Cruciferous vegetables and cancer risk in a network of case-control studies.

BACKGROUND: Cruciferous vegetables have been suggested to protect against various cancers, though the issue is open to discussion. To further understand their role, we analyzed data from a network of case-control studies conducted in Italy and Switzerland. PATIENTS AND METHODS: The studies included a total of 1468 cancers of the oral cavity/pharynx, 505 of the esophagus, 230 of the stomach, 2390 of the colorectum, 185 of the liver, 326 of the pancreas, 852 of the larynx, 3034 of the breast, 367 of the endometrium, 1031 of the ovary, 1294 of the prostate, 767 of the kidney, and 11,492 controls. All cancers were incident, histologically confirmed; controls were subjects admitted to the same network of hospitals as cases for a wide spectrum of acute nonneoplastic conditions. RESULTS: The multivariate odds ratio (OR) for consumption of cruciferous vegetables at least once a week as compared with no/occasional consumption was significantly reduced for cancer of the oral cavity/pharynx (OR=0.83), esophagus (OR=0.72), colorectum (OR=0.83), breast (OR=0.83), and kidney (OR=0.68). The OR was below unity, but not significant, for stomach (OR=0.90), liver (OR=0.72), pancreatic (OR=0.90), laryngeal (OR=0.84), endometrial (OR=0.93), ovarian (OR=0.91), and prostate (OR=0.87) cancer. CONCLUSION: This large series of studies provides additional evidence of a favorable effect of cruciferous vegetables on several common cancers.

The temporal effect of a wild blueberry (Vaccinium angustifolium)-enriched diet on vasomotor tone in the Sprague-Dawley rat.

BACKGROUND AND AIMS: We have previously reported that wild blueberry (Vaccinium angustifolium)-enriched diets (WB) attenuate aortic adrenergic response through endothelial-mediated pathways. The duration of dietary intervention necessary to induce the positive changes on vasomotor tone has not been studied to date. Thus, our objective was to investigate the temporal effect of WB consumption on vascular function and reactivity in Sprague-Dawley (SD) rat aorta after 4 and 7 weeks of dietary treatment. METHODS AND RESULTS: Forty male SD rats were randomly assigned to a control (AIN-93) (C) or a WB diet for 4 or 7 weeks. Vascular ring studies were conducted in 3-mm isolated rat aortic rings to investigate vasoconstriction induced by six doses of the α(1)-adrenergic agonist, L-phenylephrine (Phe, 10(-8)-3×10(-6) M) alone or in the presence of the NOS inhibitor, L-N(G)-monomethyl-arginine (L-NMMA, 10(-4)M). The maximum force of contraction (F(max)) and vessel sensitivity (pD(2)) were determined. Analysis of variance revealed no significant differences on F(max) after 4 weeks of the WB diet but only a significant increase in pD(2) in the absence of L-NMMA. Seven week WB consumption significantly attenuated contraction in response to L-Phe and resulted in lower pD(2). Inhibition of NOS induced a significant increase in the constrictor response in both diet groups at both time periods, with the WB group fed for 7 weeks having the greater response. CONCLUSION: Thus wild blueberries incorporated into the diet at 8% w/w positively affect vascular smooth muscle contractility and sensitivity but these effects are evident only after 7 weeks of WB consumption.

Blood orange juice inhibits fat accumulation in mice.

