Characterization of starch from tubers of yam bean (Pachyrhizus ahipa).

Detailed studies of the starch present in tubers of six accessions of Pachyrhizus ahipa (ahipa) have been carried out using starches from tubers of P. erosus (Mexican yam bean) and seeds of ahipa and wheat for comparison. Starch accounted for 56-58% of the tuber dry weight with granules occurring in a range of geometric forms and in sizes from below 5 microm to about 35 microm (mean about 10 microm in all accessions except two). The amylose content ranged from 11.6 to 16.8% compared with 16.9% in P. erosus tubers and over 23% in the seed starches. X- ray diffraction analysis showed A-type or C(A)-type diffraction patterns. The chain-length distribution of the amylopectin after enzyme debranching showed a peak at DP11 similar to that of wheat starch, but had a less marked shoulder at DP 21-22 and contained a higher proportion of longer chains. Differential scanning calorimitry showed an endothermic peak corresponding to gelatinization with T(max) ranging from 59 to 63 degrees C, which was similar to the T(max) of wheat (about 64 degrees C). The composition of the ahipa starch may mean that it is suitable for food applications that require low amylose content and low retrogradation after processing.     

Issues in assessing the validity of nutrient data obtained from a food-frequency questionnaire: folate and vitamin B12 examples

OBJECTIVE: To compare methods used to assess the validity of nutrient intake data obtained from a food-frequency questionnaire (FFQ), using folate and vitamin B12 as nutrient examples. DESIGN: Cross-sectional sample from a population cohort. SETTING: Two postcode areas west of Sydney, Australia. SUBJECTS: In total, 2895 people aged 49 years and older provided dietary data using a semi-quantitative FFQ (79% of 3654 subjects examined). The validity of the FFQ was assessed against three 4-day weighed food records (WFRs) completed by 78 people (mean age 70 years). RESULTS: Folate and vitamin B12 validity data were assessed using different methods. The Spearman ranked correlations (energy-adjusted) were 0.66 for folate and 0.38 for vitamin B12. Using the Bland-Altman method, following loge transformation, no linear trend existed between the differences and means for folate and vitamin B12. Large differences existed between the FFQ and WFR in individual cases, particularly for vitamin B12. Finally, data were divided into quintile categories for the test and reference method: 79% classified folate within one quintile, 65% classified vitamin B12 within one quintile; there was no gross misclassification for folate and only 3% misclassification for vitamin B12. CONCLUSIONS: Different methods of analysis provided different information about the validity of the FFQ. Correlation coefficients should not be used alone to assess the validity of nutrient data, but should be used in conjunction with Bland-Altman analyses. Depending on the use of the data, additional assessment of classification categories is recommended. This worked example demonstrates that absolute intakes of folate and vitamin B12 should be used with caution.     

Broth from canned clams is suitable for use as an excellent source of free vitamin B12

Vitamin B(12) was assayed and characterized in the broth of canned clams (boiled plain). The broth contained considerable amounts of vitamin B(12) (2.7-14.1 μg/100 g, 1.3-6.7 μg/can). HPLC and LC/ESI-MS/MS chromatograms demonstrated that the clam broth contained true vitamin B(12). Gel filtration experiments indicated that most (72%) of the vitamin B(12) found in the broth was recovered in free vitamin B(12) fractions. These results indicate that the clam broth would be suitable for use an excellent source of free vitamin B(12) for elderly persons with food-bound vitamin B(12) malabsorption.     

Determination of vitamin B12 in dry feeds by atomic absorption spectrophotometry.

Vitamin B12 was determined in dry feeds by atomic absorption spectrophotometry (AAS). Samples containing B12 were extracted with an assay solution, 5 g EDTA was added to the filtrate, the pH was adjusted to 7 with NH4OH, and 5 g charcoal was added. The charcoal was removed by filtering through ashless paper which was then placed in a beaker and ashed at 600 degrees C. After dissolving the cobalt oxide from the ash in 5N HNO3, cobalt content was determined by using AAS. To determine mg B12/lb feed, ppm cobalt in the feed is multiplied by 10.43. The sensitivity of the proposed procedure is 1 mg vitamin B12/lb. The procedure is rapid and precise, and results compare favorably with AOAC method 43.109.     

