Skip to main content
Top
Published in: Urolithiasis 6/2012

01-12-2012 | Original Paper

A simple and rapid colorimetric method for determination of phytate in urine

Authors: Antonia Costa-Bauza, F. Grases, I. Gomila, A. Rodriguez, R. M. Prieto, F. Tur

Published in: Urolithiasis | Issue 6/2012

Login to get access

Abstract

Phytate is a natural product present in urine and biological fluids that is associated with health benefits, such as the prevention of calcium renal stone formation. The available methods for phytate analysis in urine all require elaborate instrumentation and cannot be routinely applied in clinical laboratories. Here, we describe a simple procedure for urinary phytate determination, employing colorimetric detection. Our method requires purification and preconcentration of phytate via solid-phase extraction prior to colorimetric detection employing Fe(III)–thiocyanate. The working linear range of the assay is 0–5 μM phytate. The limit of detection is 0.055 μM. The relative standard deviation obtained upon assay of samples containing 2 μM phytate was 3.5 %. Several urine samples were analyzed using an alternative method based on the detection of phosphorus; the results of the two assays were comparable. Our novel method of phytate analysis in human urine is simple, rapid (3 h for 10 samples), accurate, precise, reliable, and highly sensitive. The assay can be run in most analytical laboratories and does not require sophisticated instrumentation.
Literature
1.
go back to reference McCance RA, Widdowson EM (1942) Mineral metabolism of healthy adults on white and brown bread dietaries. J Physiol 101:44–85PubMed McCance RA, Widdowson EM (1942) Mineral metabolism of healthy adults on white and brown bread dietaries. J Physiol 101:44–85PubMed
2.
go back to reference McCance RA, Walsham CM (1948) The digestibility and absorption of the calories, proteins, purines, fat and calcium in wholemealwheaten bread. Br J Nutr 2:26–41PubMedCrossRef McCance RA, Walsham CM (1948) The digestibility and absorption of the calories, proteins, purines, fat and calcium in wholemealwheaten bread. Br J Nutr 2:26–41PubMedCrossRef
3.
go back to reference Halsted JA, Ronaghy HA, Abadi P, Haghshenass M, Amirhakemi GH, Barakat RM, Reinhold JG (1972) Zinc deficiency in man. Am J Med 53:277–284PubMedCrossRef Halsted JA, Ronaghy HA, Abadi P, Haghshenass M, Amirhakemi GH, Barakat RM, Reinhold JG (1972) Zinc deficiency in man. Am J Med 53:277–284PubMedCrossRef
4.
go back to reference Graf E, Empson KL, Eaton JW (1987) Phytic acid—a natural antioxidant. J Biol Chem 262:11647–11650PubMed Graf E, Empson KL, Eaton JW (1987) Phytic acid—a natural antioxidant. J Biol Chem 262:11647–11650PubMed
5.
go back to reference Shamsuddin AM (1995) Inositol phosphates have novel anticancer function. J Nutr 125:725S–732SPubMed Shamsuddin AM (1995) Inositol phosphates have novel anticancer function. J Nutr 125:725S–732SPubMed
6.
go back to reference Grases F, Costa-Bauzá A (1999) Phytate (IP6) is a powerful agent for preventing calcifications in biological fluids: usefulness in renal lithiasis treatment. Anticancer Res 19:3717–3722PubMed Grases F, Costa-Bauzá A (1999) Phytate (IP6) is a powerful agent for preventing calcifications in biological fluids: usefulness in renal lithiasis treatment. Anticancer Res 19:3717–3722PubMed
7.
go back to reference Grases F, Perelló J, Prieto RM, Simonet BM, Torres JJ (2004) Dietary myo-inositol hexaphosphate prevents dystrophic calcifications in soft tissues: a pilot study in Wistar rats. Life Sci 75:11–19PubMedCrossRef Grases F, Perelló J, Prieto RM, Simonet BM, Torres JJ (2004) Dietary myo-inositol hexaphosphate prevents dystrophic calcifications in soft tissues: a pilot study in Wistar rats. Life Sci 75:11–19PubMedCrossRef
8.
go back to reference Grases F, Sanchis P, Perelló J, Isern B, Prieto RM, Fernández-Palomeque C, Torres JJ (2007) Effect of crystallization inhibitors on vascular calcifications induced by vitamin D: a pilot study in Sprague-Dawley rats. Circ J 71:1152–1156PubMedCrossRef Grases F, Sanchis P, Perelló J, Isern B, Prieto RM, Fernández-Palomeque C, Torres JJ (2007) Effect of crystallization inhibitors on vascular calcifications induced by vitamin D: a pilot study in Sprague-Dawley rats. Circ J 71:1152–1156PubMedCrossRef
