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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

Buprenorphine and Norbuprenorphine

Simultaneous LCMSMS Method Buprenorphine and Norbuprenorphine
Analyte	Lower Limit of Quantitation (mg/L)	Acceptable Anticoagulants	Required PlasmaVolume	CV% low QC
Buprenorphine	0.000250	EDTA Plasma (K)	Minimal 1 mL*	10
Norbuprenorphine	0.000250	EDTA Plasma (K)	Minimal 1 mL*	8.8

Samples are prepared by solid phase extraction prior to chromatography. Deuterated internal standards are added to 0.5 mL of sample mixed with HCL solution prior to application onto a preconditioned Waters Oasis� MCX cartridge. The cartridge is rinsed with 5% methanol in 0.1N HCl. The samples are then eluted with 1% ammonium hydroxide in methanol and evaporated to dryness. The samples are reconstituted with a mix of mobile phase A and B. Forty percent of the reconstituted samples are injected in the LCMSMS system.

Samples are analyzed on a Agilent HPLC system coupled to a Sciex API3000 triple quadupole mass analyzer used in the positive MRM mode. A gradient chromatography method separates the compounds from other components on a Waters Symmetry� C18 column. Buprenorphine is detected using MS/MS pair 468/396; NBUP is detected as the protonated species only, 414.7. A slow collection of ions at the detector allows for some gains in sensitivity. Each sample analysis requires 6 minutes.