Food Analytical Methods

, Volume 9, Issue 5, pp 1391–1397 | Cite as

Evaluation of Phytic Acid Content of Some Tea and Nut Products by Reverse-Phase High Performance Liquid Chromatography/Visible Detector

  • Kenan DostEmail author
  • Gülçin Karaca


Phytic acid contents of nine different types of nut and six different types of tea and steeped tea were analysed by reverse-phase high performance liquid chromatography with visible detector. The extraction method was based on hydrochloric acid extraction, and the analysis method was based on metal replacement reaction of phytic acid from coloured complex (iron(III)-thiocyanate), separation on CN column and monitoring the absorbance at 460 nm. The retention time for the monitored iron(III)-thiocyanate peak was achieved less than 3 min. The proposed HPLC/Vis procedure shows good linearity over the concentration range of 1–150 mg L−1 with a correlation coefficient value of 0.9938. The effectiveness of metal replacement reaction was presented in terms of relative standard deviation that was 0.62 and 0.88 % for 5 and 50 mg L−1 of phytic acid, respectively. Repeatability of the analytical method was ranging between 1.58–7.88 % (n = 10, for 50 mg L−1) and 0.98–4.63 % (n = 10, for 5 mg L−1) in terms of relative standard deviation. Accuracy of the method is good, ranging relative error between 4.52 and 8.00 % (n = 10, for 50 mg L−1). Phytic acid content is in the range of 1.54 to 9.74 mg g−1 in nuts, 27.67 to 28.82 mg g−1 in green teas, and 20.49 to 21.96 mg g−1 in pocketed roasted teas.


Phytic acid HPLC Iron(III)-thiocyanate Iron(III)-phytate Nuts and teas 



The authors are grateful to the Scientific Research Projects Department, Celal Bayar University, for their financial support.

Compliance with Ethical Standards


This study was funded by the Celal Bayar University Scientific Research Projects Department (grant number BAP 2012-077).

Conflict of Interest

Kenan Dost declares that he has no conflict of interest. Gülçin Karaca declares that she has no conflict of interest.

Ethical Approval

This article does not contain any studies with human participants or animal performed by any of the authors.

Informed Consent

Not applicable.


