Background: Hyperlipidemia is one of the most common chronic diseases encountered globally, and atorvastatin (ATV) is mainly metabolized into two major active metabolites. Methodology: Hence, we aimed to estimate ATV and ezetimibe (EZE) simultaneously in the presence of ATV major and active metabolites using a validated LC–MS/MS method. Conclusion: The proposed method was linear (r² >0.99), accurate (92.02–109.94%) and precise (CV% ≤14) over the concentration range of 0.50–120 ng/ml, 0.20–48 ng/ml, 0.50–120 ng/ml and 0.20–48 ng/ml for ATV, EZE, 2-hydroxy ATV and 4-hydroxy ATV, respectively. The applied liquid–liquid extraction gave rise to reliable extraction recoveries of 84.91 ± 1.14%, 85.20 ± 1.62%, 85.46 ± 0.41% and 105.46 ± 2.35% for ATV, EZE, 2-hydroxy ATV and 4-hydroxy ATV, respectively.

Keywords:

Papers of special note have been highlighted as: • of interest

References

1. Jain KS, Kathiravan M, Somani RS, Shishoo CJ. The biology and chemistry of hyperlipidemia. Bioorg. Med. Chem. 15(14), 4674–4699 (2007).
Medline CASGoogle Scholar
2. Navar-Boggan AM, Peterson ED, D'Agostino RB Sr, Neely B, Sniderman AD, Pencina MJ. Hyperlipidemia in early adulthood increases long-term risk of coronary heart disease. Circulation 131(5), 451–458 (2015).
Medline CASGoogle Scholar
3. Meex RC, Watt MJ. Hepatokines: linking nonalcoholic fatty liver disease and insulin resistance. Nat. Rev. Endocrinol. 13(9), 509–520 (2017).
Medline CASGoogle Scholar
4. Samuel VT, Shulman GI. The pathogenesis of insulin resistance: integrating signaling pathways and substrate flux. J. Clin. Invest. 126(1), 12–22 (2016).
MedlineGoogle Scholar
5. Catapano A. Ezetimibe: a selective inhibitor of cholesterol absorption. Eur. Heart J. Suppl. 3(Suppl. E), e6–e10 (2001).
CASGoogle Scholar
6. Patel J, Sheehan V, Gurk-Turner C. Ezetimibe (Zetia): a new type of lipid-lowering agent. Presented at: Baylor University Medical Center Proceedings. (2003).
Google Scholar
7. Van Heek M, Davis H. Pharmacology of ezetimibe. E. Heart J. Suppl. 4(Suppl. J), J5–J8 (2002).
CASGoogle Scholar
8. Stancu C, Sima A. Statins: mechanism of action and effects. J. Cell. Mol. Med. 5(4), 378–387 (2001).
Medline CASGoogle Scholar
9. Zahra Bathaie S, Ashrafi M, Azizian M, Tamanoi F. Mevalonate pathway and human cancers. Curr. Mol. Pharmacol. 10(2), 77–85 (2017).
MedlineGoogle Scholar
10. Ghosh C, Jain I, Gaur S, Patel N, Upadhyay A, Chakraborty BS. Simultaneous estimation of atorvastatin and its two metabolites from human plasma by ESI–LC–MS/MS . Drug Test. Anal. 3(6), 352–362 (2011).
Medline CASGoogle Scholar
11. Lennernäs H. Clinical pharmacokinetics of atorvastatin. Clin. Pharmacokinet. 42(13), 1141–1160 (2003). • Clinical pharmacokinetics of atorvastatin.
MedlineGoogle Scholar
12. Bays HE, Chen E, Tomassini JE, McPeters G, Polis AB, Triscari J. Fixed-dose combination ezetimibe+ atorvastatin lowers LDL-C equivalent to co-administered components in randomized trials: use of a dose-response model. Fundam. Clin. Pharmacol. 29(2), 209–218 (2015).
Medline CASGoogle Scholar
13. Patiño-Rodríguez O, Torres-Roque I, Martínez-Delgado M, Escobedo-Moratilla A, Pérez-Urizar J. Pharmacokinetic non-interaction analysis in a fixed-dose formulation in combination of atorvastatin and ezetimibe. Front. Pharmacol. 5, 261 (2014). • Pharmacokinetic analysis of atorvastatin and ezetimibe.
MedlineGoogle Scholar
14. Elkady EF, Aboelwafa AA. A rapid and optimized LC–MS/MS method for the simultaneous extraction and determination of sofosbuvir and ledipasvir in human plasma. J. AOAC Int. 99(5), 1252–1259 (2016).
Medline CASGoogle Scholar
