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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

Doxorubicin, Doxorubicinol, and Cyclophosphamide

Simultaneous LCMSMS Method for Doxorubicin, Doxorubicinol, and Cyclophosphamide
Analyte	Lower Limit of Quantitation (mg/L)	Acceptable Anticoagulants	Required PlasmaVolume	CV% low QC
Doxorubicin	0.00304	Heparinized Plasma (Na)	Minimal 1 mL*	5
Doxorubicinol	0.0072			5
Cyclophosphamide	0.44			10

Samples are prepared by solid phase extraction prior to chromatography. Internal standards are added to 0.4 mL of sample mixed with HCL solution prior to application onto a preconditioned Waters Oasis� HLB cartridge. The cartridge is rinsed with 5% methanol in HPLC grade water. The samples are then eluted with methanol and evaporated to dryness. The samples are reconstituted with a 75:25 mix of mobile phase A and acetonitrile, respectively. Twenty 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. Each sample analysis requires 11 minutes.