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Core Analytical Laboratory

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University at Buffalo, the State University of New York School of Pharmacy and Pharmaceutical Sciences

LCMSMS

Buprenorphine and Norbuprenorphine

Protease Inhibitor

Cortisol, Corticosteriods, Mycophenolic Acid and Mycophenolic Acid Glucuronide

Midazolam, Omeprazole and their Hydroxy Metabolites

Doxorubicin, Doxorubicinol, and Cyclophosphamide

Naltrexone and Naltrexol

Tenofovir

Protease

Simultaneous LCMSMS Method for Protease Inhibitors Plus Efavirenz
Analyte	Lower Limit of Quantitation(ug/L)	Acceptable Anticoagulants	Required PlasmaVolume	MSMSMode
Amprenavir (APV)	16.3	K3-EDTA Heparin	Minimal 0.6 mL
Atazanavir (ATV)	41.0
Efavirenz (EFV)	205
Indinavir (IDV)	16.3
Lopinavir (LPV)	16.3
Nelfinavir (NFV)	16.3
Nelfinavir'sActive Metabolite	8.19
Ritonavir (RTV)	51.2
Saquinavir	16.3

After the addition of three internal standard solutions and base, drugs within plasma samples are extracted via liquid-liquid extraction using a 1:1 hexane: ethyl acetate mixture. The organic supernatant is collected and evaporated to dryness. Then, the samples are reconstituted in mobile phase and injected into the LCMS system. A reversed phase chromatography method, using gradient elution, separates the compounds, which are then detected using MS/MS technology. Thus, after the drug molecule is identified, it is isolated and fragmented. The mass of the charged, unfragmented molecule and its corresponding primary fragment are monitored by the instrument. Since the drugs analyzed here form positively charged ions in all cases except EFV, the LCMS is operated in such a manner as to detect cations. Quantitation of EFV is accomplished through a separate injection, with an isocratic HPLC method and negative ion mass spectrometric detection. The lower limit of quantitation for these eight drugs range from 8 to 200 ng/ml.