Detection of Chlormezanone in Blood Using HPLC-MS/MS Method

doi:10.12116/j.issn.1004-5619.2024.340403

Abstract

Abstract:

Objective 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. Conclusion 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.

Key words: forensic medicine, toxicological analysis, chlormezanone, blood, high performance liquid chromatography-tandem mass spectrometry （HPLC-MS/MS）

CLC Number:

Yue LIU, Weiwei LIANG, Huanhui ZHU, Tianfu HE, Yuanyuan TIAN, Cong PENG, Songcai WANG. Detection of Chlormezanone in Blood Using HPLC-MS/MS Method[J]. Journal of Forensic Medicine, 2026, 42(2): 130-134.

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Figures/Tables 6

References 14

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质量浓度/（ng·mL^-1）	准确度		精密度
质量浓度/（ng·mL^-1）	日内（n=6）	日间（n=18）	日内（n=6）	日间（n=18）
100	4.2	19.1	7.3	10.2
1 000	3.4	14.8	3.2	8.4
5 000	-0.6	6.6	2.0	6.9

质量浓度/（ng·mL^-1）	准确度		精密度
质量浓度/（ng·mL^-1）	日内（n=6）	日间（n=18）	日内（n=6）	日间（n=18）
100	4.2	19.1	7.3	10.2
1 000	3.4	14.8	3.2	8.4
5 000	-0.6	6.6	2.0	6.9

时间/h	低质量浓度（100 ng/mL）		中质量浓度（1 000 ng/mL）		高质量浓度（5 000 ng/mL）
时间/h	响应强度/cps	偏倚/%	响应强度/cps	偏倚/%	响应强度/cps	偏倚/%
0	284 101.32	0	2 700 275.89	0	12 985 162.34	0
3	282 056.54	-0.7	2 657 591.53	-1.6	13 152 382.14	1.3
6	287 985.28	1.4	2 720 039.71	0.7	13 778 935.32	6.1
9	285 343.74	0.4	2 796 894.99	3.6	13 593 984.91	4.7
12	294 527.44	3.7	2 834 354.35	5.0	14 049 303.15	8.2
15	302 279.34	6.4	2 956 726.60	9.5	14 100 833.44	8.6
18	307 858.07	8.4	3 154 050.23	16.8	14 233 764.87	9.6
21	319 625.01	12.5	3 213 727.91	19.0	14 950 949.79	15.1
24	322 671.21	13.6	3 363 848.14	24.6	15 511 575.78	19.5

时间/h	低质量浓度（100 ng/mL）		中质量浓度（1 000 ng/mL）		高质量浓度（5 000 ng/mL）
时间/h	响应强度/cps	偏倚/%	响应强度/cps	偏倚/%	响应强度/cps	偏倚/%
0	284 101.32	0	2 700 275.89	0	12 985 162.34	0
3	282 056.54	-0.7	2 657 591.53	-1.6	13 152 382.14	1.3
6	287 985.28	1.4	2 720 039.71	0.7	13 778 935.32	6.1
9	285 343.74	0.4	2 796 894.99	3.6	13 593 984.91	4.7
12	294 527.44	3.7	2 834 354.35	5.0	14 049 303.15	8.2
15	302 279.34	6.4	2 956 726.60	9.5	14 100 833.44	8.6
18	307 858.07	8.4	3 154 050.23	16.8	14 233 764.87	9.6
21	319 625.01	12.5	3 213 727.91	19.0	14 950 949.79	15.1
24	322 671.21	13.6	3 363 848.14	24.6	15 511 575.78	19.5

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