基于体外全仿生消化模型分析海带和紫菜中微量元素的生物可给性

探究海带和紫菜中6种必需元素(Cu、Fe、Mn、Zn、Co和V)与5种有害元素(As、Cd、Pb、Al和Sr)的生物可给性及胃、肠消化过程中As、Cd等有害元素的形态转化,分析加工过程对样品中元素含量、形态及生物可给性的影响,结果表明:紫菜对除Sr外10种元素的富集能力均显著高于海带,经加工,海带和紫菜中6种人体必需微...     

Development of a three‐tier in vitro system,using Caco‐2 cells,to assess the effects of lactate on iron uptake and transport from rye bread following in vitro digestion

A three‐tier Caco‐2 cell system was developed to assess simultaneously iron dialysability, uptake and transport across the Caco‐2 monolayer from an in vitro digested food matrix. The effect of lactate (0–200 mmol L⁻¹) on iron absorption from rye bread subjected to simulated peptic (pH 5.5) and pancreatic digestion (pH 6.5) was investigated to model absorption pre and post the sphincter of Oddi. Lactate increased dialysability (11.8%, P < 0.05) in peptic digests whereas it reduced it in pancreatic digests (4.9%, P < 0.001). Iron uptake from the peptic digests was in the region of 39–76 pmol mg⁻¹ protein whereas it decreased from 281 to 51 pmol mg⁻¹ protein in pancreatic digests. Iron transport was calculated for the peptic digests from [¹⁴C]polyethylene glycol movement and only at 200 mmol L⁻¹ lactate was there any detectable transcellular transport (180 pmol mg⁻¹ protein, P < 0.05). Iron absorption was positively correlated to dialysable iron for both digests (R² = 0.48 and 0.41, respectively, P < 0.01) and the effect of lactate was therefore associated mainly with iron bioaccessibility. The three‐tier system showed the potential to obtain detailed insight into each step involved in iron transport across the monolayer from a food mixture. Copyright © 2006 Society of Chemical Industry     

混合膳食组成对体外消化过程中GI与食物消解的影响

对14种单品食物与8种混合膳食展开了体外消化实验,测定了消化过程中GI的变化过程,研究膳食中不同成分对血糖生成指数的影响,并通过显微镜观察食物在不同消化过程中结构和形貌变化,研究不同成分对食物消解过程的影响。根据食物碳水化合物的含量计算含有50 g碳水化合物的食物量,用50 g葡萄糖作为参照物,利用烧杯或者锥形瓶模拟人体食用食物后在胃部和小肠的4 h消化过程中葡萄糖变化情况。相对于单独的主食来说,加入蔬菜、肉类、蛋类或水果后的混合膳食出现明显的GI下降。研究食物消解过程也发现,混合膳食中米饭的形貌与结构变化会显著慢于单独米饭的外形变化;而挂面的消解趋势与其相反。结果证实了膳食纤维、脂肪和蛋白质会抑制淀粉类食物的消化过程,减缓葡萄糖释放速率。     

Verbascosides from olive mill waste water: assessment of their bioaccessibility and intestinal uptake using an in vitro digestion/Caco-2 model system

Olive mill wastewater (OMWW) is an agricultural waste material produced in high quantities in the Mediterranean basin. OMWW may be an inexpensive source of health promoting phytochemicals with potential economic value including many low molecular weight compounds such as verbascosides. While promising as antioxidants in vitro, little information is available on the potential absorption of verbascosides by humans. The main objective of the present study was to characterize the verbascoside content and potential for their bioavailability from a partially purified phenolic fraction (IP) of OMWW. The IP was obtained after ultrafiltration step at 5000 Dalton and gel filtration low-pressure chromatography (LH20) of OMWW. RP-HPLC analysis identified several soluble phenolics compounds including verbascoside and isoverbascoside as major components of OMWW fractions. The potential for bioavailability of these polyphenols was estimated by using both in vitro digestion and Caco-2 human intestinal cell models. In vitro digestive recoveries (bioaccessibility) were found to be 35.5%± 0.55% for verbascoside and 9.2% ± 0.94% for isoverbascoside highlighting potential sensitivity of these phenolics to gastric and small intestinal digestive conditions. Accumulation of verbascosides by highly differentiated Caco-2 monolayers was linear between 10 and 100 μM of verbascoside and isoverbascoside from IP extract. Uptake of verbascoside and isoverbascoside was rapid with peak accumulation occurring after 30 min with total accumulation efficiency of 0.1% and 0.2% providing intracellular levels of 130 and 80 pmol/mg cell protein for verbascoside and isoverbascoside, respectively. Combined, these data suggest that verbascosides present in OMWW are bioaccessible and provides a rationale for subsequent in vivo studies on the bioavailability and bioactivity of OMWW components.     

