Sample preparation and liquid chromatographic determination of vitamin D in food products

Vitamin D in different fortified foods is determined by using liquid chromatography (LC). Sample preparation is described for fortified skim milk, infant formulas, chocolate drink powder, and diet food. The procedure involves 2 main steps: saponification of the sample followed by extraction, and quantitation by LC analysis. Depending on the sample matrix, additional steps are necessary, i.e., enzymatic digestion for hydrolyzing the starch in the sample and cartridge purification before LC injection. An isocratic system consisting of 0.5% water in methanol (v/v) on two 5 microns ODS Hypersil, 12 X 0.4 cm id columns is used. Recovery of vitamin D added to unfortified skim milk is 98%. The results of vitamin D determination in homogenized skim milk, fortified milk powder, fortified milk powder with soybean, chocolate drink powder, and sports diet food are given.     

Functional properties of edible agar-based and starch-based films for food quality preservation

Edible films made of agar (AG), cassava starch (CAS), normal rice starch (NRS), and waxy (glutinous) rice starch (WRS) were elaborated and tested for a potential use as edible packaging or coating. Their water vapor permeabilities (WVP) were comparable with those of most of the polysaccharide-based films and with some protein-based films. Depending on the environmental moisture pressure, the WVP of the films varies and remains constant when the relative humidity (RH) is >84%. Equilibrium sorption isotherms of these films have been measured; the Guggenheim-Anderson-de Boer (GAB) model was used to describe the sorption isotherm and contributed to a better knowledge of hydration properties. Surface hydrophobicity and wettability of these films were also investigated using the sessile drop contact angle method. The results obtained suggested the migration of the lipid fraction toward evaporation surface during film drying. Among these polysaccharide-based films, AG-based film and CAS-based film displayed more interesting mechanical properties: they are transparent, clear, homogeneous, flexible, and easily handled. NRS- and WRS-based films were relatively brittle and have a low tension resistance. Microstructure of film cross section was observed by environmental scanning electron microscopy to better understand the effect of the structure on the functional properties. The results suggest that AG-based film and CAS-based films, which show better functional properties, are promising systems to be used as food packaging or coating instead of NRS- and WRS-based films.     

Spinach cultigen variation for tissue carotenoid concentrations influences human serum carotenoid levels and macular pigment optical density following a 12-week dietary intervention

Increasing intakes of carotenoid-rich plant foods can increase serum carotenoid concentrations and macular pigment optical density (MPOD) in most, but not all, individuals. Research objectives for this study were to (1) characterize tissue lutein (L) and beta-carotene (BC) concentrations in carotenoid-rich spinach (Spinacia oleracea L.) cultigens and (2) determine serum carotenoid and MPOD responses in human subjects consuming spinach cultigens differing in tissue L and BC concentrations. Thirteen spinach cultigens were evaluated for carotenoid accumulations over two consecutive growing seasons. "Springer" (8.4 and 6.5 mg/100 g of fresh mass for L and BC, respectively) and "Spinner" (12.1 and 9.2 mg/100 g of fresh mass for L and BC, respectively) spinach cultigens were selected for a dietary intervention study and represented low- and high-L concentrations. The high-L ("Spinner") and low-L ("Springer" ) spinach treatment groups consisted of 10 subject volunteers ingesting five 50-g spinach servings/week during a 12-week intervention. Average serum L concentrations increased by 22% (P = 0.07) from baseline (0.233 micromol/L) to 12 weeks (0.297 micromol/L) for subjects consuming low-L spinach. Subjects consuming high-L spinach showed increases of 33% (P = 0.04) in serum L from baseline (0.202 micromol/L) to 12 weeks (0.300 micromol/L). Average MPOD did not change for the low-L treatment group; however, subjects in the high-L group demonstrated increases (P = 0.02) in MPOD at the 30' eccentricity between baseline (0.343) and 12 weeks (0.374). This study demonstrates that serum carotenoid and MPOD are determined by L concentrations present in the spinach matrix. Results emphasize the role of cultigen selection among vegetable crops in determining phytochemical effects on human health.     

