Rapid Quantification of Buprenorphine-Glucuronide and Norbuprenorphine-Glucuronide in Human Urine by LC-MS-MS

Abstract

A liquid chromatography-tandem mass spectrometry (LC-MS-MS) method was developed and validated for the determination of buprenorphine-glucuronide (BUP-G) and norbuprenorphine-glucuronide (NBUP-G) in human urine. The method included a dilution step followed by filtration through a Mini-Uniprep Filter and direct injection onto the LC column. The analytes were quantified in multiple reactions monitoring mode using one transition ion. Norbuprenorpine-d(3) (NBUP-d(3)) was used as the internal standard. The concentration ranges were 6-161 ng/mL for BUP-G and 12-295 ng/mL for NBUP-G. Recoveries determined after filtration for the analytes were 75%. The between-day precision of the method was in the range of 4.8-11%. The limits of quantification were found to be 4.6 ng/mL for BUP-G and 11.8 ng/mL for NBUP-G. Approximately 1000 samples from law enforcement, prison inmates, probation services, and hospitals were analyzed by the presented method. The ratios of drug glucuronides versus creatinine were calculated for a selection of samples (n = 151), where there was information on treatment with buprenorphine between 16 and 20 mg/day. The majority (86%) of the samples had a ratio of BUP-G/creatinine below 570 microg/g, and 76% of the samples had NBUP-G/creatinine lower than 1060 microg/g. The LC-MS-MS method proved to be robust and specific for the determination of BUP-G and NBUP-G in urine.

Laboratory Testing for Prescription Opioids

“…It is also not detected by morphine-specific opiate immunoassay screening tests (see Table 1). Immunoassay [30], GC-MS [75][76][77], and LC-MS [78][79][80][81][82] methods that are specific for buprenorphine are available and should be used when detection of this drug is required.…”

Determination of opiates and cocaine in urine by high pH mobile phase reversed phase UPLC–MS/MS

“…Different sample preparation procedures for opiates, cocaine and/or benzoylecgonine in urine for LC-MS and LC-MS/MS analyses are described in the literature. Gustavsson et al described a LC-MS/MS method with only dilution of the urine specimen before LC-MS/MS analysis [3], while Hegstad et al have described a method using filtration and dilution of the urine before LC-MS/MS analysis [4]. To reduce the possibility of ion suppression in the ESI source caused by matrix components, a more thorough sample preparation of biological species has been recommended [5][6][7].…”

“…Several LC-MS/MS methods for determination of opiates, cocaine and/or benzoylecgonine in human biological samples with acidic mobile phases have been published, e.g. in oral fluid [10], in plasma [11][12][13][14], in serum [11], in whole blood [11,15,16], in hair [17], in meconium [11] and urine [3,4,11,12,16,[18][19][20][21]. However, chromatographic performance of basic compounds is usually improved with high pH mobile phases as less silanol interactions, increased retention and increased loadability are achieved [22,23].…”

Buprenorphine and norbuprenorphine determination by GC/MS in putrefied biological samples: evaluation of a buprenorphine related death

“…International literature reports survey about deaths involving buprenorphine, where dosages in biological samples, obtained in LC/MS or GC/MS, are presented [1,3,7,8] pointing out the frequent mixture of buprenorphine with sedatives, neuroleptics, antidepressants and other antipsychotics drugs, or alcohol. Difficulty to evaluate the toxicological data related to therapeutic, toxic and lethal levels is also discussed [2,[4][5][6].…”