OBJECTIVE: To analyze the effect of the juice obtained from two varieties of sweet orange (Citrus sinensis L. Osbeck), Moro (a blood orange) and Navelina (a blond orange), on fat accumulation in mice fed a standard or a high-fat diet (HFD). METHODS: Obesity was induced in male C57/Bl6 mice by feeding a HFD. Moro and Navelina juices were provided instead of water. The effect of an anthocyanin-enriched extract from Moro oranges or purified cyanidin-3-glucoside (C3G) was also analyzed. Body weight and food intake were measured regularly over a 12-week period. The adipose pads were weighted and analyzed histologically; total RNA was also isolated for microarray analysis. RESULTS: Dietary supplementation of Moro juice, but not Navelina juice significantly reduced body weight gain and fat accumulation regardless of the increased energy intake because of sugar content. Furthermore, mice drinking Moro juice were resistant to HFD-induced obesity with no alterations in food intake. Only the anthocyanin extract, but not the purified C3G, slightly affected fat accumulation. High-throughput gene expression analysis of fat tissues confirmed that Moro juice could entirely rescue the high fat-induced transcriptional reprogramming. CONCLUSION: Moro juice anti-obesity effect on fat accumulation cannot be explained only by its anthocyanin content. Our findings suggest that multiple components present in the Moro orange juice might act synergistically to inhibit fat accumulation.

The total antioxidant capacity of the diet is an independent predictor of plasma beta-carotene.

OBJECTIVE: To investigate the contribution of the total antioxidant capacity (TAC) of the diet to plasma concentrations of beta-carotene. DESIGN: Cross-sectional study. SETTING: Department of Public Health and Department of Internal Medicine and Biomedical Sciences, University of Parma. SUBJECTS: A total of 247 apparently healthy adult men (n=140) and women (n=107). METHODS: A medical history, a physical exam including height, weight, waist circumference and blood pressure measurements, a fasting blood draw, an oral glucose tolerance test and a 3-day food record. RESULTS: We observe a negative trend across quartiles of plasma beta-carotene for most biological variables clustering in the insulin resistance syndrome, as well as for traditional and new risk factors for type II diabetes and cardiovascular disease (CVD), including C-reactive protein and gamma-glutamyltranspeptidase (P<0.05). Regarding dietary characteristics, energy-adjusted intake of fat, fiber, fruits, vegetables, beta-carotene, vitamin C, vitamin E and dietary TAC significantly increased with increasing plasma beta-carotene (P<0.05), whereas alcohol intake decreased (P=0.013). Adjusted geometric means (95% confidence interval) of plasma beta-carotene significantly increased across quartiles of dietary TAC, even when single dietary antioxidants were considered in the model (QI=0.087 mg/dl (0.073-0.102); QII=0.087 mg/dl (0.075-0.103); QIII=0.114 mg/dl (0.098-0.132) and QIV=0.110 mg/dl (0.093-0.130); P for linear trend=0.026). When the population was divided on the basis of alcohol consumption, this trend was also observed in subjects drinking <20 g alcohol/day (P=0.034), but not in those with higher alcohol intake (P=0.448). CONCLUSIONS: Dietary TAC is an independent predictor of plasma beta-carotene, especially in moderate alcohol drinkers. This may explain, at least in part, the inverse relationship observed between plasma beta-carotene and risk of chronic diseases associated to high levels of oxidative stress (i.e., diabetes and CVD), as well as the failure of beta-carotene supplements alone in reducing such risk.

Comparison between daidzein and genistein antioxidant activity in primary and cancer lymphocytes.

The main objective of this study was to compare the protective effect of daidzein and genistein against induced oxidative damage in Jurkat T-cell line and in peripheral blood lymphocytes of healthy subjects. After supplementation of cells with isoflavones (from 2.5 to 20micromol/L in Jurkat T-cell and from 0.01 to 2.5micromol/L in primary lymphocytes, 24h), we determined DNA damage induced by hydrogen peroxide using the comet assay and lipid peroxidation evaluating malondialdehyde (MDA) production after ferrous ion treatment. Supplementation of Jurkat cells and primary lymphocytes with both isoflavones significantly increased DNA protection from oxidative damage at concentrations between 0.1 and 5micromol/L (P<0.05), and with just daidzein, at concentrations higher than 2.5micromol/L, there was a decrease in the production of MDA (P<0.05). Our results seem to support that daidzein is just as effective as genistein in protecting cells against oxidative damage especially with respect to DNA. Moreover, since the protective effect was found at concentrations reachable in plasma after soy consumption (less than 2micromol/L), it can be assumed that the antioxidant activity of isoflavones could really contribute to the healthy properties of soy.

