Microchimica Acta

, 185:505 | Cite as

An amino-functionalized mesoporous silica (KIT-6) as a sorbent for dispersive and ultrasonication-assisted micro solid phase extraction of hippuric acid and methylhippuric acid, two biomarkers for toluene and xylene exposure

  • Mohammad BehbahaniEmail author
  • Saman Bagheri
  • Fariborz Omidi
  • Mostafa Mohammadpour Amini
Original Paper


The authors described a new application of amino-functionalized KIT-6 for dispersive ultrasonication-assisted micro solid phase extraction of hippuric acid (HA) and methyl hippuric acid (MHA) from human urine and water samples. In the first step, an amino-functionalized mesoporous silica of type KIT-6 was synthesized. It was characterized by field emission scanning electron microscopy, Fourier transform infrared spectrometry, nitrogen adsorption-desorption analysis, thermogravimetry and X-ray diffraction. Following sorption and desorption with 1.0 mL methanol/NH4OH (1%; v/v), HA and MHA were quantified by HPLC with UV detection. Various important parameters were optimized by Box-Behnken design. Under optimized conditions, The limit of detections (LOD) were calculated by a signal-to-noise ratio of 3, which were 0.5 μg L−1 and 0.2 μg L-1 for HA and MHA, respectively, and the calibration plot is linear in the 1–1000 μg L−1 concentration range. Only small matrix effects were found. The method was successfully implemented for the sensitive determination of HA and MHA in (spiked) human urine samples.

Graphical abstract

Schematic of a sorbent for dispersive micro solid phase extraction coupled with ultrasonic power. It consists of amino functionalized KIT-6 and was used for the simultaneous preconcentration and determination of low levels of hippuric and methyl hippuric acid in urine and water samples.


Urine sample High performance liquid chromatography Box-Behnken design Simultaneous detection Metabolites Occupational health Surface functionalization 



We gratefully acknowledge Iranian National Science Foundation (INSF).

Compliance with ethical standards

The author(s) declare that they have no competing interests.

Supplementary material

604_2018_3038_MOESM1_ESM.docx (258 kb)
ESM 1 (DOCX 258 kb)


