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Current Pharmaceutical Analysis

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ISSN (Print): 1573-4129
ISSN (Online): 1875-676X

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Research Article

Ultrafiltration-based Extraction and LC-MS/MS Quantification of Phenylalanine in Human Blood Sample for Metabolite Target Analysis

Author(s): Mustafa Çelebier*, Tuba Reçber, Emirhan Nemutlu and Sedef Kır

Volume 17, Issue 1, 2021

Published on: 15 July, 2019

Page: [81 - 86] Pages: 6

DOI: 10.2174/1573412915666190715095300

Price: $65

Abstract

Background: Phenylalanine is a significant biomarker for various diseases like phenylketonuria, gastric cancers, and ischemic stroke according to recent studies.

Method: In the present study; a simple, sensitive, selective and novel analytical method was validated by using an ultrafiltration-based extraction and LC-MS/MS quantification of phenylalanine in human plasma using 13C phenylalanine heavy isotope. Amicon® Ultra Centrifugal Filter was used for ultrafiltration. Parameters affecting LC separation and MS/MS detection were investigated and optimized. Chromatographic separation was achieved on a Merck SeQuant ZIC-HILIC (100x4.6 mm, 5 μm) at a column temperature of 40°C using a mobile phase of mixture of acetonitrile containing 0.1% formic acid and water containing 0.1% formic acid (50:50 v/v) at a flow rate of 0.35 mL/min. The transitions m/z 167→121 for 13C phenylalanine, m/z 166→120 for phenylalanine itself were monitored using the MRM mode.

Result: The assay was linear concentration range of 0.0025 μg/mL to 1.20 μg/mL (R2=0.999). The developed method was validated according to FDA guidelines. The method was found linear, sensitive, precise, accurate, and selective.

Keywords: LC-MS/MS, metabolomics, phenylalanine, quantification, ultrafiltration, blood.

