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Analysis of phytic acid in foods by HPLC
Journal of Food Science
Abstract
ABSTRACTA quantitative HPLC method for the analysis of phytic acid in foods was developed based on the precipitation of phytic acid with ferric chloride followed by conversion to sodium phytate before injection onto a C18 reversed-phase column. Standard food grade wheat bran samples were analyzed by the method of standard addition and recovery of phytic acid ranged from 99 – 103%. 3% H2SO4 was found to be as effective as 3% TCA in the extraction of phytic acid. AAS was shown to be potentially valuable as a metal specific detector for the HPLC of phytate-metal complexes.
... Being a vital player of the biosynthetic pathway among the species, this gene can be targeted for gene silencing and genome editing approaches. Identification and characterization of biosynthetic pathway enzymes help in the development of low phytic acid mutant varieties [10]. Earlier reports for IPK gene have been studied in soybean which can be the reference for other legumes [10]. ...
... Identification and characterization of biosynthetic pathway enzymes help in the development of low phytic acid mutant varieties [10]. Earlier reports for IPK gene have been studied in soybean which can be the reference for other legumes [10]. Molecular characterization of IPK2 gene in the Pusa variety of soybean by in-silico technologies could serve as a potential candidate for developing low phytic acid mutants in legumes [97]. ...
... In many cereals like maize, rice, and barley, low phytate mutant lines have been identified which serve as genetic resources for producing the seeds in order to develop the mutant lines with significantly reduced phytic acid accumulation [10,21]. Manipulation of Ins(3)p1 synthase gene in rice was done in order to develop the low phytate crops. ...
Phytic acid is considered as one of the most abundant storage forms of phosphorous in the plant seed tissues. It accumulates throughout seed development until maturity and accounts for 60–70% of the total phosphorus in legume seed. Its amount ranges from 0.40 to 2.06% in legumes. It is a strong cationic chelator and negatively impacts the bioavailability of essential micronutrients like iron (Fe) and Zinc (Zn). It has very limited digestibility in humans and other non-ruminants which lack phytases. Besides, it also acts as an inhibitor of salivary and gastric enzymes during digestion. On the contrary, the beneficial effects of phytic acid include its role as an antioxidant, anti-carcinogen, and regulator of blood glucose and cholesterol levels. Several biotechnological strategies have been employed to reduce the phytic acid content in plants especially legumes for better absorption of essential micronutrients and in turn for bio-fortification to address the malnutrition challenges across the world. This review focuses primarily on the mechanisms governing phytic acid biosynthesis and its regulation/ reduction using various biotechnological, mutation breeding and computational approaches. This will help us in understanding the molecularity of the phytic acid development and will also contribute towards developing newer and more efficient strategies aimed at regulating phytic acid content in legumes.
... Metoda HPIC (High performance ion chromatography) se koristi kao specifi čna za određivanje inozitol fosfata (Pontoppodian et al. 2007, Camire & Clydesdale 1982 i kao referentna metoda za poređenje rezultata sa indirektnim metodama, kao što je modifi kacija Haug & Lantzsch (1983) (Reicwald & Hatzack 2008). Precizno određivanje sadržaja fi tinske kiseline, na koju ne utiče proteinski sadržaj, omogućeno je ultrafi ltracijom na membrani (30kDa) i ispiranjem proteinskih anjona sa kolone pre inozitol fosfata i fi tinske kiseline. ...
... Autori Febles et al. (2001) su utvrdili da je koncentracija fi tinske kiseline u rafi niranom pšeničnom brašnu iznosila 2-4 mg g -1 , dok je za brašno dobijeno od celog zrna bila 6-10 mg g -1 . Camire & Clydesdale (1982) su utvrdili da sadržaj fi tinske kiseline kod mekinja dobijenih od durum pšenice crvenog zrna iznosi 68,8 ± 1,85 mg g -1 , dok je kod mekinja dobijenih od hlebnih pšenica belog zrna iznosila 50,27 ± 1,45 mg g -1 . ...
Interest in wheat breeding for low phytic acid content arised from its roll as antinutrient factor which chelates mineral elements (Ca, Zn, Fe, Mn, Cu and P), leading to their inadequate use. Excretion of unused P in phytic acid complex through non-ruminant animals such as poultry, swine and fish causes water eutrophication. Numerous indirect methods (e.g. spectrophotometric) and direct methods (HPLC - High Performance Liquid Chromatography) were developed for fast and accurate phytic acid determination in wheat. It typically represents 50-85% of seed total phosphorus and one to several percents of dry seed weight. Phytic acid content and phytate phosphorus genetic variability have been determined for wheat cultivars and lines under different environmental conditions. Wheat mutant (Triticum aestivum L) for low phytic acid content Js-12-LPA was created through breeding efforts.
... O procedimento se baseia na extração ácida das amostras seguida de purificação em coluna de troca aniônica forte (SAX), secagem do eluato até resíduo e dissolução do resíduo na fase móvel antes da injeção. O método de detecção mais freqüentemente utilizado é o índice de refração embora a espectrofotometria de absorção atômica também tenha sido usada [9]. Neste caso, a análise de complexos metal-fitato foi executada acoplando-se o cromatógrafo líquido a um espectrômetro de absorção atômica junto à saída do detector de índice de refração e utilizando como padrão fitato monoférrico. ...
... Desde as primeiras pesquisas baseadas na cromatografia de fase reversa [9,22,23], foi demonstrada a eficiência da coluna aniônica nos processos de purificação e concentração das amostras. LEHRFELD [24] e SANDBERG et al. [40] já haviam demonstrado o uso de par iônico na fase reversa para identificar e quantificar este composto e seus derivados desfosforilados, concluindo que suas separações e quantificações foram eficientes. ...
Considerando a composição da multimistura distribuída em Belo Horizonte/MG (trigo, fubá, casca de ovo e folha de mandioca), as metodologias de detecção e/ou quantificação dos fitatos e oxalatos foram validadas. A purificação em coluna de troca aniônica forte do ácido fítico e seus derivados desfosforilados extraídos da multimistura apresentou baixos valores de recuperação (49%) nas concentrações menores do fitato adicionado, em relação às mais elevadas (101%), sugerindo a interferência de minerais presentes em altas concentrações, principalmente cálcio. Dos métodos avaliados para a determinação do ácido oxálico, a cromatografia de exclusão iônica foi a que apresentou os melhores resultados, com boas recuperações nos níveis mais altos de adição (88%), apresentando, contudo, recuperação mais baixa nos níveis inferiores de adição (61%). Os teores de ácido fítico nas amostras variaram de 1,61 g/100g a 2,25g/100g e, para o ácido oxálico, de 0,044 g/100g a 0,057 g/100g. Os critérios de validação dos métodos analíticos por cromatografia líquida de alta eficiência foram considerados satisfatórios quanto a seletividade e especificidade, linearidade, repetitividade e limites de detecção e quantificação. Contudo, o de recuperação não foi atendido satisfatoriamente, visto que as porcentagens obtidas foram inferiores ao esperado, levando-se em conta a concentração adicionada nas amostras.
... The results also showed that 5% of H 2 SO 4 gave the highest amount of phytic acid as compared to 10% of TCA and 0.8 M of HCl. This finding is consistent with that of Camire and Clydesdale (1982) which reported that H 2 SO 4 gave the highest content of phytic acid in wheat bran. They also reported that 3% of TCA was more effective than 3% of H 2 SO 4 in the extraction of phytic acid from wheat protein concentrate. ...
... The length of extraction time used were similar as used by Averill and King (1986) that analyzed a large number of foodstuffs using 3 h extraction period for their analyses. Studies by Tangendjaja Camire and Clydesdale (1982) and Cilliers and Niekerk (1986) stated 2 h of extraction time with the introduction of HPLC to the analysis of phytate. Also, when the anion-exchange concept was incorporated into quantitative phytate analysis, Harland and Oberleas (1986) considered 2 h extraction period length appropriate, while Graf and Dintzis (1982) used 2 h extraction time by using 0.5 N of HCl. ...
Phytic acid is one of the bioactive compounds that are being intensively studied to evaluate their effects on health. This study was carried out to determine the optimum condition for phytic acid extraction from rice bran. Three main parameters were considered to optimize the condition for phytic acid extraction; different types of extracting solvent, length of extraction time and influence of different pH adjustment. Three acidic solutions were selected and were trichloroacetic acid (TCA), hydrochloric acid (HCl) and sulphuric acid (H 2 SO 4). The results showed that 5% of H 2 SO 4 in pH 0.6 and after 30 min of extraction time gave the highest amount of phytic acid compared to 10% of TCA and 3% of HCl. The content of phytic acid in rice bran ranged from 0.22 to 2.22% for the different parameters optimized. Different methods produced different content of phytic acid from rice bran. Therefore, 5% of H 2 SO 4 in pH 0.6 and 30 min of extraction time was the best condition for the optimum production of phytic acid.
