## 1 Introduction
The determination of the metabolic stability of DVB in HLMs is vital for understanding the kinetics of its metabolic reactions and elimination [14]. The stability of a pharmaceutical drug to metabolic enzymes refers to its susceptibility to undergo metabolic processes, and is estimated by its intrinsic clearance (Cl\({}_{\text{int}}\)) and in vitro half-life (t\({}_{1/2}\)) [15; 16]. The phrase \"half-life\" denotes the time required for the 50% metabolism of the DVB. The intrinsic clearance (Cl\({}_{\text{int}}\)) generally refers to the hepatic capacity to metabolize the drug (DVB in the current investigation). It is a term used to designate enzyme activity, and does not depend on hepatic blood flow and protein binding [17]. The objective of this work was to develop a fast, selective, ecologically green, and sensitive UHPLC-MS/MS method for determining the metabolic stability of DVB in HLMs. The data's validity was verified by doing in vitro testing using HLMs incubationsand performing in silico analyses with the help of the StarDrop software package. Green analytical chemistry (GAC) has gained increasing significance in recent years. Its objective is to indicate the presence of dangerous chemicals, reduce waste production, and reduce energy consumption through diverse analytical approaches [18; 19; 20]. Different measurement methods have been used to determine the degree of ecological sustainability depending on different analytical discoveries, with the aim of accomplishing these objectives. The collection includes a series of methodologies known as the Analytical Green-ness Metric Approach (AGREE), Analytical Eco-Scale (AES), Green Analytical Procedures Index (GAPI), the National Environmental Methods Index (NEMI), and Red-Green-Blue (RGB) [18]. The GAPI, AES, RGB, and NEMI approaches show a reliance on specific GAC values, as explained. The study employed the \"AGREE\" approach to determine the level of ecological sustainability by considering 12 Greenness Assessment Criteria (GAC) and conveying values to each criterion [21]. The UHPLC-MS/MS method employed a constant mobile phase and had a running time of 1 min, representing an exceptionally rapid analytical technique. Introducing a flow rate of 0.5 mL/min and the use of low ACN% (45%) significantly enhanced the ecologically friendly features of the existing system. Furthermore, the methodology employed in the current research showed a linear relationship in the range from 1 to 3000 ng/mL. The verification of the importance of the necessity to develop an UHPLC-MS/MS approach for estimating DVB in HLMs matrix was achieved by undertaking software assessments of the metabolic stability of DVB, employing the WhichP450 module (StarDrop's software) before beginning with the incubation of DVB with HLMs [10; 22; 23; 24]. The current strategy was introduced with the aim of minimizing time and reducing costs. The current research used the UHPLC-MS/MS technology to estimate the Cl\({}_{\text{int}}\) and in vitro t\({}_{1/2}\) of DVB, as previously described in other investigations [25; 26; 27; 28]. These methods can be utilized to compute the metabolic rate in living organisms using three different models: parallel tube, dispersion, and venous equilibrium. An in vitro technique was used to estimate the t\({}_{1/2}\) and Cl\({}_{\text{int}}\) of DVB, based on a well-stirred model [29; 30]. The usage of this model is mostly noted in research on drug metabolism due to its simplicity and straightforwardness of execution.