Determination of polysorbates in foods by colorimetry with confirmation by infrared spectrophotometry, thin-layer chromatography, and gas chromatography

A method is presented for the detection of polysorbates (PSs) in 8 kinds of processed foods by colorimetric and thin-layer chromatographic (TLC) techniques. The PSs are extracted from processed foods with a mixture of methylene chloride and ethanol by using an Extrelut column. The extract is further purified by using a silica gel column. The PS extract is complexed with cobalt-thiocyanate (Cothiocyanate) reagent and is determined spectrophotometrically at 620 nm. The recoveries and coefficients of variation for 8 kinds of processed foods fortified with 0.1% PS 80 were 67.9-94.6% and 4.0-11.3%, respectively. The detection limit of TLC corresponded to 50 mg PS 80/kg. PS identity was confirmed by infrared spectrophotometry of PS extract, and gas chromatography of fatty acids and thin layer chromatography of POE-sorbitan residues after saponification.     

Simple and rapid spectrophotometric method for determination of adrenaline and isoprenaline

A simple, rapid, and specific method for determination of adrenaline bitartrate and isoprenaline sulfate was developed. The method is based on the oxidation reaction in aqueous solution of either adrenaline bitartrate or isoprenaline sulfate in the presence of silver oxide to give a red aminochrome measurable at 490 nm. The color is stable for 2 h. Beer's law is valid within a concentration range of 5-80 micrograms/mL for each drug. All variables were studied to optimize the reaction conditions. The method is specific for catecholamine drugs having a secondary amine in the side chain. Other catecholamines such as orciprenaline and noradrenaline do not interfere, and no interference was observed in the presence of common pharmaceutical adjuvants. Interference due to sodium metabisulfite and sodium chloride was circumvented. The validity of the method was tested by analyzing adrenaline injections and isoprenaline tablets. Good recoveries were obtained for these preparations. The results were comparable to those obtained by official procedures. The proposed method is also recommended as a stability indicating assay for oxidative degradation of both drugs.     

Comparison of postcolumn derivatization-liquid chromatography with thin-layer chromatography for determination of aflatoxins in naturally contaminated corn

Quantitation of aflatoxins by liquid chromatography with postcolumn iodine derivatization (LC-PCD) and fluorescence detection was compared with quantitation by the AOAC CB method, 968.22. Thirty-seven naturally contaminated corn samples were ground and then divided. One portion was extracted, and the extract was cleaned up and analyzed by thin-layer chromatography according to the CB method. The second portion was extracted and cleaned up in a similar fashion, but quantitation was by the LC-PCD method. For aflatoxin B1 concentrations ranging from 0 to 150 ng/g, results obtained by the 2 methods were fitted to a linear equation with the LC-PCD results as the dependent variable. The correlation coefficient was 0.99, the intercept was near 0, and the slope was near 1. For aflatoxin B2, the correlation coefficient was 0.97, and the intercept was near 0. However, the slope of the equation relating LC-PCD concentration to TLC concentration was only 0.5. We believe that this lack of equivalence between the methods for determination of aflatoxin B2 is due to overestimation by the TLC method because the low levels present are near the TLC detection limit for B2.     

Detection and separation of fenpropathrin, flucythrinate, fluvalinate, and PP 321 by thin-layer chromatography

Four synthetic pyrethroids having alpha-cyano ester groups, i.e., fenpropathrin, flucythrinate, fluvalinate, and PP 321, are separated by thin-layer chromatography and detected by a new set of chromogenic reagents. Synthetic pyrethroids containing the alpha-cyano group react with sodium hydroxide to liberate cyanide which forms pink spots with o-dinitrobenzene and p-nitrobenzaldehyde. The detection limit is 0.1 microgram and the method can be applied for identification and confirmation of these synthetic pyrethroids in vegetables.     

