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Buprenorphine Detection in Urine Using Liquid Chromatography-High-Resolution Mass Spectrometry: Comparison With Cloned Enzyme Donor Immunoassay (ThermoFisher) and Homogeneous Enzyme Immunoassay (Immunalysis)

Buprenorphine Detection in Urine Using Liquid Chromatography-High-Resolution Mass Spectrometry: Comparison With Cloned Enzyme Donor Immunoassay (ThermoFisher) and Homogeneous Enzyme Immunoassay (Immunalysis)

Sarah L Belsey, Lewis Couchman, Robert J Flanagan

J Anal Toxicol. 2014 Sep;38(7):438-43.

PMID: 24925983

Abstract:

A sensitive liquid chromatographic-high-resolution mass spectrometric (LC-HR-MS) assay for buprenorphine and its urinary metabolites has been developed that requires minimal sample preparation. The results obtained have been compared with those given by (i) cloned enzyme donor immunoassay (CEDIA) and (ii) homogeneous enzyme immunoassay (HEIA) in the analysis of patient urines submitted for buprenorphine analysis. Centrifuged urine (100 µL) was diluted with internal standard solution (25 µL) + LC eluent (875 µL), and 50 µL of the prepared sample were analyzed (Accucore Phenyl-Hexyl column). MS detection was in alternating positive and negative mode using heated electrospray ionization (ThermoFisher Q Exactive). Intra- and inter-assay accuracy and precision were 104-128 and <11%, respectively, at 5 µg/L. Limits of detection were 1.3 µg/L (buprenorphine, norbuprenorphine and buprenorphine glucuronide) and 2.5 µg/L (norbuprenorphine glucuronide). Immunoassay sensitivity and selectivity were 97 and 100% (HEIA) and 99 and 84% (CEDIA), respectively, compared with LC-HR-MS. In 120 patient urines, norbuprenorphine glucuronide was easily the most abundant analyte except when adulteration with buprenorphine had occurred. The median immunoreactive buprenorphine species present (unhydrolysed urine) were 7.5 and 13% for HEIA and CEDIA, respectively. However, codeine, dihydrocodeine, morphine and morphine-3-glucuronide did not interfere in the HEIA assay.

Chemicals Related in the Paper:

Catalog Number	Product Name	CAS Number	Price
AP101224220	Buprenorphine glucuronide solution	101224-22-0	Price
AP469887294	Norbuprenorphine glucuronide solution	469887-29-4	Price
AS271861	Norbuprenorphine glucuronide-D3 solution		Price

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