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Simultaneous quantitation of hydroxychloroquine and its metabolites in mouse blood and tissues using LC-ESI-MS/MS: An application for pharmacokinetic studies

Chhonker et al., Journal of Chromatography B, Analytical Technologies in the Biomedical and Life Sciences, 22 Nov 2017, 1072:320-327 doi:10.1016/j.jchromb.2017.11.026
Nov 2017  
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HCQ for COVID-19
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Presents a method for quantification of HCQ in mouse blood and tissues. They show a lung concentration significantly higher than other organs, and about 30 times the blood concentration.
Chhonker et al., 23 Nov 2017, peer-reviewed, 5 authors.
This PaperHCQAll
Simultaneous quantitation of hydroxychloroquine and its metabolites in mouse blood and tissues using LC–ESI–MS/MS: An application for pharmacokinetic studies
Yashpal S Chhonker, Richard L Sleightholm, Jing Li, David Oupický, Pharm. D Daryl J Murry
Journal of Chromatography B, doi:10.1016/j.jchromb.2017.11.026
Hydroxychloroquine (HCQ) has been shown to disrupt autophagy and sensitize cancer cells to radiation and chemotherapeutic agents. However, the optimal delivery method, dose, and tumor concentrations required for these effects are not known. This is in part due to a lack of sensitive and reproducible analytical methods for HCQ quantitation in small animals. As such, we developed and validated a selective and sensitive liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) method for simultaneous quantitation of hydroxychloroquine and its metabolites in mouse blood and tissues. The chromatographic separation and detection of analytes were achieved on a reversed phase Thermo Aquasil C 18 (50 × 4.6 mm, 3μ) column, with gradient elution using 0.2 % formic acid and 0.1% formic acid in methanol as mobile phase at a flow rate of 0.5 mL/min. Simple protein precipitation was utilized for extraction of analytes from the desired matrix. Analytes were separated and quantitated using MS/MS with an electrospray ionization source in positive multiple reaction monitoring (MRM) mode. The MS/MS response was linear over the concentration range from 1-2000 ng/mL for all analytes with a correlation coefficient (R 2 ) of 0.998 or better. The within-and between-day precision (relative standard deviation, % RSD) and accuracy were within the acceptable limits per FDA guidelines. The validated method was successfully applied to a preclinical pharmacokinetic mouse study involving low volume blood and tissue samples for hydroxychloroquine and metabolites.
Author Manuscript Chhonker et al. Page 17 Table 1 Summary of MS/MS parameters: precursor ion, fragment ions, voltage potential (Q1), collision energy (CE) and voltage potential (Q3) for analytes. Table 3 Mean extraction recoveries of the HCQ, DCQ, BDCQ and DHCQ from mouse blood and tissues
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Chhonker, Page 22 Table 6 Blood and tissues concentration (ng/mL) and tissue to blood concentration ratio (Kp) of HCQ, DCQ, BDCQ and DHCQ after 5 mg/kg Intra venous administration of HCQ (mean ± SD, n=5)
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