High-throughput method for the quantitation of total folate in whole blood using LC-MS/MS

A high-throughput liquid chromatography tandem mass spectrometry (HT LC-MS/MS) method for red blood cell (RBC) folate analysis was developed from a previously described manual (M) LC-MS/MS method. The HT LC-MS/MS method used 96-well plates in which RBC folates were hydrolyzed with concentrated HCl in the presence of the [13C6]pABA internal standard (IS). The pH of the hydrolysate was adjusted to 5.0 before cleanup using 96-well plate OASIS HLB SPE cartridges. The analyte and IS were eluted with ethyl acetate/hexane (95:5, v/v) and methylated with methanol and trimethylsilyldiazomethane. The methylated analyte and IS were quantified with LC-MS/MS as previously described. The HT LC-MS/MS method was validated by determining the recovery of six different folate vitamers, which were quantitatively recovered (84-105% with CV<9.0%). RBC folate concentrations in whole blood samples correlated between HT and M LC-MS/MS methods (r=0.922, p<0.0001 for n=43 samples) and between the HT LC-MS/MS method and a chemiluminescence assay (r=0.664, p<0.001 for n=325 samples). Comparison of the results between HT LC-MS/MS and chemiluminescence methods with Bland-Altman difference plots and by ROC curve analysis indicates that the chemiluminescence assay underreports RBC folate concentrations. The HT LC-MS/MS method allows for high-throughput sample preparation for the analysis of RBC folate.     

Quantitation of total folate in whole blood using LC-MS/MS

An accurate method for measuring whole blood total folate using liquid chromatography with tandem mass spectrometry is described and compared to GC/MS and a chemiluminescence assay. Whole blood from normal adults (n = 15) was fortified with a [(13)C(6)]para-aminobenzoic acid (pABA) internal standard and treated with 12.1 N hydrochloric acid at 110 degrees C for 4 h to hydrolyze all folates to pABA. Contaminants in the hydrolysate were adsorbed onto a C18 SPE cartridge. The eluate containing the folate catabolite pABA was partitioned into ethyl acetate and methylesterified with trimethylsilyldiazomethane. The methyl-pABA derivatives were quantified by positive-ion atmospheric pressure chemical ionization (APCI)LC-MS/MS. An isocratic mobile phase of acetonitrile-water (70:30) (v/v) on a C18 analytical column was used with a postcolumn reagent of 0.025% formic acid. The limit of quantitation for folate was 56.6 nmol/L RBC, and the limit of detection was 22.6 nmol/L RBC. Folate levels as determined by LC-MS/MS correlated well with the chemiluminescence assay and a GC/MS method. This new LC-MS/MS method provides enhanced sample throughput (n = 36 per day) as compared to GC/MS methods. LC-MS/MS will enable accurate measurements of red blood cell (RBC) folate in nutrition surveys and clinical trials.     

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.     

Performance of a short tool to assess dietary intakes of fruits and vegetables, percentage energy from fat and fibre

OBJECTIVES: We describe the methods used to develop and score a 17-item 'screener' designed to estimate intake of fruit and vegetables, percentage energy from fat and fibre. The ability of this screener and a food-frequency questionnaire (FFQ) to measure these exposures is evaluated. DESIGN: Using US national food consumption data, stepwise multiple regression was used to identify the foods to be included on the instrument; multiple regression analysis was used to develop scoring algorithms. The performance of the screener was evaluated in three different studies. Estimates of intakes measured by the screener and the FFQ were compared with true usual intake based on a measurement error model. SETTING: US adult population. SUBJECTS: For development of instrument, n=9323 adults. For testing of instrument, adult men and women in three studies completing multiple 24-hour dietary recalls, FFQ and screeners, n=484, 462 and 416, respectively. RESULTS: Median recalled intakes for examined exposures were generally estimated closely by the screener. In the various validation studies, the correlations between screener estimates and estimated true intake were 0.5-0.8. In general, the performances of the screener and the full FFQ were similar; estimates of attenuation were lower for screeners than for full FFQs. CONCLUSIONS: When coupled with appropriate reference data, the screener approach described may yield useful estimates of intake, for both surveillance and epidemiological purposes.     