9.
go back to reference Jariwalla RJ, Sabin R, Lawson S, Herman ZS (1990) Lowering of serum cholesterol and triglycerides and modulation of divalent cations by dietary phytate. J Appl Nutr 42:18–28 Jariwalla RJ, Sabin R, Lawson S, Herman ZS (1990) Lowering of serum cholesterol and triglycerides and modulation of divalent cations by dietary phytate. J Appl Nutr 42:18–28
10.
go back to reference Lee S, Park H, Chun H, Cho S, Jung H, Cho S, Kim D, Kang M, Lillehoj H (2007) Dietary phytic acid improves serum and hepatic lipid levels in aged ICR mice fed a high-cholesterol diet. Nutr Res 27:505–510CrossRef Lee S, Park H, Chun H, Cho S, Jung H, Cho S, Kim D, Kang M, Lillehoj H (2007) Dietary phytic acid improves serum and hepatic lipid levels in aged ICR mice fed a high-cholesterol diet. Nutr Res 27:505–510CrossRef
11.
go back to reference Schlemmer U, Frølich W, Prieto RM, Grases F (2009) Phytate in foods and significance for humans: food sources, intake, processing, bioavailability, protective role and analysis. Mol Nutr Food Res 53:S330–S375PubMedCrossRef Schlemmer U, Frølich W, Prieto RM, Grases F (2009) Phytate in foods and significance for humans: food sources, intake, processing, bioavailability, protective role and analysis. Mol Nutr Food Res 53:S330–S375PubMedCrossRef
12.
go back to reference Grases F, Simonet BM, Prieto RM, March JG (2001) Phytate levels in diverse rat tissues: influence of dietary phytate. Br J Nutr 86:225–231PubMedCrossRef Grases F, Simonet BM, Prieto RM, March JG (2001) Phytate levels in diverse rat tissues: influence of dietary phytate. Br J Nutr 86:225–231PubMedCrossRef
13.
go back to reference Grases F, Perelló J, Isern B, Prieto RM (2004) Determination of myo-inositol hexakisphosphate (phytate) in urine by inductively coupled plasma atomic emission spectrometry. Anal Chim Acta 510:41–43CrossRef Grases F, Perelló J, Isern B, Prieto RM (2004) Determination of myo-inositol hexakisphosphate (phytate) in urine by inductively coupled plasma atomic emission spectrometry. Anal Chim Acta 510:41–43CrossRef
14.
go back to reference Muñoz JA, Valiente M (2003) Determination of phytic acid in urine by inductively coupled plasma mass spectrometry. Anal Chem 75:6374–6378PubMedCrossRef Muñoz JA, Valiente M (2003) Determination of phytic acid in urine by inductively coupled plasma mass spectrometry. Anal Chem 75:6374–6378PubMedCrossRef
15.
go back to reference Chen Y, Chen J, Ma K, Cao S, Chen X (2007) Fluorimetric determination of phytic acid in urine based on replacement reaction. Anal Chim Acta 605:185–191PubMedCrossRef Chen Y, Chen J, Ma K, Cao S, Chen X (2007) Fluorimetric determination of phytic acid in urine based on replacement reaction. Anal Chim Acta 605:185–191PubMedCrossRef
16.
go back to reference March JG, Simonet BM, Grases F (2001) Determination of phytic acid by gas chromatography-mass spectroscopy: application to biological samples. J Chromatogr B Biomed Sci Appl 757:247–255PubMedCrossRef March JG, Simonet BM, Grases F (2001) Determination of phytic acid by gas chromatography-mass spectroscopy: application to biological samples. J Chromatogr B Biomed Sci Appl 757:247–255PubMedCrossRef
17.
go back to reference Perello J, Isern B, Munoz JA, Valiente M, Grases F (2004) Determination of phytate in urine by high-performance liquid chromatography-mass spectrometry. Chromatographia 60:265–268CrossRef Perello J, Isern B, Munoz JA, Valiente M, Grases F (2004) Determination of phytate in urine by high-performance liquid chromatography-mass spectrometry. Chromatographia 60:265–268CrossRef
18.
go back to reference Latta M, Eskin M (1980) A simple and rapid colorimetric method for phytate determination. J Agric Food Chem 28:1313–1315CrossRef Latta M, Eskin M (1980) A simple and rapid colorimetric method for phytate determination. J Agric Food Chem 28:1313–1315CrossRef
19.
go back to reference Cilliers JJL, Niekerk PJ (1986) HPLC determination of phytic acid in food by postcolumn colorimetric detection. J Agric Food Chem 34:680–683CrossRef Cilliers JJL, Niekerk PJ (1986) HPLC determination of phytic acid in food by postcolumn colorimetric detection. J Agric Food Chem 34:680–683CrossRef
20.