... Determination of phytates. Phytic acid was analyzed by reversephase high performance liquid chromatography (HPLC) with slight modification (Camire & Clydesdale, 1982). Precisely 5 g of the ground sample were extracted using 3% H 2 SO 4 , and ferric chloride was used to precipitate the phytic acid. ...
... The phytic acid content in the insect sample was analyzed using HPLC based on a method described earlier by (Camire and Clydesdale, 1982) with slight modification. First, 5 g of the blended insect sample was mixed with 3% sulfuric acid and subsequently, phytic acid precipitated using iron (III) chloride, centrifuged at 2,500 rpm for 10 min after which the supernatant was discarded. ...
The prevailing global market demands locally produced, sustainable oils for biomedical applications. This study focused on evaluating the quality of cricket-derived oils and meals from Scapsipedus icipe Hugel, Tanga, and Gryllus bimaculatus De Geer common delicacy in Africa, following standard methods for physicochemical properties, fatty acid composition, and phytochemicals (oxalates, phytates, tannins, and polyphenols). The cricket oils physicochemical properties aligned with Codex Alimentarius standards for edible oils, including low solidification temperature (< 2 °C), a high refractive index (1.46), and a specific gravity of 0.88. Notably, peroxide values (1.9 to 2.5 mg mEq O2/kg), acid values (1.1 to 2.2 mg KOH/g), and saponification values (234–246 mg KOH/g) all are indicative of lightness and unsaturated fatty acids. Nutritionally, cricket powder was rich in protein (56.8–56.9% -) and fat (31.7–33.5% -of dry matter), with significant amounts of essential omega-3 and omega-6 fatty acids. Predominant saturated and monounsaturated fatty acids were palmitic (23.9–31.2 mg/100 g-) and oleic acids (10.9–11.4 mg/100 g- of oil), respectively. Antioxidant values (48.0 to 65.0 mg/100 g), inferred from total polyphenols, suggests a stable oil with long shelf-life. These results highlight the promising and sustainable potential of cricket-derived oils for applications in the food and pharmaceutical industries.
... The phytate content was determined using high-performance liquid chromatography (HPLC) (Camire and Clydesdale, 1982). The level of phytates was quantified using an HPLC apparatus (Model C-R7A plus, Shimadzu Corp., Kyoto, Japan) fitted with a 50377 RP-18 (5 μm) column cartridge at an oven temperature of 30°C, and a refractive index detector (RID) (Model RID-6A, Shimadzu Corporation, Kyoto, Japan) for identification. ...
Fortification with micronutrient powders (MNPs) is recommended as a strategy for increasing the micronutrient content in complementary foods. However, plant-based diets commonly consumed in developing countries are rich in phytates and tannins, which decrease the micronutrient bioavailability. The present work analysed the relationship between the antinutrient content, and also iron and zinc bioaccessibility, in home-made MNP-fortified complementary feeding porridges refined with white rice, maize, white sorghum, finger millet, pearl millet, Irish potato, and banana samples, which were obtained from the local market and milled into flour. Porridges were prepared using the flour, cooled to 50°C, fortified with MNPs, and subjected to in vitro digestion. Total and bioaccessible zinc and iron were quantified using atomic absorption spectrometry. Tannins and phytates were analysed using Folin-Denis and high-performance liquid chromatography methods, respectively. Porridges were classified as having poor bioavailability if their phytate-zinc and phytate-iron molar ratios were > 15 and > 0.4, respectively. Only pearl millet and soybeans showed tannin levels higher than the recommended values. The lowest phytate level was observed in refined white rice (0.11 ± 0.04 g/100 g), and the highest was in pearl millet (2.83 ± 0.10 g/100 g). Zinc bioaccessibility ranged from 7.31% (finger millet) to 26.05% (corn-soy blend). Only pearl millet porridge was classified as having poor zinc bioavailability. Iron bioaccessibility ranged from 20.73% (refined white rice) to 0.62% (pearl millet). Refined white rice and Irish potato were the only foods with the phytate-iron ratio within the recommended range. Iron bioaccessibility decreased significantly with an increase in both tannin (r = -0.31, p = 0.045) and phytate (r = -0.39, p = 0.01) contents. Zinc bioaccessibility showed a significant positive relationship with tannin levels (r = 0.41, p = 0.008), but an insignificant inverse relationship with phytate levels (r = -0.072, p = 0.700). Iron bioaccessibility was adversely affected by phytate and tannin levels. To improve iron and zinc bioavailability in complementary foods, strategies for lowering the phytate and tannin contents at the household level are recommended.
... Phytate content was determined using the methodology described by Camire [22]. Ten milliliters of 3% H 2 SO 4 was used to extract 0.5 g of the sample. ...
... Phytates were extracted using the method of Camire and Clydesdale (2006) with modification. About 0.5 g of milled bean flours was extracted. ...
Common beans (Phaseolus vulgaris L.) are nutritious and confer numerous health benefits. However, they are also high raffinose family oligosaccharides (RFOs) and antinutrients. Appreciable amounts of RFOs and antinutrients remain after soaking and cooking, causing flatulence and lowered mineral bioavailability to bean consumers. Fermentation has been shown to lower RFOs and antinutrients in bean flours and milk. However, beans are majorly consumed as whole grains. The purpose of this study was to develop a protocol for fermenting whole common beans. We fermented boiled whole red haricot beans and evaluated their effect on RFOs, tannins, and phytates. A factorial research design was used. Beans were sorted, soaked for 15 h, and boiled for 1 h. The beans were then fermented in 2% salt–sugar solution (SSF) and 2% salt‐only solution (SOF) for 120 h. Microbial growth and pH were monitored every 24 h during fermentation. After fermentation, the beans were dried, milled, and the flours subjected to biochemical analysis. Fermentation favored the growth of lactic acid bacteria (LAB), lowering the pH to 3.88 and 5.26 in SSF and SOF batches, respectively. Tannin content reduced significantly by 64.70% and 73.19% in the SSF and SOF batches, respectively. Phytates reduced by 58.88% and 68.85%, respectively. Raffinose reduced significantly by 96.40% and 95.01%, respectively, whereas stachyose reduced by 95.92% and 94.11%, respectively. The highest reduction of antinutrients and RFOs occurred between 24 and 72 h of fermentation. Higher antinutrient losses occurred in the SOF batch, whereas higher RFO losses occurred in the SSF batch.
... Phytate content was determined according to [42] using the HPLC tannin, and saponin content was determined colorimetrically [43] with a UV-Vis spectrophotometer (Shimadzu model UV-1601 PC, Kyoto, Japan). ...
Complementary foods must be adequate to satisfy the nutritional needs of the growing child together with breastfeeding. This study
was aimed at evaluating the nutritional composition, microbial safety, and sensory quality of extruded complementary foods
developed from blends of staple grains and insect bee larva (Apis mellifera). Teff, maize, soybean, and bee larva samples were
milled to flour and blended before extrusion as follows: ComF01 (57% maize, 29% teff, and 14% soybean) and ComF02 (58%
maize, 29% teff, and 13% bee larvae) using NutriSurvey software (version, 2007). Nutrient composition, microbial, and sensory
analyses of developed flour blends were conducted using standard methods. The proximate composition of moisture, fat, fiber,
carbohydrate, and energy was significantly different between the developed and commercial wean-mix foods. ComF02 recorded
the highest fat content (14.3 g/100 g), energy (427.18 kcal/100 g), and vitamins A (706 μg/100 g), B3 (8.2 mg/100 g), and B9
(86.7 mg/100 g) while ComF01 had the highest protein content (12.56 g/100 g). Iron (40.94 mg/100 g) and calcium
(68.20 mg/100 g) were the minerals with the highest content in ComF02. Both ComF01 and ComF02 met the recommended
dietary allowance of nutrients for infants aged 6-12 months. Overall, the present study showed that bee larvae can be used to
develop complementary foods that are nutritionally adequate, microbiologically safe, and sensory acceptable meeting the dietary
allowance of infants at an acceptable level compared to conventional cereal-based foods.
... In the early 80s, the development of different separation techniques brought also the first applications of high-performance liquid chromatography (HPLC) for the analysis of food samples, as reported by Tangendjaja et al. [60] (1980) who determined phytic acid content in rice bran. As reported also by food studies of Camire and Clydesdale [61], Knuckles et al. [62], and Graf and Dintzis [63] (all in 1982) the separation was performed on the reversed-phase columns and a differential refractive index (RI) detector was used for the analysis of different inositol phosphates. However, the applicability of these methods was limited due to the difficulties with separation of inositol phosphates and quantification as the solvent front coincided with the phytic acid peak. ...
