Measurement of hydroxychloroquine in blood from SLE patients using LC-HRMS—evaluation of whole blood, plasma, and serum as sample matrices

Abstract

Background Hydroxychloroquine (HCQ) is the standard of care in the treatment of systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), and other inflammatory rheumatic diseases and potentially for the treatment in COVID-19 patients. Determination of HCQ for therapeutic drug monitoring (TDM) can be performed in whole blood (WB), serum, and plasma. Direct comparisons of WB, serum, and plasma levels of HCQ in patients with SLE have not previously been reported. We describe a method for the determination of HCQ in human blood using liquid chromatography-high-resolution mass spectrometry (LC-HRMS) and compare the suitability of the three sample matrices. Methods A method for the determination of HCQ in human blood using LC-HRMS was developed, validated, and applied for the determination of HCQ levels in WB, serum, and plasma from 26 SLE patients. The reproducibility of the method, in the three matrices, was evaluated using quality control samples and repeated preparations and measurements of patient samples. The performance of the developed method for HCQ measurement in serum was further evaluated by comparison with two previously reported extraction methods. Results The performance of the presented method demonstrated high accuracy and precision. A large range of HCQ concentrations was observed for the SLE patients in all three matrices (WB, serum, and plasma). The mean levels in WB were approximately two-fold the levels in serum and plasma (813 ng/mL compared to 436 ng/mL and 362 ng/mL, respectively). Spiked quality controls showed high reproducibility for all matrices (coefficient of variation, CV, approx. 5%), whereas in patient samples, equally high-precision was only found using WB as the matrix (CV 3%). The CV for serum and plasma was 14% and 39%, respectively. Two alternative methods applied to serum samples did not demonstrate improved precision. Conclusions A LC-HRMS method for the measurement of HCQ in human blood was developed and validated. Whole blood was found to be the superior sample matrix in terms of sample reproducibility. Thus, whole blood samples should be used for HCQ analysis when patients are monitored for HCQ treatment effects. The assay is in clinical use to monitor levels of HCQ in patients.

Multi-Compartment Lymph-Node-on-a-Chip Enables Measurement of Immune Cell Motility in Response to Drugs

“…In this study, we sought to demonstrate the ability of our newly developed LN-on-a-chip to support cellular motility and immune-drug interaction studies in an in vivo like LN microenvironment using HCQ as a model drug. When administered, HCQ is found widely distributed in the human body accumulating in tissues and blood cells [ 72 , 73 , 74 , 75 , 76 ]. We show, through continuous real time monitoring, that T and B immune cells freely move in the biomimetic LN-on-a-chip.…”

Pharmacokinetic Basis of the Hydroxychloroquine Response in COVID-19: Implications for Therapy and Prevention

“…The antiviral effectiveness of the drug ( ɛ ) was herein simulated at (0, 33, 62, 70, 78%, adopted in the system as 0, 0.33, 0.62, 0.7, 0.78, assuming a 100% drug efficacy of 1 and 0% drug efficacy of 0), on the basis of clinical pharmacokinetic results [ 7 , 18 , 26 ] (Figs. 1 , 2 ).…”

“…Unfortunately, no blood concentrations were reported by Gautret et al, but these could be proportionally derived from the standard curve in (Fig. 3 ) and the standard curves for conversion of serum to whole blood and serum to plasma provided in [ 26 ]. This calculation led to a prediction of drug efficacy using Eq.…”

Accurate and sensitive determination of hydroxychloroquine sulfate used on COVID-19 patients in human urine, serum and saliva samples by GC-MS

“…Generally, hydroxychloroquine is qualified and quantified by hyphenated and electroanalytical techniques such as high performance liquid chromatography-ultraviolet detection (HPLC-UV) [ 8 ], high performance liquid chromatography-fluorescence detection (HPLC-FLD) [ 9 ], liquid chromatography-tandem mass spectrometry (LC-MS/MS) [ 10 ], liquid chromatography-high resolution mass spectrometry (LC-HRMS) [ 11 ], potentiometry [ 12 ], differential pulse voltammetry [ 13 ], and square-wave voltammetry [ 14 ]. Gas chromatographs with an appropriate detector are also an important analytical instrument for drug analyses [ 15 , 16 ].…”