LC-MS/MS method for simultaneous Doxorubicin and Baicalein estimation: formulation and pharmacokinetic applications

<b>Aim &amp; objective:</b> Combinatorial delivery of Doxorubicin (DOX) and Baicalein (BAC) has a potential to improve breast cancer treatment by mitigating the cardiotoxicity induced by DOX. The nanoformulation has been optimized and subjected to pharmacokinetic studies using LC-MS/MS. <b>Materials &amp; methods:</b> Nanoformulation bearing DOX and BAC was optimized using quality by design approach and method validation was done following USFDA guidelines. <b>Results:</b> The particle size, PDI and zeta potential of developed nanoformulation were 162.56 ± 2.21 nm, 0.102 ± 0.03 and -16.5 ± 1.21 mV, respectively. DOX-BAC-SNEDDs had a higher AUC_0-t values of 6128.84 ± 68.71 and 5896.62 ± 99.31 ng/mL/h as compared with DOX-BAC suspension. <b>Conclusion:</b> These findings hold promise for advancing breast cancer treatment and facilitating therapeutic drug monitoring. Mechanistic scheme of DOX-BAC-loaded SNEDDs development and its characterization using LC-MS/MS. Baicalein (BAC) lowers cardiac oxidative stress which reduces the cardiotoxicity associated with Doxorubicin (DOX). Combination therapies enhance efficacy by targeting multiple disease pathways simultaneously, improving treatment outcomes and reducing drug resistance. The Quality by Design approach was employed in optimizing the self-nanoemulsifying drug delivery system. Currently, there is no published bioanalytical method utilizing either LC-MS/MS or HPLC for simultaneous determination of both DOX and BAC. Both DOX and BAC were resolved using mobile phase of 0.1% v/v FA in milli-Q water and 0.1% v/v FA in MeOH (20:80%v/v) with a flow rate of 0.3 mL/min. USFDA guidelines were employed for assessing several validation factors, including recovery, stability studies, specificity, accuracy and percent relative standard deviation. The co-estimation of DOX and BAC in rat plasma in order to generate an oral P.K. profile was successfully demonstrated using the developed LC-MS/MS.