15. Mowaka S, Elkady EF, Elmazar MM, Ayoub BM. Enhanced LC–MS/MS determination of alogliptin and metformin in plasma: application to a pharmacokinetic study. Microchem. J. 130, 360–365 (2017).
CASGoogle Scholar
16. Elkady EF, Aboelwafa AA. Rapid bioanalytical LC–MS/MS method for the simultaneous determination of sofosbuvir and velpatasvir in human plasma-application to a pharmacokinetic study in Egyptian volunteers. J. Chromatogr. B Analyt. Technol. Biomed. Life Sci. 1102, 116–124 (2018).
MedlineGoogle Scholar
17. Elkady EF, Fouad MA, Alshoba N, Mahmoud ST. Validated LC–MS/MS method for the determination of some prescribed CNS drugs: application to an in vivo pharmacokinetic study of drug-herb metabolic interaction potential of khat. Microchem. J. 158, 105261 (2020).
CASGoogle Scholar
18. Elkady EF, Mandour AA, Algethami FK, Aboelwafa AA, Farouk F. Sequential liquid–liquid extraction coupled to LC–MS/MS for simultaneous determination of amlodipine, olmesartan and hydrochlorothiazide in plasma samples: application to pharmacokinetic studies. Microchem. J. 155, 104757 (2020). • Sequential liquid–liquid extraction coupled to LC–MS/MS.
CASGoogle Scholar
19. Van De Merbel NC. Protein quantification by LC–MS: a decade of progress through the pages of Bioanalysis. Bioanalysis 11(07), 629–644 (2019).
Google Scholar
20. Needham SR, Valaskovic GA. Microspray and microflow LC–MS/MS: the perfect fit for bioanalysis. Bioanalysis 7(9), 1061–1064 (2015).
CASGoogle Scholar
21. Côté C, Bergeron A, Mess J-N, Furtado M, Garofolo F. Matrix effect elimination during LC–MS/MS bioanalytical method development. Bioanalysis 1(7), 1243–1257 (2009).
CASGoogle Scholar
22. Arnold DW, Needham SR. Micro-LC–MS/MS : the future of bioanalysis. Bioanalysis 5(11), 1329–1331 (2013).
CASGoogle Scholar
23. Ewles M, Goodwin L. Bioanalytical approaches to analyzing peptides and proteins by LC–MS/MS . Bioanalysis 3(12), 1379–1397 (2011).
CASGoogle Scholar
24. Haneef J, Shaharyar M, Husain A et al. Application of LC–MS/MS for quantitative analysis of glucocorticoids and stimulants in biological fluids. J. Pharm. Anal. 3(5), 341–348 (2013).
Medline CASGoogle Scholar
25. Partani P, Verma SM, Gurule S, Khuroo A, Monif T. Simultaneous quantitation of atorvastatin and its two active metabolites in human plasma by liquid chromatography/(-) electrospray tandem mass spectrometry. J. Pharm. Anal. 4(1), 26–36 (2014). • Simultaneous quantitation of atorvastatin and its two active metabolites in human plasma.
MedlineGoogle Scholar
26. Polagani SR, Pilli NR, Gajula R, Gandu V. Simultaneous determination of atorvastatin, metformin and glimepiride in human plasma by LC–MS/MS and its application to a human pharmacokinetic study. J. Pharm. Anal. 3(1), 9–19 (2013).
Medline CASGoogle Scholar
27. Gajula R, Pilli NR, Ravi VB et al. Simultaneous determination of atorvastatin and aspirin in human plasma by LC–MS/MS: its pharmacokinetic application. Sci. Pharm. 80(4), 923–940 (2012).
Medline CASGoogle Scholar
28. Pilli NR, Inamadugu JK, Mullangi R, Karra VK, Vaidya JR, Seshagiri Rao J. Simultaneous determination of atorvastatin, amlodipine, ramipril and benazepril in human plasma by LC–MS/MS and its application to a human pharmacokinetic study. Biomed. Chromatogr. 25(4), 439–449 (2011).
Medline CASGoogle Scholar
29. Bichala PK, Atmakuri LR, Kotra V. Development and validation of an LC–MS/MS method for determination of atorvastatin in human plasma. Asian J. Medicine and Health Sciences 4, 255 (2021).
Google Scholar
30. Le J, Liao Y, Li S, Chen X, Hong Z. High-throughput LC–MS/MS method for simultaneous determination of pantoprazole and atorvastatin in rat plasma: application to a pharmacokinetic interaction study. Curr. Drug Metab. (2021). https://www.ingentaconnect.com/content/ben/cdm/2021/00000022/00000006/art00007
