Croubels, Siska

De Clercq, Nathalie

Delahaut, Philippe

Vanden Bussche, Julie

Vanhaecke, Lynn

Ghent University Academic Bibliography

Plackett-­‐Burman	  design	  as	  a	  tool	  to	  develop	  a	  generic	  extraction	  for	  U-­‐HPLC-­‐Orbitrap-­‐MS	  analysis	  of	  glucocorticoids	  in	  livestock	  urine.	  	  Nathalie	  De	  Clercq1,	  Julie	  Vanden	  Bussche1,	  Siska	  Croubels2,	  Philippe	  Delahaut3	  and	  Lynn	  Vanhaecke1	  	  1Ghent	  University,	   Faculty	   of	   Veterinary	  Medicine,	   Research	   group	   of	   Veterinary	   Public	   Health	   and	  Zoonoses,	  Laboratory	  of	  Chemical	  Analysis,	  Salisburylaan	  133,	  B-­‐9820	  Merelbeke,	  Belgium.	  2Ghent	   University,	   Faculty	   of	   Veterinary	   Medicine,	   Department	   of	   Pharmacology,	   Toxicology	   and	  Biochemistry,	  Salisburylaan	  133,	  B-­‐9820	  Merelbeke,	  Belgium.	  3CER	  Groupe,	  Département	  Santé,	  Rue	  du	  Point	  du	  Jour	  8,	  B-­‐6900	  Marloie,	  Belgium.	  	  Keywords:	  prednisolone,	  methylprednisolone,	  urine,	  Plackett-­‐Burman,	  U-­‐HPLC,	  Orbitrap-­‐MS	  	  The	   natural	   glucocorticoids,	   cortisol	   and	   cortisone,	   are	   steroid	   hormones	   secreted	   by	   the	   adrenal	  cortex.	  Their	  anti-­‐inflammatory	  properties	  have	   led	  to	  the	  development	  of	  synthetic	  glucocorticoids	  exerting	   even	   higher	   inflammatory	   activities,	   including	   prednisolone	   and	   methylprednisolone,	  nowadays	   commonly	   used	   as	   therapeutic	   drugs.	   Besides,	   corticosteroids	   are	   known	   to	   display	  growth-­‐promoting	   effects,	   which	   has	   led	   to	   their	   illegal	   abuse	   in	   livestock.	   In	   order	   to	   protect	   the	  consumer	   against	   potential	   harmful	   residues	   present	   in	   the	   derived	   animal	   products,	   their	   use	   for	  growth-­‐promoting	   purposes	   in	   cattle	   has	   been	   prohibited	   within	   the	   European	   Union	   (EC	   Council	  directive	   96/22).	   Recent	   developments	   within	   the	   field	   of	   analytical	   chemistry	   and	   the	   resulting	  lowering	   limits	   in	   detection,	   have	   resulted	   in	   the	   regular	   detection	  of	   prednisolone-­‐positive	   bovine	  urine	   samples.	   In	   order	   to	   determine	   the	   origin	   of	   this	   prednisolone,	   a	   general	   extraction	   protocol	  was	  developed.	  The	   sample	  preparation	  was	  based	  on	  a	   two-­‐step	   liquid-­‐liquid	  extraction	  with	   tert-­‐butyl	  methylether	  and	  was	  developed	  by	  using	  a	  Plackett-­‐Burman	  design.	  This	  experimental	  design	  identifies	   the	  most	   significant	   independent	   factors	   in	   a	  minimal	   number	   of	   experiments.	   The	   time	  saving	   approach	   allows	   estimating	   all	  main	   effects	   clear	   of	   any	   confounded	   two-­‐factor	   interactions	  and	   does	   not	   describe	   interaction	   between	   factors.	   Analysis	   was	   executed	   on	   an	   ultra-­‐high	  performance	   liquid	   chromatograph	   coupled	   to	   an	   Orbitrap	   mass	   spectrometer	   and	   successfully	  validated	   accordingly	   2002/657/EC.	   The	   limit	   of	   quantification	   (LOQ)	   for	   dihydrocortisone,	   cortisol	  and	  cortisone	  ranged	  from	  0.3-­‐0.8	  μg	  L-­‐1.	  CCβs	  for	  prednisolone,	  prednisone	  and	  methylprednisolone	  were	  respectively	  0.4	  μg	  L-­‐1,	  0.3	  μg	  L-­‐1	  and	  0.8	  μg	  L-­‐1.	  Recovery	  ranged	  from	  85%-­‐105%,	  repeatability	  (RSD)	  and	  within-­‐laboratory	  reproducibility	  (RSD)	  were	  lower	  than	  13.9%	  and	  16.4%	  respectively.	  This	  method	  allowed	  the	  simultaneous	  and	  accurate	  analysis	  of	  the	  endogenous	  (cortisol,	  cortisone)	  and	  exogenous	  (prednisone,	  prednisolone,	  methylprednisolone)	  glucocorticoids	  and	  potential	  metabolites	  or	   derivates	   thereof	   in	   urine	   and	   demonstrates	   the	   usefulness	   of	   a	   Plackett-­‐Burman	   experimental	  design	  in	  generic	  extraction	  development.	   

Plackett-Burman design as a tool to develop a generic extraction for U-HPLC-Orbitrap-MS analysis of glucocorticoids in livestock urine

https://biblio.ugent.be/publication/4246614/file/4246634

Plackett-Burman design as a tool to develop a generic extraction for U-HPLC-Orbitrap-MS analysis of glucocorticoids in livestock urine

Abstract

Similar works

Full text

Available Versions

Ghent University Academic Bibliography