Enhanced selectivity in alpelisib quantification in human plasma: a validated LC-MS/MS method with deuterated internal standard

Venkateswararao Agraharapu; Venkata Rao  Vutla; Vidyadhara Suryadevara

doi:10.69857/joapr.v14i4.2268

Authors

Venkateswararao Agraharapu School of Pharmaceutical Sciences and Technologies, Jawaharlal Nehru Technological University, Kakinada. Andhra Pradesh, Indi
Venkata Rao Vutla Department of Pharmaceutical Analysis, Chebrolu Hanumaiah Institute of Pharmaceutical Sciences, Chandramoulipuram, Chowdavaram, Andhra Pradesh, India.
Suryadevara Vidyadhara Department of Pharmaceutics, Chebrolu Hanumaiah Institute of Pharmaceutical Sciences, Chandramoulipuram, Chowdavarm, Guntur-522019, Andhra Pradesh, India

DOI:

https://doi.org/10.69857/joapr.v14i4.2268

Keywords:

Alpelisib, LC-MS/MS, bioanalysis, validation of methods, monitoring of therapeutic drugs, human plasma

Abstract

Background: To enhance efficacy and manage dose-limiting toxicities such as hyperglycemia, therapeutic drug monitoring of alpelisib (ALB), a phosphatidylinositol 3-kinase alpha (PI3Kα) inhibitor used in advanced breast cancer, is essential. This study aimed to develop and validate a selective, sensitive, and high-throughput liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the quantification of ALB in human plasma to support pharmacokinetic studies and therapeutic drug monitoring. Methodology: Alpelisib-D3 was used as a deuterated internal standard (IS). Analytes were extracted from 50 µL of plasma by liquid-liquid extraction using n-hexane and methyl tert-butyl ether (20:80, v/v). Chromatographic separation was performed on an X-Terra RP8 column using an isocratic mobile phase of 5 mM ammonium acetate buffer (pH 5.00) and acetonitrile (10:90, v/v) at 0.4 mL/min. Detection was carried out in positive electrospray ionization mode using multiple reaction monitoring of m/z 442.15→141.07 for ALB and m/z 445.36→144.05 for the IS. Validation included linearity, precision, accuracy, recovery, matrix effect, carryover, dilution integrity, and stability. Results and discussion: The method was linear over 50.0–10,000 ng/mL. Intra- and inter-day precision (CV%) was <4.2%, and accuracy ranged from 94.5% to 106.2%. Mean extraction recovery exceeded 90%, with no significant matrix effects. Carryover, stability, and dilution integrity also met acceptance criteria. Conclusion: A reliable, sensitive, and selective LC-MS/MS method was successfully developed and validated. The method is suitable for high-throughput pharmacokinetic studies and therapeutic drug monitoring of ALB in human plasma.

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References

André F, Ciruelos E, Rubovszky G, Campone M, Loibl S, Rugo HS, et al. Alpelisib for PIK3CA-mutated, hormone receptor-positive advanced breast cancer. N. Engl. J. Med., 380(20), 1929–1940 (2019) https://doi.org/10.1056/NEJMoa1813904

Tan A, Fanaras JC. Use of high-pH (basic/alkaline) mobile phases for LC-MS or LC-MS/MS bioanalysis. Biomed. Chromatogr., 33(1), e4409 (2019) https://doi.org/10.1002/bmc.4409

Peleg Hasson S, Rubinek T, Ryvo L, Wolf I. Endocrine resistance in breast cancer: Focus on the phosphatidylinositol 3-kinase/Akt/mammalian target of rapamycin signaling pathway. Breast Care, 8(4), 248–255 (2013) https://doi.org/10.1159/000354757

Markham A. Alpelisib: First global approval. Drugs, 79(11), 1249–1253 (2019) https://doi.org/10.1007/s40265-019-01161-6

Rugo HS, André F, Yamashita T, Cerda H, Toledano I, Stemmer SM, et al. Time course and management of key adverse events during the randomized phase III SOLAR-1 study of PI3K inhibitor alpelisib plus fulvestrant in patients with HR-positive advanced breast cancer. Ann. Oncol., 31(8), 1001–1010 (2020) https://doi.org/10.1016/j.annonc.2020.05.001

