Skip to main content
Log in

Determination of pipecolic acid following trimethylsilyl and trifluoroacyl derivatisation on plasma filter paper by stable isotope GC-MS for peroxisomal disorders

  • Research Articles
  • Drug Actions
  • Published:
Archives of Pharmacal Research Aims and scope Submit manuscript

Abstract

If early diagnosis is not made, patients with peroxisomal disorders rapidly progress to sudden death, physical defect or mental retardation resulted in storage of the toxic material into the brain. Therefore, it is necessary to develop the analytical method for rapid screening and/or correct confirmation diagnosis. The method utilizes [2H9]pipecolic acid as internal standard. The formation of trimethylsilyl derivative (TMS) of the carboxylic functional group was performed by adding MSTFA. And then 5 μL of methyl orange was added until the color of methyl orange was to yellow. Consecutively, the trifluoroacyl (TFA)-derivative of the -NH functional group was produced by adding MBTFA. GC-MS was set with specific ions (m/z 282, m/z 297) of the TMSTFA derivative of pipecolic acid for selected ion monitoring. The linearity of pipecolic acid in pooled plasma spots showed 0.9999 in the range of 10–150 ng investigated. The precision and accuracy was within S.D. of 5% (RSD, within 5%) for intra-day and inter-day assay. Normal control value has been determined in plasma spots of infant and adults aged 0–30 (including newborn). The utility of the method was demonstrated by quantifying various concentration of fortified pipecolic acid on a filter plasma spot. The new method was simple with just two step derivatisation, time and labor saving without SPE or liquid-liquid extraction, and convenience of delivery owing to the use of filter paper. The described method could be used for routine analysis, monitoring, and clinical diagnostic application of peroxisomal disorders on dietary therapy.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Danks, D. M., Tippett,. P, Adams, C., and Campbell, P., Cerebro-hepipecolic acidto-renal syndrome of Zellweger. A report of eight cases with comments upon the incidence, the liver lesion, and a fault in pipecolic acid metabolism. J. Pediatr., 86, 382–387(1975).

    Article  PubMed  CAS  Google Scholar 

  • Datta, N. S., Wilson, G. N., and Hajra, A. K., Deficiency of enzymes catalyzing the biosynthesis of glycerol-ether lipids in Zellweger syndrome. A new category of metabolic disease involving the absence of peroxisomes. N. Engl. J. Med., 311, 1080–1083 (1984).

    Article  PubMed  CAS  Google Scholar 

  • Hutzler, J. and Dancis, J., The determination of pipecolic acid: method and results of hospital survey. Clin. Chim. Acta, 128, 75–82 (1983).

    Article  PubMed  CAS  Google Scholar 

  • Kim, J. S. and Giacobini, E., Quantitative determination and regional distribution of pipecolic acid in rodent brain. Neurochem. Res., 9, 1559–1569 (1984).

    Article  PubMed  CAS  Google Scholar 

  • Kok, R. M., Kaster, L., de Jong, A. P., Poll-The, B., Saudubray, J. M., and Jakobs, C., Stable isotope dilution analysis of pipecolic acid in cerebrospinal fluid, plasma, urine and amniotic fluid using electron capture negative ion mass fragmentography. Clin. Chim. Acta, 168, 143–152 (1987).

    Article  PubMed  CAS  Google Scholar 

  • Lam, S. and Karmen, A., Stereoselective D- and L-amino acid analysis by high-performance liquid chromatography. J. Chromatogr., 289, 339–345 (1984a).

    Article  PubMed  CAS  Google Scholar 

  • Lam, S., Azumaya, H., and Karmen, A., High-performance liquid chromatography of amino acids in urine and cerebrospinal fluid. J. Chromatogr., 302, 21–29 (1984b).

    Article  PubMed  CAS  Google Scholar 

  • Lazarow, P. B., Black, V., Shio, H., Fujiki, Y., Hajra, A. K., Datta, N. S., Bangaru, B.S. and Dancis, J., Zellweger syndrome: biochemical and morphological studies on two pipecolic acidpatients treated with clofibrate. Pediatr. Res., 19, 1356–1364 (1985).