Determination of folate concentrations in diverse potato germplasm using a trienzyme extraction and a microbiological assay

Folate deficiency is a leading cause of birth defects and is implicated in several other diseases. We are interested in how much folate concentrations vary among potato germplasm. We determined total folate concentrations of potato tubers from 67 cultivars, advanced breeding lines, or wild species. Folates were extracted by a tri-enzyme treatment and analyzed by using a Lactobacillus rhamnosus microbiological assay. Folate concentrations varied from 521 +/- 96 to 1373 +/- 230 ng/g dry weight and were genotype and location dependent. The highest folate concentrations were mostly found in color-fleshed potatoes. Variations of folate concentrations within either color- or white-fleshed tubers were similar ( approximately 2-fold). Skin contained approximately 30% higher folate concentrations than flesh. Storage of tubers for 7 months generally led to an increase in folate contents. Semiquantitative RT-PCR analyses showed that higher folate contents were correlated with lower mRNA expression of some folate genes.     

Comparative validation of standard, picture-sort and meal-based food-frequency questionnaires adapted for an elderly population of low socio-economic status

OBJECTIVE: To compare the validity of a modified Block food-frequency questionnaire (FFQ), a picture-sort administration of the FFQ (PSFFQ) and a meal pattern-based questionnaire (MPQ) in a multi-ethnic population of low socio-economic status (SES). DESIGN: Participants completed six 24-hour dietary recalls (24HR) over six months; the FFQ, PSFFQ and MPQ were completed in random order in the subsequent month. Instruments were interviewer-administered. The PSFFQ and MPQ were developed in formative research concerning difficulties for older adults in responding to standard food-frequency instruments. SETTING: Rural North Carolina, USA.Subjects One hundred and twenty-two African American, Native American and white adults aged > or = 65 years, with approximately one-third in each ethnic group. Inclusion criteria included education < or = 12 years and income < or = 150% of national poverty level or Medicaid recipient. RESULTS: Comparing median intakes from the average of the 24HR with the three diet assessment instruments, the MPQ tended to overestimate intakes compared with the FFQ and PSFFQ. Correlations among nutrients obtained by the 24HR and the other three instruments were generally statistically significant and positive. Across nutrients, the PSFFQ was most highly correlated with the 24HR for women, while the FFQ was most highly correlated with the 24HR for men. CONCLUSIONS: Dietary assessments using 24HR and FFQ were similar to results reported elsewhere, although correlations between 24HR and FFQ were somewhat lower. Interviewer-administered dietary assessments should be used with caution to evaluate dietary intake among older adults with low SES. Gender differences and the lower correlations should be investigated more thoroughly to assist in choosing dietary assessment instruments for this population.     

Frequency of soy food consumption and serum isoflavone concentrations among Chinese women in Shanghai

OBJECTIVE: The food-frequency questionnaire (FFQ) can be an efficient tool to evaluate dietary intake in large, population-based studies, especially for specific foods. The objective of this study was to validate the assessment of soy and isoflavone (daidzein and genistein) intakes, measured by an FFQ, by comparing intakes with serum isoflavone concentrations. DESIGN AND SETTING: Soy and isoflavone intakes and serum isoflavone concentrations were determined as part of a case-control study of dietary factors and risks of benign breast disease and breast cancer. The FFQ, administered during an in-person interview, included six soy-specific line items. Blood was drawn within one week of FFQ completion. SUBJECTS: In total, 1823 women living in Shanghai, People's Republic of China. RESULTS: In this population, soybean milk, fresh bean curd and other bean foods were eaten once per week, and fermented bean curd, fried bean curd puff and soybeans were eaten less than once per week. A significant linear trend (P<0.01) in serum isoflavone concentrations across increasing categories of soy and isoflavone intakes was observed, indicating that soy and isoflavone intakes, measured by the FFQ, well distinguished serum isoflavone concentrations. Linear trends were also observed in both case and control groups in stratified analyses, suggesting little differential bias by case-control status. CONCLUSIONS: The results suggest that the FFQ provides a useful marker of soy food consumption and isoflavone exposure in this population.     

