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  6. Simultaneous Analysis of Chlorzoxazone and its Metabolites

Simultaneous Analysis of Chlorzoxazone and its Metabolites

Ingested drugs are typically metabolized by cytochrome P450 (CYP). CYP is a superfamily of enzymes that convert foreign substances taken up in the body into a form(s) that can easily be excreted. There is a large number of CYP isoforms. Among these, nine isoforms (CYP1A2, 2A6, 2B6, 2C8, 2C9, 2C19, 2D6, 2E1, and 3A4) are involved in the metabolism of drugs in human. This sheet describes an analysis of chlorzoxazone 6-hydroxylation, which is a marker activity of CYP2E1.

* This reaction is one of the probe activities in human liver microsomes (HLM).

Detection of chlorzoxazone and its metabolites


Incubation mixture including HLM and chlorzoxazone

* Metabolite: 6-Hydroxychlorzoxazone

* Chlorzoxazone: Central muscle relaxant


Sample preparation method


System configuration

5110 Pump
5210 AutoSampler
5310 Column Oven
5420 UV-VIS Detector
Empower2 Data Processing System


Column HITACHI LaChrom C18 (5 µm) (4.6 mmI.D. x 150 mm)
HITACHI LaChrom C18 (3 µm) (4.6 mmI.D. x 100 mm)
Elute (A) 50 mM KH2PO4(pH 4.5)
*(A)/(B) = 80/20
Flow rate 1.0 mL/min
Column Temperature 35°C
Detection 295 nm
Injection vol. 10 µL

Results of analysis


* This analysis sample was provided by Miki Nakajima, Associate Professor of College of Medical, Pharmaceutical,and Health Science School of Pharmacy at Kanazawa University.

These data are an example of measurement; the individual values cannot be guaranteed.
The system is for research use only, and is not intended for any animal or human therapeutic or diagnostic use.

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