Armaghani AJ, Han HS. Alpelisib in the treatment of breast cancer: A short review on the emerging clinical data. Breast Cancer Targets Ther., 12, 251–258 (2020) https://doi.org/10.2147/BCTT.S219436

Miricescu D, Totan A, Stanescu-Spinu II, Badoiu SC, Stefani C, Greabu M. PI3K/AKT/mTOR signaling pathway in breast cancer: From molecular landscape to clinical aspects. Int. J. Mol. Sci., 22(1), 173 (2021) https://doi.org/10.3390/ijms22010173

Srivastava S, Upadhye V J, Prasad M E, Singh P. Applications of bioactive compounds of traditional Chinese medicine in breast cancer management. Journal of Applied Pharmaceutical Research, 13(4), 1-15 (2025) https://doi.org/10.69857/joapr.v13i4.935

Tankova T, Mrinakova B, Frolova M. Management strategies for hyperglycemia associated with the α-selective PI3K inhibitor alpelisib for the treatment of breast cancer. Cancers, 14(7), 1598 (2022) https://doi.org/10.3390/cancers14071598

Costa RLB, Han HS, Gradishar WJ. Targeting the PI3K/AKT/mTOR pathway in triple-negative breast cancer: A review. Breast Cancer Res. Treat., 169(3), 397–406 (2018) https://doi.org/10.1007/s10549-018-4697-y

James A, Hazell K, Trandafir L, et al. Absorption, distribution, metabolism, and excretion of BYL719 (alpelisib) in healthy male volunteers. Cancer Chemother. Pharmacol., 76(4), 751–760 (2015) https://doi.org/10.1007/s00280-015-2835-9

De Buck SS, Jakab A, Boehm M, et al. Population pharmacokinetics and pharmacodynamics of BYL719, a phosphoinositide 3-kinase antagonist, in adult patients with advanced solid malignancies. Br. J. Clin. Pharmacol., 78(3), 543–555 (2014) https://doi.org/10.1111/bcp.12354

Seo SW, Kim JM, Han DG, Geum D, Yun H, Yoon IS. A sensitive HPLC-FLD method for the quantification of alpelisib in rat plasma: Drug metabolism and pharmacokinetic evaluation in vitro and in vivo. J. Chromatogr. B, 1163, 122508 (2021) https://doi.org/10.1016/j.jchromb.2020.122508

Wang Q, Lan X, Zhao Z. Characterization of alpelisib in rat plasma by a newly developed UPLC-MS/MS method: Application to a drug-drug interaction study. Front. Pharmacol., 12, 725384 (2021) https://doi.org/10.3389/fphar.2021.725384

Lee S, Kim MS, Jeong JW. Bioanalysis of alpelisib using liquid chromatography–tandem mass spectrometry and application to pharmacokinetic study. J. Anal. Sci. Technol., 13, 31 (2022) https://doi.org/10.1186/s40543-022-00340-7

James AD, Marvalin C, Luneau A, et al. Comparison of ^19F NMR and ^14C measurements for the assessment of ADME of BYL719 (alpelisib) in humans. Drug Metab. Dispos., 45(8), 900–907 (2017) https://doi.org/10.1124/dmd.117.075051

Cai HL, Deng Y, Fang PF, Cao S, Hou ZY, Wu YQ, et al. A sensitive LC-MS/MS method for analysis of pericyazine in presence of 7-hydroxypericyazine and pericyazine sulphoxide in human plasma and its application to a comparative bioequivalence study in Chinese healthy volunteers. J. Pharm. Biomed. Anal., 135, 67–74 (2017) https://doi.org/10.1016/j.jpba.2016.12.007

Tiwari G, Tiwari R. Bioanalytical method validation: An updated review. Pharm. Methods, 1(1), 25–38 (2010) https://doi.org/10.4103/2229-4708.72226

Kaza M, Karaźniewicz-Łada M, Kosicka K, Siemiątkowska A, Rudzki PJ. Bioanalytical method validation: New FDA guidance vs. EMA guideline. Better or worse? J. Pharm. Biomed. Anal., 165, 381–385 (2019) https://doi.org/10.1016/j.jpba.2018.12.030