Longitudinal changes in dietary intake in Scottish women around the menopause: changes in dietary pattern result in minor changes in nutrient intake

Objective: To examine dietary change that has occurred over 5 to 6 years.Subjects: A cohort of Scottish women (n=898) with a mean age of 47.5 years (range 45-54 years) at baseline.Design: Dietary intake was assessed by validated food-frequency questionnaire (FFQ) and analysed using the UK Composition of Foods database.Results: Since the first dietary assessment, mean daily energy intake had decreased from 8.2+/-2.3 to 7.9+/-2.2 MJ. The degree of low energy reporting (defined as ratio of energy intake to basal metabolic rate <1.1) had increased from 18.7% at baseline to 25.6% at follow-up. Low energy reporters were significantly heavier than 'normal' energy reporters (mean weight at follow-up, 68.9+/-12.6 vs. 66.8+/-11.3 kg) and could be deliberately restricting intake rather than underreporting. Overall there were decreases in intakes of red meat, processed meat and cheese, but increases in poultry and non-oily fish consumption. Consumption of bread, biscuits and cakes had gone down and there was an increase in cereal and rice/pasta consumption. Intake of potatoes had decreased whereas fruit intake had increased. There were small but statistically significant differences in intakes for most nutrients (<8% change). Nutrient intakes at both visits were similar across menopausal status and usage groups of hormone replacement therapy. Modifications to the computer version of the McCance and Widdowson nutrient database, which differed from the published version, were noted. These changes altered the original baseline values for our study.Conclusions: The menopause per se is not a period of marked change in nutrient intake. Caution is advised when using computer databases of food compositions for longitudinal studies.     

Changes of folate and other potential health-promoting phytochemicals in legume seeds as affected by germination

Folate deficiency associated with low dietary intake is a well-documented public health problem, resulting in serious health and socioeconomic burdens. Therefore, optimization of the germination process of different cultivars of legume seeds in relation to the content and composition of folate, vitamin C, and total phenolics and total antioxidant capacity was carried out to maximize the health-promoting properties. The content and composition of folate, vitamin C, and total phenolic and total antioxidant capacities varied between species, among cultivars, and with germination time. During germination, total folate content was maximum at 815.2 μg/100 g fresh weight in soybean sprout and at 675.4 μg/100 g fresh weight in mungbean sprout on the fourth day, which were equivalent to, respectively, 3.5- and 3.9-fold increases in the seed's content, and total folate content strongly decreased thereafter. 5-CH(3)-H(4)folate was the most abundant folate species in legume sprouts and reached a maximum on the fourth day. Vitamin C was not detected in raw seeds, and its content increased sharply in soybean and mungbean sprouts and reached a maximum at the fourth day of germination (29 and 27.7 mg/100 g fresh weight, respectively). Germination of soybean and mungbean for 4 days provided the largest amount of total folate as well as the more stable species 5-CH(3)-H(4)folate and also brought about large amounts of vitamin C and total phenolics and substantial antioxidant capacities.     

Total folate content and retention in rosehips (Rosa ssp.) after drying

Folate concentrations in rosehips and commercial rosehip products and factors affecting folate retention during drying were investigated. On the basis of the raw material studied during 3 years, rosehips were shown to be a rich folate source, 400-600 microg/100 g based on dry matter and 160-185 microg/100 g based on the fresh weight (edible part). Rosehips are not often consumed fresh; therefore, drying to produce stable semimanufactures is a crucial step. The degradation of folate was shown to be dependent on the drying time until the water activity was below 0.75. The required drying time was reduced by cutting the rosehips in slices and to some extent also by increasing the temperature. Retention of folate and ascorbic acid was affected by the same factors, and high content of ascorbic acid could provide a possible protection for folate degradation.     