Multiresidue screen for cardiotoxins by two-dimensional thin-layer chromatography

A two-dimensional thin-layer chromatographic method was developed for the qualitative determination of the cardiotoxins oleandrin, gitoxin, digitoxin, gitoxigenin, and grayanotoxins I, II, and III in gastrointestinal contents (stomach, rumen, colon, and cecum contents), feces, and plant material. The cardiotoxins were extracted with dichloromethane. The extract was cleaned up by charcoal and reverse phase solid-phase extraction columns. Analysis was performed by two-dimensional thin-layer chromatography on silica gel plates and visualized by aluminum chloride followed by chloramine T spray. The method detection limits were 0.05 microg/g for oleandrin, 0.1 microg/g for gitoxin, and 0.2 microg/g for the other toxicants in gastrointestinal contents and feces and were 5 times higher in plant material. Four replicate fortifications of bovine rumen contents, bovine feces, and alfalfa at these levels were all well recovered. The diagnostic utility of the method was tested by analyzing samples submitted to the veterinary toxicology laboratory.     

Simple, rapid method for determination of total extractable fat in canned pet foods

The rapid column method described, unlike AOAC method 7.056, determines both neutral ("crude") and total fat in canned pet foods, and uses nonflammable solvent mixtures and simple laboratory equipment. Neutral fat values are obtained by eluting the column with dichloromethane, whereas total fat values are determined by using dichloromethane-methanol (9 + 1). For 7 samples analyzed in triplicate, fat ranged from 2.9 to 10.8%. Neutral fat values by the dry column method were significantly lower (P less than 0.05) than were those by 7.056 (6.29 vs 6.49), although these differences were practically unimportant. Total fat determinations by the dry column method and by 7.056 yielded overall means of 7.40 and 6.49%, respectively. The 0.91% mean difference is significant (P less than 0.01) and represents a more complete extraction of polar lipids by the proposed method.     

Determination of amphetamine by Schiff base formation and quantitative gas-liquid chromatography.

A procedure was developed to determine amphetamine salts in solid dosage forms. Amphetamine was extracted from the solid matrix with dilute hydrochloric acid and reacted with cyclohexanone in a strongly basic aqueous methanolic solution. The Schiff base reaction product was extracted with hexane for gas chromatographic determination. Reaction time and optimum conditions were studied. Phenethylamine, similarly treated, was used as an internal standard. Results compared favorably with those obtained by using USP methods.     

Simple saponification method for the quantitative determination of carotenoids in green vegetables

A simple, reliable, and gentle saponification method for the quantitative determination of carotenoids in green vegetables was developed. The method involves an extraction procedure with acetone and the selective removal of the chlorophylls and esterified fatty acids from the organic phase using a strongly basic resin (Ambersep 900 OH). Extracts from common green vegetables (beans, broccoli, green bell pepper, chive, lettuce, parsley, peas, and spinach) were analyzed by high-performance liquid chromatography (HPLC) for their content of major carotenoids before and after action of Ambersep 900 OH. The mean recovery percentages for most carotenoids [(all-E)-violaxanthin, (all-E)-lutein epoxide, (all-E)-lutein, neolutein A, and (all-E)-beta-carotene] after saponification of the vegetable extracts with Ambersep 900 OH were close to 100% (99-104%), while the mean recovery percentages of (9'Z)-neoxanthin increased to 119% and that of (all-E)-neoxanthin and neolutein B decreased to 90% and 72%, respectively.     

Evaluation of a hand held reflectometer for rapid quantitative determination of nitrate

Abstract

An inexpensive hand held reflectometer was used to estimate NO₃ concentration with Merckoquant NO₃ indicator strips. For KNO₃ solutions a fourfold improvement in precision was obtained over the visual method of colour comparison. The instrument calibration was stable over several days and not affected by temperature in the range 8–34^o C. The reflectometer showed quantitatively that increasing temperature (8–34^oC) reduced the optimal time for reading the strips from 120s to 40s. With standard NO₃ solutions a simple hyperbolic transformation of the reflectometer readings produced a linear calibration of good fit (r²=0.99). NO₃ could be estimated in different soil types within half an hour. The results agreed well with values from ion chromatography (r² =0.97, slope=1.07). Plant sap analysis was moderately precise (r² =0.92) but inaccurate and underestimated by more than 60% values obtained by ion chromatography. This discrepancy was not caused by the reflectometer but seemed to be due to factors within the plant sap that interfered with the NO₃ colour reaction in the Merckoquant strips.     