From the early precipitation-based techniques, introduced more than a century ago, to the latest development of enzymatic bio- and nano-sensor applications, the analysis of phytic acid and/or other inositol phosphates has never been a straightforward analytical task. Due to the biomedical importance, such as antinutritional, antioxidant and anticancer effects, several types of methodologies were investigated over the years to develop a reliable determination of these intriguing analytes in many types of biological samples; from various foodstuffs to living cell organisms. The main aim of the present work was to critically overview the development of the most relevant analytical principles, separation and detection methods that have been applied in order to overcome the difficulties with specific chemical properties of inositol phosphates, their interferences, absence of characteristic signal (e.g., absorbance), and strong binding interactions with (multivalent) metals and other biological molecules present in the sample matrix. A systematical and chronological review of the applied methodology and the detection system is given, ranging from the very beginnings of the classical gravimetric and titrimetric analysis, through the potentiometric titrations, chromatographic and electrophoretic separation techniques, to the use of spectroscopic methods and of the recently reported fluorescence and voltammetric bio- and nano-sensors.
... This result supported the view that wheat and bran were rich sources of phytic acid. It was stated that phytic acid was found mostly in the outer layers of the wheat grain, and that different wheat fractions obtained by milling process had different phytic acid content [25]. Moreover, it was indicated that the bran of wheat and rice contains a considerable amount of phytic acid (myo-inositol hexaphosphate) which acts as an antinutrient substance [10]. ...
Introduction. The aim of the study was to the examination of effects of the milling process which were applied differently to cereals for commercially flour production, on the phytic acid, microelement, and in-vitro digestibility. Materials and methods. The nutritional consequences of the milling processes of the cereals (wheat, barley, rye, oat, paddy) were evaluated by examining protein, ash, phytic acid, mineral contents, and protein/mineral digestibility rates. Mineral contents were measured by inductively coupled plasma optical emission spectroscopy (ICP-OES), protein content by Kjeldahl method, and mineral digestibility and phytic acid were determined by in-vitro assays. Results and discussion. The dehulling/debranning process increased the protein digestibility rate over to 65% and decreased phytic acid content significantly (p<0.05) of the cereal grains except for rye. The mineral digestibility raised for all grain samples, but it was evaluated statistically insignificant (p>0.05). The rice flour was the sample that had the lowest phytic acid content (921.87 mg/100g) and the highest mineral digestibility rate (58.35%). Although the contents of total ash and some of the minerals (Na, Ca, K, Mg, Zn, Fe, Ba, and P) decreased in flours especially in wheat and paddy, the rate of some minerals (Na, Mg, Zn, Fe, and Al) increased due to their distribution and localization in the grain layers, especially in rye and oat. The result of the study has shown that the level of the minerals changed depending on the bran/hull content of the grains, and the milling process was more effective on phytic acid content and on protein digestibility than on the mineral digestibility. While the phytic acid content was located on the outer layer of the grain, it was decreased in the kernel. Conclusions. Although the phytic acid content which affects the digestibility of the cereals were reduced by the milling process, the combination of different treatments such as soaking, fermentation could be suggested for improving the nutritional quality of the cereal.
... Phytates content was determined using HPLC as described by (Camire, & Clydesdale. (2006). To achieve this, about 0.5 g of each sample was extracted with 10 mL of 3% H 2 SO 4 and shaken at the automatic shaker for 45minutes. The contents were filtered and the filtrate was transferred to a boiling water bath to heat for 5 minutes followed by addition of 3 mL of FeCl 3 solution (6 mg ferric iron per mL in 3% H 2 SO 4 ). The co ...
This study was carried out to evaluate the effects of boiling and wet frying on nutritional and antinutrients content of Amaranth hybridus, Moringa oleifera, Bidens pilosa (black jack), Corchorus olitorius (Jute mallow) and Ipomea batatas (sweet potato) leaves. The edible portions of the vegetables were either boiled or wet fried for ten minutes then dried alongside the raw vegetables under the shade. Crude fats, minerals, vitamins and antinutrients were determined in the dried materials. Wet frying increased the oil content of the vegetables by a range of 15.49% to 28.40 % and was hence associated with lower % ash and mineral contents. Wet frying significantly reduced (P≤0.05) beta-carotene in all the vegetables except in jute mallow. Boiling had no significant effect on beta-carotene in most of the vegetables. Boiling significantly reduced (P≤0.05) ascorbic acid in all the vegetables while wet frying preserved ascorbic acid in all the vegetables. Both boiling and wet frying significantly reduced (P≤0.05) oxalates in all the vegetables except in black jack. Both boiling and wet frying significantly (P≤0.05) reduced the concentration of phytates in most of the vegetables. However, boiling was more effective in reducing the amount of phytates. Boiling reduced higher concentrations of tannins in all the vegetables as compared to wet frying. Boiling was associated with better retention of minerals and beta-carotene, and greater reduction of antinutrients in most of the vegetables. Wet frying was more advantageous in retaining vitamin C. The different species showed differences in retention of various minerals and vitamins.
... The drug release study of microsphere formulation was carried out using similar procedure adopted earlier for drug release in normal colonic fluid of pH 7.4. The samples withdrawn for analysis were further centrifuged, filtered using 0.45 μm membrane filter, and diluted suitably with PBS if required to determine the IP6 content at 520 nm using UV-visible spectroscopy (14,19,23,(28)(29)(30)(31)(32)(33)(34)(35)(36)(37)(38)(39). ...
Colorectal cancer has become the third most frequent reason of cancer death in men and women. Currently, natural compounds are being looked up to, for subversion and deterrence of cancers. Inositol hexaphosphate (IP6) is one such naturally occurring phosphorylated carbohydrate present in most legumes and cereals which acts as a potential antineoplastic agent and can be used effectively to prevent and treat colon carcinomas. Despite the immense potential, due to the prevalence of high charge and ability to form salts and chelates with various divalent metals, it gets excreted out quickly from the body. On reaching the colon in its original form, it can serve as an effective anticancer agent. Therefore, a suitable dosage form that can prevent the drugs from being absorbed from the upper gastrointestinal tract is required to be prepared, to target it to the colon. Thus, microspheres of IP6 using a biodegradable polymer that degrades in the colon were attempted using the solvent evaporation method. The formulation was investigated for percentage yield, encapsulation efficiency, particle size distribution modification, and release rate. Optimized formulation showed particle size of 92 ± 0.76 μm, entrapment efficiency of 67.26% ± 0.75, percent drug loading of 15.74%, and in vitro drug release 82.36 ± 0.51. The results of the in vivo study divulged that IP6 loaded pectin microspheres showed significant positive modulation of biomarker levels and restoration of colonic architecture to almost normal as observed through histopathology and scanning electron microscopy studies in DMH-induced colon tumors in Albino Wistar rats.
... Phytic acid content in the bread roll was determined by the HPLC (High Performance Liquid Chromatography)according to the method described by Camire and Clydesdale [23]. ...
Iron deficiency is the most common form of malnutrition. Factors responsible for this so-called “hidden hunger” include poor diet, increased micronutrient needs and health problems such as diseases and infections. Body iron status can be increased by the intake of dietary supplements and fortified food. The aim of the present study was to compare iron bioaccessibility from commercial nutritional supplements and iron microcapsules. A comparison study was performed under conditions mimicking gastric and gastrointestinal digestion. A preparation of encapsulated ferrous sulphate or lactate and vitamin C, in a formula, showed bioaccessibility factors of up to 100% when digested individually, and around 60% in the presence of a food matrix. The degree of oxidation of the ferrous ions differed, depending on the type of preparation, the presence of vitamin C and the food matrix. The highest percentage content of ferrous ion, in the soluble fractions after gastrointestinal digestion, was shown by the preparation containing microencapsulated ferrous lactate or ferrous sulphate and vitamin C. Encapsulation seems to limit the interaction of iron with the food matrix and protect it against oxidation, thus making it more accessible for intestinal uptake.
... PA in wheat grain was extracted using the method described by Dost and Tokul (2006) and Camire and Clydesdale (1982) with slight modification. Briefly, about 0.2 g of grain sample was weighed and 10 mL HCl (0.5 mol L −1 ) was added into a centrifuge tube. ...