Validity of protein, retinol, folic acid and n-3 fatty acid intakes estimated from the food-frequency questionnaire used in the Danish National Birth Cohort

OBJECTIVE: To validate intakes of protein, folic acid, retinol and n-3 fatty acids estimated from a food-frequency questionnaire in week 25 of pregnancy (FFQ-25). DESIGN: Validation was done against a 7-day weighed food diary (FD) and biomarkers of the nutrients in gestation week 32-38. SUBJECTS AND SETTING: The FFQ-25 to be validated was used in the Danish National Birth Cohort comprising 101 042 pregnant Danish women, of whom 88 participated in the present validation study. RESULTS: Estimated intakes of protein, retinol and folic acid did not differ significantly between the two dietary methods, but intake of n-3 fatty acids was one third larger when estimated from the FFQ-25. The intakes estimated from the two dietary methods were all significantly correlated, ranging from 0.20 for retinol intake to 0.57 for folic acid intake. Sensitivities of being correctly classified into low and high quintiles were between 0.22 and 0.77, and specificities were between 0.62 and 0.89. Urinary protein content did not correlate significantly with protein estimated from the FFQ (r = 0.17, P > 0.05), but did with intake estimated from the FD (r = 0.56, P < 0.0001). Erythrocyte folate correlated significantly with the estimated total intake from the FFQ (r = 0.55, P < 0.0001) and the FD (r = 0.52, P < 0.0001). No correlations with plasma retinol were found. Erythrocyte eicosapentaenoic acid (C20:5n-3) correlated significantly with n-3 fatty acids intake estimated from both the FFQ-25 (r = 0.37, P < 0.001) and the FD (r = 0.62, P < 0.0001). CONCLUSION: The FFQ-25 gives reasonable valid estimates of protein, retinol and folic acid intakes, but seems to overestimate intake of n-3 fatty acids.     

What do babies eat? Evaluation of a food frequency questionnaire to assess the diets of infants aged 6 months

OBJECTIVE: To evaluate the relative validity of a food frequency questionnaire (FFQ) for assessing nutrient intakes in 6-month-old infants. DESIGN AND SETTING: The FFQ was developed to assess the diets of infants born to women in the Southampton Women's Survey (SWS), a population-based survey of young women and their offspring. The energy and nutrient intakes obtained from an interviewer-administered FFQ were compared with those obtained from a 4-day weighed diary. SUBJECTS AND METHODS: A sub-sample of 50 infants aged 6 months from the SWS had their diets assessed by both methods. The FFQ recorded the frequencies and amounts of milks, baby foods, regular foods and drinks consumed by the infants over the previous seven days. The diaries recorded the weights of all foods and drinks consumed by the infants on four separate days within 15 days following FFQ completion. RESULTS: Spearman rank correlation coefficients for intakes of energy, macronutrients and 18 micronutrients, determined by the two methods, ranged from r = 0.39 to 0.86; adjustment for energy intake tended to increase the correlation coefficients, range r(a) = 0.55 to 0.89. Bland-Altman statistics showed that mean differences between methods were in the range of -12.5% to +12.5% except for vitamin B12 (-18.9%). CONCLUSION: Although there were differences in absolute energy and nutrient intakes between methods, Spearman rank correlation coefficients indicated reasonable agreement in the ranking of intakes. The interviewer-administered FFQ is a useful tool for assessing energy and nutrient intakes of healthy infants aged about 6 months.     

