Quenched phosphorescence as alternative detection mode in the chiral separation of methotrexate by electrokinetic chromatography

Quenched phosphorescence was used, for the first time, as detection mode in the chiral separation of methotrexate (MTX) enantiomers by electrokinetic chromatography. The detection is based on dynamic quenching of the strong emission of the phosphorophore 1-bromo-4-naphthalene sulfonic acid (BrNS) by MTX under deoxygenated conditions. The use of a background electrolyte with 3 mg/mL 2-hydroxypropyl-β-cyclodextrin and 20% MeOH in 25 mM phosphate buffer (pH 7.0) and an applied voltage of 30 kV allowed the separation of l-MTX and its enantiomeric impurity d-MTX with sufficient resolution. In the presence of 1 mM BrNS, a detection limit of 3.2 × 10−7 M was achieved, about an order of magnitude better than published techniques based on UV absorption. The potential of the method was demonstrated with a degradation study and an enantiomeric purity assessment of l-MTX. Furthermore, l-MTX was determined in a cell culture extract as a proof-of-principle experiment to show the applicability of the method to biological samples

Technique for the measurement of intrinsic pulse-shape discrimination for organic scintillators using tagged neutrons

Fast-neutron/gamma-ray pulse-shape discrimination has been performed for the organic liquid scintillators NE 213A and EJ 305 using a time-of-flight based neutron-tagging technique and waveform digitization on an event-by-event basis. Gamma-ray sources and a Geant4-based simulation were used to calibrate the scintillation-light yield. The difference in pulse shape for the neutron and gamma-ray events was analyzed by integrating selected portions of the digitized waveform to produce a figure-of-merit for neutron/gamma-ray separation. This figure-of-merit has been mapped as a function of detector threshold and also of neutron energy determined from time-of-flight. It shows clearly that the well-established pulse-shape discrimination capabilities of NE 213A are superior to those of EJ 305. The extra information provided by the neutron-tagging technique has resulted in a far more detailed assessment of the pulse-shape-discrimination capabilities of these organic scintillators