Bioavailability of carotenoids from spinach and tomatoes.

BACKGROUND AND AIM: Few published studies have described the bioavailability of the different carotenoids in spinach. This was designed to evaluate the effects on plasma carotenoid concentrations of a daily consumption of spinach (rich in lutein and beta-carotene), alone or together with lycopene-rich tomato puree. METHODS AND RESULTS: Nine healthy young women consumed a standard low-carotenoid diet during the pre-study phase, the spinach diet (standard diet plus 150 g spinach: 9 mg lutein, 4 mg beta-carotene) from day 0 to day 21, and then, after a wash-out period, the spinach-tomato diet (standard diet plus 150 g spinach and 25 g tomato puree: 9 mg lutein, 4.3 mg beta-carotene and 7 mg lycopene) from day 35 to day 56. The spinach and spinach-tomato supplements were consumed together with 10 g olive oil. Fasting blood samples were collected on day -7, and every week thereafter. Plasma carotenoid concentrations significantly decreased during the standard low-carotenoid diet. Lutein levels gradually increased after spinach consumption from 0.36+/-0.05 to 1.59+/-0.19 micromol/L (p<0.0001), decreased during the wash-out period from 1.59+/-0.19 to 0.62+/-0.07 micromol/L (p<0.001), and rose again after the intake of spinach-tomato puree from 0.62+/-0.07 to 1.55+/-0.17 micromol/L (p<0.0001). beta-carotene levels also increased during both dietary supplementation periods. Lycopene decreased during the spinach diet from 0.20+/-0.03 to 0.07+/-0.01 micromol/L (p<0.001) and increased during the spinach-tomato diet from 0.05+/-0.01 to 0.52+/-0.06 micromol/L (p<0.0001). CONCLUSIONS: The results of this study confirm that a regular intake of selected vegetables leads to a progressive increase in plasma carotenoid concentrations. The addition of tomato puree to spinach does not decrease lutein plasma concentrations. Furthermore, baseline plasma levels of lutein and lycopene are important variables affecting the relative increase in their levels after supplementation: ie more depleted subjects are expected to have a greater percent rise in plasma carotenoid concentrations.

Lycopene and vitamin C concentrations increase in plasma and lymphocytes after tomato intake. Effects on cellular antioxidant protection.

OBJECTIVE: This study seeks to verify whether the regular consumption of small amounts of tomato products can protect lymphocyte DNA and lipids from oxidative damage. DESIGN: Standardized dietary intervention. SUBJECTS: Twelve healthy female subjects (mean age 25.2 y). INTERVENTION: Subjects were instructed to follow a standardized diet for 1 week, followed by 3 weeks consumption of the same diet enriched with small amounts of different tomato products providing as a mean 8 mg lycopene, 0.5 mg beta-carotene and 11 mg vitamin C per day. Plasma and lymphocyte concentrations of carotenoids, vitamin C and vitamin E were analysed. Ex vivo protection of lymphocyte DNA from oxidative injury produced by iron ions was evaluated by means of the Comet assay, and lipid peroxidation by HPLC analysis of malondialdehyde (MDA). RESULTS: Dietary intervention with tomato products increased lycopene concentration both in plasma (P < 0.001) and lymphocytes (P < 0.01). Vitamin C concentrations increased by approximately 35% in plasma (P < 0.05) and by approximately 230% in lymphocytes (P < 0.005). Vitamin E decreased significantly in plasma (P < 0.0001) but not in lymphocytes. Finally, there was an improved protection from DNA oxidative damage (P < 0.05) with no significant effect on MDA levels. CONCLUSIONS: Our results suggest that tomato products are not only good sources of lycopene but also sources of bioavailable vitamin C. A Regular intake of small amounts of tomato products can increase cell protection from DNA damage induced by oxidant species. This effect may originate from the synergism of different antioxidants present in tomatoes.

Black tea extract supplementation decreases oxidative damage in Jurkat T cells.

