Determination of Isomers of Inositol Mono- to Hexaphosphates in Selected Foods and Intestinal Contents Using High-Performance Ion Chromatography

Skoglund, Erika; Carlsson, Nils‐Gunnar; Sandberg, Ann‐Sofie

doi:10.1021/jf9603238

Cited by 119 publications

(115 citation statements)

References 21 publications

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“…However, Skoglund et al (1997a) showed that the concentration of InsP 1 -P was lower than the concentration of the other lower inositol phosphates in ileal digesta. Therefore, the accumulation of InsP 1 -P was expected to be negligible in this study.…”

Section: Discussionmentioning

confidence: 92%

Heat-treatment, phytase and fermented liquid feeding affect the presence of inositol phosphates in ileal digesta and phosphorus digestibility in pigs fed a wheat and barley diet

Blaabjerg

Jørgensen

Tauson

et al. 2010

Animal

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The aim was to evaluate the effect of heat-treatment, microbial phytase addition and feeding strategy (dry feeding v. fermented liquid feeding) on degradation of phytate (myo-inositol hexakisphosphate, InsP 6 ) and formation and further degradation of lower inositol phosphates (myo-inositol pentakisphosphate-myo-inositol bisphosphate, InsP 5 -InsP 2 ) at the distal ileum of pigs. Furthermore, the apparent ileal digestibility/degradability (AID) of phosphorus (P), InsP 6 -P and calcium (Ca) and the apparent total tract digestibility (ATTD) of P and Ca were studied. Pigs were fitted with a T-shaped ileal cannula for total collection of digesta at 2 h intervals during an 8 h sampling period after feeding the morning meal. Each period lasted for 2 weeks: 8 days of adaptation followed by 3 days of total collection of faeces and 3 days of total collection of ileal digesta. The experiment was designed as a 4 3 4 Latin square with four pigs fed four diets. A basal wheat/barley-based diet was fed either as nonheat-treated or heat-treated (steam-pelleted at 908C). The heat-treatment resulted in an inactivation of plant phytase below detectable level. Diet 1 (non-heat-treated basal diet fed dry); diet 2 (heat-treated basal diet fed dry); diet 3 (as diet 2 but with microbial phytase (750 FTU/kg as fed) fed dry); diet 4 (as diet 3 fed liquid (fermented for 17.5 h nighttime and 6.5 h daytime at 208C with 50% residue in the tank)). Chromic oxide (Cr 2 O 3 ) was included as marker and ATTD was determined both by total collection of faeces (ATTD Total ) and Cr 2 O 3 (ATTD Cr ). InsP 6 was completely degraded in diet 4 before feeding resulting in no InsP 6 -P being present in ileal digesta. InsP 6 -P concentration in ileal digesta decreased with increasing dietary levels of plant or microbial phytase in pigs fed the dry diets. Consequently, AID and ATTD of P and Ca were greatest for pigs fed diet 4 followed by diets 3, 1 and 2. The ATTD of P depended on the used method as ATTD Total of P was 72%, 61%, 44% and 34%, whereas ATTD Cr of P was 65%, 52%, 38% and 23% for diets 4, 3, 1 and 2, respectively. In all pigs the ileal concentration of InsP 5 -InsP 2 -P was extremely small, and thus unimportant for maximisation of ATTD of plant P. In conclusion, fermented liquid feeding with microbial phytase seems to be an efficient approach to improve ATTD of plant P compared with dry feeding. This opens up for further reductions in P excretion.

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Section: Discussionmentioning

confidence: 92%

Heat-treatment, phytase and fermented liquid feeding affect the presence of inositol phosphates in ileal digesta and phosphorus digestibility in pigs fed a wheat and barley diet

Blaabjerg

Jørgensen

Tauson

et al. 2010

Animal

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“…The isomers of inositol penta-and tetraphosphates in fecal samples were determined employing the method of Skoglund et al (1997). The latter method includes extraction of samples with HCl, separation of the inositol phosphates from the crude extract by anionexchange chromatography, separation on high-performance ion-exchange columns with gradient elution and detection using post-column reaction and UV detection.…”

Section: Methodsmentioning

confidence: 99%

Phytate hydrolysis in pigs fed a barley-rapeseed meal diet treated with Aspergillus niger phytase or steeped with whey

Skoglund¹,

Näsi²,

Sandberg³

1998

Can. J. Anim. Sci.