  1. Abeywickrama KRW, Ratnasooriya WD, Amarakoon AMT (2011) Oral hypoglycaemic, antihyperglycaemic and antidiabetic activities of Sri Lankan Broken Orange Pekoe Fannings (BOPF) grade black tea (Camellia sinensis L.) in rats. J Ethnopharmacol 135:278–286CrossRefGoogle Scholar
  2. Amaro R, Murillo M, Gonzalez Z, Escalona A, Hernandez L (2009) Optimization of the treatment of wheat samples for the determination of phytic acid by HPLC with refractive index detection. J AOAC Int 92:873–878Google Scholar
  3. Arcan I, Yemenicioglu A (2009) Antioxidant activity and phenolic content of fresh and dry nuts with or without the seed coat. J Food Compos Anal 22:184–188CrossRefGoogle Scholar
  4. Bunkova R, Marova I, Nemec M (2005) Antimutagenic properties of green tea. Plant Food Hum Nutr 60:25–29CrossRefGoogle Scholar
  5. Cai Y, Luo Q, Sun M, Corke H (2004) Antioxidant activity and phenolic compounds of 112 traditional Chinese medicinal plants associated with anticancer. Life Sci 74:2157–2184CrossRefGoogle Scholar
  6. Cao S, Dong N, Chen J (2011) Synchronous fluorescence determination of phytic acid in foodstuffs and urine based on replacement reaction. Phytochem Anal 22:119–123CrossRefGoogle Scholar
  7. Carneiro JMT, Zagatto EAG, Santos JLM, Lima JLFC (2002) Spectrophotometric determination of phytic acid in plant extracts using a multi-pumping flow system. Anal Chim Acta 474:161–166CrossRefGoogle Scholar
  8. Chaudhuri AKN, Karmakar S, Roy D, Pal S, Pal M, Sen T (2005) Antiinflammatory activity of Indian black tea (Sikkim variety). Pharmacol Res 51:169–175CrossRefGoogle Scholar
  9. Chen QC (2004) Determination of phytic acid and inositol pentakisphosphates in foods by high-performance ion chromatography. J Agric Food Chem 52:4604–4613CrossRefGoogle Scholar
  10. Ciarmiello LF, Mazzeo MF, Minasi P, Peluso A, De Luca A, Piccirillo P, Siciliano RA, Carbone V (2014) Analysis of different European hazelnut (Corylus avellana L.) cultivars: authentication, phenotypic features, and phenolic profiles. J Agric Food Chem 62:6236–6246CrossRefGoogle Scholar
  11. Costa-Bauza A, Grases F, Gomila I, Rodriguez A, Prieto RM, Tur F (2012) A simple and rapid colorimetric method for determination of phytate in urine. Urol Res 40:663–669CrossRefGoogle Scholar
  12. Dost K, Tokul O (2006) Determination of phytic acid in wheat and wheat products by reverse phase high performance liquid chromatography. Anal Chim Acta 558:22–27CrossRefGoogle Scholar
  13. El-Beshbishy HA, Tork OM, El-Bab MF, Autifi MA (2011) Antioxidant and antiapoptotic effects of green tea polyphenols against azathioprine-induced liver injury in rats. Pathophysiology 18:125–135CrossRefGoogle Scholar
  14. Evans WJ, McCourtnay EJ, Shrager RI (1982) Titration studies of phytic acid. J Am Oil Chem Soc 59:189–191CrossRefGoogle Scholar
  15. Febles CI, Arias A, Hardisson A, Rodriguez-Alvarez C, Sierra A (2002) Phytic acid level in wheat flours. J Cereal Sci 36:19–23CrossRefGoogle Scholar
  16. Grases F, March P (1989) A study about some phosphate derivatives as inhibitors of calcium-oxalate crystal-growth. J Cryst Growth 96:993–995CrossRefGoogle Scholar
  17. Grases F, Garcia-Ferragut L, Costal-Bauza A (1996) Study of the early stage of renal stone formation: experimental model using urothelium of pig urinary bladder. Urol Res 24:305–311CrossRefGoogle Scholar
  18. Grases F, Perello J, Isern B, Prieto RM (2005) Determination of myo-inositol hexakisphosphate (phytate) in urine by inductively coupled plasma atomic emission spectrometry. Anal Chim Acta 510:41–43CrossRefGoogle Scholar
  19. Helfrich A, Bettmer J (2004) Determination of phytic acid and its degradation products by ion-pair chromatography (IPC) coupled to inductively coupled plasma-sector field-mass spectrometry (ICP-SF-MS). J Anal Atom Spectr 19:1330–1334CrossRefGoogle Scholar
  20. Hix DK, Klopfenstein CF, Walker CE (1997) Physical and chemical attributes and consumer acceptance of sugar-snap cookies containing naturally occurring antioxidants. Cereal Chem 74:281–283CrossRefGoogle Scholar
  21. Hurrel RF (2004) Phytic acid degradation as a means of improving iron absorption. Int J Vitam Nutr Res 74:445–452CrossRefGoogle Scholar
  22. Khokhar S, Magnusdottir SGM (2002) Total phenol, catechin, and caffeine contents of teas commonly consumed in the United Kingdom. J Agric Food Chem 50:565–570CrossRefGoogle Scholar
  23. Kornsteiner M, Wagner K, Elmadfa I (2006) Tocopherols and total phenolics in 10 different nut types. Food Chem 98:381–387CrossRefGoogle Scholar
  24. Latta M, Eskin M (1980) A simple and rapid colorimetric method for phytate determination. J Agric Food 28:1313–1315CrossRefGoogle Scholar
  25. Lee SH, Park HJ, Chun HK, Cho SY, Cho SM, Lillehoj HS (2006) Dietary phytic acid lowers the blood glucose level in diabetic KK mice. Nutr Res 26:474–479CrossRefGoogle Scholar
