Microwave-assisted rapid determination of vitamins a and e in beverages

A new rapid procedure for the determination of vitamins A and E in beverages has been developed and validated. Key steps include a microwave-assisted saponification of the sample and a single-step extraction of the vitamins prior to HPLC analysis. All sample preparation steps are carried out consecutively in the same vial. The vitamins are determined using normal-phase (Si-60) HPLC with fluorescence detection. The method is applicable to beverages with a content of all-trans-retinol >0.14 mg/L and/or a content of alpha-tocopherol >1 mg/L. Recoveries determined by spiking experiments ranged from 91.3 to 106.3%. The precision of the method is characterized by relative standard deviations of <2% for alpha-tocopherol and <5% for all-trans-retinol.     

Quantification of tocopherols and tocotrienols in portuguese olive oils using HPLC with three different detection systems

Three different HPLC detection systems were compared for the determination of tocopherols and tocotrienols in olive oil: fluorescence and diode array connected in series, ultraviolet, and evaporative light scattering. The best results were obtained with the fluorescence detector, which was successfully applied in the quantification of tocopherols and tocotrienols in 18 samples of Portuguese olive oils. To support the validity of the method, the parameters evaluated were linearity, detection limits, repeatability, and recovery. All of the studied samples showed similar qualitative profiles with six identified compounds: alpha-T, beta-T, gamma-T, delta-T, alpha-T3, and gamma-T3. Alpha-tocopherol (alpha-T) was the main vitamin E isomer in all samples ranging from 93 to 260 mg/kg. The total tocopherols and tocotrienols ranged from 100 to 270 mg/kg. Geographic origin did not seem to influence the tocopherol and tocotrienol composition of the olive oils under evaluation.     

DC polarography of cephradine and its application to capsules.

A polarographic method has been developed for the quantitative analysis of cephradine and its dosage forms. Direct determinations on capsules are carried out; excipients and coloring matter do not interfere in the determination. The electroactive product is formed by acidic hydrolysis with 5.0N HCl and heating at 80 degrees C for 60 min. Two polarographic waves are obtained: I = -0.46 V and II = -0.78 v vs. SCE. Both reduction waves are diffusion controlled. Wave I is preferred for analytical purposes. The precise chemical identity of the electroactive product has not been determined, but UV spectral data and the TLC Rf value are reported. A linear relation is established for levels of cephradine between 10(-2) and 10(-5)M in 5.0N HCl.     

Disposable cartridge extraction of retinol and alpha-tocopherol from fatty samples

A new approach is proposed for liquid/solid extraction of retinol and alpha-tocopherol from samples, using a disposable kieselguhr cartridge. The substitution of the mixture methanol-ethanol-n-butanol (4 + 3 + 1) for methanol in the alkaline hydrolysis solution makes it now possible to process fatty samples. Methanol is necessary to solubilize the antioxidant ascorbic acid, and a linear chain alcohol such as n-butanol is necessary to reduce the size of soap micelles so that they can penetrate into the kieselguhr pores. In comparisons of the proposed method with conventional methods on mineral premixes and fatty feedstuffs, recovery and accuracy are at least as good by the proposed method. Advantages are increased rate of determinations and the ability to hydrolyze and extract retinol and alpha-tocopherol together from the same sample.     

Simultaneous HPLC analysis of alpha-tocopherol and cholesterol in fresh pig meat

A method has been developed for simultaneously determining alpha-tocopherol and cholesterol in fresh pig meat by HPLC. It allows a reduction in the number of analyses and brings savings in time and materials. The unsaponifiable fraction is extracted following the modified method of Liu et al. (Liu, Q.; Scheller, K. K.; Schaefer, D. M. Technical note: A simplified procedure for vitamin E determination in beef muscle. J. Anim. Sci. 1996, 74, 2406-2410). The modifications introduced are the use of nitrogen atmosphere during the extraction, the addition of an antioxidant in the organic extraction phase, and the use of alpha-tocopherol itself as an internal standard. There is then a chromatographic analysis which allows the separation of the two compounds in question. To identify and quantify, two different detectors are used in series: the first is a fluorescence detector (alpha-tocopherol), and the second is a light-scattering detector (cholesterol). The technique shows sufficient sensitivity to determine the normal levels of alpha-tocopherol and cholesterol in meat, with recovery percentages of 78% and 97%, respectively. The average amount of alpha-tocopherol and cholesterol in samples from pig Longissimus dorsi muscle analyzed using this method is 1.8 and 620 mg/kg of fresh meat, respectively.     