体外消化/Caco-2细胞模型分析发酵对面粉铁生物利用率的影响

中国居民膳食以谷物为主食,谷物中铁吸收率低是造成营养性贫血的主要原因,适当的加工方式可缓解谷物中植酸、多酚等物质对铁生物利用率(Fe bioavailability,FeBV)的影响。为考察酵母发酵对面粉FeBV的影响,采用体外消化/Caco-2细胞模型。结果表明,面团在发酵后pH值呈下降趋势,酸度呈上升趋势;多酚、植酸含量降低,植酸酶活性升高,以上各指标在发酵前后差异均具有统计学意义(P0.05)。发酵后面粉样品的FeBV增加约5%~38%,大多数面粉FeBV发酵前后比较,差异具有统计学意义(P0.05)。结论:发酵可降低面粉的pH值,增加酸度,有利于面粉多酚和植酸的降解以及植酸酶活力的升高,可有效提高面粉FeBV。     

In vitro β‐Carotene Bioaccessibility and Lipid Digestion in Emulsions: Influence of Pectin Type and Degree of Methyl‐Esterification

Citrus pectin (CP) and sugar beet pectin (SBP) were demethoxylated and fully characterized in terms of pectin properties in order to investigate the influence of the pectin degree of methyl‐esterification (DM) and the pectin type on the in vitro β‐carotene bioaccessibility and lipid digestion in emulsions. For the CP based emulsions containing β‐carotene enriched oil, water and pectin, the β‐carotene bioaccessibility, and lipid digestion were higher in the emulsions with pectin with a higher DM (57%; “CP57 emulsion”) compared to the emulsions with pectin with a lower DM (30%; “CP30 emulsion”) showing that the DM plays an important role. In contrast, in SBP‐based emulsions, nor β‐carotene bioaccessibility nor lipid digestion were dependent on pectin DM. Probably here, other pectin properties are more important factors. It was observed that β‐carotene bioaccessibility and lipid digestion were lower in the CP30 emulsion in comparison with the CP57, SBP32, and SBP58 emulsions. However, the β‐carotene bioaccessibility of CP57 emulsion was similar to that of the SBP emulsions, whereas the lipid digestion was not. It seems that pectin type and pectin DM (in case of CP) are determining which components can be incorporated into micelles. Because carotenoids and lipids have different structures and polarities, their incorporation may be different. This knowledge can be used to engineer targeted (digestive) functionalities in food products. If both high β‐carotene bioaccessibility and high lipid digestion are targeted, SBP emulsions are the best options. The CP57 emulsion can be chosen if high β‐carotene bioaccessibility but lower lipid digestion is desired.     

Assessment of iron bioavailability from twenty elite late‐maturing tropical maize varieties using an in vitro digestion/Caco‐2 cell model

An in vitro digestion/Caco‐2 cell model was used to assess iron bioavailability of twenty elite late‐maturing tropical maize varieties grown in three diverse agroecologies in West and Central Africa (WCA). Kernel‐iron concentration of the varieties, averaged across locations, varied from 19.2 to 24.4 mg kg^?1, while mean kernel‐zinc concentration ranged between 19.4 and 24.6 mg kg^?1. Significant differences in iron bioavailability were observed among varieties, but the environment had no significant effect. Mean bioavailable iron ranged between 14% below and 43% above the reference control variety, TZB‐SR. Variety DMR‐LSR‐Y with the highest concentrations of kernel‐iron and ‐zinc of 24–25 mg kg^?1 across the three locations had a similar quantity of bioavailable iron as the reference control, TZB‐SR. In the long run this variety could be potentially effective in reducing iron deficiency because of its high kernel‐iron. The most promising varieties were Mid‐altitude STR synthetic and ACR91SUWAN‐1‐SRC1. They had kernel‐iron and ‐zinc levels of 22–24 mg kg^?1 and bioavailable iron 24–36% higher than the reference control, TZB‐SR. Additional research is necessary to determine if the increases in kernel‐iron concentration and bioavailable iron observed in this study can significantly improve the iron status of individuals in WCA at risk for iron deficiency. Copyright © 2004 Society of Chemical Industry     

Effect of Acylglycerol Composition and Fatty Acyl Chain Length on Lipid Digestion in pH‐Stat Digestion Model and Simulated In Vitro Digestion Model