Although winter wheat (Triticum aestivum L) are commonly colonized by arbuscular mycorrhizal fungi (AMF) under field conditions, the effects and related mechanisms of the fungi on concentrations and bioavailability of Zn in grain is still unclear. A pot trial was done to survey the influence of two species of AMF (Funneliformis mosseae BGC HEB02 and Rhizophagus intraradices BGC HEB07D) on grain yield, concentrations of phytic acid and Zn in winter wheat (T. aestivum cv. Xiaoyan22) in Zn-spiked soils under two levels of P application. The present study showed that mycorrhizal inoculation apparently improved P nutrition in wheat and significantly (p < 0.01) increased grain yield, especially under low soil P conditions. Increasing levels of P and Zn in soils significantly (p < 0.01) enhanced phytic acid concentrations in wheat grain, while inoculation of AM fungi had insignificant effect on grain phytic acid concentrations. Grain Zn concentrations under the mycorrhizal treatments were 1.13–2.76 times of those without inoculation, especially for R. intraradices. The present study revealed that concentrations and bioavailability of Zn in wheat grain can be significantly increased by inoculation of AMF, indicating the potential of AMF to cope with Zn deficiency for residents living on wheat as their staple food.
... While the amounts of phytic acid were found to be 42 mg / g-54 mg / g and 46 mg / g-67 mg / g, in hard and soft wheat brans respectively with colorimetric methods (Camire andClydesdale 1982, Knuckles et al 1982) ), in a study carried out by Kasim ve Edwards (1998) with the HPLC method the amount of phytic acid in the wheat barn was reported as 39.5 mg / g. Harland and Oberleas (1986) reported that phytic acid values in bran varied between 34 mg / g and 47 mg / g. ...
Being a phosphorous storage and energy source, phytic acid is an important organic compound. Phytic acid has many benefits in terms of health, such as being an antioxidant. Even though phytic acid is beneficial source, it can be dangerous due to its ability related to binding minerals. At this point, it is important to know the effect of phytic acid. The present book will be of use for researchers and students keen on studying the structure, sources, and functions of phytic acid. The book explains history and physiological functions of phytates, chemical structure and sources of phytic acid, and finally the effects of some technological processes on the concentration of phytic acid in details.
... The protein percent in the lablab beans was multiplied by 3.47, fats multiplied by 8.37 and carbohydrates by 4.07. Determination of phytic acid: Phytic acid was determined using HPLC according to the method of Camire and Clydesdale (1982). About 0.5g of the sample was extracted with 25 mL of 3% H2S04 in Erlenmeyer flasks for 30 min. ...
... The tannin content of the amaranth-sorghum product was determined using the modified vanillin-HCl method (Makokha et al. 2002adapted from Price et al. 1978and Burns 1963. The phytate content was determined by the Camire and Clydesdale (1982) method. In vitro-protein digestibility was determined by the Mertz et al. (1984) method. ...
Thin porridge from cereals and starchy tubers is a common complementary food in Sub Saharan Africa. It may be high in antinutrients, low in energy, and nutrient density hence inadequate in providing infants' high energy and nutrients requirements per unit body weight. Consequently, undernourishment levels among children under 5 years are high. Therefore, there is need to avail nutrient‐dense complementary foods especially for children in low‐resource settings. The study was aimed at developing a nutrient‐dense complementary food from amaranth and sorghum grains. Amaranth grain, a pseudocereal, though rarely used as a complementary food in Kenya has a higher nutritional quality than other staples. Plant‐based foods are known to have high levels of antinutrients. Steeping and germination were used to reduce the levels of antinutrients and enhance the bioavailability of minerals in the grains. Various steeped and germinated amaranth and sorghum grains formulations were made to find the ratio with the highest nutrient content and lowest antinutrient levels. The 90% amaranth‐sorghum grains formulation had significantly (F = 32.133, P < 0.05) higher energy (5 kcal per g on dry weight basis) than the other formulations and a protein content of 14.4%. This is higher than the estimated protein needs from complementary foods even for a 12–23 months child of low breast milk intake (9.1 g/d). Antinutrients could not be detected which could imply enhanced nutrient bioavailability. Therefore, a nutrient‐dense complementary food product was developed from steeped and germinated amaranth and sorghum grains with 90% amaranth grain. In ready to eat form, it would give an energy content of 1.7 kcal per g (dilution of 1:2 amaranth‐sorghum flour to water) and 1.2 kcal per g (dilution of 1:4 amaranth‐sorghum flour to water). It can be used as a nutrient‐dense complementary food and for other vulnerable groups.
... Garcia-Estepa et al. (1999) reported that the phytic acid content in wheat brans was in the range of 25-58 mg/g. Furthermore, phytic acid content in hard and soft wheat brans was 42-54 and 46-67 mg/g, respectively (Camire and Clydesdale 1982;Knuckles et al. 1982). Phytic acid cannot be digested by nonruminants, and the utilization of bran-rich wheat milling by-products as animal feed results in high phytic acid content in animal waste. ...
... This was done by HPLC method of phytic acid according to Camire and Clydesdale (1982). Approximately 50 mg of sample was weighed into a 125 ml Erlenmeyer flask and 10 ml of 3% H 2 SO 4 added. ...
Sorghum and millet and their products require specialized treatment in order to improve their nutritive value, organoleptic properties and shelf-life. They contain anti-nutrients which are the major phytochemicals which negatively affects their nutritive values. The phytochemicals of concern include tannins and phytates, which interfere with mineral absorption, palatability and protein digestibility. Malting and fermentation treatments were applied to reduce the anti-nutrients, improve protein digestibility, and acidity to increase the products shelf life. The effects of malting and fermentation on the cereals nutritive value and anti-nutrient reduction were studied and evaluated for a period of 8 days. A treatment combining malting for 3 days and fermentation for 2 days respectively both at room temperatures (25°C) was employed. Tannins and phytates were significantly reduced (p ? 0.05) by malting and fermentation. Protein digestibility was significantly (p ? 0.05) improved by malting and fermentation treatments; malted cereals digestibility ranged between 34.5-68.1% while the fermented flours protein digestibility range was 97.4-98.3%. The pH values were lowered to below 4.0, a level at which they could effectively inhibit spoilage microorganisms at the end of the fermentation period. A combination of optimum time treatments of malting and fermentation for 3 days and 2 days respectively were effective in reducing tannins and phytates and improving protein digestibility of the cereals.
... For sample extraction, trichloroacetic acid, H 2 SO 4 , and HCl are the reagents commonly used (Tangendjaja et al., 1980;Camire and Clydesdale, 1982;Graf and Dintzis, 1982). These acids may interfere with detection of IPs, so need to be removed or neutralized before loading extracted samples into the column separation system. ...
... Tannin content was determined using the Vanillin-Hydrochloric acid method [6], [7] [8]. Phytates were determined the method by Camire and Clydesdale [9]. Protein digestibility was carried out using Mertz et al [10] procedure with slight modification. ...
According to Kenya Demographic Health Survey, 7% of children under five years were wasted with 16% of them being underweight probably an indication of poor and inappropriate feeding practices. The children suffer from protein energy malnutrition (PEM) and micro-nutrient deficiencies which may lead to physical, mental and motor development retardation. Children are most at risk of PEM during the introduction of complementary foods usually thin porridge prepared predominantly from cereals and starchy tubers. Such porridge is low in energy and nutrient density, and may be high in anti-nutrients, despite the fact that infants at this stage of rapid development have high requirements of energy and nutrients per unit body weight. There is need therefore to develop appropriate nutrient-dense complementary foods that could be used by low income families. Amaranth grain has high biological value proteins and a better amino acid profile than nearly all cereals. It is also rich in essential fatty acids. However it is not commonly used as a complementary food in Kenya. The main objective was to determine the optimum steeping and germination time for amaranth grain. The grains were steeped and germinated for various time periods. The dry matter loss, proximate composition and some antinutrient levels were determined. Dry matter loss was least in amaranth grain steeped for 5 hours and germinated for 24 hours. At p<0.05, there were no significant differences in ash, fat and protein contents with respect to steeping and germination time. The crude fiber content and the invitro protein digestibility varied with different steeping and germination time. The tannin and phytate contents could not be detected after steeping and germination. Based on dry matter loss and reduction in antinutrient levels, steeping amaranth grain for 5 hours and germinating for 24 hours were the optimum processing times. INTRODUCTION National level estimates show that 35% of children in Kenya underfive years old are stunted, 7% are wasted and 16% are undernourished [1]. The children fail to reach their full potential growth and development, and suffer long term deprivation of energy and nutrients and consequently chronic PEM, often accompanied by micronutrient deficiencies. KDHS (2010) also reported that the most commonly used foods given to breastfeeding children under age 3 include food made from grains (72%), vitamin A rich fruits and vegetables (53%) and other milk (51%). The most commonly used first complementary food for babies in Kenya is porridge [2]. Most families often depend on inadequately processed traditional foods consisting mainly of unsupplemented cereal porridges made from maize, sorghum and millet. These staples may not contain adequate energy and nutrients. These staples are plant based. Plant-based diets are often associated with micronutrient deficits, exacerbated in part by poor micronutrient bioavailability [3]. Therefore the children may develop PEM and micro-nutrient deficiencies.
