Determination of a Nucleotide Diphosphate in Monkey and Rabbit Tissues
Instrument: Precellys® 24
Sample: Monkey and rabbit tissue from liver, lung, spleen, kidney and skin
Protocol:
- Lysing beads were not used to prevent absorption or degradation of the analyte.
- Samples: Monkey and rabbit tissue from liver, lung, spleen, kidney and skin (30 mg each).
- Solvent: 430 μl of 70:30 methanol:water with 30 μl of internal standard.
- Run the Precellys 24 at 6500rpm, 1x30 sec.
- The Precellys was used at full capacity (24 samples at a time), followed by centrifugation at 13000 rpm for 5 minutes. The supernatant was filtered through a Sirocco Protein Precipitation plate and transferred to a 96-well collection block for LC-MS/MS analysis.
Results:
The Precellys 24 allowed validation of this method over a linear range of 5 to 1000 pg/mg. Full validations were performed for monkey and rabbit liver tissue. Validation experiments included intra- and inter-day precision and accuracy, selectivity/specificity, recovery, matrix effect, lower and upper limit of quantification and stability tests.
The accuracy for monkey liver ranged from 97.7% to 101% and the precision ranged from 4.3% to 8.2% (CV). For rabbit liver, the accuracy range was 101% to 102% and the precision range was 4% to 7.5% (CV).
