Fast and simultaneous analysis of two immunosuppressants in whole blood using high-performance liquid chromatography

Yongpeng Huang, Hui Tang, Xiangyan Meng, Hui Zhong, Yunyang Song, Bo Chen, Zhiyun Zou

Article ID: 2120
Vol 5, Issue 1, 2024
DOI: https://doi.org/10.54517/aas.v5i1.2120
Received: 04 March 2024; Accepted: 25 April 2024; Available online: 15 May 2024;
Issue release: 30 June 2024

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Abstract

Cyclosporine A and sirolimus are commonly used immunosuppressants in organ transplantation, known for their complementary and synergistic mechanisms of action. However, due to their narrow therapeutic index and critical dosing requirements, maintaining their concentrations within a specific blood range is essential for optimal efficacy and safety. This study systematically compared the chromatographic behaviors of cyclosporine A and sirolimus using a biological liquid chromatography (BioLC) column and traditional liquid chromatography (TraLC) columns under identical conditions. The results indicated that the BioLC column, specifically ZORBAX 300SB C8 (250 mm × 4.6 mm, 5.0 μm), provided the highest peak heights and the narrowest peak widths for both drugs. The number of theoretical plates for both drugs significantly increased on this column when acetonitrile in the mobile phase exceeded 70%. Retention times on both column types were minimally affected by formic acid and trifluoroacetic acid in the mobile phases. Additionally, the ZORBAX 300SB C8 column exhibited a higher number of theoretical plates and a distinct relationship between retention factor and column temperature compared to the TraLC columns. For analysis, a 50 μL whole blood sample was prepared through protein precipitation with 1 mol/L sodium hydroxide, extracted into 500 μL ether-methanol (95:5, v/v), and then centrifuged. The organic layer was evaporated under nitrogen at 50 ℃, and the residue was reconstituted in 200 μL methanol. Cyclosporine A and sirolimus were then separated via isocratic elution on the ZORBAX 300SB C8 column


Keywords

high performance liquid chromatography (HPLC); immunosuppressant; cyclosporine A; sirolimus; biological liquid chromatography column


References

1.

1.         Komeili M, Noorbakhsh F, Esmaili J, et al. Combination therapy of phosphatidylserine liposome with cyclosporine A improves nephrotoxicity and attenuates delayed-type hypersensitivity response. Life Sciences. 2021; 265: 118780. doi: 10.1016/j.lfs.2020.118780

2.

2.         Pınar SG, Canpınar H, Tan Ç, et al. A new nanosuspension prepared with wet milling method for oral delivery of highly variable drug Cyclosporine A: development, optimization and in vivo evaluation. European Journal of Pharmaceutical Sciences. 2022; 171: 106123. doi: 10.1016/j.ejps.2022.106123

3.

3.         Ye Q, Ling S, Jiang G, et al. Sirolimus-based immunosuppression improves the prognosis of liver Transplantation Recipients with low TSC1/2 expression in hepatocellular carcinoma beyond the Milan Criteria. European Journal of Surgical Oncology. 2021; 47(10): 2533-2542. doi: 10.1016/j.ejso.2021.04.001

4.

4.         Lindholm A, Säwe J. Pharmacokinetics and Therapeutic Drug Monitoring of Immunosuppressants. Therapeutic Drug Monitoring. 1995; 17(6): 570-573. doi: 10.1097/00007691-199512000-00004

5.

5.         Stenton SB, Partovi N, Ensom MHH. Sirolimus. Clinical Pharmacokinetics. 2005; 44(8): 769-786. doi: 10.2165/00003088-200544080-00001

6.

6.         Noreikaitė A, Saint-Marcoux F, Kaduševičius E, et al. Cyclosporine therapeutic window evaluation by Chebyshev’s inequality method in kidney recipients. Medicina. 2014; 50(1): 37-43. doi: 10.1016/j.medici.2014.05.008

7.

7.         Anlamlert W, Sermsappasuk P. Pomegranate Juice does not Affect the Bioavailability of Cyclosporine in Healthy Thai Volunteers. Curr Clin Pharmacol. 2020; 15: 145. doi: 10.2174/1574884715666200110153125

8.

