Analysis of amino acid neurotransmitters from rat and mouse spinal cords by liquid chromatography with fluorescence detection.

Analysis of amino acid neurotransmitters from rat and mouse spinal cords by liquid chromatography with fluorescence detection.

Journal of pharmaceutical and biomedical analysis (2015-01-18)

Nurullah Şanlı, Sarah E Tague, Craig Lunte

ABSTRACT

A RP-LC-FL detection method has been developed to identify and quantitate four amino acid neurotransmitters including glutamic acid, glycine, taurine and γ-aminobutyric acid in rat and mouse spinal cord tissue. 3-(4-carboxybenzoyl)-2-quinolinecarboxaldehyde (CBQCA) was employed for the derivatization of these neurotransmitters prior to RP-LC-FL analysis. Different parameters which influenced separation and derivatization were optimized. Under optimum conditions, linearity was achieved within the concentration ranges of 0.50-50.00 μM for all analytes with correlation coefficients from 0.9912 to 0.9997. The LODs ranged from 0.03 μM to 0.06 μM. The proposed method has been successfully applied to the determination of amino acid neurotransmitters in biological samples such as rat and mouse spinal cord with satisfactory recoveries.

MATERIALS

Product Number

Brand

Product Description

50439

Supelco

Perchloric acid, 0.01 M HClO₄ in water (0.01N), eluent for IC

673439

Sigma-Aldrich

Dimethyl sulfoxide solution, 50 wt. % in H₂O

07-4875

Sigma-Aldrich

Dimethyl sulfoxide, ≥99.5%

24-1010

Sigma-Aldrich

Perchloric acid, SAJ super special grade, 60.0-62.0%

03-2910

Sigma-Aldrich

Boric acid, SAJ first grade, ≥99.5%

07-4872

Sigma-Aldrich

Dimethyl sulfoxide, ≥99.0%, suitable for absorption spectrum analysis

07-4860

Sigma-Aldrich

Dimethyl sulfoxide, JIS special grade, ≥99.0%

07-4870

Sigma-Aldrich

Dimethyl sulfoxide, SAJ first grade, ≥99.0%

07-4871

Sigma-Aldrich

Dimethyl sulfoxide, suitable for HPLC

311421

Sigma-Aldrich

Perchloric acid, 70%, 99.999% trace metals basis

471267

Sigma-Aldrich

Dimethyl sulfoxide, ≥99.6%, ReagentPlus^®

721956

Sigma-Aldrich

Methanol solution, suitable for NMR (reference standard), 4% in methanol-d₄ (99.8 atom % D), NMR tube size 3 mm × 8 in.

339067

Sigma-Aldrich

Boric acid, 99.97% trace metals basis

24-1060

Sigma-Aldrich

Perchloric acid, 0.1 M

32900

Supelco

Benalaxyl-M, PESTANAL^®, analytical standard

229946

Sigma-Aldrich

Sodium tetraborate, 99.998% trace metals basis

221732

Sigma-Aldrich

Sodium tetraborate, 99%

276855

Sigma-Aldrich

Dimethyl sulfoxide, anhydrous, ≥99.9%

202878

Sigma-Aldrich

Boric acid, 99.999% trace metals basis

D2438

Sigma-Aldrich

Dimethyl sulfoxide, sterile-filtered, BioPerformance Certified, meets EP, USP testing specifications, suitable for hybridoma

41639

Sigma-Aldrich

Dimethyl sulfoxide, BioUltra, Molecular Biology, ≥99.5% (GC)

74875

Sigma-Aldrich

8-Octanoyloxypyrene-1,3,6-trisulfonic acid trisodium salt, suitable for fluorescence, ≥90% (HPCE)

94563

Supelco

Dimethyl sulfoxide, analytical standard

15663

Sigma-Aldrich

Boric acid, BioUltra, Molecular Biology, ≥99.5% (T)

B7901

Sigma-Aldrich

Boric acid, suitable for electrophoresis, ≥99.5%

B2645

Sigma-Aldrich

Boric acid, tablet, 1 g boric acid per tablet

B6768

Sigma-Aldrich

Boric acid, BioReagent, Molecular Biology, suitable for cell culture, suitable for plant cell culture, ≥99.5%

D2650

Sigma-Aldrich

Dimethyl sulfoxide, Hybri-Max^™, sterile-filtered, BioReagent, suitable for hybridoma, ≥99.7%

D4540

Sigma-Aldrich

Dimethyl sulfoxide, ≥99.5% (GC), suitable for plant cell culture

D9170

Sigma-Aldrich

Dimethyl sulfoxide, PCR Reagent