MedlineGoogle Scholar
31. Xia B, Li Y, Zhang Y et al. UHPLC–MS/MS method for determination of atorvastatin calcium in human plasma: application to a pharmacokinetic study based on healthy volunteers with specific genotype. J. Pharm. Biomed. Anal. 160, 428–435 (2018).
Medline CASGoogle Scholar
32. Varghese SJ, Kochupappy Ravi T. Liquid chromatography/mass spectrometry method for the simultaneous determination of atorvastatin and telmisartan in human plasma. Analytical Chemistry Letters 9(4), 552–563 (2019).
CASGoogle Scholar
33. Rezk MR, Badr KA. Quantification of amlodipine and atorvastatin in human plasma by UPLC–MS/MS method and its application to a bioequivalence study. Biomed. Chromatogr. 32(7), e4224 (2018).
MedlineGoogle Scholar
34. Bae J-W, Choi C-I, Park S-H, Jang C-G, Lee S-Y. Analytical LC–MS/MS method for ezetimibe and its application for pharmacokinetic study. J. Liq. Chromatogr. Relat. Technol. 35(1), 141–152 (2012).
CASGoogle Scholar
35. Yong S, Dong P, Suk H et al. Development and validation of simple and rapid LC–MS/MS method for ezetimibe in human plasma and its application to bioequivalence study. J. Pharm. Biol. Sci. 12(4), 1–8 (2017).
Google Scholar
36. Nishida C, Matsumoto Y, Fujimoto K et al. The bioequivalence and effect of food on the pharmacokinetics of a fixed-dose combination tablet containing rosuvastatin and ezetimibe in healthy Japanese subjects. Clin. Transl. Sci. 12(6), 704–712 (2019).
Medline CASGoogle Scholar
37. Migoya E, Bergman A, Hreniuk D et al. Bioequivalence of an ezetimibe/simvastatin combination tablet and coadministration of ezetimibe and simvastatin as separate tablets in healthy subjects. Int. J. Clin. Pharmacol. Ther. 44(2), 83 (2006).
Medline CASGoogle Scholar
38. Uçaktürk E, Özaltin N. Optimization of a gas chromatography–mass spectrometry method using chemometric techniques for the determination of ezetimibe in human plasma. Turk. J. Chem. 37(5), 734–745 (2013).
Google Scholar
39. Li S, Liu G, Jia J, Li X, Yu C. Liquid chromatography–negative ion electrospray tandem mass spectrometry method for the quantification of ezetimibe in human plasma. J. Pharm. Biomed. Anal. 40(4), 987–992 (2006).
Medline CASGoogle Scholar
40. Gowda KV, Rajan DS, Mandal U et al. Bioequivalence study of fixed dose combination of atorvastatin and ezetimibe tablet in healthy volunteers by LC–MS/MS method. Asian J. Chemistry 19(2), 1293 (2007).
CASGoogle Scholar
41. Abdelbary G, Nebsen M. Application of a novel UPLC–MS/MS method for the pharmacokinetic/bioequivalence determination of atorvastatin and ezetimibe in human plasma. J. Pharm. Res. 7(1), 24–32 (2013). • Ultra-high-performance LC–MS/MS for pharmacokinetic estimation of atorvastatin and ezetimibe.
CASGoogle Scholar
42. El-Bagary RI, Elkady EF, El-Sherif ZA, Kadry AM. LC–MS-MS simultaneous determination of atorvastatin and ezetimibe in human plasma. J. Chromatogr. Sci. 52(8), 773–780 (2014).
Medline CASGoogle Scholar
43. Courlet P, Spaggiari D, Desfontaine V et al. UHPLC–MS/MS assay for simultaneous determination of amlodipine, metoprolol, pravastatin, rosuvastatin, atorvastatin with its active metabolites in human plasma, for population-scale drug-drug interactions studies in people living with HIV. J. Chromatogr. B Analyt. Technol. Biomed. Life Sci. 1125, 121733 (.2019).
Medline CASGoogle Scholar
44. Yacoub M, Alawi M, Arafat T. Simultaneous determination of amlodipine and atorvastatin with its metabolites; ortho and para hydroxy atorvastatin; in human plasma by LC–MS/MS . J. Chromatogr. B Analyt. Technol. Biomed. Life Sci. 917, 36–47 (2013).
MedlineGoogle Scholar