In this study, a pH‐stat digestion model and a simulated in vitro digestion model were employed to evaluate the digestion degree of lipids depending on different acylglycerols and acyl chain length (that is, diacylglycerol [DAG] compared with soybean oil representing long‐chain triacylglycerol compared with medium‐chain triacylglycerol [MCT]). In the pH‐stat digestion model, differences were observed among the digestion degrees of 3 oils using digestion rate (k), digestion half‐time (t_1/2), and digestion extent (Φmax). The results showed the digestion rate order was MCT > soybean oil > DAG. Accordingly, the order of digestion half‐times was MCT < soybean oil < DAG. In simulated in vitro digestion model, digestion rates (k′) and digestion half‐times (t′_1/2) were also obtained and the results showed a digestion rate order of MCT (k′ = 0.068 min^?1) > soybean oil (k′ = 0.037 min^?1) > DAG (k′ = 0.024 min^?1). Consequently, the order of digestion half‐times was MCT (t′_1/2 = 10.20 min) < soybean oil (t′_1/2 = 18.74 min) < DAG (t′_1/2 = 29.08 min). The parameters obtained using the 2 models showed MCT was digested faster than soybean oil, and that soybean oil was digested faster than DAG.     

体外消化/Caco-2 细胞模型评价面粉锌生物利用率

采用体外消化/Caco-2 细胞模型对10 种小麦粉进行锌生物利用率比较。以锌吸收率(Caco-2 细胞中的锌含量占加入到细胞的面粉消化液总锌量比例)反映锌生物利用率(Zn bioavailability,Zn BV)。发现不同面粉间锌生物利用率差异具有统计学意义,出粉率78% 的安阳中优9507 是面粉锌吸收率和可吸收锌量都高的品种,它将进入下一步人体实验进行营养功效的验证。     

Bioavailability of zinc from infant foods by in vitro methods (solubility,dialyzability and uptake and transport by Caco‐2 cells)

The aim of this study was to evaluate the bioavailability of zinc from infant foods (adapted, follow‐up and toddler milk‐based formulas and fruit juices containing milk and cereals, FMC) using solubility, dialyzability and a model combining simulated gastrointestinal digestion and zinc uptake and transport by Caco‐2 cells. The greater solubility of zinc from infant formulas compared with fruit juices (FMC) could be due to the greater casein phosphopeptide content resulting from casein hydrolysis. The highest zinc dialysis percentage corresponded to FMC, which on the other hand had the lowest zinc contents of the analyzed samples. The presence of organic acids in samples of this kind favors the formation of soluble low molecular weight complexes with zinc, thereby increasing the solubility of the latter. Bifidobacterium addition exerted no effect upon zinc bioavailability. Transport and uptake efficiency in Caco‐2 cells were significantly greater for toddler formulas, which presented the highest casein contents. The greater efficiency in zinc transport and uptake from the powdered toddler formula compared with the liquid formulation could be explained by the effect of Maillard reaction products. Copyright © 2006 Society of Chemical Industry     

基于体外模拟消化/Caco-2细胞模型测定大米中铅的生物有效性

为考察铅污染大米对人体的潜在健康风险,研究建立体外模拟消化/Caco-2细胞模型来测定大米中铅的生物有效性。结果表明：在模拟大米中铅的胃肠消化过程中,生米和熟米在胃消化阶段铅的生物利用率为61.34%～70.59%和39.69%～47.48%,肠消化阶段分别为24.39%～41.79%和13.57%～15.13%,经胃肠消化后籼米中铅的生物利用率均高于粳米,生米经胃、肠消化生物有效性程度均高于蒸煮后的大米。建立并利用成熟Caco-2细胞模型对大米中铅的生物有效性进行测定,使用四乙基铅与无机铅对大米加标,生米的生物有效性为3.32%和7.0%,熟米的生物有效性为2.65%和5.7%。体外模拟消化/Caco-2细胞模型是一种评价大米中铅生物有效性的有效手段。     

Bioavailability of iron from a heat treated glucose‐lysine model food system: assays in rats and in Caco‐2 cells

The influence of Maillard reaction products (MRP) from glucose–lysine on iron bioavailability was investigated. Equimolar glucose–lysine mixtures (GL, 40% moisture) were heated (150 °C, for 30, 60 and 90 min) to prepare samples GL30, GL60 and GL90, respectively, and were used to study the influence of MRP on iron solubility, iron uptake and transport in Caco‐2 cells and iron balance in rats. After an initial increase with GL30, in vitro iron solubility decreased in the presence of most of the heated samples. The Caco‐2 cell experiments showed that Fe cell content increased in the presence of heated mixtures with respect to GL, whereas transport was less affected. Iron balance was determined in rats fed diets containing the GL30 and GL90 samples (3%) for a 21 day period and compared with that measured in a control group. Consumption of glucose–lysine heated mixtures increased total iron retention, although hemoglobin values decreased. Iron accumulations in organs such as the spleen, kidney and small intestine were observed. Copyright © 2004 Society of Chemical Industry     

