Amino Acids and Catecholamines Analysis in Microdialysates
High speed separation of primary amino acids and catecholamines
This is the fastest LC separations of amino acids using fluorescence detection at such low concentrations, opening the way to high throughput amino acids and catecholamines analyses in neuropharmacology.
Primary amino acids and catecholamines separate in a single run
High speed method: 8 minutes for a run
Limit of detection :
- Glutamate: 7.8 x 10-10 mol/L (7.8 x 10-15 moles)
- Gaba: 1.6 x 10-9 mol/L (1.6 x 10-14 moles)
- Aspartate: 8.1 x 10-10 mol/L (8.1 x 10-15 moles)
(LOD calculated for a S/N ratio of 3)
Sample injection of 10 µl
(microdialysate diluted 1/10)
NDA labelling reagent:
NDA is a fluorogenic dye which fluoresces weakly in its native form but has a good fluorescent yield when reacted with CN- and a primary amine. Such derivatives are excitable at 410 nm or 442 nm, using respectively diode or HeCd lasers.
Derivatization procedure: refer to TN002
Instruments | Waters® ACQUITY Ultra Performance LC™ System Picometrics ZETALIF LASER detector Laser: 410 nm, 25 mW |
Samples | Standards : Amino acids at 10-7 M and Histamine, Noradrenaline, Dopamine at 5 x 10-7 M Microdialysates : diluted 10 times |
Method | Flowrate: 0.2 ml/min T= 30°C Column: Acquity UPLC BEH C18 1.7µm 2.1x100mm Capillary: 150 µm ID Phases : A : Sodium citrate buffer 5mM pH=3.1 B : Acetonitrile |