Competitive enzyme-linked immunoassay for sialoglycoprotein of edible bird's nest in food and cosmetics

The proliferation of fake and inferior edible bird's nest (EBN) products has recently become an increasingly serious concern. To identify and classify EBN products, a competitive enzyme-linked immunoassay (ELISA) was developed to quantitate sialoglycoprotein in EBN used in food and cosmetic applications. The characteristic sialoglycoprotein in EBN was found, extracted, purified, and analyzed. Sialoglycoprotein, considered the main carrier of sialic acid in EBN, consisted of 106 and 128 kDa proteins. A monoclonal antibody that could recognize both proteins was prepared. The heat-treated process did not change the affinity of sialoglycoprotein with the antibody. An optimized ELISA method was established with a cross-reactivity of less than 0.1% and an IC(50) of 3.3 μg/mL. On the basis of different food and cosmetic samples, the limits of detection (LOD) were 10-18 μg/g. Recoveries of fortified samples at levels of 20 and 80 μg/g ranged from 81.5 to 96.5%, respectively. The coefficients of variation were less than 8.0%.     

Semiautomated method for riboflavin in food products: collaborative study

A collaborative study was conducted comparing a semiautomated riboflavin method with a manual riboflavin method for 10 food products. Six laboratories provided results from the semiautomated method and 16 laboratories used the manual technique. The semiautomated method was more repeatable within a laboratory than was the manual method. The semiautomated method results compared favorably with the manual method for all 10 products.     

Rapid determination of polyphenols and vitamin C in plant-derived products

Polyphenols, widely spread in our diet by the consumption of plant food products, are commonly determined using Folin-Ciocalteu reagent that interacts with other different reducing nonphenolic substances and leads to an overestimation of polyphenol content. In this paper we report an optimized Folin-Ciocalteu method to specifically determine the contents of total polyphenols and vitamin C. After the optimal conditions for the colorimetric assay were set, solid-phase extraction (Oasis HLB (hydrophilic-lipophilic balance)) was carried out to eliminate the water-soluble reducing interferences including vitamin C. Colorimetric correction was thus performed by subtracting interfering substances contained in the water washing extract from the raw extract. Moreover, vitamin C present in the water washing extract can be destroyed by heating and thus colorimetrically deduced. This procedure was set up with synthetic solutions and validated on different extracts from fruit products.     

Determination of total dietary fiber in foods and food products: collaborative study

A collaborative study was conducted to determine the total dietary fiber (TDF) content of food and food products, using a combination of enzymatic and gravimetric procedures. The method was basically the same as published earlier (J. Assoc. Off. Anal. Chem. (1984) 67, 1044-1052), with changes in the concentration of alcohol and buffers, time of incubation, sample preparation, and some explanatory notes, all with the intent of decreasing the coefficient of variation (CV) of the method. Duplicate blind samples of soy isolate, white wheat flour, rye bread, potatoes, rice, wheat bran, oats, corn bran, and whole wheat flour were analyzed by 9 collaborators. TDF was calculated as the weight of the residue minus the weight of protein and ash. CV values of the data from all laboratories for 7 of the samples ranged from 1.56 to 9.80%. The rice and soy isolate samples had CV values of 53.71% and 66.25%, respectively; however, each sample contained only about 1% TDF. The enzymatic-gravimetric method for determining TDF has been adopted official first action.     

Interaction of nanoparticles with edible plants and their possible implications in the food chain

The uptake, bioaccumulation, biotransformation, and risks of nanomaterials (NMs) for food crops are still not well understood. Very few NMs and plant species have been studied, mainly at the very early growth stages of the plants. Most of the studies, except one with multiwalled carbon nanotubes performed on the model plant Arabidopsis thaliana and another with ZnO nanoparticles (NPs) on ryegrass, reported the effect of NMs on seed germination or 15-day-old seedlings. Very few references describe the biotransformation of NMs in food crops, and the possible transmission of the NMs to the next generation of plants exposed to NMs is unknown. The possible biomagnification of NPs in the food chain is also unknown.     

