Quantitation of the nitrosamine 2-ethylhexyl-4-(N-methyl-N-nitrosoamino) benzoate (NPABAO) in sunscreen products.

We have devised a method to quantitate the nitrosamine, 2-ethylhexyl-4-(N-methyl-N-nitrosoamino) benzoate (NPABAO), in commercial products containing the sunscreen ingredient, Padimate O. The method involves a minimum of cleanup steps to afford a nonaqueous extract from product emulsions suitable for analysis by a liquid chromatograph interfaced to a thermal energy analyzer (LC/TEA). The method is applicable to lotions, creams, and gels. Oils are normally soluble in the mobile phase and can be analyzed directly on the LC/TEA without additional cleanup procedures. The method has a minimum detectable limit of about 30 ppb and yields greater than 80% recovery. It is highly reproducible and generates no NPABAO artifactually prior to quantitation on the LC/TEA. Application of the method to 22 different commercial product formulas disclosed that the level of NPABAO in each of the products is below 250 ppb, with 18 of the products containing less than 100 ppb. Of interest was the observation that musk ketone, a common fragrance constituent, produces a false-positive TEA response that can interfere with accurate analysis of NPABAO content in typical commercial products.     

The problem of pretreatment and unintentional variations in the fumigation-extraction method for time-course measurements in the field

A control soil stored at 4°C was analyzed 38 times by fumigation-extraction during a period of 11 months to correct for variations caused by the analytical procedure. The difference in extractable C between fumigated and unfumigated samples oscillated around the average without a positive or negative trend. When data from contemporaneously extracted field samples were corrected with control soil data the variations were lowered. The deviations between corrected and uncorrected biomass C values had maxima of ±12%. Data obtained for seven dates using pre-extraction, wet-sieving, and centrifuging were compared with data obtained by the conventional procedure without any pretreatment. A negative difference from data obtained without pretreatment was found when the soil water content was decreased to 6%. The largest positive difference (+38%) was found in May during the period of highest root growth.     

Effects of lactoferrin, phytic acid, and EDTA on oxidation in two food emulsions enriched with long-chain polyunsaturated fatty acids

The influence of the addition of metal chelators on oxidative stability was studied in a milk drink and in a mayonnaise system containing highly polyunsaturated lipids. Milk drinks containing 5% (w/w) of specific structured lipid were supplemented with lactoferrin (6-24 muM) and stored at 2 degrees C for up to 9 weeks. Mayonnaise samples with 16% fish oil and 64% rapeseed oil (w/w) were supplemented with either lactoferrin (8-32 muM), phytic acid (16-124 muM), or EDTA (16-64 muM) and were stored at 20 degrees C for up to 4 weeks. The effect of the metal chelators was evaluated by determination of peroxide values, secondary volatile oxidation products, and sensory analysis. Lactoferrin reduced the oxidation when added in concentrations of 12 muM in the milk drink and 8 muM in the mayonnaise, whereas it was a prooxidant at higher concentrations in both systems. In mayonnaise, EDTA was an effective metal chelator even at 16 muM, whereas phytic acid did not exert a distinct protective effect against oxidation. The differences in the equimolar effects of the metal chelators are proposed to be due to differences in their binding constants to iron and their different stabilities toward heat and low pH.     

Comparison of Closely Related Orthopoxvirus Isolates by Random Amplified Polymorphic DNA and Restriction Fragment Length Polymorphism Analysis

The reliability and reproducibility of random amplified polymorphic DNA analysis (RAPD) was compared with restriction fragment length polymorphism (RFLP) by analysing three virus strains isolated from zoo animals in Berlin and three isolates which were cultivated from pets from Northern Germany. The RAPD technique was evaluated as a reliable tool with good reproducibility of the patterns for each virus strain investigated. Problems of interpretation due to inconsistent intensity of bands in different polymerase chain reaction runs may arise for less experienced personnel. The RAPD analysis can be performed within one working day and needs less DNA compared with RFLP so costs will be reduced. The obvious advantage of RFLP is that the pattern can be traced to the recognition site of the restriction enzyme whereas the RAPD primer sequence is not present in the orthopoxvirus genome at all. To the authors knowledge, the RAPD technique has never been applied in DNA viruses before and they conclude that this technique is a useful tool for the discrimination of closely related cowpoxviruses.     

Immune response in pigs to Aujeszky's disease viruses defective in glycoprotein g1 or gX.

Two Aujeszky's disease virus glycoprotein genes, gX and g1, have been used to produce deletion mutants which have then been developed into vaccines. These deletions then allow differentiation between pigs infected with wild type virus and those given the vaccine. It is not clear whether the glycoproteins encoded for by these genes are needed to induce a full protective immune response, in which case deletion mutants would suffer from lack of potency. To test this, commercially available Aujeszky's virus vaccines which lacked either gX or g1 were compared and isogenic constructs were made which differed only in the absence or presence of gX and, or, g1. These constructs and vaccines were used to vaccinate the natural host of Aujeszky's disease, the pig, and potency was measured using challenge with wild type virus. In all cases vaccines which lacked g1 performed significantly less well than those in which g1 was present, whereas deletions of gX had no significant effect on vaccine performance.     

Adrenocortical suppression associated with topical otic administration of glucocorticoids in dogs

Commercial otic preparations that contained dexamethasone or triamcinolone acetate were applied twice daily to both ears of 2 groups of dogs (n = 8). Marked adrenocortical suppression, reflected by low serum cortisol concentrations, was observed in all dogs. Results of ACTH response tests were blunted after 7 days of treatment. Twenty-one days after treatment, serum cortisol concentrations still were suppressed in all dogs, compared with pretreatment control concentrations. Fourteen days after cessation of otic treatment, 5 of 8 dogs still had inadequate release of cortisol in response to ACTH.     

Eel acetylcholinesterase inhibition studies with heteroarylphosphinates

The inhibition of eel acetylcholinesterase by the 4-nitrophenyl esters of 2-furyl(methyl)-, methyl(2-thienyl)-, di-2-furyl-, and di-2-thienylphosphinic acid (I, II, III, and IV, respectively) was investigated at pH 6.90 in 0.067 M phosphate buffer (25.0°C) using stopped-flow instrumentation and automated data processing. Our evaluation of the dissociation constant, K_d, the unimolecular bonding rate constant, k₂, and the bimolecular reaction constant, k_i, are the first reported values for these constants for alkyl/heteroaryl and diheteroaryl esters of phosphinic acids. The largest k_i value (19,330 M^?1 sec^?1) was observed for the reaction of I with the enzyme. The order for the remaining three is II > IV > III. There is no direct relationship between the hydrolysis rates of the esters and their anticholinesterase activities on eel acetylcholinesterase. Likewise, there is no direct relationship between their anticholinesterase activities and the LD₅₀ values in rats.