... To neutralize the sample, FeCl3 was added to the IP6, and the mixture was placed in a boiling water bath to allow complete precipitation of the ferric phytate complexes. Ferric phytate was then heated with NaOH and centrifuged to obtain ferum (III), hydroxide (pellet), and sodium phytate (supernatant) [24]. Finally, the supernatant was collected and freeze dried. ...
Nutritional or dietary factors have drawn attention due to their potential as an effective chemopreventive agent, which is considered a more rational strategy in cancer treatment. This study was designed to evaluate the effect of IP6 extracted from rice bran on azoxymethane- (AOM-) induced colorectal cancer (CRC) in rats. Initially, male Sprague Dawley rats were divided into 5 groups, with 6 rats in each group. The rats received two intraperitoneal (i.p.) injections of AOM in saline (15 mg/kg body weight) over a 2-week period to induce CRC. IP6 was given in three concentrations, 0.2% (w/v), 0.5% (w/v), and 1.0% (w/v), via drinking water for 16 weeks. The deregulation of the Wnt/ β -catenin signaling pathway and the expression of cyclooxygenase (COX)-2 have been implicated in colorectal tumorigenesis. β -Catenin and COX-2 expressions were analysed using the quantitative RT-PCR and Western blotting. Herein, we reported that the administration of IP6 markedly suppressed the incidence of tumors when compared to the control. Interestingly, the administration of IP6 had also markedly decreased β -catenin and COX-2 in colon tumors. Thus, the downregulation of β -catenin and COX-2 could play a role in inhibiting the CRC development induced by IP6 and thereby act as a potent anticancer agent.
... They determined the phytic acid content of the wheat bran obtained from different varieties using the same method used in the current study and reported that most of the samples had a phytic acid content in the range of 34-47 mg/g. In other studies the phytic acid content was in the range of 42-54 mg/g as determined by colorimetric method (Harland and Oberleas 1986), and 46-67 mg/g as determined by HPLC method (Camire and Clydesdale 1982). ...
With the aim of reducing phytic acid content of wheat bran, particle size reduction (from 1,200 to 90 μm), hydrothermal (wet steeping in acetate buffer at pH 4.8 at 55 °C for 60 min) and fermentation (using bakery yeast for 8 h at 30 °C) and combination of these treatments with particle size reduction were applied and their effects on some properties of the bran were studied. Phytic acid content decreased from 50.1 to 21.6, 32.8 and 43.9 mg/g after particle size reduction, hydrothermal and fermentation, respectively. Particle size reduction along with these treatments further reduced phytic acid content up to 76.4 % and 57.3 %, respectively. Hydrothermal and fermentation decreased, while particle size reduction alone or in combination increased bran lightness. With reducing particle size, total, soluble and insoluble fiber content decreased from 69.7 to 32.1 %, 12.2 to 7.9 % and 57.4 to 24.3 %, respectively. The highest total (74.4 %) and soluble (21.4 %) and the lowest insoluble fiber (52.1 %) content were determined for the hydrothermaled bran. Particle size reduction decreased swelling power, water solubility and water holding capacity. Swelling power and water holding capacity of the hydrothermaled and fermented brans were lower, while water solubility was higher than the control. The amount of Fe+2, Zn+2 and Ca+2 decreased with reducing particle size. Fermentation had no effect on Fe+2and Zn+2 but slightly reduced Ca+2. The hydrothermal treatment slightly decreased these elements. Amongst all, hydrothermal treatment along with particle size reduction resulted in the lowest phytic acid and highest fiber content.
... (Adeyeye et al., 2000; Adeyeye and Fagbohun, 2005; Aletor et al., 2007; Aletor, 1995; Al-Wahsh et al., 2005; Amrein et al., 2003; Atukorala and Waidyanatha, 1987; Becker et al., 1981; Beleia et al., 1990; Bhatty and Cherdkiatgumchai, 1990; Bieleski et al., 1997; Bressani et al., 2002; Calhoun et al., 1958; Calhoun et al., 1960; Camire and Clydesdale, 1982; Carlavilla et al., 2006; Cataldi et al., 1998; Chan et al., 2007; Chen, 2004 Fredlund et al., 1997; Gao et al., 2007; Garcia-Estepa et al., 1999; García Estepa et al., 1999; Greiner et al., 2001; Haraldsson et al., 2004; Harland and Frolich, 1989; Harland and Oberleas, 1987; Harland et al., 2004; Helfrich and Bettmer, 2004; Hooda and Jood, 2003; Horbowicz and Obendorf, 1994; Ikawa et al., 1968; Isiguro et al., 2005; Jährig et al., 2000; Kirby and Nelson, 1988; Koziol, 1992; Le Francois, 1988; Lee, 1990; Lehrfeld, 2002; Lestienne et al., 2005; Liu and Chang, 2004; Lolas et al., 1976; Lorenz and Wright, 1984; Lott et al., 2000; Ma et al., 2005; Macfarlane et al., 1988; Mameesh and Tomar, 1993; Masuda et al., 2003; Matsunaga et al., 1989; Mckenzie, 1984; Mckenzie-Parnell and Guthrie, 1986; Michalska et al., 2007; Morris and Hill, 1995; Morris and Hill, 1996; Nayini and Markakis, 1983; Ola and Oboh, 2001; Olguin et al., 2003; Oomah et al., 1996; Oyetayo and Akindahunsi, 2004; Pattee et al., 2000; Phillippy et al., 2003; Phillippy et al., 1988; Phillippy and Wyatt, 2001; Plaami and Kumpulainen, 1995; Pradeep et al., 1993; Proskova, 1998; Ramirez-Cardenas et al., 2008; Ravindran et al., 1994; Repo-Carrasco Valencia et al., 2009; Sanz et al., 2004; Schweizer et al., 1978; Scriban, 1970; Seifert, 1972; Simonet et al., 2003; Somsub et al., 2008; Soria et al., 2009; Sosulski et al., 1982; Souci et al., Khan, 1998; Thompson and Erdman Jr., 1982; Toda et al., 2006; Trugo et al., 1999; Udosen and Ukpanah, 1993; Umeta et al., 2005; Valencia et al., 1999; Venkatachalam and Sathe, 2006; Vitali et al., 2008; Yadav and Sehgal, 2003) ...
... In recent years, some HPLC methods for the determination of phytate have been developed (Tangendjaja et al., 1980;Camire and Clydesdale, 1982;Knuckles et al., 1982;Graf and Dint&, 1982;Lee and Abendroth, 1983). Myo-inositol monophosphates have been determined by HPLC (Hallcher and Sherman, 1980) but not the inositol 2-5 phosphates. ...
ABSTRACTA HPLC method was developed for separation and quantitative determination of inositol tri-, tetra-, penta-, and hexaphosphates. The method included extraction of inositol phosphates with HCI, separation of the inositol phosphates from the crude extract by ion-exchange chromatography, and ion-pair C18 reverse phase HPLC analysis using formic acid/methanol and tetrabutylammonium hydroxide in the mobile phase. The inositol 3–6 phosphates of raw and extruded bran, soy flour, and intestinal contents were determined by HPLC and compared to phytate determinations by two iron precipitation methods. Inositol 3–5 phosphates were found in extruded products and intestinal contents. The HPLC method was rapid and gave reproducible values, which differed from those obtained by the precipitation methods in some samples.
Due to the significant role of bread in the food basket of people in the world, it is necessary consider its quality and nutritional attributes. Due to the significance of alpha-amylase and phytase enzymes in bread quality and the sensitivity of these enzymes to some environmental conditions, it was attempted to optimize and model the encapsulation conditions of the combination of two enzymes in beeswax. The parameters of amylase and phytase, beeswax amount, tween 20, and mixing speed were regarded as independent parameters and their effect on the encapsulation efficiency of alpha-amylase and phytase enzymes was studied. The results indicated that the concentration of enzymes and the amount of beeswax had a direct and inverse effect on the encapsulation efficiency of both enzymes, respectively. The proposed model for the encapsulation efficiency of type 2 alpha-amylase and phytase and the fitting coefficients of the models were reliable. Finally, the optimal encapsulation conditions due to the maximum encapsulation efficiency were 156.53 μL of alpha-amylase enzyme, 178.28 mg of phytase enzyme, 1.87 g of beeswax, 1.19% by weight/ volume of tween 20 and mixing speed of 1081-89 r/min were determined with a utility factor of 0.897.