Disposable cartridge extraction of retinol and alpha-tocopherol from feedstuffs

Automated determination of fat-soluble vitamins by modern methods such as liquid chromatography is hampered by the initial extraction step. A simple technique is proposed that allows an appreciable increase in the actual rates of determination. Feedstuff samples are first hydrolyzed in an aqueous alcohol (mainly methanol)-potassium hydroxide solution. Instead of extracting retinol and alpha-tocopherol from the hydrolysis solution by an organic solvent, an aliquot of the solution is mixed with a small volume of a strong antioxidant solution (ascorbic acid) and pipetted onto a kieselguhr disposable cartridge where it is adsorbed. Retinol and alpha-tocopherol are eluted with isooctane at normal pressure. The proposed method has been compared with conventional techniques on many feed samples.     

Comparison of Colorimetric and ICP Methods of Phosphorus Determination in Soil Extracts

The traditional method for quantifying phosphorus (P) in Manitoba soil extracts is the molybdate blue–ascorbic acid colorimetric method. The shift from this traditional method to newer and more sophisticated analytical methods such as inductively coupled plasma (ICP) optical emission spectroscopy for P determination in soil extract could have serious implications on agronomic and environmental P management. Thus, the objectives of this study were to compare P determination by colorimetric and ICP methods in four extractants, namely Olsen, Mehlich 3, CaCl₂, and water extraction methods and to evaluate the possibility of developing conversion equations for P determination for the two methods in Manitoba soils. A laboratory experiment was conducted to establish relationships between P determination by colorimetric and ICP methods. Sixty surface soil samples (30 manured and 30 nonmanured) were collected from across Manitoba and extracted with Mehlich 3 reagent, Olsen solution, calcium chloride (CaCl₂) solution, and deionized water. Extractable P in the extract was determined by colorimetric (Col-P) and ICP (ICP-P) methods. The concentrations of P measured by the two methods were statistically analyzed. Mean comparison showed that P amounts determined by ICP in Mehlich 3, water, and CaCl₂ solutions were significantly greater than those determined by colorimetric method (P < 0.05) in the study. The differences between P determinations by the two analytical methods in the extractants were probably due to the presence of organic P, which was included in ICP determination but not in colorimetric determination. The influence of other factors such as the presence of colloidal particles on the P that was determined by the two methods could not be ruled out. However, Olsen P determined by the colorimetric method was not significantly different from the values determined by ICP (P > 0.05) probably because the alkaline nature of this extractant enhanced the hydrolysis of organic P in the extract, thus including organic P in the colorimetric determination of P. There were significant correlations between the two methods of P determination in the various extracting solutions with correlation coefficients ranging between 0.94 and 1.00. The two methods of P determination were linearly related for all the extracting solutions.     

Molecular mechanism of antioxidant synergism of tocotrienols and carotenoids in palm oil

During repeated deep-fat frying of potato slices at 163 degrees C in yellow or red palm olein of comparable fatty acid profiles, the oxidative stability (peroxide value and anisidine value) of the palm oleins was similar, and in yellow palm olein, the rate of antioxidant depletion decreased in the order gamma-T3 > alpha-T3 > delta-T3 (T3, tocotrienol). In red palm olein, which had a total tocopherol/tocotrienol content of 1260 vs 940 ppm in yellow palm olein and a corresponding longer induction period in the Rancimat stability test at 120 degrees C, only depletion of gamma-T3 was significant among the phenols during frying and slower as compared to that in yellow palm olein. The carotenes in the red palm olein were depleted linearly with the number of fryings, apparently yielding an overall protection of the phenols. In antioxidant-depleted palm olein and in phospholipid liposomes with added increasing concentrations of phenols, gamma-T3 was found to be a better antioxidant than alpha-T3. alpha-T3 and alpha-T (T, tocopherol) had a similar antioxidant effect in antioxidant-depleted palm olein in the Rancimat stability test, while in the liposomes the ordering as determined by induction period for the formation of conjugated dienes was gamma-T3 > alpha-T3 > alpha-T. The addition of 100-1000 ppm beta-carotene to antioxidant-depleted palm olein or liposomes (lycopene also tested) did not provide any protection against oxidation. In the liposomes, synergistic interactions were observed between beta-carotene or lycopene and alpha-T, alpha-T3, or gamma-T3 for carotene/phenol ratios of 1:10 and 1:2 but not for 1:1. In chloroform, carotenes were regenerated by tocopherols/tocotrienols from carotene radicals generated by laser flash photolysis as shown by transient absorption spectroscopy, suggesting that carotenes rather than phenols are the primary substrate for lipid-derived radicals in red palm olein, in effect depleting carotenes prior to phenols during frying. Regeneration of carotenes by the phenols also explains the synergism in liposomes. In the laser flash photolysis experiments, gamma-T3 was also found to be faster in regenerating carotenes than alpha-T3 and alpha-T.     