Semiautomated method for niacin and niacinamide in food products: collaborative study.

A collaborative study was conducted comparing a semiautomated colorimetric niacin method with a manual colorimetric and a microbiological method for 10 food products. Seven laboratories used the microbiological method, 7 laboratories used the manual colorimetric method, and 6 laboratories used the semiautomated method. The semiautomated method was more repeatable within a laboratory and more reproducible between laboratories than was either of the other methods. The semiautomated method results compared favorably with both the microbiological and manual colorimetric method results.     

Automated analysis of vitamin C in pharmaceutical products.

The determination of total vitamin C in the form of both l-ascorbic acid (AA) and dehydroascorbic acid (DHAA) present in pharmaceutical preparations has been automated. Total vitamin C (completely oxidized to DHAA) was determined by reaction with 2,4-dinitrophenylhydrazine while blanks utilized the same reagent after reducing all DHAA to AA. The automated method was applicable to a variety of multivitamin preparations including those containing iron and copper. The mean recovery of L-ascorbic acid added to 11 multivitamin preparations was 99.4% with a coefficient of variation of 2.5%. In the analysis of these products, results obtained by the automated method were essentially the same as those obtained by the original manual method. For preparations containing no copper salts, the results were also comparable to those obtainable by titration with 2,6-dichloroindophenol except in 1 product which contained some DHAA.     

Survey of food products for volatile N-nitrosamines.

A variety of food products containing nitrite were analyzed for 14 volatile N-nitrosamines by using a method demonstrated to be sensitive to 10 ppb. A total of 121 food samples were screened for volatile N-nitrosamine content. N-Nitrosopyrrolidine was confirmed in fried bacon at levels up to 139 ppb. N-Dimethylnitrosamine, N-nitrosopyrrolidine, and N-nitrosopiperidine were also confirmed in spice-cure mixtures at levels ranging from 50 to 2000 ppb.     

Determination of epsilon-N-pyrrolylnorleucine in fresh food products.

epsilon-N-Pyrrolylnorleucine was determined in different fresh food products to study its presence as a normal component of food proteins. Twenty-two different products were screened: cod, cuttlefish, salmon, sardine, trout, beef, chicken, pork, broad bean, broccoli, chickpea, garlic, green pea, lentil, mushroom, soybean, spinach, sunflower, almond, hazelnut, peanut, and walnut. Foods were homogenized, their proteins were precipitated with trichloroacetic acid and hydrolyzed with 2 N NaOH for 20 h, and the epsilon-N-pyrrolylnorleucine content was determined by capillary electrophoresis. The epsilon-N-pyrrolylnorleucine, which was identified by HPLC/MS in sardine muscle hydrolysate, ranged in the 22 foods analyzed from 0.24 to 6.36 micromol/g. This concentration was correlated with the protein content of the food (r = 0.687, p = 0.00041). In addition, the epsilon-N-pyrrolylnorleucine/lysine ratio was found to be a function of the lipid, iron, and protein contents of the food (r = 0.881, p < 0.0001) and was directly correlated with lipid and iron contents and inversely correlated with the protein content. These results are in agreement with the oxidative stress origin proposed for epsilon-N-pyrrolylnorleucine and suggest that the epsilon-N-pyrrolylnorleucine/lysine ratio is a characteristic of each food. In addition, epsilon-N-pyrrolylnorleucine seemed to be a normal component of many fresh food products, in which it may be acting as a natural antioxidant.     

