Determination of propofol UDP-glucuronosyltransferase (UGT) activities in hepatic microsomes from different species by UFLC-ESI-MS.
Propofol O-glucuronidation has been used as probe reaction to phenotype UGT1A9 activity in human liver, thus a sensitive and specific method for determination of propofol O-glucuronide (PG) is urgently desirable. In the current study, a new LC-ESI-MS method for determination of PG in hepatic microsomes from human (HLM), monkey (CyLM), dog (DLM), minipig (PLM), rat (RLM) and mouse (MLM) was developed and validated using 4-methylumbelliferyl-β-d-glucuronide as an internal standard (IS). PG and IS was separated by a Shim-pack XR-ODS column (100 mm × 2.0mm, 2.2 μm, Shimadzu) under gradient conditions with the mobile phase of acetonitrile and water containing 0.2% acetic acid (v/v). The mass spectrometric detection was performed under selected ion monitoring (SIM) for PG at m/z 353 and IS at m/z 351. The assay exhibited linearity over the range 0.05-30 μM for PG with the correlation coefficient of 0.9995. The intra- and inter-day precision was less than 7.2%, with accuracy in the range 93.8-107.5%. The developed method was successfully used for characterizing interspecies and human individual differences in the O-glucuronidation activity towards propofol, as well as investigating inhibitory effects of androsterone and phenylbutazone on propofol O-glucuronidation in HLM.
Liang
Si-Cheng
SC
Laboratory of Pharmaceutical Resource Discovery, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhong-shan Road, Dalian, China.
Ge
Guang-Bo
GB
Liu
Hui-Xin
HX
Shang
Hai-Tao
HT
Wei
Hong
H
Fang
Zhong-Ze
ZZ
Zhu
Liang-Liang
LL
Mao
Yu-Xi
YX
Yang
Ling
L
D019342:Acetic Acid / Q000737:chemistry; D000818:Animals; D002138:Calibration; D004285:Dogs; D014453:Glucuronosyltransferase / Q000032:analysis*; D000882:Haplorhini; D006801:Humans; D006923:Hymecromone / Q000031:analogs & derivatives / Q000737:chemistry; D007700:Kinetics; D013058:Mass Spectrometry / Q000379:methods*; D051379:Mice; D008862:Microsomes, Liver / Q000378:metabolism*; D008956:Models, Chemical; D015742:Propofol / Q000737:chemistry; D051381:Rats; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
D006923:Hymecromone; C034888:4-methylumbelliferyl glucuronide; D014453:Glucuronosyltransferase; D019342:Acetic Acid; D015742:Propofol