The purpose of this study was to investigate the protective effect of black tea (BT) extract against induced oxidative damage in Jurkat T-cell line. Cells supplemented with 10 or 25 mg/L BT were subjected to oxidation with ferrous ions. Malondialdehyde (MDA) production as marker of lipid peroxidation, DNA single strand breaks as marker of DNA damage, and modification of the antioxidant enzyme activity, glutathione peroxidase (GPX) were measured. Results show the efficacy of BT polyphenols to decrease DNA oxidative damage and to affect GPX activity (P<0.05), while no effect was shown on MDA production. The succeeding investigation of the activity of caffeine and epigallocatechin gallate demonstrated their antioxidant potential with respect to the cellular markers evaluated. In conclusion, this study supports the protective effect of BT against ferrous ions induced oxidative damage to DNA and the ability of BT to affect the enzyme antioxidant system of Jurkat cells.

Malondialdehyde production in Jurkat T cells subjected to oxidative stress.

OBJECTIVE: We investigated the relation between membrane lipid peroxidation, as evaluated by malondialdehyde (MDA), and oxidative stimuli in the Jurkat T-cell line and designed a cellular model to assess the antioxidant potential of compounds. METHODS: Jurkat T cells were subjected to different concentrations of Fe(2+) ions (from 25 to 150 micromol/L) or H(2)O(2) (from 0.1 to 5 mmol/L), and MDA was determined after separation in high-performance liquid chromatography of the adduct with thiobarbituric acid. MDA production also was investigated in cells supplemented with epigallocatechin gallate and genistein and subjected to Fe(2+) oxidative treatment. RESULTS: MDA production increased with the concentration of Fe(2+), whereas H(2)O(2) had no effect at any concentration. Oxidative stress for 15 min or 2 h produced similar MDA levels. The supplementation of epigallocatechin gallate partly prevented MDA production (about 40%, P < 0.05), whereas genistein exerted no preventive effect on lipid peroxidation. CONCLUSION: We propose this cellular model, consisting of Jurkat T cells subjected to 100 micromol/L of Fe(2+) for 15 min, to study the protective effect of antioxidant supplementation against membrane lipid peroxidation.

Oxidative stress signalling in the apoptosis of Jurkat T-lymphocytes.

The pathways of transduction of oxidative stress signals have been studied using the Jurkat T cell model. The oxidative stress was induced by exposure of the cells to 100 microM H(2)O(2). DNA damage was detected within 15 min after commencement of treatment. DNA damage repair occurred within about 1 h in cells exposed to oxidative stress for 15 min. In continuous exposure to stress, DNA repair was slower and control levels of DNA integrity were not reached. DNA repair did not involve gene transcription. H(2)O(2) at 100 microM caused cell death by necrosis as well as by apoptosis. Both these processes were induced by 15 min exposure to the stress stimulus. However, some important differences were found between necrosis and apoptosis. Necrosis was more rapid, began within an hour of treatment and continued to increase during the full duration of the experiment. But apoptosis was seen after 4 h from treatment and was conspicuous between 6 and 20 h after the start of treatment. The necrotic phase preceded apoptosis, although these did show an overlap. In the necrotic phase, Bcl-2, Caspase 8 genes were down regulated. The 6-20 h phase characterised by a marked increase in apoptosis is accompanied by the up regulation of both Bcl-2 and Caspase genes. Expression of the Fas and p53 genes was not altered in either phase. We also analysed the levels of expression of the scavenging genes whose gene products are involved in detoxification. No modulation of the antioxidant enzymes, catalase, Cu/Zn superoxide dismutase and glutathione peroxidase was detectable.

Nutritional evaluation of some processed catering foods.

Five prepared catering dishes were analysed to evaluate the proximate composition and the fatty acids, vitamin E, thiamine and riboflavin content. The correspondent values were calculated from actually available food composition tables (two from Italy, one from the UK and one from the USA). When using more than one database to calculate the composition of a complex recipe the average values were similar to the analytical ones despite the wide range reported for some variables. However, there was no significant difference in the statistical analyses between the analytical values and databases, or among the databases themselves. Therefore if the composition of a specific recipe is required, analyses would be advisable, but the available databases are quite adequate if the evaluation is for groups of people, even allowing for the seasonal variability of ingredients.