Development and simulated validation of a food-frequency questionnaire for the Colombian population

OBJECTIVE: To develop a food-frequency questionnaire (FFQ) useful for ranking of nutrient intakes. DESIGN: Subjects consuming their regular diet completed 7 days of weighed intake registry (7-WIR). Foods for the FFQ were selected by stepwise multiple regression. The FFQ was then completed for each subject using data on individual food consumption from the 7-WIR. The correlation and agreement between the extrapolated FFQ and the 7-WIR data were assessed using Spearman's rank correlation coefficients (rS) and Bland and Altman's limits of agreement (LOA). SETTING: Bucaramanga, Colombia. SUBJECTS: We studied 97 randomly selected 20-40-year-old subjects. RESULTS: Sixty foods were selected for the FFQ. The 7-WIR and the extrapolated FFQ intake estimates correlated well. rS was 0.58 for energy, 0.53 for carbohydrate, 0.50 for total fat, and 0.48 for protein. For micronutrients, rS varied from 0.46 (manganese) to 0.71 (vitamin B12). FFQ average intake estimates were 83%, 80%, 86.2% and 86.4% of 7-WIR estimates for energy, carbohydrate, total fat and protein, respectively. LOA for these nutrients ranged between 45% and 165%. FFQ micronutrient intakes were on average 96% (median) of those from the 7-WIR, and the median lower and upper LOA were 50% and 203%. However, there was no indication that the degree of agreement varied with the level of intake. CONCLUSIONS: According to our simulated validation, this FFQ may be useful to rank subjects by nutrient intake. Its validity against standard independent measurements and its applicability to other subsets of the Colombian population should be carefully considered.     

Comparison of blood folate levels among pregnant Chinese women in areas with high and low prevalence of neural tube defects

OBJECTIVE: To characterise plasma and red-blood-cell (RBC) folate status among pregnant women in an area with an extremely high prevalence of neural tube defects, and to compare them with those of women from a low prevalence area. DESIGN: A cross-sectional survey conducted in 2003. SETTING: One county and one city from each of the high prevalence area and the low prevalence area in China. SUBJECTS: Five hundred and sixty-two women in their first trimester of pregnancy in the high prevalence area and 695 pregnant women in the low prevalence area. RESULTS: Women in the high prevalence area had less than half the plasma and RBC folate concentrations (12.2 and 440.0 nmol l- 1, respectively) of women in the low prevalence area (33.5 and 910.4 nmol l- 1, respectively). In the high prevalence area, 40% of rural women were deficient in RBC folate and 50% were deficient in plasma folate; 20% of urban women were deficient in RBC folate and 30% deficient in plasma folate. In contrast, only 4% (RBC folate) and 6% (plasma folate) of rural women, and 2% (RBC folate) and 1% (plasma folate) of urban women, were folate-deficient in the low prevalence area. Less than 10% of rural and about 26% of urban women in the high prevalence area took folic acid periconceptionally, compared with 70% and 60% of women in the low prevalence area. CONCLUSIONS: Blood folate deficiency is highly prevalent among pregnant women in an area of China with a very high prevalence of neural tube defects.     

The influence of fruit and vegetable intake on the nutritional status and plasma homocysteine levels of institutionalised elderly people

OBJECTIVE: To determine the difference in the nutritional status of elderly people depending on their consumption of fruits and vegetables, and to study the possible association between the consumption of these foods and different cardiovascular risk factors, especially total plasma homocysteine (t-Hcys) levels. DESIGN, SETTING AND SUBJECTS: A cross-sectional study in 152 institutionalised older people from Madrid aged > or = 65 years. Food and nutrient intakes were recorded over 7 days using the 'precise individual weighing' method. The weight, height, and waist and hip circumferences of all subjects were recorded, as were their alpha-erythrocyte glutathione reductase, serum B6, B12 and folate levels, erythrocyte folate levels, t-Hcys levels, serum lipids and blood pressure. The experimental population was then divided into tertiles depending on the serving intake of fruit and vegetables (T1, < 2.29 servings day(-1); T2, 2.29-2.79 servings day(-1); and T3, >2.79 servings day(-1)). RESULTS: Compared with T1 subjects, T3 subjects showed consumptions of cereals, pulses, meat, fish and eggs closer to those recommended (P < 0.05). In addition, the contribution of their diet towards covering the recommended daily intake of vitamin B1, niacin, vitamin B6, folic acid, vitamin C, B12, vitamin A, and P, Mg, Zn and Fe was higher. The intake of fibre increased with consumption of fruit and vegetables (r = 0.6839, P < 0.001). T3 subjects also had better serum and erythrocyte folate levels than T1 and T2 subjects (P < 0.05). A positive correlation was found between the consumption of fruit and vegetables and serum folate (r = 0.2665, P < 0.01) and with erythrocyte folate levels (r = 0.2034, P < 0.05), and a negative correlation with t-Hcys (r = -0.2493, P < 0.01). CONCLUSIONS: Greater consumption of fruit and vegetables is associated with better food habits, increased vitamin and mineral intakes and lower t-Hcys levels. Considering that the fruit and vegetable intake in Spanish elderly people is very low, it is recommended that the consumption of fruits and vegetables by elderly people be increased.     

