Methods
In Part 2 of the study, the following adaptations were made to the study
methodology:
- Mixing was conducted in a deactivated glass HPLC vial to minimise the
possibility of surface interactions and accommodate limited sample
quantities.
- Test mixtures were prepared in 0.9%w/v saline only as no differences
between saline and Ringer’s Lactate were observed in the first part of
the study.
- Only OXY concentration was assayed as Part 1 results had provided no
indication that TXA concentration is compromised by co-mixing.
1.0 mL of 0.9%w/v saline solution was accurately pipetted into a
deactivated glass HPLC vial. An accurately pipetted aliquot of OXY
injection solution was added to the saline solution in the HPLC vial and
the solution vortex mixed. The volume of OXY injection solution added
was 20µL for experiments using OXY 1 (5 IU/mL) and 10µL for OXY 2 (10
IU/mL) to accommodate the different concentration strengths of the two
products. Following the addition of OXY, a 100µL aliquot of TXA
injection solution was accurately measured into the HPLC vial and the
solution vortex mixed. This provides solution concentrations for both
active compounds comparable to those used in Part 1 of the study. On
completion of mixing, the HPLC vial was placed immediately into the
LC-MS equipment and the assay process started with samples analysed at
t=0 minutes and every 10 minutes thereafter over a 1-hour period.
In total, 19 combinations of OXY and TXA in saline solution were
evaluated. All 18 TXA products were assessed in combination with the OXY
1 product, while a combination of TXA 8 and OXY 2 was assessed to
evaluate a specific combination of interest to the E-MOTIVE Trial.