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[Detection of Chlormezanone in Blood Using HPLC-MS/MS Method]
Yue Liu1, Weiwei Liang1, Huanhui Zhu1
1Guangzhou Forensic Science Institute, Guangzhou 510442, China.
Objectives:
To establish a high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method for detecting chlormezanone in blood samples.
Methods:
Acetonitrile was added to the samples to precipitate proteins. After vortex and ultrasonication, the mixture was centrifuged, and the supernatant was collected. It was then filtered using a 0.22 μm polytetrafluoroethylene membrane. Separation was performed using an InfinityLab Poroshell 120 EC-C18 column (100 mm×4.6 mm, 2.7 μm). The mobile phase consisted of phase A (0.1% formic acid) and phase B (acetonitrile), with gradient elution at a flow rate of 0.4 mL/min. The mass spectrometer was operated with an electrospray ionization in positive ion mode and multiple reaction monitoring mode.
Results:
Chlormezanone in blood samples showed good linearity within the tested range, with the correlation coefficients (r) all greater than 0.999. The limit of detection and the limit of quantitation of chlormezanone were 22.83 ng/mL and 100 ng/mL, respectively. The matrix effects were 2.1%-3.9% and recoveries were 88.8%-92.3%. Using this method, the mass concentration mass of chlormezanone detected in a positive sample was 1 217.23 ng/mL.
Conclusions:
This method requires simple sample preparation and a small sample volume, and offers a wide linear range, making it suitable for the detection of chlormezanone in blood.
