Intake of myo-inositol hexaphosphate and urinary excretion of inositol phosphates in Wistar rats: Gavage vs. oral administration with sugar

Autoři: F. Grases aff001;  A. Costa-Bauzá aff001;  F. Berga aff001;  R. M. Gomila aff002;  G. Martorell aff002;  M. R. Martínez-Cignoni aff003
Působiště autorů: Laboratory of Renal Lithiasis Research, University Institute of Health Sciences Research (IUNICS-IdISBa), University of Balearic Islands, Ctra Valldemossa, Palma de Mallorca, Spain aff001;  Serveis Cientificotècnics, University of Balearic Islands, Ctra Valldemossa, Palma de Mallorca, Spain aff002;  Grup de Metabolisme Energètic i Nutrició, Dept. Biologia Fonamental i Ciències de la Salut, University Institute of Health Sciences Research (IUNICS-IdISBa), University of Balearic Islands, Ctra Valldemossa, Palma de Mallorca, Spain aff003
Vyšlo v časopise: PLoS ONE 14(10)
Kategorie: Research Article
doi: 10.1371/journal.pone.0223959



To evaluate the urinary levels of inositol phosphates (InsPs) in rats that received different salts of myo-inositol hexaphosphate (InsP6) by gavage or by oral administration.


Thirty rats received AIN-76A diet (in which InsPs are undetectable) for 15 days. Then, 12 rats received InsP6 by gavage as a Na salt or a Ca/Mg salt; after 4 days, the Na or Ca/Mg InsP6 was administered with water containing 15 g/L sucrose and urine samples were collected. The other 18 rats received oral InsP6, in which 0.5 g of sugar was combined with InsP6 as a Na salt, a Ca/Mg salt, or a Na salt with CaCO3; daily urine samples were collected. Urine levels of InsPs were determined using a nonspecific method and a specific method (polyacrylamide gel electrophoresis, PAGE), and different InsPs were identified by mass spectroscopy (MS).


After 15 days of the InsP6-free diet, the non-specific method detected no urinary InsPs, and MS detected only InsP2. After administration of Na-InsP6 by gavage, the non-specific method indicated more urinary InsPs than the amount of InsP6 determined by PAGE. MS indicated the presence of urinary InsP2, InsP3, InsP4, InsP5, and InsP6 in these rats, with notable variations among animals. Use of the same treatment to administer Ca/Mg-InsP6 led to a lower overall content of urinary InsPs and a lower level of InsP6. Oral administration of InsP6 as a sugar pill led to lower urinary levels of InsPs than administration of InsP6 by gavage, and administration as a Ca/Mg pill or a Ca/Mg pill with CaCO3 led to lower levels than administration as a Na pill.


Administration of InsP6 to rats leads to the excretion of a mixture of different InsPs. Rats more effectively absorb InsP6 when supplied without dietary components that interfere with its uptake, such as the Ca ion and sugar.

Klíčová slova:

Analysis of variance – Diet – Excretion – Sodium chloride – Sucrose – Urine – Polyacrylamide gel electrophoresis – Oral administration


1. Schlemmer U, Frølich W, Prieto RM, Grases F. Phytate in foods and significance for humans: Food sources, intake, processing, bioavailability, protective role and analysis. Mol Nutr Food Res 2009; 53 (S2): S330–S375.

2. Grases F, Costa-Bauza A, Berga F, Rodriguez A, Gomila RM, Martorell G, et al. Evaluation of inositol phosphates in urine after topical administration of myo-inositol hexaphosphate to female Wistar rats, Life Sci 2018: 192, 33–37. doi: 10.1016/j.lfs.2017.11.023 29155299

3. Chakraborty A, Kim S, Snyder SH. Inositol pyrophosphates as mammalian cell signals. Sci Signal 2011: 4(188), re1. doi: 10.1126/scisignal.2001958 21878680

4. Tuytten R, Lemiere F, Witters E, Van Dongen W, Slegers H, Newton RP, et al. Stainless steel electrospray probe: A dead end for phosphorylated organic compounds?, J Chromatogr A 2006:1104 (1–2), 209–221. doi: 10.1016/j.chroma.2005.12.004 16378618

5. Sakamaki H, Uchida T, Lim LW, Takeuchi T, Evaluation of column hardware on liquid chromatography–mass spectrometry of phosphorylated compounds, J. Chromatogr. A 2015: 1381, 125–131. doi: 10.1016/j.chroma.2014.12.088 25604270

6. McIntyre CA, Arthur CJ, Evershed RP, High-resolution mass spectrometric analysis of myo-inositol hexakisphosphate using electrospray ionisation Orbitrap, Rapid Commun Mass Spectrom 2017: 31(20), 1681–1689. doi: 10.1002/rcm.7935 28696018

7. Tur F, Tur E, Lentheric I, Mendoza P, Encabo M, Isern B, et al. Validation of an LC-MS bioanalytical method for quantification of phytate levels in rat, dog and human plasma, J Chromatogr B Anal. Technol Biomed Life Sci 2013: 928, 146–154.

8. Rougemont B, Fonbonne C, Lemoine J, Bourgeaux V, Salvador A. Liquid chromatography coupled to tandem mass spectrometry for the analysis of inositol hexaphosphate after solid-phase extraction, J Liq Chromatogr Relat Technol 2016: 39(8), 408–414.

9. Ito M, Fujii N, Wittwer C, Sasaki A, Tanaka M, Bittner T, et al. Hydrophilic interaction liquid chromatography–tandem mass spectrometry for the quantitative analysis of mammalian-derived inositol poly/pyrophosphates, J Chromatogr A 2018: 1573, 87–97. doi: 10.1016/j.chroma.2018.08.061 30220429

10. Corbett A, McGowin A, Sieber S, Flannery T, Sibbitt B, A method for reliable voluntary oral administration of a fixed dosage (mg/kg) of chronic daily medication to rats, Lab Anim 2012: 46 (4), 318–324. doi: 10.1258/la.2012.012018 22969146

11. Diogo LN, Faustino IV, Afonso RA, Pereira SA, Monteiro EC, Santos AI, Voluntary Oral Administration of Losartan in Rats. J Am Assoc Lab Anim Sci 2015: 54(5), 549–56. 26424254

12. Costa-Bauzá A, Grases F, Fakier S, Rodriguez A, A novel metal-dye system for urinary phytate detection at micro-molar levels in rats, Anal. Methods 2013: 5, 3016–3022.

13. Wilson MSC, Bulley SJ, Pisani F, Irvine RF, Saiardi A, A novel method for the purification of inositol phosphates from biological samples reveals that no phytate is present in human plasma or urine, Open Biol 2015: 5(3), 150014–150014. doi: 10.1098/rsob.150014 25808508

14. Losito O, Szijgyarto Z, Resnick AC, Saiardi A, Inositol Pyrophosphates and Their Unique Metabolic Complexity: Analysis by Gel Electrophoresis, PLoS One 2009: 4(5), e5580. doi: 10.1371/journal.pone.0005580 19440344

15. Thomas WC, Tilden MT, Inhibition of mineralization by hydrolysates of phytic acid., Johns Hopkins Med J 1972: 131(2), 133–42. 4340528

16. Van Den Berg CJ, Hill LF, Stanbury SW, Inositol phosphates and phytic acid as inhibitors of biological calcification in the rat, Clin Sci 1972: 43, 377–383. 5077515

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2019 Číslo 10