Tomato consumption does not affect the total antioxidant capacity of plasma.

This study was performed to evaluate the effect of tomato intake on total antioxidant activity of plasma measured by the radical trapping antioxidant parameter assay in 11 healthy female subjects. After 7 d of a diet low in carotenoids and free from lycopene, subjects ate 25 g tomato puree daily (containing 7.0 mg lycopene and 0.25 mg beta-carotene) for 14 consecutive days. At the beginning and end of tomato supplementation, the carotenoid plasma concentration and the total antioxidant activity of plasma were assessed. Before tomato puree consumption, mean +/- SE total lycopene and beta-carotene plasma concentrations were 0.13 +/- 0.02 micromol/L and 0.24 +/- 0.04 micromol/L, respectively. After tomato puree supplementation, both concentrations increased significantly (0.57 +/- 0.06 micromol/L, P < 0.0001 for total lycopene, and 0.31 +/- 0. 04 micromol/L, P = 0.0036 for beta-carotene); however, total plasma antioxidant capacity values did not change significantly. From our results, intake of a food rich in carotenoids does not seem to modify the antioxidant capacity of plasma as evaluated by the radical trapping antioxidant parameter assay.

Lymphocyte lycopene concentration and DNA protection from oxidative damage is increased in women after a short period of tomato consumption.

Several epidemiologic studies have suggested a role of tomato products in protecting against cancer and chronic diseases. In nine adult women, we evaluated whether the consumption of 25 g tomato puree (containing 7 mg lycopene and 0.3 mg beta-carotene) for 14 consecutive days increased plasma and lymphocyte carotenoid concentration and whether this was related to an improvement in lymphocyte resistance to an oxidative stress (500 micromol/L hydrogen peroxide for 5 min). Before and after the period of tomato intake, carotenoid concentrations were analyzed by HPLC and lymphocyte resistance to oxidative stress by the Comet assay, which detects DNA strand breaks. Intake of tomato puree increased plasma (P <0.001) and lymphocyte (P<0.005) lycopene concentration and reduced lymphocyte DNA damage by approximately 50% (P<0.0001). Beta-carotene concentration increased in plasma (P<0.05) but not in lymphocytes after tomato puree consumption. An inverse relationship was found between plasma lycopene concentration (r = -0.82, P<0.0001) and lymphocyte lycopene concentration (r = -0.62, P<0.01) and the oxidative DNA damage. In conclusion, small amounts of tomato puree added to the diet over a short period can increase carotenoid concentrations and the resistance of lymphocytes to oxidative stress.

Supplementation of Jurkat T cells with green tea extract decreases oxidative damage due to iron treatment.

Regular tea consumption has been associated with a reduced risk of cancer. As demonstrated in vitro, green tea contains catechins with antioxidant properties. We evaluated the effect of the supplementation of the Jurkat T-cell line with green tea extract on oxidative damage. Cells grown in medium with or without green tea extract (10 mg/L) were treated with Fe(2+) (100 micromol/L) as an oxidative stimulus for 2 h. Cell membrane lipid peroxidation was evaluated by fatty acids pattern analysis and malondialdehyde production in alpha-linolenic acid-loaded cells. Furthermore, oxidative DNA damage (single strand breaks) was detected in cells by the Comet assay and quantified as relative tail moment (RTM). Supplementation with green tea extract significantly decreased malondialdehyde production (1.6 +/- 0.3 vs. 0.6 +/- 0.1 nmol/mg protein, P < 0.05) and DNA damage (0.32 +/- 0.07 vs. 0.12 +/- 0.04 RTM, P < 0.05) after Fe(2+) oxidative treatment. In control cells, there was no effect on membrane distribution of (n-3) fatty acids due to Fe(2+) treatment. Cell enrichment with alpha-linolenic acid increased total membrane (n-3) fatty acids. However, the oxidative treatment did not modify the distribution of polyunsaturated fatty acids. It is likely that the observed protective effects can be attributed to epigallocatechin gallate, which is present mainly (670 g/kg) in green tea extract; however, we cannot exclude contributions by other catechins. These data support a protective effect of green tea against oxidative damage.