    Article  PubMed  CAS  Google Scholar 

  • Mihalik, S. J., Moser, H. W., Watkins, P. A., Danks, D. M., Poulos, A., and Rhead, W. J., Peroxisomal L-pipecolic acid oxidation is deficient in liver from Zellweger syndrome pipecolic acidpatients. Pediatr. Res., 25, 548–552 (1989).

    Article  PubMed  CAS  Google Scholar 

  • Okano, Y., Kataoka, M., Miyata, T., Morimoto, H., Takahama, K., Hitoshi, T., Kas, Y., Matsumoto, I., and Shinka, T., Simultaneous analysis of pipecolic acid with proline in the brain by selected ion-monitoring technique. Anal. Biochem., 117, 196–202 (1981).

    Article  PubMed  CAS  Google Scholar 

  • Okano, Y., Kadota, T., Nagata, J., Matsuda, A., Ijima, S., Takahama, K., and Miyata, T., Quantification by selected ion monitoring of pipecolic acid, proline, gamma-aminobutyric acid and glycine in rat brain. J. Chromatogr., 310, 251–259 (1984).

    Article  PubMed  CAS  Google Scholar 

  • Pduto, A., Baumgartner, M. R., Verhoeven, N. M., Rabier, D., Spipecolic acidda, M., Nassogne, M. C., Poll-The, B. T., Bonetti, G., Jakobs, C., and Saudubray, J. M., Hyperpipecolic acidaemia: a diagnostic tool for peroxisomal disorders. Mol. Genet. Metab., 82, 224–230 (2004).

    Article  CAS  Google Scholar 

  • Poulos, A., Pollard, A. C., Mitchell, J. D., Wise, G., and Mortimer, G., Pipecolic acidtterns of Refsum’s disease. Phytanic acid oxidase deficiency. Arch. Dis. Child., 59, 222–229 (1984).

    Article  PubMed  CAS  Google Scholar 

  • Schutgens, R. B., Heymans, H. S., Wanders, R. J., van den Bosch, H., and Tager, J, M., Peroxisomal disorders: a newly recognised group of genetic diseases. Eur. J. Pediatr., 144, 430–440 (1986).

    Article  PubMed  CAS  Google Scholar 

  • Steinberg, S., Jones, R., Tiffany, C., and Moser, A., Investigational methods for peroxisomal disorders. Curr. Protoc. Hum. Genet., Chapter 17, Unit 17.6 (2008).

  • van den Berg, G. A., Breukelman, H., Elzinga, H., Trijbels, J. M., Monnens, L. A., and Muskiet, F. A., Determination of pipecolic acid in urine and plasma by isotope dilution mass fragmentography. Clin. Chim. Acta, 159, 229–237 (1986).

    Article  PubMed  Google Scholar 

  • Verhoeven, N. M., Kulik, W., van den Heuvel, C. M., and Jakobs, C., Pre- and postnatal diagnosis of peroxisomal disorders using stable-isotope dilution gas chromatographymass spectrometry. J. Inherit. Metab. Dis., 18Suppl 1, 45–60 (1995).

    Article  PubMed  Google Scholar 

  • Wanders, R. J., Kos, M., Roest, B., Meijer, A. J., Schrakamp, G., Heymans, H. S., Tegelaers, W. H., van den Bosch, H., Schutgens, R. B., and Tager, J. M., Activity of peroxisomal enzymes and intracellular distribution of catalase in Zellweger syndrome. Biochem. Biophys. Res. Commun., 123, 1054–1061 (1984).

    Article  PubMed  CAS  Google Scholar 

  • Yoon, H. R., Two step derivatization for the analyses of organic, amino acids and glycines on filter paper plasma by GC-MS/SIM. Arch. Pharm. Res., 30, 387–395 (2007).

    Article  PubMed  CAS  Google Scholar 

  • Zee, T., Stellaard, F., and Jakobs, C., Analysis of pipecolic acid in biological fluids using capillary gas chromatography with electron-capture detection and [2H11]pipecolic acid as internal standard. J. Chromatogr., 574, 335–339 (1992).

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Hye-Ran Yoon.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Yoon, HR., An, YW. Determination of pipecolic acid following trimethylsilyl and trifluoroacyl derivatisation on plasma filter paper by stable isotope GC-MS for peroxisomal disorders. Arch. Pharm. Res. 33, 317–323 (2010). https://doi.org/10.1007/s12272-010-0218-1

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12272-010-0218-1

Key words

Navigation