Comparison of validated HPLC methods using two derivatizing agents for gamma-aminobutyric acid quantification
Abstract
Gamma-aminobutyric acid (GABA) has many advantages in physiological functions and skin benefits. Therefore, a feasible and simple method is needed for its determination. The purpose of this study was to validate and compare two HPLC analyses of GABA after derivatization with 2-hydroxynaphthaldehyde (HN) or with o-phthaladehyde (OPA) and 3-mercaptopropionic acid (MPA) in terms of reproducibility, sensitivity and stability of two derivatives. Reliability resulted in satisfaction for the HPLC methods in each derivatizing agent. Linear correlation was obtained in the concentration ranges of 40-600 μg/ml and 0.2-0.9 μg/ml for GABA-HN and GABA-OPA/MPA, respectively. The low limit of detection (LOD) and the limit of quantification (LOQ) of GABA-HN were 1 μg/ml and 5 μg/ml while LOD and LOQ of GABA-OPA/MPA were 0.004 μg/ml and 0.02 μg/ml. Validation results of two methods with chosen derivatizing agents showed that the methods were reproducible and precise on quantification of GABA. The method with GABA-OPA/MPA derivatization revealed higher sensitivity compared to one with GABA-HN derivatization. However, GABA-OPA/MPA derivative was less stable. In conclusion, both validated methods could be used for sensitive and accurate analysis of GABA.
Full Text:
PDFReferences
Hayakawa K, Kimura M, Kasaha K, et al. Effect of a γ-aminobutyric acid-enriched dairy product on the blood pressure of spontaneously hypertensive and normotensive Wistar–Kyoto rats. Br J Nutr 2004; 92: 411-417.
Mody I, De Koninck Y, Otis TS, et al. Bridging the cleft at GABA synapses in the brain. Trends Neurosci 1994; 17: 517-525.
Takahashi H, Sumi M, Koshino F. Effect of gamma-aminobutyric acid (GABA) on normotensive or hypertensive rats and men. Jpn J Physiol 1961; 11: 89-95.
Roberts E, Frankel S. γ-Aminobutyric acid in brain: its formation from glutamic acid. J Biol Chem 1950; 187: 55-63.
Awapara J, Landua AJ, Fuerst R, et al. Free γ-aminobutyric acid in brain. J Biol Chem 1950; 187: 35-39.
Diana M, Quílez J, Rafecas M. Gamma-aminobutyric acid as a bioactive compound in foods: A review. J Funct Foods 2014; 10: 407-420.
Ishikawa A, Oka H, Hiemori M, et al. Development of a method for the determination of γ-aminobutyric acid in foodstuffs. J Nutr Sci Vitaminol (Tokyo) 2009; 55: 292-295.
Varanyanond W, Tungtrakul P, Surojanametakul V, et al. Effect of water soaking on gamma-aminobutyric acid (GABA) in germ of different Thai rice varieties. Kasetsart Journal (Natural Science) 2005; 39: 411-415.
Abdou AM, Higashiguchi S, Horie K, et al. Relaxation and immunity enhancement effects of γ-aminobutyric acid (GABA) administration in humans. Biofactors 2006; 26: 201-208.
Amihăesei I, Mungiu O. Main neuroendocrine features and therapy in primary sleep troubles. Rev Med Chir Soc Med Nat Iasi 2011; 116: 862-866.
Oh CH, Oh SH. Effects of germinated brown rice extracts with enhanced levels of GABA on cancer cell proliferation and apoptosis. J Med Food 2004; 7: 19-23.
Miura D, Ito Y, Mizukuchi A, et al. Hypocholesterolemic action of pre-germinated brown rice in hepatoma-bearing rats. Life Sci 2006; 79: 259-264.
Hagiwara H, Seki T, Ariga T. The effect of pre-germinated brown rice intake on blood glucose and PAI-1 levels in streptozotocin-induced diabetic rats. Biosci Biotechnol Biochem 2004; 68: 444-447.
Adeghate E, Ponery AS. GABA in the endocrine pancreas: cellular localization and function in normal and diabetic rats. Tissue Cell 2002; 34: 1-6.
