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Improving Compound Extraction Efficiency using the Precellys 24

Instrument: Precellys® 24

Sample:  Deramatomed human abdominal skin

Protocol:

  • Precellys kits: MK28-R_2mL (KT03961-1-008.2) for use with a Biocap® decapper
  • Samples: Deramatomed human abdominal skin (1.5 cm2, 500 μm thickness, and <300 mg) after a 6-hr skin penetration study
  • Solvent: 500 μl of 1:1 methanol:water + 1% formic acid
  • Run the Precellys 24 at 6500 rpm, 1x30 sec (epidermis) and 6500 rpm, 2x60 sec each, 30 sec pause (dermis)
  • Implemented automation methods by using a decapper, customized tubes, and liquid handling for sample preparation prior to LC-MS/MS
  • LE: Samples placed in glass vials with 8mL of solvent were incubated for 15 or 48 hours before LC-MS/MS analysis.

Results:

The dermis and epidermis values were compared between the three different extraction methods. The five minute Precellys homogenization procedure showed increased extraction of compound SB-275833 from the dermis samples for all three formulations compared to LE (Figure 1A). For epidermis samples, similarly higher extraction was observed for formulations B and C (Figure 1B).

 
Dermal (A) and epidermal (B) amount (μg) of SB-275833 after a 15 and 48 hour liquid extraction (LE) versus homogenization using the Precellys.