Enzymatic hydrolysis of biological and environmental samples as pretreatment for analysis.

Four commercially available proteases were tested, in conjunction with a lipase, for efficacy in hydrolyzing 3 tissue substrates: cod fillet, chicken egg, and bovine liver. Enzymatic hydrolysis of tissues minimizes the formation of emulsions during liquid-liquid extraction and does not accelerate the decomposition of acid- or base-labile analytes. Recovery of hexane and benzene phases from the hydrolysates was also evaluated. Protease from Streptomyces griseus combined with lipase from Candida cylindracea (available commercially) produced the highest percent hydrolysis (relative to classical acid hydrolysis) in all 3 tested tissues (60-95%) and the greatest recovery of hexane (100%) and benzene (92-100%) solvent phases.

Gas chromatographic determination of polysaccharide gums in foods after hydrolysis and derivatization.

A gas chromatographic method was evaluated for the determination of food grade gums in dairy products, salad dressings and meat sauces. The gums studied were tragacanth, karaya, ghatti, carob, guar, arabic and xanthan gum. The extraction method included removal of fat followed by starch degradation then precipitation of protein. The isolated gums were hydrolysed with trifluoroacetic acid and the resulting neutral monosaccharides converted to their aldonitrile acetate derivatives for determination by gas chromatography. Recoveries from thirteen different commodities averaged 85%. However, the recovery of guar gum from ice cream and cold pack cheese was 42 and 50%, respectively. In a comparison of enzyme hydrolysis and iodine complexation for the removal of starch the former was simpler and provided cleaner extracts than the iodine treatment. Both gave similar results.

Preparation of deoxynivalenol (vomitoxin) from field-inoculated corn.

A process was developed for production of gram quantities of deoxynivalenol (DON) from corn that had been inoculated in the field with Fusarium graminearum and was estimated to contain 400-500 mg DON/kg. Steps in the purification procedure included extraction with methanol-water (1 + 1), partition from an aqueous solution into ethyl acetate by using a hydrophilic matrix, defatting, Florisil column chromatography, methylene chloride-water partition, semipreparative liquid chromatography (LC), and crystallization from ethyl acetate. The average yield of crystalline DON produced by this method (or with minor variations) was 281 mg/kg.

The determination of formaldehyde in beer and soft drinks by HPLC of the 2,4-dinitrophenylhydrazone derivative.

A simple procedure was developed for the determination of formaldehyde in samples of beer and soft drinks. A volume of sample was distilled and the aqueous distillate containing the formaldehyde collected directly in a solution of 2,4-dinitrophenylhydrazine. The resulting hydrazone was extracted and analysed by reversed-phase high-performance liquid chromatography with UV absorbance detection. Levels of formaldehyde found were in the low mg/kg range. Detection limits were less than 0.1 mg/kg of sample. Results were confirmed by mass spectrometry (probe) of the derivatives after fraction collection from the HPLC.

Gravimetric determination of ethoxylated mono- and diglycerides in bread.

A gravimetric method is described for the quantitative determination of ethoxylated mono-and diglycerides (EMGs) in bread at levels as low 0.42% (dry sample). The air-dried pulverized samples are Soxhlet-extracted 22 h with an azeotropic mixture of n-propanol-water. The contents are evaporated to dryness, and then treated with 5% HCl in methanol to trans-esterify the fatty acids present which are then removed by petroleum ether extraction. The EMGs are subsequently precipitated from aqueous solution with phosphomolybdic acid in the presence of barium ions. The precipitate is weighed and compared with known amounts of standard carried through the same procedure.

Gas-liquid chromatographic determination of monosodium glutamate in soups and soup bases.

A method is given for determining monosodium glutamate (MSG) in soups and soup bases by gas-liquid chromatography (GLC) of the trimethylsilyl ether derivative of glutamic acid. This method compared favorably with existing methods including GLC of the trifluoroacetate (TFA)/butyl ester derivative, and analysis on an amino acid analyzer. In addition, some samples were analyzed by GLC/mass spectrometry. The MSG content of various products ranged from approximately 0.2% in some condensed soups to 13.1% on bouillon cubes. The method, which can detect as little as 0.05%, requires only a 10 min single step derivatization at room temperature and is preferred to the TFA/butyl ester technique.

Recent studies in Canada on the analysis and occurrence of volatile and non-volatile N-nitroso compounds in foods.

The present paper briefly summarizes the research activities of the Food Research Laboratories, Canadian Health Protection Branch, during the past two years. Data presented indicate that traces of nitrosamines, mainly nitrosodimethylamine and nitrosodiethylamine, can occur in cheese. Several new analytical methods are presented for the determination of non-volatile nitroso compounds, such as nitrosohydroxypyrrolidine, nitrosodiethanolamine, nitrosoproline, nitrosohydroxyproline and nitrosoglyphosphate (a nitrosated herbicide). It is shown that all of the above compounds, even the very polar nitrosoglyphosphate, can be converted to volatile derivatives and then analysed by GLC-MS or by conventional GLC using a specific (such as P-photometric) detector. Traces of nitrosohydroxypyrrolidine have been detected in a few samples of cooked bacon. Contrary to previously reported findings, no significant levels of nitrosoproline were detected in the 6 samples of raw bacon analysed.

Nitrosamines in cured meat products.

One hundred samples of specially selected spiced meat products (sausages, salami, bologna, wieners, meat loaf, canned luncheon mean, etc) were analysed for nitrate, nitrite and volatile nitrosamines. None of the samples contained high levels of nitrosamines, but many contained traces, generally in the range from 2-50 mug/kg. Some contained as many as four nitrosamines, namely, NDMA, NDEA, NPip and NPy. In a few cases the samples were reanalysed after two weeks' storage at 4 or -20 degrees C, but no significant change in the nitrosamine levels could be detected. The identity of the nitrosamines was confirmed by GLC-high-resolution-MS.