HPLC-MS/MS analytical method was developed at the ERDC laboratories for the determination of DPX in human plasma.24 (link) Assay linearity was verified for DPX at 2–500 ng/mL with a regression coefficient (R2=0.994). The HPLC-MS/MS system consisted of HPLC Agilent series 1200 (Agilent Technologies Deutschland GmbH, Waldbronn, Germany) system with a mass spectrometer detector (Agilent 6420) Triple Quad G1311A quaternary pump, G1329A autosampler, G1322A vacuum degasser, and mass hunter software. A mixture of 75% acetonitrile: 25% buffer (ammonium formate 20 mM +0.2 % (v/v) formic acid in water) was used as a mobile phase at a flow rate of 0.35 mL/min. The reversed-phase column Intersil ODS −3 (4.6 mm x 50 cm, dp 5 µm Sigma–Aldrich, USA) was temporized at 25 °C. Sildenafil was used as an internal standard (IS).
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HPLC-MS/MS Quantification of DPX in Plasma
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Corresponding Organization :
Other organizations : King Abdulaziz University, Al-Azhar University, Minia University, Military Medical Academy, Virginia Commonwealth University
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Variable analysis
independent variables
- Mobile phase composition (75% acetonitrile: 25% buffer)
- Mobile phase flow rate (0.35 mL/min)
- Column temperature (25 °C)
- Determination of DPX in human plasma
- HPLC-MS/MS analytical method
- Agilent 1200 HPLC system with Agilent 6420 Triple Quad mass spectrometer detector
- Reversed-phase column Intersil ODS-3 (4.6 mm x 50 cm, dp 5 µm)
- Sildenafil as internal standard (IS)
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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