Development of a new colorimetric method determining the yield of microencapsulation of alpha-tocopherol

Microencapsulation of alpha-tocopherol effectively protects alpha-tocopherol from oxidation and produces high-value-added and long-shelf-stable foods. High-performance liquid chromatography (HPLC) has been applied to measure the yield of microencapsulated alpha-tocopherol with high accuracy; however, it takes long analysis time. An alternative method is required to determine the yield of microencapsulated alpha-tocopherol in food industry. A new, easy, and sensitive colorimetric method using 5% cupric acetate pyridine and oleic acid was developed. Correlation coefficient (r) of colorimetric method on alpha-tocopherol in microencapsulation system and of results between colorimetric method and HPLC were +0.996 and +0.989, respectively, which indicates that this novel colorimetric method can be successfully applied to evaluate the yield of microencapsulated alpha-tocopherol instead of HPLC. The optimum storage temperature and pH of microencapsulated alpha-tocopherol for 7-day storage were 25 degrees C and pH 9, respectively, determined by this new colorimetric method.     

Glutathione peroxidase activity, TBARS, and alpha-tocopherol in meat from chickens fed different diets.

This study investigated the effect of feeding broilers with diets differing in dietary fat source (lard, sunflower oil, olive oil) and vitamin E (basal vs supplemented with 200 mg of alpha-tocopheryl acetate/kg) on meat lipid oxidative stability. The diets differed by their degree of unsaturation and included the natural antioxidant alpha-tocopherol (vitamin E). Glutathione peroxidase (GSHPx) activity was measured in raw meat and ranged from 3.62 to 8.06 nmol NADPH/min/mg protein. The enzyme activity was influenced by the degree of unsaturation of the diet. Capillary gas chromatography analyses showed that dietary alpha-tocopherol accumulated in the muscle tissue and contributed to a better oxidative stability of the raw and cooked meat. Thigh meat alpha-tocopherol levels ranged from 2.73 to 3.62 microg/g in unsupplemented chickens whereas levels from 8.69 to 13.37 microg/g were observed in the thigh meat from alpha-tocopherol supplemented animals. The inclusion of olive oil and alpha-tocopherol in the animal diet gave lower thiobarbituric acid reactive substance (TBARS) values and lower GSHPx activity. High correlations were found between the parameters studied. The results suggest that the glutathione peroxidase activity could be used as an indicator of the meat oxidative stability. A negative relationship was observed between GSHPx activity and tissue alpha-tocopherol levels, and a positive relationship was evidenced between TBARS and antioxidant enzyme activity.     

DETERMINATION OF LEAD IN AEROSOL SAMPLES COLLECTED ON GLASS FIBER FILTERS BY AN IMPROVED ATOMIC ABSORPTION SPECTROMETRY METHOD

This paper presents a simple, reliable, economical, safe, accurate and reproducible method for atmospheric aerosol lead determination in glass fiber filters, consisting on an acid digestion procedure and atomic absorption spectrometry quantification. The acid digestion is accelerated by the use of a microwave oven with capped Teflon PFA vessels, and a two steps power and time program. The mixture of 10 mL HNO₃ and 1 mL HF was selected between many tries, for both economic and environmental reasons. The use of direct standards for quantification is proposed instead of added standards on filters, using background correction (deuterium lamp). The filter lead content quantification was carried out through blank analyses. Lead determinations were carried out then in 2629 samples of atmospheric aerosol at three sampling points in the city of Cartagena (Spain), from 1990 to 1994. We present the annual average of these values for each year and sampling location.     