Total and reactive lysine contents in selected cereal-based food products

The aim of the study was to determine and compare reactive and total lysine contents in a range of breakfast cereal products. Crude fiber, fat, ash, and crude protein contents of 20 breakfast cereal products ranged from 4 to 38, 14 to 144, 7 to 32, and 52 to 253 g/kg, respectively. The concentrations of glutamic acid (18.7-32.1 g/100 g protein) and proline (4.7-10.8 g/100 g protein) were high while those of the amino acids methionine (1.2-2.0 g/100 g protein) and histidine (1.2-3.3 g/100 g protein) were relatively low. There was a strong relationship between reactive lysine determined using the guanidination and fluorodinitrobenzene methods (R = 0.99). The total lysine content, determined after conventional acid hydrolysis, ranged from 0.8 to 3.7 g/100 g protein, while the reactive lysine content (guanidination) ranged from 0.4 to 2.8 g/100 g protein. Reactive lysine was 20-54% lower than total lysine in the cereal products. The large differences between total and reactive lysine suggest a considerable loss of lysine in the breakfast cereals tested.     

Development of an indirect competitive ELISA for ciprofloxacin residues in food animal edible tissues

An indirect competitive enzyme-linked immunosorbent assay (ELISA) was developed to detect ciprofloxacin (CPFX) in food animal edible tissues. CPFX was converted by an active ester method into conjugates CPFX-bovine serum albumin (CPFX-BSA) and CPFX-human serum albumin (CPFX-HSA), which both allowed production of CPFX-specific rabbit antisera. In the ELISA, CPFX-HSA was coated onto the microtiter plate, followed by incubation with standard CPFX and anti-CPFX antibody. The indirect competitive ELISA revealed that the antisera have no cross-reactivity with penicillin, gentamicin, neomycin, sulfadiazine, and chlortetracycline. The antisera cross-reacted with enrofloxacin and norfloxacin about 69.8 and 44.6% as much as they did with CPFX. This ELISA was highly sensitive (0.32 ng/mL) to CPFX determination. Recovery of CPFX at 40 microg/kg was 75.58% in pork, 81.29% in chicken, and 84.30% in milk. The coefficients of variation varied from 3.7 to 9.2% over the range of CPFX concentrations studied. The linear detection range was between 1.6 and 1000 ng/mL. The results suggest that this ELISA is a specific, accurate, and convenient method for the detection of CPFX residues in food animal edible tissues.     

High pressure liquid chromatographic determination of carbohydrates in food products: evaluation of method

A high pressure liquid chromatographic (HPLC) method for determining 5 common carbohydrates in food products was evaluated. Reproducibility data were generated showing a relative standard deviation of 2.8%. Recovery studies on a variety of foods gave an average recovery of 98.8%. The HPLC data for 3 varieties of ready-to-eat cereals were compared with data from 4 independent laboratories using current AOAC chemical methods. The HPLC mean values differed from the chemical mean values by 3.2%.     

Glucose oxidase-mediated gelation: a simple test to detect glucose in food products

This paper reports a simple, rapid, and sugar-selective method to induce gelation from glucose-containing samples. This method employs glucose oxidase (GOx) to selectively "recognize" and oxidize glucose to generate gluconic acid, which acts to solubilize calcium carbonate and release calcium ions. The release of calcium ions triggers gelation of the calcium-responsive polysaccharide alginate to form a calcium-alginate hydrogel. Rheological measurements confirm that gel formation is triggered by glucose but not fructose or sucrose (consistent with GOx's selectivity). Vial inversion tests demonstrate that gel formation can be readily observed without the need for instrumentation. Proof-of-concept studies demonstrate that this gel-forming method can detect glucose in food/beverage products sweetened with glucose or high-fructose corn syrups. These results indicate that the enzyme-induced gelation of alginate may provide a simple means to test for sweeteners using components that are safe for use on-site or in the home.     

The transfer of cadmium from sewage-sludge amended soils into the edible components of food crops

Water, Air, & Soil Pollution - The application of sewage sludge to soils used for the production of food crops represents a potential means by which human dietary exposures to Cd may be...