Liquid chromatography/electrospray ionization mass spectrometric characterization of flavonol glycosides in tomato extracts and human plasma.

Flavonoids continue to attract wide attention as possible very useful agents for combating free radical pathologies, i.e. the pathological states associated with free radical overproduction. Commonly used methods for the analysis of plant flavonoids include high performance liquid chromatography (HPLC) and capillary electrophoresis (CE). On the other hand, the soft-ionization approach based on electrospray ionization (ESI-MS) permits highly selective analysis of complex matrices. In this work, we examined firstly the ESI-MS behaviour of representative aglycones and glycosides of flavonols, flavones and isoflavones with the aim of suggesting a possible relationship between structure and mass spectra. Using HPLC coupled to a diode array detector (DAD) for on-line UV spectra acquisition, and in parallel to ESI-MS for mass spectra (LC/DAD-ESI-MS), we have developed methodology to observe flavonols directly in tomato puree extract. In this way, it has been possible to detect intact flavonol glycosides in tomato extracts and to characterize a flavonol trisaccharide. For the first time, using LC/ESI-MS, it has been possible to detect intact flavonol glycosides in plasma of healthy volunteers and to provide further evidence on the absorption of flavonoid glycosides after consumption of common vegetables like tomatoes.

Does tomato consumption effectively increase the resistance of lymphocyte DNA to oxidative damage?

BACKGROUND: Lycopene, the main carotenoid in tomato, has been shown to be a potent antioxidant in vitro. However, there is no significant evidence of its antioxidant action in vivo. OBJECTIVE: We evaluated the effect of tomato intake on plasma carotenoid concentrations and lymphocyte resistance to oxidative stress. DESIGN: Ten healthy women (divided into 2 groups of 5 subjects each) ate a diet containing tomato puree (providing 16.5 mg lycopene) and a tomato-free diet for 21 d each in a crossover design. Before and after each diet period, plasma carotenoid concentrations and primary lymphocyte resistance to oxidative stress (evaluated by means of single-cell gel electrophoresis) were analyzed. RESULTS: After the first 21-d experimental period, total plasma lycopene concentrations increased by 0.5 micromol/L (95% CI: 0.14, 0.87) in the group that consumed the tomato diet and decreased by 0.2 micromol/L (95% CI: -0.11, -0.30) in the group that consumed the tomato-free diet (P < 0.001). Tomato consumption also had an effect on cellular antioxidant capacity: lymphocyte DNA damage after ex vivo treatment with hydrogen peroxide decreased by 33% (95% CI: 0.8%, 61%; P < 0.05) and by 42% (95% CI: 5.1%, 78%; P < 0.05) in the 2 groups of subjects after consumption of the tomato diet. CONCLUSION: The consumption of tomato products may reduce the susceptibility of lymphocyte DNA to oxidative damage.

Absorption of lycopene from single or daily portions of raw and processed tomato.

To study the relationship between lycopene intake and plasma concentration, ten healthy female subjects were given one or more portions of tomato purée or fresh raw tomato containing 16.5 mg total lycopene (all-trans + cis forms). In Expt 1 subjects (n 9) were randomly assigned the single portions of the two tomato products and blood samples were collected to follow the change in plasma carotenoid concentrations within the first 12 h and on each of the following 5 d (104 h). In Expt 2 subjects (n 10) were divided into two groups of five each receiving daily dietary portions of tomato purée or fresh raw tomato containing 16.5 mg total lycopene for 7 d. Fasting blood samples were collected daily. In Expt 1 the plasma total lycopene (all-trans + cis forms) concentration, after the single portions of tomato purée and raw tomato, varied significantly over time, with a first peak reached after 6 h, a further increase after 12 h and a slow decrease until 104 h. In Expt 2, when the tomato products were given daily, there was a day-by-day increase in the plasma total lycopene concentration, and through the following week of a diet without tomato there was a gradual decrease. However, values did not return to basal concentrations. Plasma total lycopene concentration was higher after the tomato purée intake than after the raw tomato in both the first (F(1,8) 7.597; P < 0.025) and the second experiments (F(1,8) 12.193; P < 0.01) demonstrating a significant effect of food matrix on absorption.