Simple and rapid determination of thiol- and organic disulfide-sulfur in foliage and humic soil horizons

A new method for rapid determination of thiol‒ (R‒SH) and disulfide (R‒S‒S‒R′) sulfur in soil and foliage samples is presented. Using a silver sulfide electrode, the thiol sulfur content of a sample is determined by potentiometric titration with AgNO₃. After reduction with a mixture of NaOH and ascorbic acid also its disulfide S content can be quantified subsequent to neutralization of the reductive solution with citric acid. The method was tested with eight organic standards, six humic soil samples, and three Norway spruce needle samples. Disulfide S from standards with abstrictable H‒atoms in α‒ or β‒position to the disulfide bond could — with one exception — be detected completely. For the aromatic disulfides which allow only direct nucleophilic attack, recovery was at 75%. For the soil samples, 32 to 60% of the carbon‒bonded S consisted of disulfide S, for the spruce needle samples 26 to 33%. The method provides satisfactory results for most studied standards and is applicable to various natural substances after adequate sample preparation.     

Rapid quantitative method for simultaneous determination of benzoic acid, sorbic acid, and four parabens in meat and nonmeat products by liquid chromatography

A liquid chromatographic (LC) method for the simultaneous determinations of benzoic acid, sorbic acid, and methyl, ethyl, propyl, and butyl parabens (methyl, ethyl, propyl, and butyl-p-hydroxybenzoates) in meat and nonmeat products was developed. Benzoic acid, sorbic acid, and parabens were extracted from meat and nonmeat products with 70% ethanol. After filtration, extracts were analyzed by reverse phase liquid chromatography. Homogeneously ground samples of fresh sausage and hamburger were fortified with benzoic acid, sorbic acid, and each paraben at 5 different concentrations. Average recovery (after discarding outliers) for each preservative at all 5 levels was greater than 95% with a coefficient of variation less than 5%.     

Simple and rapid determination of thiabendazole,imazalil, and o-phenylphenol in citrus fruit using flow-injection electrospray ionization tandem mass spectrometry

A simple and rapid analytical method for thiabendazole (TBZ), imazalil (IMA), and o-phenylphenol (OPP) in citrus fruit has been developed by using flow-injection electrospray ionization tandem mass spectrometry for the first time. The method involves the combined use of stable isotopically labeled internal standards (thiabendazole-(13)C(6), imazalil-d(5), and p-phenylphenol-d(9)) and a multiple reaction monitoring technique. The average recoveries for the fungicides at the tolerance levels (TBZ and OPP, 10 mg/kg; IMA, 5 mg/kg) ranged from 77 to 101%, with the coefficients of variation (CVs) ranging from 0.7 to 4.2% (n = 5). At half the tolerance levels (TBZ and OPP, 5 mg/kg; IMA, 2.5 mg/kg), the average recoveries ranged from 62 to 112%, with the CVs ranging from 0.7 to 8.4% (n = 5). The CVs of the average recoveries, obtained from lemon samples fortified with three fungicides at the tolerance levels, obtained on three different days over two weeks, ranged within 2%. The analysis time, including sample preparation and determination, is only 15 min.     

Simultaneous determination of oxytetracycline, tetracycline, and chlortetracycline in milk by liquid chromatography

A simple, rapid procedure is described for the simultaneous quantification of 3 tetracycline drugs in bovine milk. Samples are prepared by dilution with an EDTA/phosphate buffer solution and filtration through a molecular weight cutoff filter. Analytes are concentrated on-column using a reverse-phase gradient elution system of oxalic acid, acetonitrile, and methanol. The limits of quantitation are approximately 15-50 ng/mL and the limits of detection are 10-20 ng/mL, depending on the compound. For oxytetracycline, over the range 50-1200 ng/mL, the average recovery and intralaboratory coefficient of variation were 97% and 4.1%, respectively. Over the same range, these parameters were, respectively, 97% and 5.0% for tetracycline, and 89% and 6.4% for chlortetracycline. The applicability of this procedure is demonstrated by separation and detection of incurred tetracycline residues in milk from treated animals.