materials is a significant challenge to analytical chemists. Goldenseal (Hydrastis canadensis L.), which has been chosen for toxicity evaluation by NIEHS, is among the top 15 herbal supplements currently on the market and contains a complex mixture of indigenous components ranging from carbohydrates and amino acids to isoquinoline alkaloids. One key component of herbal supplement production is botanical authentication, which is also recommended prior to initiation of efficacy or toxicological studies. To evaluate material available to consumers, goldenseal root powder was obtained from three commercial suppliers and a strategy was developed for characterization and comparison that included Soxhlet extraction, HPLC, GC-MS, and LC-MS analyses. HPLC was used to determine the weight percentages of the goldenseal alkaloids berberine, hydrastine, and canadine in the various extract residues. Palmatine, an isoquinoline alkaloid native to Coptis spp. and other common goldenseal adulterants, was also quantitated using HPLC. GC-MS was used to identify non-alkaloid constituents in goldenseal root powder, whereas LC-MS was used to identify alkaloid components. After review of the characterization data, it was determined that alkaloid content was the best biomarker for goldenseal. A 20-min ambient extraction method for the determination of alkaloid content was also developed and used to analyze the commercial material. All three lots of purchased material contained goldenseal alkaloids hydrastinine, berberastine, tetrahydroberberastine, canadaline, berberine, hydrastine, and canadine. Material from a single supplier also contained palmatine, coptisine, and jatrorrhizine, thus indicating that the material was not pure goldenseal. Comparative data for three commercial sources of goldenseal root powder are presented.     

Estimates of minimum viable population sizes for vertebrates and factors influencing those estimates

Population size is a major determinant of extinction risk. However, controversy remains as to how large populations need to be to ensure persistence. It is generally believed that minimum viable population sizes (MVPs) would be highly specific, depending on the environmental and life history characteristics of the species. We used population viability analysis to estimate MVPs for 102 species. We define a minimum viable population size as one with a 99% probability of persistence for 40 generations. The models are comprehensive and include age-structure, catastrophes, demographic stochasticity, environmental stochasticity, and inbreeding depression. The mean and median estimates of MVP were 7316 and 5816 adults, respectively. This is slightly larger than, but in general agreement with, previous estimates of MVP. MVPs did not differ significantly among major taxa, or with latitude or trophic level, but were negatively correlated with population growth rate and positively correlated with the length of the study used to parameterize the model. A doubling of study duration increased the estimated MVP by approximately 67%. The increase in extinction risk is associated with greater temporal variation in population size for models built from longer data sets. Short-term studies consistently underestimate the true variances for demographic parameters in populations. Thus, the lack of long-term studies for endangered species leads to widespread underestimation of extinction risk. The results of our simulations suggest that conservation programs, for wild populations, need to be designed to conserve habitat capable of supporting approximately 7000 adult vertebrates in order to ensure long-term persistence.