The UK Women's Cohort Study: comparison of vegetarians, fish-eaters and meat-eaters

BACKGROUND: This paper describes the development of the UK Women's Cohort Study and presents cohort baseline characteristics. METHODS: In total, 35,372 women, aged 35-69 years at recruitment, were selected to ensure a wide range of dietary intakes. Diet was assessed by a 217-item food-frequency questionnaire (FFQ). Detailed lifestyle information was collected by postal questionnaire. Vegetarians, fish-eaters and meat-eaters were compared. RESULTS: The cohort women are mainly white, well-educated, middle-class and married with children. They are health-conscious with only 11% current smokers and 58% taking dietary supplements. Twenty-eight per cent of subjects self-report as being vegetarian and 1% as vegan. However, only 18% are defined as 'vegetarian' from the FFQ. Fat provides 32% of energy; vitamin and mineral intakes are high, with a broad range of intakes. Meat-eaters are older, with a higher body mass index (BMI) and the lowest intakes of carbohydrate, fibre, vitamin C, folate, iron and calcium. Other fish-eaters are similar to vegetarians. Vegetarians have the lowest intakes of protein, fat and saturated fat. Oily fish-eaters have the lowest BMI; are the least likely to smoke or use full-fat milk; and are the most likely to use dietary supplements and consume the most fruit and vegetables. Oily fish-eaters have the highest total energy intake and vegetarians the lowest. Semi-skimmed milk, bread, potatoes, wine, bananas and muesli are important contributors to energy for all groups. CONCLUSION: A large cohort of middle-aged women has been created encompassing a wide range of different eating patterns, including diets currently of interest to research into protection against cancer and coronary heart disease. Participants will be followed up to study the effects of different food and nutrient intakes on long-term health outcomes.     

Folate content in strawberries (Fragaria x ananassa): effects of cultivar,ripeness, year of harvest,storage, and commercial processing

Folate concentrations in strawberries and folate retention during storage and commercial processing of strawberries were investigated. No previous study has focused on the effects of cultivar, ripeness, and year of harvest of strawberries with respect to the folate content. This study showed the folate concentration in strawberries to significantly depend on all of these different factors. Total folate was quantified using a modified and validated radioprotein-binding assay with external calibration (5-CH(3)-H(4)folate). Folate content in 13 different strawberry cultivars varied from 335 microg/100 g of dry matter (DM) for cv. Senga Sengana to 644 microg/100 g of DM for cv. Elsanta. Swedish harvests from 1999 and 2001 yielded higher folate concentrations than did the harvest from 2000, and the grade of ripeness affected the folate content in strawberries. This study indicated high folate retention in intact berries during storage until 3 or 9 days at 4 degrees C (71-99%) and also in most tested commercial products (79-103%). On the basis of these data fresh strawberries as well as processed strawberry products are recommended to be good folate sources. For instance, 250 g (fresh weight) of strawberries ( approximately 125 microg of folate) supplies approximately 50% of the recommended daily folate intake in various European countries (200-300 microg/day) or 30% of the U.S. recommendation (400 microg/day).     