Determination and confirmation of 5-hydroxyflunixin in raw bovine milk using liquid chromatography tandem mass spectrometry

A method was developed and validated to determine 5-hydroxyflunixin in raw bovine milk using liquid chromatography tandem mass spectrometry (LC/MS/MS). The mean recovery and percentage coefficient of variation (%CV) of 35 determinations for 5-hydroxyflunixin was 101% (5% CV). The theoretical limit of detection was 0.2 ppb with a validated lower limit of quantitation of 1 ppb and an upper limit of 150 ppb. Accuracy, precision, linearity, specificity, ruggedness, and storage stability were demonstrated. A LC/MS/MS confirmatory method using the extraction steps of the determinative method was developed and validated for 5-hydroxyflunixin in milk from cattle. Briefly, the determinative and confirmatory methods were based on an initial solvent (acetone/ethyl acetate) precipitation/extraction of acidified whole milk. The solvent precipitation/extraction effectively removed incurred ((14)C) residues from milk samples. The organic extract was then purified by solid phase extraction (SPE) using a strong cation exchange cartridge (sulfonic acid). The final SPE-purified sample was analyzed using LC/MS/MS. The methods are rapid, sensitive, and selective and provide for the determination and confirmation of 5-hydroxyflunixin at the 1 and 2 ppb levels, respectively.     

Reversed phase ion-pair liquid chromatographic determination of nicotine in commercial tobacco products. 1. Moist snuff.

The official methods for the determination of nicotine in commercial tobacco products (AOAC, CORESTA) are based on approaches that are not selective for nicotine (colorimetric measurement, steam distillation, perchloric acid titration), and the availability of published methods based on state-of-the-art chromatographic methods is limited. Reversed phase ion-pair liquid chromatography has been established as a viable alternative for the analysis of basic analytes. A reversed phase ion-pair liquid chromatographic method for the determination of nicotine in commercial tobacco products was developed and optimized in separate experiments (Ciolino, L. A.; Turner, J. A.; McCauley, H. A.; Smallwood, A. W.; Yi, T. Y. J. Chromatogr. 1999a, 852 (2), 451-463). An extensive within-laboratory performance study of the optimized method was subsequently conducted, and results are presented here for the determination of nicotine in commercial moist snuff. Results for the determination of nicotine in commercial cigarettes are presented in a subsequent paper.     

An automatic flow procedure for the determination of 3-hydroxybutyrate in animal serum and plasma

An automatic flow procedure based on the multicommutation concept, comprising three-way solenoid valves, for the spectrophotometric determination of 3-hydroxybutyrate in animal serum and plasma is proposed. The 3-hydroxybutyrate was enzymatically converted to acetoacetate with the reduction of NAD+ to NADH monitored at 340 nm. It was possible to carry out up to 600 determinations without a significant decrease in the analytical signal, with 5 mg of 3-hydroxybutyrate dehydrogenase immobilized on porous silica beads and packed in a column. The system enabled 60 determinations/h of 3-hydroxybutyrate in the range of 10-150 mg L(-1), with a consumption of 0.9 mg of NAD+ and 200 microL of sample per determination. A detection limit of 2 mg L(-1) for both animal serum and plasma and coefficients of variation of 1.4% and 1.2% (n = 17), respectively, were determined. Animal serum and plasma samples were analyzed without previous treatment, the results of which agreed with those obtained using the conventional method (UV kit, Sigma).     

Analysis of pesticide residues by chemical derivatization. II. N-methylcarbamates in natural water and soils.

A method for the quantitative determination of several N-methylcarbamates in natural waters and the applicability of the derivative to soil samples using a previously published extraction procedure are described. After extraction of the carbamates from the substrate, the carbamates are hydrolyzed in a 10% methanol-potassium hydroxide solution to form the phenolic hydrolysis products, which are isolated and derivatized with pentafluorobenzyl (PFB) bromide to produce the PFB ether derivatives. The PFB derivatives are cleaned up and fractionated on a silica gel microcolumn and determined by electron capture gas-liquid chromatography (GLC). Eight organophosphate pesticides and 2 phthalate acid esters that hydrolyze to phenols or phthalic acid were evaluated as potential interferences and were found not to interfere with any of the carbamates tested. Quantitative determinations of 0.1 mug carbofuran and 3-ketocarbofuran and 0.5 mug carbaryl, metmercapturon, and Mobam in a 1 L water sample are possible. Propoxur was not determined at levels less than 1 mug/L due to the short GLC retention time of the derivative and interferences from the reagents at the lower levels.     