8.         Bardelmeijer HA, Ouwehand M, Beijnen JH, et al. Determination of cyclosporin A in human and mouse plasma by reversed-phase high-performance liquid chromatography. Journal of Chromatography B: Biomedical Sciences and Applications. 2001; 763: 201. doi: 10.1016/S0378-4347(01)00389-9

9.

9.         Watanabe T, Tanaka R, Ono H, et al. Sensitive, wide-range and high-throughput quantification of cyclosporine in whole blood using ultra-performance liquid chromatography coupled to tandem mass spectrometry and comparison with an antibody-conjugated magnetic immunoassay. Biomed Chromatogr. 2021; 35: e5128.

10.

10.      Krnáč D, Reiffová K, Rolinski B. A new HPLC-MS/MS method for simultaneous determination of Cyclosporine A, Tacrolimus, Sirolimus and Everolimus for routine therapeutic drug monitoring. Journal of Chromatography B. 2019; 1128: 121772. doi: 10.1016/j.jchromb.2019.121772

11.

11.      Antunes NJ, Kipper K, Couchman L, et al. Simultaneous quantification of cyclosporin, tacrolimus, sirolimus and everolimus in whole blood by UHPLC–MS/MS for therapeutic drug monitoring. Biomedical Chromatography. 2021; 35(6). doi: 10.1002/bmc.5071

12.

12.      Kvamsøe MM, Hansen KR, Skadberg Ø, et al. Salting Out-Assisted Liquid–Liquid Extraction for Liquid Chromatography–Tandem Mass Spectrometry Measurement of Tacrolimus, Sirolimus, Everolimus, and Cyclosporine a in Whole Blood. Therapeutic Drug Monitoring. 2020; 42(5): 695-701. doi: 10.1097/ftd.0000000000000794

13.

13.      Mika A, Stepnowski P. Current methods of the analysis of immunosuppressive agents in clinical materials: A review. Journal of Pharmaceutical and Biomedical Analysis. 2016; 127: 207-231. doi: 10.1016/j.jpba.2016.01.059

14.

14.      Baldelli S, Zenoni S, Merlini S, et al. Simultaneous determination of everolimus and cyclosporine concentrations by HPLC with ultraviolet detection. Clinica Chimica Acta. 2006; 364(1-2): 354-358. doi: 10.1016/j.cca.2005.07.019

15.

15.      Vosough M, Tehrani SM. Development of a fast HPLC-DAD method for simultaneous quantitation of three immunosuppressant drugs in whole blood samples using intelligent chemometrics resolving of coeluting peaks in the presence of blood interferences. Journal of Chromatography B. 2018; 1073: 69-79. doi: 10.1016/j.jchromb.2017.12.012

16.

16.      Bowers LD, Mathews SE. Investigation of the mechanism of peak broadening observed in the high-performance liquid chromatographic analysis of cyclosporine. J Chromatogr. 1985; 333(1): 231. doi: 10.1016/S0021-9673(01)87348-9

17.

17.      Hatsis P, Volmer DA. Evaluation of a cyano stationary phase for the determination of tacrolimus, sirolimus and cyclosporin A in whole blood by high-performance liquid chromatography–tandem mass spectrometry. J Chromatogr B. 2004; 809: 287. doi: 10.1016/S1570-0232(04)00516-1

18.

18.      Deprez S, Stove CP. Fully Automated Dried Blood Spot Extraction coupled to Liquid Chromatography-tandem Mass Spectrometry for Therapeutic Drug Monitoring of Immunosuppressants. Journal of Chromatography A. 2021; 1653: 462430. doi: 10.1016/j.chroma.2021.462430

19.

19.      Matarashvili I, Kobidze G, Chelidze A, et al. The effect of temperature on the separation of enantiomers with coated and covalently immobilized polysaccharide-based chiral stationary phases. Journal of Chromatography A. 2019; 1599: 172-179. doi: 10.1016/j.chroma.2019.04.024

20.

20.      Asnin LD, Stepanova MV. Van’t Hoff analysis in chiral chromatography. Journal of Separation Science. 2018; 41(6): 1319-1337. doi: 10.1002/jssc.201701264

21.

21.      Horváth S, Németh G. Hysteresis of retention and enantioselectivity on amylose tris(3,5-dimethylphenylcarbamate) chiral stationary phases in mixtures of 2-propanol and methanol. Journal of Chromatography A. 2018; 1568: 149-159. doi: 10.1016/j.chroma.2018.07.033

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