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[1]
Bracewell-Milnes, T.; Saso, S.; Abdalla, H.; Nikolau, D.; Norman-Taylor, J.; Johnson, M.; Holmes, E.; Thum, M.Y. Metabolomics as a tool to identify biomarkers to predict and improve outcomes in reproductive medicine: a systematic review. Hum. Reprod. Update, 2017, 23(6), 723-736.
[http://dx.doi.org/10.1093/humupd/dmx023] [PMID: 29069503]
[2]
Mastrangelo, A.; Armitage, E.G.; García, A.; Barbas, C. Metabolomics as a tool for drug discovery and personalised medicine. A review. Curr. Top. Med. Chem., 2014, 14(23), 2627-2636.
[http://dx.doi.org/10.2174/1568026614666141215124956] [PMID: 25515755]
[3]
Yi, L.; Dong, N.; Yun, Y.; Deng, B.; Ren, D.; Liu, S.; Liang, Y. Chemometric methods in data processing of mass spectrometry-based metabolomics: A review. Anal. Chim. Acta, 2016, 914, 17-34.
[http://dx.doi.org/10.1016/j.aca.2016.02.001] [PMID: 26965324]
[4]
Siddiqui, M.R.; AlOthman, Z.A.; Rahman, N. Analytical techniques in pharmaceutical analysis: a review. Arab. J. Chem., 2017, 10(1), 1409-1421.
[http://dx.doi.org/10.1016/j.arabjc.2013.04.016]
[5]
AlOthman, Z.A.; Rahman, N.; Siddiqui, M.R. Review on pharmaceutical impurities, stability studies and degradation products: an analytical approach. Rev. Adv. Sci. Eng., 2013, 2(2), 155-166.
[http://dx.doi.org/10.1166/rase.2013.1039]
[6]
Al Hafid, N.; Christodoulou, J. Phenylketonuria: a review of current and future treatments. Transl. Pediatr., 2015, 4(4), 304-317.
[PMID: 26835392]
[7]
Bilder, D.A.; Noel, J.K.; Baker, E.R.; Irish, W.; Chen, Y.; Merilainen, M.J.; Prasad, S.; Winslow, B.J. Systematic review and meta-analysis of neuropsychiatric symptoms and executive functioning in adults with phenylketonuria. Dev. Neuropsychol., 2016, 41(4), 245-260.
[http://dx.doi.org/10.1080/87565641.2016.1243109] [PMID: 27805419]
[8]
Waisbren, S.E.; Noel, K.; Fahrbach, K.; Cella, C.; Frame, D.; Dorenbaum, A.; Levy, H. Phenylalanine blood levels and clinical outcomes in phenylketonuria: a systematic literature review and meta-analysis. Mol. Genet. Metab., 2007, 92(1-2), 63-70.
[http://dx.doi.org/10.1016/j.ymgme.2007.05.006] [PMID: 17591452]
[9]
Ormstad, H.; Verkerk, R.; Sandvik, L. Serum phenylalanine, tyrosine, and their ratio in acute ischemic stroke: on the trail of a biomarker? J. Mol. Neurosci., 2016, 58(1), 102-108.
[http://dx.doi.org/10.1007/s12031-015-0659-6] [PMID: 26423306]
[10]
Pasieka, A.; Morgan, J. Quantitative determination of phenylalanine on paper chromatograms. Anal. Chem., 1956, 28(12), 1964-1966.
[http://dx.doi.org/10.1021/ac60120a040]
[11]
McCaman, M.W.; Robins, E. Fluorimetric method for the determination of phenylalanine in serum. J. Lab. Clin. Med., 1962, 59(5), 885-890.
[12]
Girotti, S.; Ferri, E.; Ghini, S.; Budini, R.; Carrea, G.; Bovara, R.; Piazzi, S.; Merighi, R.; Roda, A. Bioluminescent flow sensor for L-phenylalanine determination in serum. Talanta, 1993, 40(3), 425-430.
[http://dx.doi.org/10.1016/0039-9140(93)80255-P] [PMID: 18965647]
[13]
Nabi, S.A.; Usmani, S.; Rahman, N. Paper-electrophoretic studies of α-amino acids in citrate–phosphate buffer and sodium dodecyl sulphate mixed systems. Acta Chromatogr., 1998, 8, 175-189.
[14]
Nishiumi, S.; Shima, K.; Azuma, T.; Yoshida, M. Evaluation of a novel system for analyzing hydrophilic blood metabolites. J. Biosci. Bioeng., 2017, 123(6), 754-759.
[http://dx.doi.org/10.1016/j.jbiosc.2017.01.015] [PMID: 28291661]
[15]
Carrozzo, M.M.; Cannazza, G.; Pinetti, D.; Di Viesti, V.; Battisti, U.; Braghiroli, D.; Parenti, C.; Baraldi, M. Quantitative analysis of acetylcholine in rat brain microdialysates by liquid chromatography coupled with electrospray ionization tandem mass spectrometry. J. Neurosci. Methods, 2010, 194(1), 87-93.
[http://dx.doi.org/10.1016/j.jneumeth.2010.09.016] [PMID: 20888860]
[16]
Beger, R.D. A review of applications of metabolomics in cancer. Metabolites, 2013, 3(3), 552-574.
[http://dx.doi.org/10.3390/metabo3030552] [PMID: 24958139]
[17]
Guasch-Ferré, M.; Hruby, A.; Toledo, E.; Clish, C.B.; Martínez-González, M.A.; Salas-Salvadó, J.; Hu, F.B. Metabolomics in prediabetes and diabetes: a systematic review and meta-analysis. Diabetes Care, 2016, 39(5), 833-846.
[http://dx.doi.org/10.2337/dc15-2251] [PMID: 27208380]
[18]
Breier, M.; Wahl, S.; Prehn, C.; Fugmann, M.; Ferrari, U.; Weise, M.; Banning, F.; Seissler, J.; Grallert, H.; Adamski, J.; Lechner, A. Targeted metabolomics identifies reliable and stable metabolites in human serum and plasma samples. PLoS One, 2014, 9(2)e89728
[http://dx.doi.org/10.1371/journal.pone.0089728] [PMID: 24586991]
[19]
Simó, C.; Ibáñez, C.; Gómez-Martínez, A.; Ferragut, J.A.; Cifuentes, A. Is metabolomics reachable? Different purification strategies of human colon cancer cells provide different CE-MS metabolite profiles. Electrophoresis, 2011, 32(13), 1765-1777.
[http://dx.doi.org/10.1002/elps.201100019] [PMID: 21626520]
[20]
Çelebier, M.; Ibáñez, C.; Simó, C.; Cifuentes, A. A foodomics approach: CE-MS for comparative metabolomics of colon cancer cells treated with dietary polyphenols.Protein Electrophoresis; Springer, 2012, pp. 185-195.
[http://dx.doi.org/10.1007/978-1-61779-821-4_15]
[21]
Tiziani, S.; Emwas, A-H.; Lodi, A.; Ludwig, C.; Bunce, C.M.; Viant, M.R.; Günther, U.L. Optimized metabolite extraction from blood serum for 1H nuclear magnetic resonance spectroscopy. Anal. Biochem., 2008, 377(1), 16-23.
[http://dx.doi.org/10.1016/j.ab.2008.01.037] [PMID: 18312846]

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