植物叶面积测定方法的比较研究

以两个不同品种的大豆叶片为研究对象，采用了不同的测定方法测定叶面积，分析并指出各种测定方法的优缺点，为在不同测定条件和实验目的要求下选择不同的叶面积测定方法提供依据，也为研究人员提供更加准确、快速、简便和经济的测定方法和依据。     

NIR spectroscopy and partial least-squares regression for determination of natural alpha-tocopherol in vegetable oils

Near-infrared (NIR) spectroscopy and partial least-square regression were used for determination of alpha-tocopherol in edible oils after extraction with ethanol. The standard error of calibration and the standard error of prediction were calculated for evaluation of the calibration models. The chemometric calibration model was prepared in spectral region 6500-4500 cm(-1) for standard alpha-tocopherol solutions (0.54-53.54 mg/mL). Obtained mean concentrations of natural alpha-tocopherol in different types of oils varied from 17.53 to 57.10 mg/100 g. Net analyte signal calculation was used to estimate detection limit (DL = 0.12 mg/mL), quantification limit (QL = 0.40 mg/mL), sensitivity (SEN = 0.045 mg/mL), and selectivity (SEL ranged between 0.24 and 0.54% of the measured reflectance signal) of the proposed NIR method. The comparable precision (RSD = 0.68-2.80% and 0.79-3.06%) and accuracy (recovery, 97.2-102.4% and 96.8-103.2%) for the proposed NIR and standard HPLC methods, demonstrate the benefit of the NIR method in the routine analysis of alpha-tocopherol in vegetable oils.     

Methods for determinging carbohydrates, hydroxymethylfurfural, and proline in honey: collaborative study.

A modification of the official selective absorption method for honey carbohydrates, 31.124--31.133, was studied collaboratively; the determinations of sucrose, total monosaccharides, disaccharides, and higher sugars by this procedure were satisfactory and were adopted by the AOAC. High pressure liquid chromatography of glucose, fructose, and sucrose in honey showed better precision than the modified official method and gave concordant results; it was also adopted. Two methods for hydroxymethylfurfural do not qualify. A method for proline was also adopted.     

Liquid chromatographic determination of polydextrose in food matrixes

A liquid chromatographic (LC) method has been developed to determine the content of polydextrose, a water-soluble 1 calorie/g bulking agent, in food matrixes such as cookies, cakes, fruit spreads, and chocolate toppings. This analysis, which requires use of a blank matrix, provides a feasible means to control the manufacture of foods containing this additive and provides a component for the accurate determination of the caloric value of a particular food product. The method involves aqueous extraction of the polydextrose from the food matrix followed by separation on a carbohydrate analysis column. The LC system uses a mobile phase of 0.005M CaSO4.2H2O and a refractive index detector for quantitation. Polydextrose recoveries from the food matrixes varied from 91.5 to 100.9% with assay precision, expressed as coefficient of variation, ranging from 0.7 to 4.3%. Each error estimate was derived from 5 parallel determinations. The present methodology is precise and selective in contrast to the modified classical phenol-sulfuric acid colorimetric method for assaying carbohydrates, which had been used for polydextrose determination in food matrixes in the past. Because the coefficient of variation frequently exceeded 10%, replicate analyses were necessary to achieve quantitation.     

Colorimetric determination of poly(N-vinyl-2-pyrrolidone) in contact lens solutions.

A rapid colorimetric method is presented for the quantitative determination of poly(N-vinyl-2-pyrrolidone) in contact lens solutions. The method is simple, requires no sample pretreatment, and uses only a small volume of sample. The procedure is based on the measurement of the net absorbance of a poly(N-vinyl-2-pyrrolidone)- Congo red complex at 545 nm. An accuracy of greater than +/- 4% was obtained for the concetnration ranges usually found in contact lens solutions with a minimum detection level of 10 ppm. The method is useful as a screening procedure for solutions of unknown composition and as a quality assurance procedure for routine determinations.