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Since collection of 24 h urine samples is very time consuming and difficult to obtain, epidemiological studies typically only obtain spot urine samples. The aim of the present study was to evaluate whether flavonoids and enterolactone in overnight urine could substitute flavonoids and enterolactone in 24 h urine as an alternative and more feasible biomarker of fruit, vegetable and beverage intake. A total of 191 individuals in the Inter99 cohort in Denmark completed the validation study. Concentrations of nine urinary flavonoid aglycones (quercetin, isorhamnetin, tamarixetin, kaempferol, hesperetin, naringenin, eriodictyol, phloretin and apigenin) and enterolactone were determined in overnight and 24 h urine samples, and their validity as biomarkers of fruit, vegetable and beverage intake was evaluated in relation to two independent reference methods (Inter99 FFQ data and plasma carotenoids) by using the method of triads. The intakes of fruit, juice, vegetables and tea reported in the FFQ were reflected by the flavonoid biomarker both in overnight and 24 h urine samples. Validity coefficients for the flavonoid biomarker in overnight urine ranged from 0·39 to 0·49, while the corresponding validity coefficients for the biomarker in 24 h urine ranged from 0·43 to 0·66. Although the validity coefficients were lower for overnight urine than for the 24 h urine flavonoid biomarker, they were still of acceptable magnitude. In conclusion, the results indicate that flavonoids and enterolactone in overnight urine samples may be used as a more feasible biomarker than 24 h urine for the assessment and validation of fruit, juice, vegetable and tea intakes in epidemiological studies.

Objective: The main purpose of the study was to investigate the feasibility of using workplaces to increase the fruit consumption of participants by increasing fruit availability and accessibility by a minimal fruit programme. Furthermore, it was investigated whether a potential increase in fruit intake would affect vegetable, total energy and nutrient intake., Design: A 5-month, controlled, workplace study where workplaces were divided into an intervention group (IG) and a control group (CG). At least one piece of free fruit was available per person per day in the IG. Total fruit and dietary intake was assessed, using two 24 h dietary recalls at baseline and at endpoint., Setting: Eight Danish workplaces were enrolled in the study. Five workplaces were in the IG and three were in the CG., Subjects: One hundred and twenty-four (IG, n 68; CG, n 56) healthy, mainly normal-weight participants were recruited., Results: Mean daily fruit intake increased significantly from baseline to endpoint only in the IG by 112(se 35) g. In the IG, mean daily intake of added sugar decreased significantly by 10·7(se 4·4) g, whereas mean daily intake of dietary fibre increased significantly by 3·0(se 1·1) g. Vegetable, total energy and macronutrient intake remained unchanged through the intervention period for both groups., Conclusions: The present study showed that it is feasible to increase the average fruit intake at workplaces by simply increasing fruit availability and accessibility. Increased fruit intake possibly substituted intake of foods containing added sugar. In this study population the increased fruit intake did not affect total energy intake.

Background/objectives: To validate 24 h dietary recall of fruit intake by measuring the total 24 h excretion of 10 different flavonoids in 24 h urine during an intervention with free fruit at workplaces., Subjects/methods: Employees at workplaces offering a free-fruit program, consisting of daily free and easy access to fresh fruit, and controls employees at workplaces with no free-fruit program were enrolled in this validation study (n=103). Dietary intake was assessed by using a 24 h dietary recall questionnaire at baseline and approximately 5 months later. Ten flavonoids, quercetin, isorhamnetin, tamarixetin, kaempferol, hesperetin, naringenin, eriodictyol, daidzein, genistein, and phloretin, were measured using HPLC-electrospray ionization-MS., Results: The 24 h urinary excretion of total flavonoids and the estimated intake of fruits were significantly correlated (r (s)=0.31, P<0.01). The dietary intake of citrus fruits and citrus juices was significantly correlated with total excretion of citrus specific flavonoids (r (s)=0.28, P<0.01), and orange was positively correlated with naringenin (r (s)=0.24, P<0.01) and hesperetin (r (s)=0.24, P<0.01). Phloretin in urine was correlated with apple intake (r (s)=0.22, P<0.01) and also with overall estimated intake of fruit (r (s)=0.22, P<0.01)., Conclusions: This study shows that a 24 h dietary recall can be used as a valid estimate of the intake of fruits in agreement with an objective biomarker of fruit intake in free fruit at workplace interventions.

To gain better insight into the potential health effects of fruits and vegetables, reliable biomarkers of intake are needed. The main purpose of this study was to investigate the ability of flavonoid excretion in both 24-h and morning urine samples to reflect a low intake and moderate changes in fruit and vegetable consumption. Furthermore, the urinary excretions of 4-pyridoxic acid (4-PA) and potassium were investigated as other potential biomarkers of fruit and vegetable intake. The study was designed as a 5-d randomized, controlled crossover study. On d 1-3, the men (n = 12) consumed a self-restricted flavonoid-free diet. On d 4, they were provided a strictly controlled diet containing no fruits or vegetables (basic diet). On d 5, they consumed the basic diet supplemented with 300 or 600 g of fruits and vegetables. The total excretion of flavonoids in 24-h urine samples increased linearly with increasing fruit and vegetable intakes (r(s) = 0.86, P < 1 x 10(-6)). The total excretion of flavonoids in morning urine also increased, but the association was weaker (r(s) = 0.59, P < 0.0001). Urinary 4-PA in 24-h and morning urine samples increased significantly only with the 600-g increase in fruit and vegetable intake, whereas the excretion of potassium in urine did not reflect the changes in fruit and vegetable intake. We conclude that the total excretion of flavonoids in 24-h urine may be used as a new biomarker for fruit and vegetable intake.