Spent engine oil is hazardous to the environment. Indiscriminate disposal of spent engine oil drain from engines after servicing has been found to affect the environment. An experiment was carried out to determine the effect of spent engine oil pollution on soil characteristics and the ability of Amaranthushybridus to thrive in the soil supplemented with varying concentrations of spent engine oil ranging from 50- 300 mL. Soil pH was slightly increased due to spent engine oil pollution. Nitrogen, phosphorus and potassium were reduced in the polluted soils and the soil organic carbon was increased. Soil samples polluted with spent engine oil showed increased bulk and particle densities and also decreased water holding capacity and porosity. The spent engine oil pollution affected the phytochemicals and resulted in the increased concentration of anti- nutrient tannin and decreased concentrations of nutrients like alkaloids, flavonoids, etc. The results of this study suggest that spent engine oil at any concentration seriously affects the soil properties and phytochemical analysis showed the inhibitory effects of spent engine oil on Amaranthushybridus .
In the previous edition, this chapter was part of the diet formulation and manufacture chapter. In this edition, a separate chapter is devoted to the topic. There are two reasons for this change. First, alternative ingredients increasingly used in fish feeds are less familiar, so identifying appropriate chemical tests to assess quality of alternative ingredients is needed. Second, as fishmeal levels in feeds are being reduced, fish feeds must be formulated to more precisely meet nutritional needs of fish, i.e., precision formulation. This makes it crucial to have tools to assess quality of feeds and ensure that available levels of essential nutrients are provided. The first portion of the chapter explains which chemical tests are appropriate to use to measure ingredient quality. Different ingredients present different potential quality problems. Interpretation of the results of chemical testing allows feed producers to select high-quality ingredients. In addition to chemical tests, microscopic examination of feed ingredients, somewhat of a lost art, is a valuable tool to detect adulteration of ingredients or heat damage. In the second portion of the chapter, in vivo methods of diet evaluation are presented. This includes digestibility measurements and calculations, feeding trials and their evaluation, and approaches to assess effects of diet on fish quality and safety to consumers. Finally, approaches used for economic evaluation of feeds are discussed.
Exploring an effective sensor to detect Fe³⁺ and phytic acid (PA) is of great importance because the levels of Fe³⁺ and PA have a vital impact on human health. Herein, we report a metal-free fluorescent nanoprobe for sensitive and selective sensing of Fe³⁺ and PA based on the sulfur quantum dots (SQDs). The SQDs with high fluorescence quantum yield (17.6%) and low cytotoxicity can be directly synthesized from cheap elemental sulfur by one-pot reaction. The SQDs display responsive and selective fluorescence “on-off-on” behavior towards Fe³⁺ and PA based on the introduction and elimination of electron transfer effect. The addition of Fe³⁺ can cause non-radiative electron transfer associated with fluorescence quenching of SQDs, while the addition of PA enables SQDs/Fe³⁺ complex release Fe³⁺ to recover the fluorescence of SQDS. By utilizing these SQDs as a fluorescent nanoprobe, it shows good linear relationship for Fe³⁺ and PA in the range of 0.05-120 µM and 0-12 µM, respectively. Moreover, its limit of detection (LOD) for Fe³⁺ and PA can reach as low as 102 and 73.5 nM, which compares favorably to many reported fluorescent probes. Additionally, the use of this SQDs-based sensor to detect PA in fresh carrot and lettuce samples was examined with satisfactory performance.
Background
The problem of micronutrient malnutrition is affecting millions of infants in the developing countries. One of the major issues that aggravates the problem is lack of appropriately processed complementary foods in which the bioavailability of the major micronutrients is improved.
Methods
Teff, soybean and orange-fleshed sweet potato were separately processed into their respective flours and blended in a ratio of 70:20:10, respectively, to prepare household- and industrial-level complementary foods. The ingredients and developed complementary foods were analysed for their vitamin A, calcium, iron, zinc and phytate contents. Moreover, phytate: mineral molar ratios of calcium, iron and zinc in the complementary foods were calculated to determine their bioavailability.
Results
The vitamin A values obtained in the complementary foods were appreciable; they were in the range of 91.89 to 160.97 µgRE/100 g. Phytate content of teff and soybean was significantly (p ≤ 0.05) reduced by the household practices employed for processing them. However, the reduction was not significant (p > 0.05) in the household-level complementary foods because of the small quantity of germinated teff flours used. Calcium, iron and zinc compositions of all complementary foods closely met the recommended levels for 6 to 8 month-old infants. Phytate: mineral molar ratios for calcium and zinc in all complementary foods were below maximum recommended limits indicating their good bioavailability. In case of iron, these ratios were above the critical limit except that of the industriallevel complementary foods.
Conclusion
Generally, complementary foods with improved compositions and bioavailability of the micronutrients analysed were developed from the teff-soybean-orange-fleshed sweet potato formulations.
By products of wheat have been used for a long time in Algeria for the poultry production. Therefore, the chemical composition of 18 samples of bran of wheat produced in Algeria was evaluated and their metabolisable energy content was measured in growing chickens. The results of chemical analysis were used to establish prediction equations of the ME value. The results of chemical composition, show that wheat bran produced locally had more starch than those published by institutions. The mean EM value of the local wheat bran is 9,04 MJ/kg DM. The chemical components and EM value of the studied wheat bran were statistically (P < 0, 001) affected by the factor sample. Crude fiber is a good predictor of the ME value (R = 0, 95; rsd = 0,19 MJ/kg DM; P < 0,05).
The influence of natural phytic acid extracted from wheat bran, and exogenous phytic acid on oxidative stability in chicken O/W emulsion was investigated. The antioxidant activity of phytic acid was evaluated by the thiobarbituric acid reactive substances (Tbars) values, oxygen absorption and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging methods. Phytic acid extracted from wheat bran (pae) was highly effective in reducing Tbars values (68% in comparison to control) and preventing lipid oxidation during storage of chicken O/W emulsion at 40°c for 2 weeks. Such antioxidant activity of exogenous phytic acid depends on its concentration in chicken O/W emulsion. Ascorbic acid at concentration 0.1% inhibited tbars formation by 56% compared to the control. Oxygen in the headspace of vials containing emulsion without phytic acid (control) was consumed faster than where phytic acid was added. Assuming a zero time reaction, the rate of oxidation without phytic acid (control) was 0.61% o 2/day, while that with phytic acid extract was 0.055% o 2/day or about 12 times slower (significantly different). Phytic acid extract and exogenous phytic acid (4 mm) were more effective in preventing lipid oxidation and oxygen absorption than ascorbic acid and BHA. Radical scavenging activity of phytic acid extract and pure phytic acid (4 mm) were 40.96±0.55 and 40.89±0.32%, respectively. Free radical scavenging ability of phyticacid was lower than other common antioxidants (ascorbic acid and bha). It can be concluded that phytic acid may substantially inhibit malondialdehyde formation, oxygen uptake, as well as warmed-over flavor development during storage of chicken meat or chicken meat products.
Underutilized local vegetables (ULVs) in Vihiga County within the Lake Victoria Basin that could provide micronutrients to fight hidden hunger contain phytate and oxalate that reduce bioavailability. These can be reduced through appropriate traditional food processing techniques adopted by households. The aim of this study was to determine the content of phytate and oxalate in formulated ULV recipes. This was an experimental design with eleven selected ULVs: spider plant (C. gynandra), pumpkin leaves (C. moschata), cowpea (V. unguiculata), amaranth (A. blitum), jute mallow (C. olitorius), sweet potatoes leaves (I. batatas), African nightshade (S. nigrum), cassava leaves (M. esculenta), Slender leaf (C. ochroleuca), vine spinach (B. alba) and kales (B. carinata). Eleven single vegetables and five vegetable combinations were blanched and divided into two lots. One lot was boiled and fermented for 48 h and the other was not fermented. The non-fermented were subjected to three treatments; cooked by boiling in water, cooked by boiling with the addition of cow's milk and lye and cooked by sautéing. Phytate and oxalate content in the obtained recipes were determined using HPLC. Independent t-test was used to compare the mean content of phytate and oxalate between fermented recipes and non-fermented recipes. One-way ANOVA was used to compare the mean content of phytate and oxalate between different methods of cooking. The phytate content in fermented ULVs ranged from 27.5-70.67 mg 100 g-1 fresh weight (FW) and 40.0- 83.44 mg 100 g-1 FW for non-fermented ones. The oxalate content ranged from 2.62- 10.17 mg 100 g-1 FW for non-fermented and 1.54-20.36 mg 100 g-1 FW for fermented. Mean levels of phytate in ULVs were higher than those of oxalate in the same vegetables. The mean phytate and oxalate levels were lower in most fermented vegetables compared to non-fermented ones (p<0.05). However, there was no significant difference in the mean phytate and oxalate levels between all fermented and all non-fermented ULVs (p>0.05) suggesting that degradation of phytate and oxalate by fermentation depends on the type of vegetable since different ULVs responded differently to fermentation. Cooking methods and microbial fermentation differently affected the levels of phytate and oxalate; in some ULVs a decrease was observed while in others there was no significant change. It is concluded that heat during cooking reduced phytate and oxalate levels in most ULVs and that microbial fermentation reduced phytate and oxalate levels in most ULVs but the degree of degradation depends on the type of vegetable sampled since different samples have a varied sample matrix. It is suggested that additives like onions and tomatoes during sautéing and milk and lye during boiling may have introduced some phytate and oxalate to the ULV recipes.