Determination of vitamin B(6) in cooked sausages

A reverse-phase high-performance liquid chromatography (HPLC) method has been described for the determination of various active forms of vitamin B(6) in meat products. Different extracting agents were tested to solubilize fully the analyte for quantification. The best data were obtained by extracting the samples with 5% (w/v) metaphosphoric acid. Separation by HPLC was performed with fluorescence detection (excitation, 290 nm; emission, 395 nm), on a 10 cm x 0.46 cm i.d. Hypersil BDS C(18) 5 microm column using a mixture of 50 mM phosphate buffer (pH 3.2) and acetonitrile (99:1, v/v) as mobile phase. Precision of the method was 0.5% (within a day) and 4.3% (between days). The detection limits were 0.020 mg/100 g for pyridoxal and pyridoxamine, 0.017 mg/100 g for pyridoxamine phosphate, 0.500 mg/100 g for pyridoxal phosphate, and 0.033 mg/100 g for pyridoxol, with a signal-to-noise ratio of 3. The recovery ranged from 92.0 to 100.0%.     

Production and characterization of a monoclonal anti-anti-idiotype antibody against fumonisin B(1).

A monoclonal anti-anti-idiotype antibody (mAb3) against fumonisin B(1) (FmB1) was produced from the hybridoma cell line 7C7F4, which was generated by the fusion of P3/NS-1/1-AG4-1 myeloma cells with spleen cells isolated from a Balb/c mouse that had been immunized with the Fab fragments of affinity-purified anti-idiotype antibodies. The mAb3 belongs to the immunoglobulin M, kappa light chain. A direct competitive enzyme-linked immunosorbent assay (dc-ELISA) and an indirect competitive ELISA (idc-ELISA) were established for antibody characterization and toxin analysis. In an idc-ELISA using FmB1-ovalbumin (OVA) as the coating antigen, the concentrations causing 50% inhibition of binding (IC(50)) of mAb3 to the solid-phase FmB1-OVA by free FmB1, FmB2, and FmB3 were found to be 75, 95, and 450 ng/mL, respectively. In the dc-ELISA, the concentration causing IC(50) of FmB1-horseradish peroxidase to the solid-phase mAb3 by free FmB1 was found to be 233 ng/mL. Analysis of 12 samples naturally contaminated with fumonisins with mAb3-based idc-ELISA and polyclonal-based dc-ELISA showed a good correlation between these two methods with a correlation coefficient of 0.76 at p < 0.02. The linear regression slope was found to be y[polyclonal ELISA] = 0.87x[mAb3 ELISA] - 52 ppb.     

Optimization of lipase-catalyzed regioselective acylation of pyridoxine (vitamin B6)

Response surface methodology was successfully applied to optimize lipase-catalyzed regioselective esterification of pyridoxine (PN). Effects of various reaction conditions, including reaction temperature, time, enzyme loading, substrate molar ratio, and water activity, were investigated. A central composite design was employed to search for the optimal conversion of PN. A quadratic polynomial regression model was used for analysis of the experimental data at a 95% level (p < 0.05). The analysis confirmed that the water activity was the most significant factor affecting the conversion of PN. It was also suggested that the conversion was strongly affected by independent variables of temperature, time, substrate molar ratio, and water activity as well as interaction terms of temperature and enzyme loading/substrate molar ratio/water activity, time and enzyme loading/substrate molar ratio, substrate molar ratio, and water activity. The coefficient of determination of the model was found to be 0.963. Three sets of optimum reaction conditions were established, and the verified experimental trials were performed for validating the optimum points. A scale-up experiment was also done under the first set of optimal conditions.     