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-S. 1998. Phytate hydrolysis in pigs fed a barley-rapeseed meal diet treated with Aspergillus niger phytase or steeped with whey. Can. J. Anim. Sci. 78: 175-180. The degradation of phytate (myo-inositol hexaphosphate) in a barley-rapeseed meal (80:20) diet due to supplemented Aspergillus niger phytase and steeping (soaking at 40°C for 3 h with feed to water ratio 1 kg:1 L) with whey was studied in eight growing pigs (initial weight 27.8 kg). Phytate and its hydrolysis products (inositol penta-, tetra-and triphosphates, abbreviated IP 5 , IP 4 and IP 3 ) in diets and feces were determined using ion-pair high-performance liquid chromatography (HPLC). Different isomeric forms of inositol tetra-and pentaphosphates were studied utilizing high-performance ion chromatography (HPIC). Supplementing the diet with microbial phytase resulted in a 47% reduction in the amount of fecal phytate. Whey steeping of the diet reduced fecal phytate by 35%. Further reduction of the amount of fecal phytate (64%) was demonstrated in pigs fed the diet both steeped with whey and supplemented with microbial phytase, compared with pigs fed the untreated diet. Identification of IP 4 and IP 5 isomers in fecal samples showed which kind of phytase enzyme was active during phytate hydrolysis in the digestive tract. From these data, it was concluded that pigs fed the basal or whey steeped diet, without supplemented microbial phytase, had higher relative fecal amounts of DL-Ins(1,2,3,4,5)P 5 , compared with pigs fed the microbial phytase supplemented diets. Adding microbial phytase to the diet increased the relative amount of DLIns(1,2,4,5,6)P 5 in feces. With whey steeping of the diet, the relative amount of DL-Ins(1,2,3,4)P 4 isomer in feces was increased and the relative amount of DL-Ins(1,2,5,6)P 4 isomer was decreased. . La quantité de phytate dans les fèces était abaissée de 47 % par l'adjonction de phytase microbienne et de 35 % par le trempage dans le petit-lait, par rapport au régime témoin. Une réduction supplémentaire (64 %) était observée chez les porcs recevant l'aliment, à la fois trempé dans le petit-lait et additionné de phytase microbienne. L'identification des isomères IP 4 et IP 5 dans les prélèvements fécaux révélait quelle sorte de phytase était en cause durant l'hydrolyse des phytates dans le tractus digestif. Ces observations nous amènent à conclure que les porcs recevant le régime témoin ou l'aliment trempé dans le petit-lait sans adjonction de phytase microbienne montraient des teneurs fécales relativement plus élevées de DL-Ins (1,2,3,4,5)P 5 que ceux recevant l'aliment additionné de phytase microbienne. En revanche, l'adjonction de phytase accroissait les teneurs relatives de DL-Ins (1,2,4,5,6)P 5 dans les matières fécales. Le trempage dans le petit-lait accroissait les quantités relatives de DL-Ins (1,2,3,4)P 4 dans les fèces et diminuait celles de DL-Ins(1,2,5,6)P 4 . Mots clés: Phosphate d'inositol, trempage, phytase d'Aspergillus niger, porcDiets fed to pigs normally contain high amounts of phosphorus in the form of ...

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“…The analysis and identification of the IP 6 degradation products IP 5 -IP 2 , was performed using the same system as for the rapid IP 6 analysis, but with a gradient elution system as described by Skoglund and co-workers [22]. Peak identification was done by analyzing a chemically hydrolyzed sodium phytate sample.…”

Section: Identification Of Phytate Degradation Products Ip 5 -Ipmentioning

confidence: 99%

“…Peak identification was done by analyzing a chemically hydrolyzed sodium phytate sample. The order of the peaks has previously been described by Skoglund et al [22]. Since the detection response for the lower inositol phosphates is lower than for IP 6 (which was used to create the standard curve), theoretical correction factors were used [22].…”

Section: Identification Of Phytate Degradation Products Ip 5 -Ipmentioning

confidence: 99%

Assessing phytase activity

2015

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Phytases are nutritionally important for increased bioavailability of dietary minerals and phosphate for monogastric animals including humans. Release of minerals and phosphate is accomplished by the enzymatic stepwise degradation of phytate (inositol hexaphosphate, IP 6 ). Activity determinations of phytase is often based on analysis of total released phosphate (P i ), but phytase activity in its purest form represents released product per time from IP 6 only. Microbial and plant preparations often also contain mixtures of phosphatases and organic phosphate compounds; hence some released phosphate in enzymatic assays may originate from non-phytase phosphatases degrading non-phytate molecules. Moreover, even purified enzyme extracts assessed via P i release may result in errors, since commercial IP 6 commonly contains contamination of lower inositol phosphates, and further, the products of phytase IP 6 hydrolysis are also substrates for the phytase. These facts motivate a quantitative comparative study. We compared enzyme activity determination in phytase assay samples at four different time points, based on analyzing the substrate IP 6 versus the product P i using different selected methods. The calculated activities varied substantially. For example, at 15 min into enzymatic assay, variations from 152 mU/ml (by IP 6 analysis on HPIC) to 275-586 mU/ml (by P i analysis using several methods) was detected. Our work emphasizes the importance of defining the type of activity assessed, showing that phytase activity based on released P i may yield false positive results and/or overestimations. We propose to differentiate between phytase activity, being the activity by which IP 6 is degraded, and total inositol phosphatase activity, corresponding to total released phosphate during the enzymatic reaction.

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Determination of Isomers of Inositol Mono- to Hexaphosphates in Selected Foods and Intestinal Contents Using High-Performance Ion Chromatography

Cited by 119 publications

References 21 publications

Heat-treatment, phytase and fermented liquid feeding affect the presence of inositol phosphates in ileal digesta and phosphorus digestibility in pigs fed a wheat and barley diet

Heat-treatment, phytase and fermented liquid feeding affect the presence of inositol phosphates in ileal digesta and phosphorus digestibility in pigs fed a wheat and barley diet

Phytate hydrolysis in pigs fed a barley-rapeseed meal diet treated with Aspergillus niger phytase or steeped with whey

Assessing phytase activity

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