go back to reference Thies W (1991) Determination of phytic acid and sinapic acid esters in rapeseed and selection of genotypes with reduced concentrations of these compounds. Lipid/Fett 93:49–52CrossRef Thies W (1991) Determination of phytic acid and sinapic acid esters in rapeseed and selection of genotypes with reduced concentrations of these compounds. Lipid/Fett 93:49–52CrossRef
21.
go back to reference García-Villanova R, García-Villanova RJ, Ruiz de Lope C (1982) Determination of phytic acid by complexometric titration of excess of iron (III). Analyst 107:1503–1506CrossRef García-Villanova R, García-Villanova RJ, Ruiz de Lope C (1982) Determination of phytic acid by complexometric titration of excess of iron (III). Analyst 107:1503–1506CrossRef
22.
go back to reference Haug W, Lantzsch HJ (1983) Sensitive method for the rapid determination of phytate in cereals and cereal products. J Sci Food Agric 34:1423–1426CrossRef Haug W, Lantzsch HJ (1983) Sensitive method for the rapid determination of phytate in cereals and cereal products. J Sci Food Agric 34:1423–1426CrossRef
23.
go back to reference Phillippy BQ, Johnston MR (1985) Determination of phytic acid in foods by ion chromatography with post-column derivatization. J Food Sci 50:541–542CrossRef Phillippy BQ, Johnston MR (1985) Determination of phytic acid in foods by ion chromatography with post-column derivatization. J Food Sci 50:541–542CrossRef
24.
go back to reference Phillippy BQ, Bland JM, Evens TJ (2003) Ion chromatography of phytate in roots and tubers. J Agric Food Chem 51:350–353PubMedCrossRef Phillippy BQ, Bland JM, Evens TJ (2003) Ion chromatography of phytate in roots and tubers. J Agric Food Chem 51:350–353PubMedCrossRef
25.
go back to reference Talamond P, Gallon G, Guyot JP, Mbome Lape I, Trèche S (1998) Comparison of high-performance ion chromatography and absorptiometric methods for the determination of phytic acid in food samples. Analusis 26:396–400CrossRef Talamond P, Gallon G, Guyot JP, Mbome Lape I, Trèche S (1998) Comparison of high-performance ion chromatography and absorptiometric methods for the determination of phytic acid in food samples. Analusis 26:396–400CrossRef
26.
go back to reference Dost K, Tokul O (2006) Determination of phytic acid in wheat and wheat products by reverse phase high performance liquid chromatography. Anal Chim Acta 558:22–27CrossRef Dost K, Tokul O (2006) Determination of phytic acid in wheat and wheat products by reverse phase high performance liquid chromatography. Anal Chim Acta 558:22–27CrossRef
27.
go back to reference Mayr GW (1988) A novel metal–dye detection system permits picomolar-range h.p.l.c. analysis of inositol polyphosphates from non-radioactively labelled cell or tissue specimens. Biochem J 254:585–591PubMed Mayr GW (1988) A novel metal–dye detection system permits picomolar-range h.p.l.c. analysis of inositol polyphosphates from non-radioactively labelled cell or tissue specimens. Biochem J 254:585–591PubMed
28.
go back to reference Laussmann T, Eujen R, Weisshuhn M, Thiel U, Vogel G (1996) Structure of diphospho-myo-inositol pentakisphosphate and bisdisphospho-myo-inositol tetrakisphosphate from Dictyostelium resolved by NMR analysis. Biochem J 315:715–720PubMed Laussmann T, Eujen R, Weisshuhn M, Thiel U, Vogel G (1996) Structure of diphospho-myo-inositol pentakisphosphate and bisdisphospho-myo-inositol tetrakisphosphate from Dictyostelium resolved by NMR analysis. Biochem J 315:715–720PubMed
29.
go back to reference Hatzack F, Hübel F, Zhang W, Hansen PE, Rasmussen SK (2001) Inositol phosphates from barley low-phytate grain mutants analysed by metal–dye detection HPLC and NMR. Biochem J 354:473–480PubMedCrossRef Hatzack F, Hübel F, Zhang W, Hansen PE, Rasmussen SK (2001) Inositol phosphates from barley low-phytate grain mutants analysed by metal–dye detection HPLC and NMR. Biochem J 354:473–480PubMedCrossRef
Metadata
Title
A simple and rapid colorimetric method for determination of phytate in urine
Authors
Antonia Costa-Bauza
F. Grases
I. Gomila
A. Rodriguez
R. M. Prieto
F. Tur
Publication date
01-12-2012
Publisher
Springer-Verlag
Published in
Urolithiasis / Issue 6/2012
Print ISSN: 2194-7228
Electronic ISSN: 2194-7236
DOI
https://doi.org/10.1007/s00240-012-0473-3

Other articles of this Issue 6/2012

Urolithiasis 6/2012 Go to the issue