Dried green and purple lavers (Nori) contain substantial amounts of biologically active vitamin B(12) but less of dietary iodine relative to other edible seaweeds

Vitamin B(12) concentrations of dried green (Enteromorpha sp.) and purple (Porphyra sp.) lavers (nori) were determined by both Lactobacillus leichmannii ATCC 7830 microbiological and intrinsic factor chemiluminescence methods. The values determined by using the microbiological method (63.58 +/- 2.90 and 32.26 +/- 1.61 microg/100 g of dry weight) were identical to those found by using the chemiluminescence method (69.20 +/- 2.21 and 25.07 +/- 0.54 microg/100 g of dry weight) in both dried green and purple lavers, respectively. A silica gel 60 thin-layer chromatography of both laver extracts shows that non-coenzyme forms (hydroxo and cyano forms) of vitamin B(12) predominate in both dried lavers. The dried lavers contained lesser amounts of dietary iodine ( approximately 4-6 mg/100 g of dry weight) relative to other seaweeds, suggesting that excessive intake of the dried lavers is unlikely to result in harmful intake of dietary iodine. These results indicate that the dried lavers (nori) are the most excellent source of vitamin B(12) among edible seaweeds, especially for strict vegetarians.     

Cloning and sequence analysis of the phytoene synthase gene from a unicellular chlorophyte, Dunaliella salina

Phytoene synthase (Psy) catalyzing the dimerization of two molecules of geranylgeranyl pyrophosphate (GGPP) to phytoene may be rate limiting for the synthesis of beta-carotene in Dunaliella salina. To elucidate the carotenogenic pathway in D. salina, the complete Psy gene was first isolated by genome walking technology and suppression PCR. Subsequently, RT-PCR was performed to obtain the Psy cDNA of 1260 bp, which codes 420 amino acids. The Psy gene of total length of 2982 bp was found to consist of five exons and four introns when compared with the cDNA sequence. The D. salina Psy amino acid has a 78-89% similarity with many other higher plants, but a phylogenic analysis shows a closer relationship between the microalgal and cyanobacterial Psy.     

Maternal serum vitamin B12, folate and homocysteine and the risk of neural tube defects in the offspring in a high-risk area of China

OBJECTIVE: To examine the association between the risk of neural tube defects (NTD) and maternal serum vitamin B12, folate and homocysteine in a high-risk area of China. DESIGN: A case-control study was carried out in Luliang mountain area of Shanxi Province.Subjects/settingA total of eighty-four NTD pregnancies and 110 matched controls were included in the study; their serum vitamin B12 and folate concentrations were measured by chemiluminescent immunoenzyme assay and total homocysteine concentrations by fluorescent polarisation immunoassay. RESULTS: Serum vitamin B12 and folate concentrations were lower in NTD-affected pregnant women than in controls (P < 0.01). Serum total homocysteine was higher in the NTD group than in controls at less than 21 weeks of gestation (P < 0.01). Adjusted odds ratios revealed that women with lower vitamin B12 (adjusted OR=4.96; 95 % CI 1.94, 12.67) and folate (adjusted OR=3.23; 95 % CI 1.33, 7.85) concentrations had a higher risk of NTD compared to controls. Based on dietary analysis, less consumption of meat, egg or milk, fresh vegetables and fruit intake would increase the risk of NTD. CONCLUSIONS: Lower serum concentrations of folate and vitamin B12 are related to the increased risk of NTD in high-risk populations. Both folate and vitamin B12 intake insufficiency could contribute to the increased risk of NTD. A dietary supplement, combining folate and vitamin B12, might be an effective measure to decrease the NTD incidence in these areas.     

Characterization of maghemite in B horizons of three soils from Southern Portugal

Maghemite with an unusual habit was identified in the sandy fraction of two Alfisols and one Ultisol by X-ray and electron diffraction. Hematite was also present. Morphological analyses by scanning and transmission electron microscopy showed needle-like particles of maghemite with average lengths of 200–500 nm depending on the degree of the crystallinity. Those particles are arranged in star-like patterns as shown by ultrathin sections. Experimental synthesis of maghemite was successfully attempted in the presence of quartz sand with the freshly-formed crystals displaying the same particular morphology. Formation of acicular maghemite catalysed by quartz surfaces is, therefore, postulated.     