Development of Food‐Grade Curcumin Nanoemulsion and its Potential Application to Food Beverage System: Antioxidant Property and In Vitro Digestion

Curcumin nanoemulsions (Cur‐NEs) were developed with various surfactant concentrations by using high pressure homogenization and finally applied to the commercial milk system. Characterization of Cur‐NEs was performed by measuring the droplet size and polydispersity index value at different Tween 20 concentrations. The morphology of the Cur‐NEs was observed by confocal laser scanning microscopy and transmission electron microscopy. Antioxidant activity and in vitro digestion ability were tested using 2,2‐diphenyl‐1‐picrylhydrazyl, 2,2′‐azino‐bis(3‐ethylbenzothiazoline‐6‐sulfonic acid) diammonium salt, pH‐stat method, and thiobarbituric acid reactive substances assays. Cur‐NEs were found to be physically stable for 1 mo at room temperature. The surfactant concentration affects particle formation and droplet size. The mean droplet size decreased from 122 to 90 nm when surfactant concentration increased 3 times. Cur‐NEs had shown an effective oxygen scavenging activity. Cur‐NEs‐fortified milk showed significantly lower lipid oxidation than control (unfortified) milk and milk containing curcumin‐free nanoemulsions. These properties make Cur‐NEs suitable systems for the beverage industry.     

Determination of S‐methyl‐L‐methionine (SMM) from Brassicaceae Family Vegetables and Characterization of the Intestinal Transport of SMM by Caco‐2 Cells

The objectives of the current study were to determine S‐methyl‐L‐methionine (SMM) from various Brassicaceae family vegetables by using validated analytical method and to characterize the intestinal transport mechanism of SMM by the Caco‐2 cells. The SMM is well known to provide therapeutic activity in peptic ulcers. The amount of SMM from various Brassicaceae family vegetables ranged from 89.08 ± 1.68 μg/g to 535.98 ± 4.85 μg/g of dry weight by using validated ultra‐performance liquid chromatography‐electrospray ionization‐mass spectrometry method. For elucidating intestinal transport mechanism, the cells were incubated with or without transport inhibitors, energy source, or a metabolic inhibitor. Phloridzin and verapamil as inhibitors of sodium glucose transport protein (SGLT1) and P‐glycoprotein, respectively, were not responsible for cellular uptake of SMM. Glucose and sodium azide were not affected by the cellular accumulation of SMM. The efflux ratio of SMM was 0.26, implying that it is not effluxed through Caco‐2 cells. The apparent coefficient permeability (P _app) of SMM was 4.69 × 10^?5 cm/s, indicating that it will show good oral absorption in in vivo .     

Iron Bioavailability from Common Raisin-containing Foods Assessed with an In Vitro Digestion/Caco-2 Cell Culture Model: Effects of Raisins

The effects of raisins on iron bioavailability from wheat bran cereal, bread, rice pudding, and granola bars were studied. Iron bioavailability was assessed with an in vitro digestion/Caco‐2 cell culture model. Raisins reduced iron bioavailability from all foods except granola bars. Raisins also reduced iron bioavailability from samples of wheat bran cereal and bread fortified with elemental iron or ferrous sulfate, but this inhibitory effect was less pronounced in samples fortified with sodium iron ethylenediaminetetraacetate (NaFeEDTA). Iron bioavailability was markedly higher for samples fortified with NaFeEDTA, suggesting that iron in the form of NaFeEDTA is more bioavailable than elemental iron or ferrous sulfate in raisin‐containing foods.     

Bioaccessibility,cellular uptake and transepithelial transport of α‐tocopherol and retinol from a range of supplemented foodstuffs assessed using the caco‐2 cell model

The bioaccessibility, or amount of a nutrient available for gastrointestinal absorption, can be determined using an in vitro digestion model, the addition of the resultant digestate to a caco‐2 transwell model system yields an approximation of nutrient bioavailability. The objective of the present study was to compare the bioaccessibility and bioavailability of α‐tocopherol and retinol from a range of digested foodstuffs. Minced pork, beef and turkey and apple sauce, bread and mayonnaise were supplemented with α‐tocopherol‐acetate and retinol‐acetate prior to being subjected to an in vitro digestion procedure. The aqueous fraction of each of the digested foodstuffs was then added to a caco‐2 transwell model and the transepithelial transport was determined. The findings of the present study indicate that α‐tocopherol and retinol are more bioaccessible from supplemented meat products than from supplemented apple sauce, bread and mayonnaise. It was found that turkey meat facilitated the highest bioaccessibility and subsequent cellular uptake and transport of retinol. The cellular uptake and secretion of α‐tocopherol was similar for all samples.