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Phytic acid is a naturally occurring polyphosphorylated carbohydrate, present ubiquitous in plants and animals. It is not only a natural antioxidant, but may also be the precursor/storage of intracellular inositol phosphates, important for various cellular functions and potential as anticancer compound. A prominent anticancer action of phytic acid has been demonstrated both in vivo and in vitro in a variety of tumor types, possibly through inhibition of tumor cell growth and differentiation. In this study, the growth inhibitory effect of phytic acid extracted from rice bran on hepatocellular cell lines (HEPG 2), cell cycle modulation and apoptosis induction were undertaken. Phytic acid prove to induce growth inhibition and differentiation in HepG 2 in a dose and time-dependent manner with IC 50 value of 17.0 µg/ml. Analysis of flow cytometry was performed for the analysis of cell cycle and apoptosis. Treatment of phytic acid against HepG 2 also resulted in cell cycle arrest in HEPG 2 cell at G 2 /M phase cell cycle arrest. Besides, Annexin V-assay and propidium iodide confirmed the apoptosis occurred early and late in the cell line. In conclusion, with the results taken from our findings, phytic acid extracted from rice bran was revealed as a potent candidate for adjuvant chemotherapy and prevention of cancer.
ABSTRACT
Legumes are one of the world´s most important sources of food supply, especially in developing countries, in terms of food energy as well as nutrients. Common beans are a good source of proteins, vitamins (thiamine, riboflavin, niacin, vitamin B6) and certain minerals (Ca, Fe, Cu, Zn, P, K and Mg). They are an excellent source of complex carbohydrates and polyunsaturated free fatty acid (linoleic, linolenic). However, common beans have several undesirable attributes, such as a long cooking time, being enzyme inhibitiors, phytates, flatus factors, and phenolic compounds, having a “beany” flavor, and being lectins and allergens., which should be removed or eliminated for effective utilization.
Grain quality of common beans is determined by factors such as acceptability by the consumer, soaking characteristics, cooking quality, and nutritive value. Acceptability characteristics include a wide variety of attributes, such as grain size, shape, color, appearance, stability under storage conditions, cooking properties, quality of the product obtained, and flavor.
Storage of common beans under adverse conditions of high temperature and high humidity renders them susceptible to a hardening phenomenon, also known as the har-to-cook (HTC) defect. Beans withs this defect are characterized by extended cooking times for cotyledon softening, are less acceptale to the consumer, and are of lower nutritive value. Mechanisms involved in the HTC defect have not been elucidated satisfactorily. Attempts to provide a definitive explanation of this phenomenon have not been successful. The most important hypotheses that have been proposed to explain the cause of bean hardening are (1) Lipid oxidation and/or polymerization, (2) Formation of insoluble pectates, (3) Lignification of middle lamella, and (4) Multiple mechanisms. Most researchers have reported that the defect develops in cotyledons. Recently, some authors have suggested that the seed coat plays a significant role in the process of common bean hardening. A better knowledge of cotyledon and seed coat microstructure may lead to better understanding of the causes of seed hardness.
In order to prevent the development of the HTC defect several procedures have been proposed: (1) Appropriate storage, (2) Controlled atmospheres, and (3) Pretreatments. Probably, the most workable solution to the hardening phenomenon may be the development of materials less prone to HTC phenomenon.
Decreasing cooking time, increasing nutritive value, and improve sensory properties of seeds with HTC defect would have great nutritional and economical impact. Furthermore, an understanding of the mechanisms leading to reversibility of this phenomenon would provide insight into the development of the defect itself and woul aid in the search for appropriate method to prevent it.
Efforts to develop technological processes are needed in order to transform the HTC beans into edible and useful products. Several economic alternatives to utilize HTC common beans have been proposed: (1) Dehulling, (2) Extrusion, (3) Solid state fermentation, (4) Quick-cooking beans, and (5) Production of protein concentrates and isolates and starch fractions.
Keywords: Legumes, common beans, cotyledon, seed coat, storage, hard-to-cook, cooking time, reversibility, technological processes
According to Kenya Demographic Health Survey, 7% of children under five years were wasted with 16% of them being underweight probably an indication of poor and inappropriate feeding practices. The children suffer from protein energy malnutrition (PEM) and micro-nutrient deficiencies which may lead to physical, mental and motor development retardation. Children are most at risk of PEM during the introduction of complementary foods usually thin porridge prepared predominantly from cereals and starchy tubers. Such porridge is low in energy and nutrient density, and may be high in anti-nutrients, despite the fact that infants at this stage of rapid development have high requirements of energy and nutrients per unit body weight. There is need therefore to develop appropriate nutrient-dense complementary foods that could be used by low income families. Amaranth grain has high biological value proteins and a better amino acid profile than nearly all cereals. It is also rich in essential fatty acids. However it is not commonly used as a complementary food in Kenya. The main objective was to determine the optimum steeping and germination time for amaranth grain. The grains were steeped and germinated for various time periods. The dry matter loss, proximate composition and some antinutrient levels were determined. Dry matter loss was least in amaranth grain steeped for 5 hours and germinated for 24 hours. At p<0.05, there were no significant differences in ash, fat and protein contents with respect to steeping and germination time. The crude fiber content and the invitro protein digestibility varied with different steeping and germination time. The tannin and phytate contents could not be detected after steeping and germination. Based on dry matter loss and reduction in antinutrient levels, steeping amaranth grain for 5 hours and germinating for 24 hours were the optimum processing times.
Soft white and hard red wheat bran were found to contain 98.8 and 99.2% water insoluble iron, respectively. As the concentration of iron added to wheat bran was increased, less of the total added iron was bound. Ascorbic acid was found to inhibit binding of ferrous iron to wheat bran. It was found that boiling for 1 hr in a boiling water bath (BWB) had no effect on the destruction of phytic acid in wheat bran, whereas toasting for 1 hr at 178°C (350°F) and boiling for 1 hr in 1N HC1 had a significant effect.
Phytic acid was quantified using a tetrabutylammonium-formate ion pair on an octadecyl column in HPLC. Unlike previous methods, phytic acid eluted significantly later than the dead volume. Good separations were possible with a linear response up to 40 micrograms and a sensitivity of 0.002 micrograms phytic acid. Results with this method are compatible with conventional methods for phytic acid.
In corn wet milling the raw material is prepared by means of steeping. Steeping is required in order to: –obtain the valuable CSL (Corn Steep Liquor),
–soften the kernel and break the cell walls.
SO 2 is added to the steepwater to inhibit bacterial growth. Depending on the type of corn, the steeping time in a semi‐continuous steeping system varies from 36 to 60 h, which is substantially longer than all subsequent steps in the process. The corn comprises phytic acid, which to a large extent ends up in the CSL and constitutes an undesirable component. It has now been found that the problem caused by this phenomenon can be avoided by using phytic acid degrading enzymes during steeping. It has further been found, that by adding phytic acid degrading enzymes together with plant cell wall degrading enzymes to the steep liquor, the steeping time can be reduced considerably in a conventional steeping system and even further in a two stage steeping process, whereby the second steeping stage is carried out after course grinding. Also in case conventional steeping times are applied higher starch yields and lower energy consumption may be obtained.
The effect of pH and heat treatments on the binding of iron, calcium, magnesium and zinc and the loss of phytic acid in defatted soy flour was investigated. The soy flour was found to bind more iron, calcium, and magnesium at pH 6.8 than at pH 5.0, but the reverse situation occurred with zinc. Boiling caused a significant increase in binding of zinc and magnesium at both pH values, but was pH dependent for iron and calcium. Toasting caused a significant increase in binding of zinc and calcium at both pH values and a pH variable effect on iron and magnesium. Phytic acid analysis under the same conditions suggested that the degree of binding of these minerals did not correlate with the presence of phytic acid.
The effect of microwave heating and gamma irradiation treatments on phosphorus compounds of soybean seeds were studied. Inorganic phosphorus was significantly (P < 0.05) increased while, phytate and phospholipids were significantly decreased when soybean seeds were microwave-heated for 9 min or more. Furthermore, gamma-irradiation treatments of 20 KGy or more significantly increased inorganic phosphorus and decreased phytate and phospholipids of the beans. Two dimensional thin-layer chromatography was employed to study the phospholipid pattern. Gamma-irradiation at doses from 40 to 100 KGy produced lysophosphatidylcholine and phosphatidic acid. The data of this study showed that, the increase of inorganic phosphorus was mainly due to the decomposition of phytic acids and phosphates of inositol.
