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. 2023 Nov 30;28(23):7877.
doi: 10.3390/molecules28237877.

Spectrophotometric Study of Charge-Transfer Complexes of Ruxolitinib with Chloranilic Acid and 2,3-Dichloro-5,6-dicyano-1,4-benzoquinone: An Application to the Development of a Green and High-Throughput Microwell Method for Quantification of Ruxolitinib in Its Pharmaceutical Formulations

Khalid A Aljaber  1 , Ibrahim A Darwish  1 , Abdullah M Al-Hossaini  1
Affiliations

Spectrophotometric Study of Charge-Transfer Complexes of Ruxolitinib with Chloranilic Acid and 2,3-Dichloro-5,6-dicyano-1,4-benzoquinone: An Application to the Development of a Green and High-Throughput Microwell Method for Quantification of Ruxolitinib in Its Pharmaceutical Formulations

Khalid A Aljaber et al. Molecules. .

Abstract

Ruxolitinib (RUX) is a potent drug that has been approved by the Food and Drug Administration for the treatment of myelofibrosis, polycythemia vera, and graft-versus-host disease. This study describes the formation of colored charge-transfer complexes (CTCs) of RUX, an electron donor, with chloranilic acid (CLA) and 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ), the π-electron acceptors. The CTCs were characterized using UV-visible spectrophotometry. The formation of CTCs in methanol was confirmed via formation of new absorption bands with maximum absorption at 530 and 470 nm for CTCs with CLA and DDQ, respectively. The molar absorptivity and other physicochemical and electronic properties of CTCs were determined. The molar ratio was found to be 1:1 for both CTCs with CLA and CTCs with DDQ. The site of interaction on RUX molecules was assigned and the mechanisms of the reactions were postulated. The reactions were employed as basis for the development of a novel green and one-step microwell spectrophotometric method (MW-SPM) for high-throughput quantitation of RUX. Reactions of RUX with CLA and DDQ were carried out in 96-well transparent plates, and the absorbances of the colored CTCs were measured by an absorbance microplate reader. The MW-SPM was validated according to the ICH guidelines. The limits of quantitation were 7.5 and 12.6 µg/mL for the methods involving reactions with CLA and DDQ, respectively. The method was applied with great reliability to the quantitation of RUX content in Jakavi® tablets and Opzelura® cream. The greenness of the MW-SPM was assessed by three different metric tools, and the results proved that the method fulfills the requirements of green analytical approaches. In addition, the one-step reactions and simultaneous handling of a large number of samples with micro-volumes using the proposed method enables the high-throughput analysis. In conclusion, this study describes the first MW-SPM, a valuable analytical tool for the quality control of pharmaceutical formulations of RUX.

Keywords: charge-transfer complex; green and high-throughput approach; microwell spectrophotometry; quality control; ruxolitinib.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
The chemical structures of ruxolitinib (RUX), chloranilic acid (CLA), and 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ).
Figure 2
Figure 2
Panel (A) absorption spectra of RUX (30 µg/mL) and reaction mixtures of RUX and CLA (RUX-CLA) containing a fixed concentration of CLA (0.025%, w/v) and varying concentrations of RUX (11.25–37.5 µg/mL). Pane (B) absorption spectra of reaction mixtures of RUX and DDQ (RUX-DDQ) containing a fixed concentration of DDQ (0.025%, w/v) and varying concentrations of RUX (7.5–22.5 µg/mL). All solutions were in methanol. Figures on the spectral lines are the RUX concentrations used to generate the spectra.
Figure 3
Figure 3
Panel (A) Tauc plots of energy (hυ) against (αhυ)2 for CTCs of RUX with CLA and DDQ in methanol solvent. Panel (B) Benesi–Hildebrand plots for formation of the CTCs of RUX with CLA and DDQ. Linear fitting equations and their determination coefficients (R2) are provided on the plot. [A], A, and [D] are the molar concentration of the acceptor reagent (CLA or DDQ), absorbance of the CTC, and molar concentration of RUX, respectively. The values are the means of 3 measurements ± SD.
Figure 4
Figure 4
Panel (A) Job’s continuous variation plots for determination of the molar ratio of the CTCs of RUX with CLA (formula image) and DDQ (formula image). The values are the means of 3 measurements ± SD. Panel (B) a molecule of RUX with atom numbers, and the arrows pointing to the potential atoms for participation in the formation of CTCs, along with their partial charges. Negative sign denotes negative electron density.
Figure 5
Figure 5
The schemes for formation of CTCs of RUX with CLA (A) and DDQ (B).
Figure 6
Figure 6
The effect of the acceptor reagent concentration (A) and time (B) on the CT reactions of RUX with CLA (formula image) and DDQ (formula image). The points are the means of 3 determinations ± SD.
Figure 7
Figure 7
The calibration curves for quantitation of RUX by the MW-SPM via formation of CTCs with CLA and DDQ. The linear regression equations and their determination coefficients (R2) are given on calibration lines.
Figure 8
Figure 8
Results of the GAPI and AGREE metric tools for assessment of the greenness of the proposed MW-SPM for RUX.
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References

    1. Geyer H.L., Knight E., Cannon K., Barr K., Tibes R., Camoriano J., Noel P., Mesa R.A. Ruxolitinib in Clinical Practice for Therapy of Myelofibrosis: Single USA Center Experience After FDA Approval. Blood. 2012;120:5061. doi: 10.1182/blood.V120.21.5061.5061. - DOI - PubMed
    1. FDA. U.S. Food & Drug Administration FDA Approves Treatment for Patients with Rare Bone Marrow Disorder. [(accessed on 7 October 2023)]; Available online: https://www.fda.gov/news-events/press-announcements/fda-approves-treatme....
    1. Jakavi (ruxolitinib) Tablets [Highlights of Prescribing Information]. Wilmington, DE: Incyte Corporation. [(accessed on 7 October 2023)]; Available online: https://www.accessdata.fda.gov/drugsatfda_docs/label/2011/202192lbl.pdf.
    1. FDA. U.S. Food & Drug Administration FDA Approves Ruxolitinib for Acute Graft-Versus-Host Disease. [(accessed on 7 October 2023)]; Available online: https://www.fda.gov/drugs/resources-information-approved-drugs/fda-appro....
    1. FDA. U.S. Food & Drug Administration FDA Approves Ruxolitinib for Chronic Graft-Versus-Host Disease. [(accessed on 7 October 2023)]; Available online: https://www.fda.gov/drugs/resources-information-approved-drugs/fda-appro....