Optima LC–MS grade methanol, acetonitrile, water, ammonium acetate, ammonium formate, and formic acid were purchased from Fisher Scientific (Hampton, NH) and used as received unless otherwise specified. Synthetic urine (Sigmatrix Urine Dilutent) was purchased from Millipore Sigma (Saint Louis, MO). All opioid and metabolite standard solution aliquots were purchased from Cerilliant (Round Rock, TX) and diluted to their stock concentrations in compatible solvents (methanol or acetonitrile). Deuterated internal standards (buprenorphine-d4, dihydrocodeine-d6, fentanyl-d5, methadone-d9, norbuprenorphine-d3, norfentanyl-d5, and oxycodone-d6) methanol solution aliquots were also purchased from Cerilliant (Round Rock, TX). Working stock solutions were prepared to a concentration of 2.5 μg/mL for all opioids and metabolites examined in this study. For flow-injection analysis, all working stock solutions were diluted to a concentration of 500 ng/mL in LC–MS grade water.
Norbuprenorphine d3
Manufactured by Cerilliant
Norbuprenorphine-d3 is a deuterated internal standard for the quantitative analysis of norbuprenorphine in biological samples using mass spectrometry. It is used as a reference compound to facilitate the identification and quantification of the target analyte in complex matrices.
Automatically generated - may contain errors
Lab products found in correlation
Optima lc ms grade methanol, by Thermo Fisher Scientific (1 mentions)
Formic acid, by Thermo Fisher Scientific (1 mentions)
Acetonitrile, by Thermo Fisher Scientific (1 mentions)
Ammonium acetate, by Thermo Fisher Scientific (1 mentions)
Ammonium formate, by Thermo Fisher Scientific (1 mentions)
Buprenorphine d4, by Cerilliant (1 mentions)
Oxycodone d6, by Cerilliant (1 mentions)
Norfentanyl d5, by Cerilliant (1 mentions)
Fentanyl d5, by Cerilliant (1 mentions)
Protease type xiv, by Merck Group (1 mentions)
Nexera uhplc, by Shimadzu (1 mentions)
Zeba spin column, by Thermo Fisher Scientific (1 mentions)
2 protocols using norbuprenorphine d3
1
Quantitative Analysis of Opioids in Urine
2
APAP-CYS Quantification in Liver
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The APAP-CYS content in liver was measured by LC-MS/MS following the previously described method71 (link). Liver protein extract was filtered through a desalting column (Zeba spin column, Thermo Fisher) pre-equilibrated with 50 mM (NH4)2HCO3 following the manufacturer’s instructions. An aliquot (45 µl) of the filtrate was mixed with 5 µl of protease type XIV (80 U/ml) (Sigma) and incubated for 24 hours at 37 °C to liberate APAP-CYS. After digestion, 5 µL of norbuprenorphine-d3 (100 µg/ml, Cerilliant Corp.) was added as an internal standard, followed by acetonitrile (300 µL) for precipitation. Dry pellet was obtained by evaporation at 13 psi, and was reconstituted in 100 µL of 2% acetonitrile buffer with 0.1% formic acid. The reconstituted solution was cleared by centrifugation and the supernatant were transferred to a clean vial for LC-MS/MS (Thermo TSQ Quantiva triple-quadrupole mass spectrometer coupled with a Shimadzu Nexera UHPLC). Free APAP-CYS (m.w. 270, Cayman #26388) was used for the quantification standard.
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