Whey fractions cor taining proteins (albumins), phytate, and minerals were prepared from black gram (Phaseolus mungo L.) cotyledons and were employee to study the interactions between protein, phytate, and minerals at pH 2.80, 6.40, and 8.40. Black gram cotyledons contained 1.7% phytate, of which 88.7% existed in watersoluble form. Phytate phosphorus represented for about 89% of total phosphorus in black gram cotyledons. Recovery of phytate in fraction I (pH 2.80), fraction II (pH 8.40), and fraction III (pH 6.40) was 45%, 69%, and 4%, respectively, after 48 hr dialysis. At pH 2.80, complexation occurred between phytate and proteins. Complexation between phytate and proteins at pH 8.40 was mediated by divalent cations such as calcium, magnesium, and zinc. Fraction II had higher concentrations of divalent cations (calcium, magnesium, and zir c) than the other two fractions I and III.
The review on the determination of phytate and inositol phosphates by Oberleas (1) indicates that most methods for the determination of phytate are derived from the method of Heubner and Stadler (2). This method is based on the principle that ferric ion forms a stable complex with phytate in dilute acid solution and is the only phosphate compound, at least in significant concentration in nature, with this property. However, the phytate values were high when we applied the procedure of Oberleas (3) to samples with high inorganic phosphate content such as rat feces or semipurified rat diets. This appeared to be a result of inorganic phosphate coprecipitating with ferric phytate.As a preliminary step to study phytate balance in rats, a modified procedure of the Oberleas (3) method was developed that appears to free the ferric phytate precipitate of inorganic phosphate. The procedure and its efficiency are reported in this communication.
The protein quantity and quality, caloric value, and overall nutrient content of oilseeds are quite good. However, oilseeds
are high in phytic acid and contain fiber and perhaps other binding agents which reduce mineral bioavailability from the seeds.
Phytic acid, the hexaphosphate of myoinositol, functions as the chief storage form of phosphate and inositol in mature seeds.
On a dry basis, whole oilseeds contain about 1.5% while some oilseed protein concentrates can contain over 7.0% of the compound.
Phytic acid is a strong chelating agent that can bind mono- and divalent metal ions to form the complex phytate. Published
results from numerous animal feeding trials suggest poor bioavailability of minerals such as zinc, calcium, magnesium, phosphorus
and possibly iron from diets containing high phytate foods. Recent studies involving the feeding of soy products to rats suggest
that zinc is the mineral of most concern as its bioavailability from some soy products is quite low. Prediction of mineral
bioavailability from phytate-containing foods is complicated by the complex interactions between the minerals and phytic acid
contained in the foods, intestinal and the meal phytase activities, previous food processing conditions (especially pH), digestibility
of the foods as well as the physiological status of the consumer of the foods. Very little is known about the chemistry of
such interactions. Therefore, most of the emphasis in controlling or reducing mineral binding in oilseed products has been
placed upon development of methodology for phytate removal.
Soy products with low, intermediate and normal phytate levels were prepared in the pilot plant for subsequent rat-feeding
experiments to evaluate zinc bioavailability. The low level (0.29%) phytate product was made by precipitation of the protein
curd at pH 5.5, whereas the normal level (1.05%) phytate product was produced by a similar process except that the phytate
previously isolated from the whey fraction was added back to the original curd as native phytate. The intermediate level (0.73%)
phytate product was also produced by acid precipitation, but at pH 4.5. The pH 5.5 precipitation process yielded a large quantity
of whey in which the ratio of water content to phytate was over 1,000 parts to 1. However, ca. 75% of the water was subsequently
removed by reverse osmosis (RO), which increased the concentration of phytate in the whey fraction and facilitated its isolation.
Protein was first removed from the whey by precipitation with trichloracetic acid, then phytate was precipitated in the supernatant
with ferric chloride. Another series of experiments was run to find optimal conditions to convert ferric phytate to the more
soluble sodium phytate form, using a minimal amount of sodium hydroxide so that the phytate could be recycled back to the
curd without causing a large increase in sodium content of the product. There were only minor differences in the protein,
lipid and mineral contents of the three products.
Destruction of phytate by yeast fermentation is compared in sponges prepared from Iranian whole wheat meals of different extraction rates. Phytate was destroyed rapidly in whole meals of 75 to 85 and 85 to 90 per cent extraction, but destruction was retarded in those of 95 to 100 per cent extraction. Production of acid-soluble phosphorus kept pace with phytate destruction in the two whole meals of lower extraction rates but was delayed with less-than-expected yield in those of 95 to 100 per cent rate. Unleavened whole meal bread contains little acid-soluble phosphorus. Leavened breads made from whole meals of slightly lower extraction rate average five times as much. Since phytate phosphorus appears to remain unavailable in the small intestine in many circumstances, dependece on unleavened whole meal bread may result in critically low intakes of available phosphorus when other sources are lacking in the diet. It is concluded that replacement of the whole meals of 95 to 100 per cent extraction rate, presently the main staple of the diet of rural Iran, by those of somewhat lower rate is an important preliminary to the introduction of leaven and fermentation into village bread-making methods.
Recent studies have shown that the iron in wheat is predominantly in the form of monoferric phytate (MFP). Unlike phytate complexed with two or more iron atoms, MFP is soluble at pH 7.0 and above and may therefore be a relatively available form of dietary iron. To examine this point, we tested iron absorption in adult dogs using a double radioisotope method and total body counting. When given without food, MFP was about one-half as available as ferrous sulphate at an iron-equivalent dose of 1.5 mg and only about one-seventh as available at a dose of 15 mg iron. When administered with food, MFP underwent complete isotopic exchange with the nonheme pool of dietary iron. When added to meals of either high or low iron availability in amounts that might be used for iron fortification, the absorption was the same for MFP iron as for the major pool of dietary inorganic iron.
The objectives of the study were to isolate and chemically characterize the iron in wheat and to determine the biological availability to the rat of the iron as the purified complex(es). Hard wheat bran contained no butanol extractable or water extractable iron, but approximately 60% of the iron was extracted by 1 to 1.2 M NaCl or ammonium acetate solution. This salt extractable iron complex was purified and identified as monoferric phytate. The purified monoferric phytate was soluble in water. Synthetic monoferric phytate was prepared from sodium phytate and ferric chloride and determined to have spectral characteristics and gel filtration chromatography behavior identical to the complex isolated from wheat bran. The butanol-water-salt extracted bran residue contained no detectable phytate and an as yet uncharacterized form of iron. The biological availability of the iron to the rat was determined by a hemoglobin depletion-repletion bioassay. The relative biological value of the iron as monoferric phytate, either isolated from wheat bran or the synthetic product, was equal to the reference compound, ferrous ammonium sulfate. In contrast, the biological availability of the iron in the bran residue was significantly lower and the low biological availability of an insoluble form of ferric phytate was confirmed. It is concluded that the major portion of the iron in wheat is monoferric phytate and has a high biological availability to the rat. Monoferric phytate in bran may be bound to cationic sites of proteins or other cellular components and utilization of the iron may be through solubilization of the monoferric phytate by ion exchange type mechanism rather than by hydrolysis of the phytate as has been postulated.
Sodium phytate corresponding to a formula of C6H6O24P6Na12 · 3H2O (formula weight 977.8) was titrated against metal ions and in each case an inflection in the titration curve was obtained when 5 moles of a divalent metal ion had been added per mole sodium phytate. At a pH of 7.4, sodium phytate formed complexes with metals in the following decreasing order: Cu++, Zn++, Ni++, Co++, Mn++ Fe+++, and Ca++.
Low-phytate wheat bran was produced by enzymatic hydrolysis and extraction. Rat bioassay methods were utilized to determine bioavailability of iron and zinc in the low-phytate brans and to study the effect of dietary phytate/zinc molar ratio on zinc bioavailability when the phytate source was bran. Endogenous phytase activity hydrolyzed 80-100% of the phytate when wheat bran was incubated in water overnight. The relative biological values of the iron in raw bran and phytate-free bran were 98 and 113, respectively, compared to 100 for ferrous ammonium sulfate in a hemoglobin repletion assay. Low-phytate brans with phytate/zinc molar ratios of 8 or less were equivalent to zinc sulfate as dietary sources of zinc for growth of rats. Rats fed diets that contained wheat bran with zinc sulfate added to reduce the dietary phytate/zinc molar ratio from 40 or 50 to 20 grew at the same rate as rats fed a phytate-free diet, but femur zinc values were lower than those in the reference group. Gel filtration chromatography of extracts of raw and low-phytate brans suggested that zinc might be associated with phytate in wheat bran.