Weight, protein, fat, and timing of preloads affect food intake.

Two foods, one rich in protein (HP) and one rich in fat (HF), were employed to evaluate the effect of macronutrients on food intake and to underline the differences that occurred when the foods were served as uniform meal, as first course of a varied meal, and as a snack 2 h before a varied meal. Our results showed that HP food always exerted a higher effect on both intrameal satiation and postingestive satiety than HF food. When a uniform meal was consumed, satiation for the specific food was reached before fullness; in this condition, sensory characteristics of foods played an important role in controlling food intake and made the uniform meal more satiating than the varied one. The consumption of a snack far from a meal did not contribute to satiety; consequently, gastric filling seems to be an important factor determining the amount consumed in a varied meal.

Determination of carotenoids in vegetable foods and plasma.

In this paper a HPLC method for the determination of lutein, zeaxanthin, beta-cryptoxanthin, alpha-carotene, beta-carotene and lycopene in mixed vegetables and fruit and in human plasma is described. The carotenoids were well separated and the separation was achieved within fifteen minutes using a HPLC system consisting of a 5 microns Vydac 201TP54C18 column, an UV detector, methanol-tetrahydrofuran (95:5 v/v) as mobile phase and a flow rate of 1.0 ml/min. The validity of the separation method was determined by evaluating the linearity of the calibration graphs of each carotenoid (between 0.1 and 1.0 microgram/ml for all compounds except lycopene between 0.1 and 0.8 microgram/ml, r = 0.999) and the accuracy of the chromatographic response (CV < 10%). The reproducibility of the retention times was also good. In the foods samples the extraction procedure was very effective whereas, the saponification step significantly damaged some of the carotenoids. In the plasma the extraction and separation of these compounds were also effective and the qualitative data obtained comparable with those reported in literature. The use of echinenone as internal standard helped to improve quality control.

Comparison of lactulose and inulin as reference standard for the study of resistant starch fermentation using hydrogen breath test.

Disaccharide lactulose is commonly used as a standard to quantitate the colonic fermentation of undigested sugars by means of H2 breath measurements. However, its high hydrogen production rate during fermentation may make it inappropriate for mimicking the fermentation of more complex carbohydrates, such as starch. Indigestible carbohydrates with a higher molecular weight might be more suitable than lactulose as a standard in H2 breath studies of starch digestibility. To test this hypothesis, we measured H2 breath in 8 healthy volunteers after a standard meal supplemented with 5 g or 10 g of lactulose or inulin, an indigestible oligosaccharide with an average degree of polymerization 4.5 times higher than that of lactulose. The results were then compared with those obtained after a standard meal containing a known amount (6.1 g) of resistant starch from high-amylose corn starch. Median H2 breath excretion per gram of reference carbohydrate was lower after the 5 g dose of inulin than after the 5 g dose of lactulose (19.1 vs 26.6 ppm x h x g-1; Wilcoxon's rank test p = 0.021) but similar after the two 10 g doses (inulin 22.4; lactulose 23.6; p = 0.234). Median H2 breath excretion per gram of resistant starch was significantly lower than that for both lactulose and inulin (p < 0.02), being 4.7 ppm x h x g-1. In vitro fermentation for 8 hrs with fecal homogenate showed similar mean hydrogen production rates for inulin and lactulose (30.5 vs 27.7 mL/mg fermented carbohydrate), and a significantly lower rate for starch (9.1 mL/mg) (n = 7; ANOVA p = 0.0007).(ABSTRACT TRUNCATED AT 250 WORDS)