Journal
JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
Volume 44, Issue 5, Pages 1139-1146Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jpba.2007.05.020
Keywords
cerebral spinal fluid; ultratiltrate; protein binding; lopinavir; chromatography coupled to mass spectrometry
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Funding
- NCRR NIH HHS [5M01-RR 00044] Funding Source: Medline
- NIDA NIH HHS [DA015024, 3R01DA015024-03S2] Funding Source: Medline
- PHS HHS [SIORRR14572] Funding Source: Medline
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A method for the determination of lopinavir (LPV) concentrations in cerebral spinal fluid (CSF) and plasma ultrafiltrate (UF) was developed and validated to analyze clinical specimens from patients receiving antiretroviral treatment with lopinavir/ritonavir. The CSF (400 mu L sample volume) final calibration range for LPV was 0.313-25.0 ng/mL. The final calibration range for UF (50 mu L sample volume) was 1.25-100 ng/mL. The samples were prepared using liquid-liquid extraction, concentrated, and analyzed using a reversed phase isocratic separation. Detection was achieved in positive mixed reaction monitoring mode on a triple quadrupole mass spectrometer. Isolation of LPV through chromatographic separation and proper selection of calibration matrix were important factors in achieving accurate results. Plasma UF was found to be an equivalent calibration matrix to CSF whereas plasma matrix produced a positive bias in samples with unknown concentrations. Artificial CSF media prepared chemically were biased and less superior than UE Sources of plasma for the UF did not affect accuracy. Several CSF sources were tested for specificity of the method and LPV concentrations were accurately produced with atmospheric pressure chemical ionization source producing more accurate results than the electrospray source. The method successfully measured LPV concentrations in CSF that were previously undetectable by HPLC as well as UF from protein binding studies. (c) 2007 Elsevier B.V. All rights reserved.
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