Characterization of (E,E,Z)-2,4,6-nonatrienal as a character impact aroma compound of oat flakes

To identify the compounds evoking the characteristic cereal-like, sweet aroma of oat flakes, an aroma extract dilution analysis (AEDA) was applied to a distillate prepared by solvent extraction/vacuum distillation from commercial oat flakes. Among the nine aroma-active compounds detected by gas chromatography-olfactometry and AEDA in the flavor dilution (FD) factor range of 4-1024, eight odorants, for example, (E)-beta-damascenone, (Z)-3-hexenal, and butanoic acid, showed only low FD factors. However, one odorant eliciting the typical cereal, sweet aroma of the flakes was detected with the highest FD factor of 1024. By mass spectrometry and nuclear magnetic resonance measurements followed by a synthesis, (E,E,Z)-2,4,6-nonatrienal, exhibiting an intense oat flake-like odor at the extremely low odor threshold of 0.0002 ng/L in air, was identified as the key odorant of the flakes. By means of a newly developed stable isotope dilution analysis using synthesized, carbon-13-labeled nonatrienal as the internal standard, a concentration of 13 mug of (E,E,Z)-2,4,6-nonatrienal per kilogram of the flakes was measured. Model studies suggested linolenic acid as the precursor of nonatrienal in oats.     

Certification of B-group vitamins (B1, B2, B6, and B12) in four food reference materials

In 1989, the Community Bureau of Reference started a research program to improve the quality of vitamin analysis in food. To achieve this task, vitamin methodology was evaluated and tested by interlaboratory studies and the preparation of certified reference materials, which will be used for quality control of vitamin measurements. The main improvements in methodology were achieved by testing and standardizing the extraction condition and enzymatic hydrolysis procedures. Results for each individual material are derived from five replicate determinations using at least two independent methods: liquid chromatography (HPLC) and microbiological assay for vitamins B1, B2, and B6; and radioprotein binding and microbiological assays for vitamin B12. The certificate of analysis for four reference materials gives mass fraction values for water-soluble vitamins. These certified values were based on the acceptable statistical agreement of results from collaborating laboratories. Certified values with uncertainties (mg/kg dry matter) for each CRM are as follows: 4.63 (0.20) and 4.10 (0.51) for vitamins B1 and B6, respectively, in CRM 121 (wholemeal flour); 6.51 (0.24), 14.54 (0.3), 6.66 (0.43), and 0.034 (0.003) for vitamins B1, B2, B6, and B12, respectively, in CRM 421 (milk powder); 3.07 (0.17) and 4.80 (0.40) for vitamins B1 and B6, respectively, in CRM 485 (lyophilized mixed vegetables), and 8.58 (0.55), 106.8 (2.8), 19.3 1.5), and 1.12 (0.044) for vitamins B1. B2, B6, and B12, respectively, in CRM 487 (lyophilized pig liver).     

Characterization of fumonisin B(1)-glucose reaction kinetics and products

The reaction of fumonisin B(1) with the reducing sugar D-glucose can block the primary amine group of fumonisin B(1) and may detoxify this mycotoxin. A method to separate hundred milligram quantities of fumonisin B(1)-glucose reaction products from the excess D-glucose with a reversed-phase C(18) cartridge was developed. Mass spectrometry revealed that there were four primary products in this chain reaction when fumonisin B(1) was heated with D-glucose at 65 degrees C for 48 h: N-methyl-fumonisin B(1), N-carboxymethyl-fumonisin B(1), N-(3-hydroxyacetonyl)-fumonisin B(1), and N-(2-hydroxy, 2-carboxyethyl)-fumonisin B(1). The N-(1-deoxy-D-fructos-1-yl) fumonisin B(1) (fumonisin B(1)-glucose Schiff's base) was detected by mass spectrometry when fumonisin B(1) was heated with D-glucose at 60 degrees C. The nonenzymatic browning reaction of fumonisin B(1) with excess D-glucose followed apparent first-order kinetics. The activation energy, E(a), was 105.7 kJ/mol. Fumonisin B(1) in contaminated corn could precipitate the nonenzymatic browning reaction with 0.1 M D-glucose at 60 and 80 degrees C.