Ito K, Tanaka K, Nishibe Y, et al. GABA-synthesizing enzyme, GAD67, from dermal fibroblasts: Evidence for a new skin function. Biochim Biophys Acta 2007; 1770: 291-296.
Di Cagno R, Mazzacane F, Rizzello C, et al. Synthesis of γ-aminobutyric acid (GABA) by Lactobacillus plantarum DSM19463: functional grape must beverage and dermatological applications. Appl Microbiol Biotechnol 2010; 86: 731-741.
Han D, Kim HY, Lee HJ, et al. Wound healing activity of gamma-aminobutyric acid (GABA) in rats. J Microbiol Biotechnol 2007; 17: 1661-1669.
Buck K, Voehringer P, Ferger B. Rapid analysis of GABA and glutamate in microdialysis samples using high performance liquid chromatography and tandem mass spectrometry. J Neurosci Methods 2009; 182: 78-84.
Eckstein JA, Ammerman GM, Reveles JM, et al. Analysis of glutamine, glutamate, pyroglutamate, and GABA in cerebrospinal fluid using ion pairing HPLC with positive electrospray LC/MS/MS. J Neurosci Methods 2008; 171: 190-196.
Sekioka N, Kato D, Kurita R, et al. Improved detection limit for an electrochemical γ-aminobutyric acid sensor based on stable NADPH detection using an electron cyclotron resonance sputtered carbon film electrode. Sens Actuators B Chem 2008; 129: 442-449.
Takeda S, Yamano N, Kawasaki N, et al. Rapid determination of 4-aminobutyric acid and L-glutamic acid in biological decarboxylation process by capillary electrophoresis-mass spectrometry. J Sep Sci 2012; 35: 286-291.
Shah AJ, Crespi F, Heidbreder C. Amino acid neurotransmitters: separation approaches and diagnostic value. J Chromatogr B 2002; 781: 151-163.
Mañes J, Gimeno MJ, Moltó JC, et al. Fluorimetric determination of hydrazine in isoniazid formulations with 2-hydroxy-1-naphthaldehyde. J Pharm Biomed Anal 1988; 6: 1023-1027.
Mohammed El-Brashy A, Mohammed Al-Ghannam S. High-performance liquid chromatographic determination of some amino acids after derivatization with 2-hydroxy-1-naphthaldehyde. Analyst 1997; 122: 147-150.
Cronin JR, Pizzarello S, Gandy WE. Amino acid analysis with o-phthalaldehyde detection: Effects of reaction temperature and thiol on fluorescence yields. Anal Biochem 1979; 93: 174-179.
Lee KS, Drescher DG. Fluorometric amino-acid analysis with o-phthaldialdehyde (OPA). Int J Biochem 1978; 9: 457-467.
Handley MK, Hirth WW, Phillips JG, et al. Development of a sensitive and quantitative analytical method for 1H-4-substituted imidazole histamine H3-receptor antagonists utilizing high-performance liquid chromatography and dabsyl derivatization. J Chromatogr B Biomed Appl 1998; 716: 239-249.
Gübitz G, Wintersteiger R, Hartinger A. Fluorescence derivatization of tertiary amines with 2-naphthyl chloroformate. J Chromatogr A 1981; 218: 51-56.
Sunol C, Artigas F, Tusell JM, et al. High-performance liquid chromatography-fluorescence detection method for endogenous γ-aminobutyric acid validated by mass spectrometric and gas chromatographic techniques. Anal Chem 1988; 60: 649-651.
International conference on harmonisation of technical requirement for registration of pharmaceuticals for human use, Q2(R1): Validation of analytical procedures: Text and methodology
Khuhawar MY, Rajper AD. Liquid chromatographic determination of γ-aminobutyric acid in cerebrospinal fluid using 2-hydroxynaphthaldehyde as derivatizing reagent. J Chromatogr B 2003; 788: 413-418.
de Freitas Silva DM, Ferraz VP, Ribeiro ÂM. Improved high-performance liquid chromatographic method for GABA and glutamate determination in regions of the rodent brain. J Neurosci Methods 2009; 177: 289-293.
Refbacks
- There are currently no refbacks.