  26. Liu X, Villalta PW, Sturla SJ (2009) Simultaneous determination of inositol and inositol phosphates in complex biological matrices: quantitative ion-exchange chromatography/tandem mass spectrometry. Rapid Commun Mass Spectrom 23:705–712CrossRefGoogle Scholar
  27. Lopez HW, Leenhardt F, Coudray C, Remesy C (2002) Minerals and phytic acid interactions: is it a real problem for human nutrition. Int J Food Sci Technol 37:727–739CrossRefGoogle Scholar
  28. Mendel S, Youdim MB (2004) Catechin polyphenols: neurodegeneration and neuroprotection in neurodegenerative diseases. Free Radic Bio Med 37:304–317CrossRefGoogle Scholar
  29. Munoz JA, Valiente M (2003) Determination of phytic acid in urine by inductively coupled plasma mass spectrometry. Anal Chem 75:6374–6378CrossRefGoogle Scholar
  30. Muraoka S, Miura T (2004) Inhibition of xanthine oxidase by phytic acid and its antioxidative action. Life Sci 74:1691–16700CrossRefGoogle Scholar
  31. Norazalina S, Norhaizan ME, Hairuszah I, Norashareena MS (2010) Anticarcinogenic efficacy of phytic acid extracted from rice bran on azoxymethane-induced colon carcinogenesis in rats. Exp Toxicol Pathol 62:259–268CrossRefGoogle Scholar
  32. Oatway L, Vasanthan T, Helm JH (2001) Phytic acid. Food Rev Int 17:419–431CrossRefGoogle Scholar
  33. Oh J, Jo H, Cho AR, Kim SJ, Han J (2013) Antioxidant and antimicrobial activities of various leafy herbal teas. Food Control 31:403–409CrossRefGoogle Scholar
  34. Perello J, Isern B, Costa-Bauza A, Grases F (2004) Determination of myo-inositol in biological samples by liquid chromatography-mass spectrometry. J Chromatogr B 802:367–370CrossRefGoogle Scholar
  35. Phillippary BQ, Lin M, Rasco B (2004) Analysis of phytate in raw and cooked potatoes. J Food Compos Anal 17:217–226CrossRefGoogle Scholar
  36. Regueiro J, Sanchez-Gonzalez C, Vallverdu-Queralt A, Simal-Gandara J, Lamuela-Raventos R, Izquierdo-Pulido M (2014) Comprehensive identification of walnut polyphenols by liquid chromatography coupled to linear ion trap-Orbitrap mass spectrometry. Food Chem 152:340–348CrossRefGoogle Scholar
  37. Rodrigues VC, de Moraes ML, Brisolari A, Soares JC, Ferreira M, Goncalves D (2011) Polypyrrole/phytase amperometric biosensors for the determination of phytic acid in standard solutions. Sensor Actuators B-Chemical 160:222–226CrossRefGoogle Scholar
  38. Rugova A, Puschenreiter M, Santner J, Fischer L, Neubauer S, Koellensperger G, Hann S (2014) Speciation analysis of orthophosphate and myo-inositol hexakisphosphate in soil- and plant-related samples by high-performance ion chromatography combined with inductively coupled plasma mass spectrometry. J Sep Sci 37:1711–1719CrossRefGoogle Scholar
  39. Sanikommu S, Pasupuleti M, Vadalkonda L (2014) Comparison of phosphate estimating methods in the presence of phytic acid for the determination of phytase activity. Prep Biochem Biotechnol 44:231–241CrossRefGoogle Scholar
  40. Saw NK, Chow K, Rao PN, Kavanagh JP (2007) Effects of inositol hexaphosphate (phytate) on calcium binding, calcium oxalate crystallization and in vitro stone growth. J Urol 177:2366–2370CrossRefGoogle Scholar
  41. 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 53:330–375CrossRefGoogle Scholar
  42. Shamsuddin AM (2002) Anticancer function of phytic acid. Int J Food Sci Technol 37:769–782CrossRefGoogle Scholar
  43. Talamond P, Doulbeau S, Rochette I, Guyot JP, Treche S (2000) Anion-exchange high-performance liquid chromatography with conductivity detection for the analysis of phytic acid in food. J Chromatogr A 871:7–12CrossRefGoogle Scholar
  44. Vucenik I, Shamsuddin AM (2006) Protection against cancer by dietary IP6 and inositol. Nutr Cancer 55:109–125CrossRefGoogle Scholar
  45. Woollard DC, Macfadzean C, Indyk HE, McMahon A, Christiansen S (2014) Determination of myo-inositol in infant formulae and milk powders using capillary gas chromatography with flame ionisation detection. Int Dairy J 37:74–81CrossRefGoogle Scholar
  46. Xu Q, Kanthasamy AG, Reddy MB (2008) Neuroprotective effect of the natural iron chelator, phytic acid in a cell culture model of Parkinson’s disease. Toxicology 245:101–108CrossRefGoogle Scholar
  47. Yang CS, Maliakal P, Meng X (2002) Inhibition of carcinogenesis by tea. Annu Rev Pharmacol Toxicol 42:25–54CrossRefGoogle Scholar
  48. Zielinski AAF, Haminiuk CWI, Alberti A, Nogueira A, Demiate IM, Granato D (2014) A comparative study of the phenolic compounds and the in vitro antioxidant activity of different Brazilian teas using multivariate statistical techniques. Food Res Int 60:246–254CrossRefGoogle Scholar
  49. Zuo YG, Chen H, Deng YW (2002) Simultaneous determination of catechins, caffeine and gallic acids in green Oolong, black and pu-erh teas using HPLC with a photodiode array detector. Talanta 57:307–316CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.Department of Chemistry, Faculty of Science and ArtCelal Bayar UniversityManisaTurkey

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