tIn this paper the development and validation of a method for the analysis of THC-COOH, THC, THC-OH,CBD and CBN in their total form in urine by LC–MS/MS is presented.Tandem hydrolysis, i.e. enzymatic and basic, has been found optimal for the simultaneous analysis of theselected analytes in urine: basic hydrolysis is more effective for the cleavage of THC-COOH glucuronidewhile enzymatic hydrolysis allows the cleavage of the conjugated cannabinoids possessing ether bonds(THC, THC-OH, CBD).The whole procedure requires a 2 h enzymatic hydrolysis using only 90 L of urine by -SPE extractiontechnique with C18tips. Clear advantages in terms of time and of enzyme reduction are obtained andthe cost of the analysis can be dramatically reduced. Satisfactory recovery values and matrix effect areobtained, and the chromatographic run, performed with a fused-core column, allowed the completeanalyte separation in only 3 min (total run 5.8 min) with a common HPLC system.Furthermore the whole procedure has been validated according to SWGTOX guidelines: LOQs arebetween 6 and 10 ppb, quite lower than the requested cut-off for urine testing; intermediate repro-ducibility of the selected analytes is below 10% and accuracy is between 85% and 113%, except for CBD,included only for semi-quantitative determination.
A μ-SPE PROCEDURE FOR THE DETERMINATION OF CANNABINOIDS AND THEIR METABOLITES IN URINE BY LC–MS/MS
SERGI, Manuel;COMPAGNONE, DARIO;
2014-01-01
Abstract
tIn this paper the development and validation of a method for the analysis of THC-COOH, THC, THC-OH,CBD and CBN in their total form in urine by LC–MS/MS is presented.Tandem hydrolysis, i.e. enzymatic and basic, has been found optimal for the simultaneous analysis of theselected analytes in urine: basic hydrolysis is more effective for the cleavage of THC-COOH glucuronidewhile enzymatic hydrolysis allows the cleavage of the conjugated cannabinoids possessing ether bonds(THC, THC-OH, CBD).The whole procedure requires a 2 h enzymatic hydrolysis using only 90 L of urine by -SPE extractiontechnique with C18tips. Clear advantages in terms of time and of enzyme reduction are obtained andthe cost of the analysis can be dramatically reduced. Satisfactory recovery values and matrix effect areobtained, and the chromatographic run, performed with a fused-core column, allowed the completeanalyte separation in only 3 min (total run 5.8 min) with a common HPLC system.Furthermore the whole procedure has been validated according to SWGTOX guidelines: LOQs arebetween 6 and 10 ppb, quite lower than the requested cut-off for urine testing; intermediate repro-ducibility of the selected analytes is below 10% and accuracy is between 85% and 113%, except for CBD,included only for semi-quantitative determination.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