Evaluation of brief dietary questions to estimate vegetable and fruit consumption - using serum carotenoids and red-cell folate

OBJECTIVE: To evaluate responses to self-administered brief questions regarding consumption of vegetables and fruit by comparison with blood levels of serum carotenoids and red-cell folate. DESIGN: A cross-sectional study in which participants reported their usual intake of fruit and vegetables in servings per day, and serum levels of five carotenoids (alpha-carotene, beta-carotene, beta-cryptoxanthin, lutein/zeaxanthin and lycopene) and red-cell folate were measured. Serum carotenoid levels were determined by high-performance liquid chromatography, and red-cell folate by an automated immunoassay system. SETTINGS AND SUBJECTS: Between October and December 2000, a sample of 1598 adults aged 25 years and over, from six randomly selected urban centres in Queensland, Australia, were examined as part of a national study conducted to determine the prevalence of diabetes and associated cardiovascular risk factors. RESULTS: Statistically significant (P<0.01) associations with vegetable and fruit intake (categorised into groups: /=4 servings per day) were observed for alpha-carotene, beta-carotene, beta-cryptoxanthin, lutein/zeaxanthin and red-cell folate. The mean level of these carotenoids and of red-cell folate increased with increasing frequency of reported servings of vegetables and fruit, both before and after adjusting for potential confounding factors. A significant association with lycopene was observed only for vegetable intake before adjusting for confounders. CONCLUSIONS: These data indicate that brief questions may be a simple and valuable tool for monitoring vegetable and fruit intake in this population.     

Validation of a food-frequency questionnaire for use in pregnancy

OBJECTIVES: As a part of an ongoing project to develop a nutritional screening tool, we evaluated the performance of a semi-quantitative food-frequency questionnaire (FFQ) in terms of validity in a Sheffield Caucasian pregnant population using two different statistical approaches--the correlation coefficient and the limits of agreement (LOA). The FFQ was designed specifically for pregnant women and previously used in a large-scale study. DESIGN: A validation study. SETTING: A community-based field study of a general population of pregnant women booked for their first antenatal appointment at the Jessop Wing, Royal Hallamshire Hospital, Sheffield, UK. SUBJECTS: One hundred and twenty-three women of different socio-economic status, aged between 17 and 43 years, provided complete dietary data. RESULTS: The validity of the FFQ was tested against a series of two 24-hour recalls. As expected, the intakes of all examined nutrients, except for iodine, carotene, vitamin E, biotin, vitamin C and alcohol, were higher when determined by the FFQ than when determined by 24-hour recall. Pearson's correlation coefficient between the two methods ranged from 0.19 (added sugar, zinc) to 0.47 (Englyst fibre). The LOA were broader for some of the nutrients, e.g. protein, Southgate fibre and alcohol, and an increasing lack of agreement between the two methods was identified with higher dietary intakes. CONCLUSIONS: The FFQ gave useful estimates of the nutrient intakes of Caucasian pregnant women and appears to be a valid tool for categorising pregnant women according to dietary intake. The FFQ performed well for most nutrients and had acceptable agreement with the 24-hour recall.     

Application of a tri-enzyme extraction for total folate determination in foods

A tri-enzyme digestion procedure using chicken pancreas conjugase, alpha-amylase, and Pronase was evaluated to determine its usefulness in the microbiological quantitation of total folate in foods. Folate values obtained by traditional conjugase digestion were compared to those obtained by the tri-enzyme method for 12 food products that represent diverse matrixes. The tri-enzyme treatment increased measurable folate from most foods when compared to levels found after conjugase digestion. Largest increases were noted for tuna fish (51%) and yogurt (33%) after tri-enzyme digestion. For the 12 foods, a mean increase of 19% in measurable folate was obtained with tri-enzyme treatment. The study shows that traditional conjugase treatment does not completely free folate from complex food matrixes before microbiological analysis. Further, as other investigations have suggested, current accepted methods for folate analysis may be underestimating folate levels in foods.