  1. 1.
    (ACGIH) (2014) A.C.o.G.I.H., Peracetic Acid Documentation of the TLVs and BEIsGoogle Scholar
  2. 2.
    Moro AM, Brucker N, Charão M, Bulcão R, Freitas F, Baierle M, Nascimento S, Valentini J, Cassini C, Salvador M, Linden R, Thiesen F, Buffon A, Moresco R, Garcia SC (2012) Evaluation of genotoxicity and oxidative damage in painters exposed to low levels of toluene. Mutat Res Genet Toxicol Environ Mutagen 746:42–48CrossRefGoogle Scholar
  3. 3.
    Kira S (1977) Measurement by gas chromatography of urinary hippuric acid and methylhippuric acid as indices of toluene and xylene exposure. Occup Environ Med 34:305–309CrossRefGoogle Scholar
  4. 4.
    De Carvalho D, Lanchote VL, Bonato PS, Queiroz RHC, Santos AC, Dreossi SAC (1991) A new derivatization procedure for the analysis of hippuric acid and m-methyl-hippuric acid by gas chromatography. Int Arch Occup Environ Health 63:33–37CrossRefGoogle Scholar
  5. 5.
    Kongtip P, Vararussami J, Pruktharathikul V (2001) Modified method for determination of hippuric acid and methylhippuric acid in urine by gas chromatography. J Chromatogr B Biomed Sci Appl 751:199–203CrossRefGoogle Scholar
  6. 6.
    Pagnotto LD, Lieberman LM (1967) Urinary hippuric acid excretion as an index of toluene exposure. Am Ind Hyg Assoc J 28:129–134CrossRefGoogle Scholar
  7. 7.
    Sakai T, Ninuma Y, Yanagihara S, Ushio K (1983) Simultaneous determination of hippuric acid and o-, m-and p-methylhippuric acids in urine by high-performance liquid chromatogaphy. J Chromatogr B Biomed Sci Appl 276:182–188CrossRefGoogle Scholar
  8. 8.
    Frederick CP, NIOSH Manual of Analytical Methods (NMNM) (2003) NIOSH 8301: National Institute for Occupational Safety and Health. Cincinnati, OHGoogle Scholar
  9. 9.
    Arabi M, Ghaedi M, Ostovan A (2017) Water compatible molecularly imprinted nanoparticles as a restricted access material for extraction of hippuric acid, a biological indicator of toluene exposure, from human urine. Microchim Acta 184:879–887CrossRefGoogle Scholar
  10. 10.
    Ghamari F, Bahrami A, Yamini Y, Ghorbani shahna F, Moghimbeigi A (2017) Hollow-fiber liquid-phase microextraction based on carrier-mediated transport for determination of urinary methyl hippuric acids. Toxicol Environ Chem 99:760–771CrossRefGoogle Scholar
  11. 11.
    Toulabi P, Daneshfar A, Sahrai R (2010) Determination of hippuric acid in biological fluids using single drop liquid–liquid-liquid microextraction. Anal Methods 2:564–569CrossRefGoogle Scholar
  12. 12.
    Vasconcelos I, Fernandes C (2017) Magnetic solid phase extraction for determination of drugs in biological matrices. TrAC Trends Anal Chem 89:41–52CrossRefGoogle Scholar
  13. 13.
    Behbahani M, Najafi M, Amini MM, Sadeghi O, Bagheri A, Salarian M (2013) Dithizone-modified nanoporous fructose as a novel sorbent for solid-phase extraction of ultra-trace levels of heavy metals. Microchim Acta 180:911–920CrossRefGoogle Scholar
  14. 14.
    Behbahani M, Abolhasani J, Amini MM, Sadeghi O, Omidi F, Bagheri A, Salarian M (2015) Application of mercapto ordered carbohydrate-derived porous carbons for trace detection of cadmium and copper ions in agricultural products. Food Chem 173:1207–1212CrossRefGoogle Scholar
  15. 15.
    Liu S, Chen J, Peng Y, Hu F, Li K, Song H, Li X, Zhang Y, Li J (2018) Studies on toluene adsorption performance and hydrophobic property in phenyl functionalized KIT-6. Chem Eng J 334:191–197CrossRefGoogle Scholar
  16. 16.
    Bagheri A, Behbahani M, Amini MM, Sadeghi O, Tootoonchi A, Dahaghin Z (2012) Preconcentration and separation of ultra-trace palladium ion using pyridine-functionalized magnetic nanoparticles. Microchim Acta 178:261–268CrossRefGoogle Scholar
  17. 17.
    Wang W, Qi R, Shan W, Wang X, Jia Q, Zhao J, Zhang C, Ru H (2014) Synthesis of KIT-6 type mesoporous silicas with tunable pore sizes, wall thickness and particle sizes via the partitioned cooperative self-assembly process. Microporous Mesoporous Mater 194:167–173CrossRefGoogle Scholar
  18. 18.
    Maria Chong, A. S., Zhao. X. S., Functionalization of SBA-15 with APTES and characterization of functionalized materials. J Phys Chem B 2003, 107, 12650–12657, Functionalization of SBA-15 with APTES and Characterization of Functionalized MaterialsCrossRefGoogle Scholar
  19. 19.
    Zhao XS, Lu GQ, Whittaker AJ, Millar GJ, Zhu HY (1997) Comprehensive study of surface chemistry of MCM-41 using 29Si CP/MAS NMR, FTIR, pyridine-TPD, and TGA. J Phys Chem B 101:6525–6531CrossRefGoogle Scholar
  20. 20.
    Gregg SJ, Sing KSW (1982) Adsorption, surface area and porosity, second edn. Academic Press, LondonGoogle Scholar
  21. 21.
    Boulaoued A, Fechete I, Donnio B, Bernard M, Turek P, Garin F (2012) Mo/KIT-6, Fe/KIT-6 and Mo–Fe/KIT-6 as new types of heterogeneous catalysts for the conversion of MCP. Microporous Mesoporous Mater 155:131–142CrossRefGoogle Scholar
  22. 22.
    Prabhu A, Kumaresan L, Palanichamy M, Murugesan V (2009) Synthesis and characterization of aluminium incorporated mesoporous KIT-6: efficient catalyst for acylation of phenol. Appl Catal A Gen 360:59–65CrossRefGoogle Scholar
  23. 23.
    Behbahani M, Omidi F, Ghanbari Kakavandi M, Hesam G (2017) Selective and sensitive determination of silver ions at trace levels based on ultrasonic-assisted dispersive solid-phase extraction using ion-imprinted polymer nanoparticles. Appl Organomet Chem 31:e3758CrossRefGoogle Scholar
  24. 24.
    Ghanbari Kakavandi M, Behbahani M, Omidi F, Hesam G (2017) Application of ultrasonic assisted-dispersive solid phase extraction based on ion-imprinted polymer nanoparticles for Preconcentration and trace determination of Lead ions in food and water samples. Food Anal Methods 10:2454–2466CrossRefGoogle Scholar
  25. 25.
    NIOSH (National Institute for Occupational Safety and Health) 2003. Manual of analytical methods (NMAM), 4th ed. Hippuric and methyl Hippuric acids in urine: method 8301. (3):1-5. Retrived from:
  26. 26.
    Ahmadi F, Asgharloo H, Sadeghi S, Gharehbagh-Aghababa V, Adibi H (2009) Post-derivatization procedure for determination of hippuric acid after extraction by an automated micro solid phase extraction system and monitoring by gas chromatography. J Chromatogr B 877:2945–2951CrossRefGoogle Scholar
  27. 27.
    Ohashi Y, Mamiya T, Mitani K, Wang B, Takigawa T, Kira S, Kataoka H (2006) Simultaneous determination of urinary hippuric acid, o-, m-and p-methylhippuric acids, mandelic acid and phenylglyoxylic acid for biomonitoring of volatile organic compounds by gas chromatography–mass spectrometry. Anal Chim Acta 566:167–171CrossRefGoogle Scholar
  28. 28.
    Wang JZ, Lu XY, Zhao NP, Cheng YY, Zeng s (2007) Simultaneous determination of phenylglyoxylic acid, mandelic acid, styrene glycol and hippuric acid in primary culture of rat hepatocytes incubate by high-performance liquid chromatography. Biomed Chromatogr 21:497–501CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2018

Authors and Affiliations

  • Mohammad Behbahani
    • 1
    Email author
  • Saman Bagheri
    • 2
  • Fariborz Omidi
    • 3
  • Mostafa Mohammadpour Amini
    • 2
  1. 1.Faculty of EngineeringShohadaye Hoveizeh University of TechnologySusangerdIran
  2. 2.Department of ChemistryShahid Beheshti UniversityTehranIran
  3. 3.Research Center for Environmental Determinants of Health (RCEDH)Kermanshah University of Medical SciencesKermanshahIran

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