45. Liu D, Jiang J, Zhou H, Hu P. Quantitative determination of atorvastatin and para-hydroxy atorvastatin in human plasma by LC–MS–MS. J. Chromatogr. Sci. 46(10), 862–866 (2008).
Medline CASGoogle Scholar
46. Zhou Y, Li J, He X et al. Development and validation of a liquid chromatography-tandem mass spectrometry method for simultaneous determination of amlodipine, atorvastatin and its metabolites ortho-hydroxy atorvastatin and para-hydroxy atorvastatin in human plasma and its application in a bioequivalence study. J. Pharm. Biomed. Anal. 83, 101–107 (2013).
Medline CASGoogle Scholar
47. Elawady T, Ibrahim F, Khedr A, Belal F. Simultaneous determination of ezetimibe, atorvastatin and simvastatin using quadrupole LC–MS: application to combined tablets and plasma after SPE. Acta Chromatogr. 33(3), 245–252 (2021).
CASGoogle Scholar
48. Serag A, Hasan MA, Tolba EH, Abdelzaher AM, Elmaaty AA. Analysis of the ternary antiretroviral therapy dolutegravir, lamivudine and abacavir using UV spectrophotometry and chemometric tools. Spectrochim. Acta A Mol. Biomol. Spectrosc. 264, 120334 (.2022).
Medline CASGoogle Scholar
49. Ibrahim AE, Elmaaty AA, El-Sayed HM. Determination of six drugs used for treatment of common cold by micellar liquid chromatography. Anal. Bioanal. Chem. 413(20), 5051–5065 (2021).
Medline CASGoogle Scholar
50. Salem WA, Elkady EF, Fouad MA, Mohammad MA-A. DoE screening and optimization of liquid chromatographic determination of nicotinic acid and six statins: application to pharmaceutical preparations and counterfeit detection. J. Chromatogr. Sci. (2021).
MedlineGoogle Scholar
51. Elkady EF, Tammam MH, El Maaty AA. Stability indicating HPLC–UV vs UPLC–DAD for estimation of atorvastatin simultaneously with aspirin, clopidogrel and their related impurities in bulk and capsules. Analytical Chemistry Letters 7(5), 596–610 (2017).
CASGoogle Scholar
52. Agency EM. Guideline on bioanalytical method validation. EMEA/CHMP/EWP/192217/2009 (2011).
Google Scholar
53. Apostolou C, Kousoulos C, Dotsikas Y et al. An improved and fully validated LC–MS/MS method for the simultaneous quantification of simvastatin and simvastatin acid in human plasma. J. Pharm. Biomed. Anal. 46(4), 771–779 (2008).
Medline CASGoogle Scholar
54. Yang Z, Hancock WS, Chew TR, Bonilla L. A study of glycoproteins in human serum and plasma reference standards (HUPO) using multilectin affinity chromatography coupled with RPLC–MS/MS . Proteomics 5(13), 3353–3366 (2005).
Medline CASGoogle Scholar
55. Barrett B, Huclova J, Bořek-Dohalský V, Němec B, Jelinek I. Validated HPLC–MS/MS method for simultaneous determination of simvastatin and simvastatin hydroxy acid in human plasma. J. Pharm. Biomed. Anal. 41(2), 517–526 (2006).
Medline CASGoogle Scholar

Vol. 14, No. 21

Supplemental Materials

Metrics

Downloaded 119 times

History

Received 21 October 2022

Accepted 15 December 2022

Published online 9 January 2023

Published in print November 2022

Information

Keywords

Supplementary data

To view the supplementary data that accompany this paper please visit the journal website at: www.future-science.com/doi/suppl/10.4155/bio-2022-0203

Acknowledgments

The authors appraise the scientific and logistic support of Pharmasolutions Contract Research Organization (New Maadi, Cairo, Egypt).

Financial & competing interests disclosure

The authors have no relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript. This includes employment, consultancies, honoraria, stock ownership or options, expert testimony, grants or patents received or pending, or royalties.

No writing assistance was utilized in the production of this manuscript.

PDF download