Simultaneous quantitation of hydroxychloroquine and its metabolites in mouse blood and tissues using LC-ESI-MS/MS: An application for pharmacokinetic studies
et al., Journal of Chromatography B, Analytical Technologies in the Biomedical and Life Sciences, 22 Nov 2017, 1072:320-327
Chhonker et al., Simultaneous quantitation of hydroxychloroquine and its metabolites in mouse blood and tissues using.., Journal of Chromatography B, Analytical Technologies in the Biomedical and Life Sciences, 22 Nov 2017, 1072:320-327
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.
Abstract: HHS Public Access
J Chromatogr B Analyt Technol Biomed Life Sci. Author manuscript; available in PMC
2019 January 01.
Published in final edited form as:
J Chromatogr B Analyt Technol Biomed Life Sci. 2018 January 01; 1072: 320–327. doi:10.1016/
Simultaneous quantitation of hydroxychloroquine and its
metabolites in mouse blood and tissues using LC–ESI–MS/MS:
An application for pharmacokinetic studies
Yashpal S. Chhonker1,*, Richard L Sleightholm2,*, Jing Li2, David Oupický2,3, and Daryl J.
of Pharmacy Practice, University of Nebraska Medical Center, Omaha, NE 68198,
for Drug Delivery and Nanomedicine, Department of Pharmaceutical Sciences, University
of Nebraska Medical Center, Omaha, NE 68198, United States
and Pamela Buffett Cancer Center, University of Nebraska Medical Center, Omaha, NE
68198, United States
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 C18 (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 (R2) 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.
Corresponding author: Daryl J. Murry, Pharm. D., Department of Pharmacy Practice, College of Pharmacy, University of Nebraska
Medical Center, Omaha, NE 68198-6025, Phone: 402-559-3790 (office), 402-559-2430 (lab), firstname.lastname@example.org.
*Authors contributed equally to this manuscript
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Conflict of interest
There is no conflict of interest to disclose.
Chhonker et al.
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