Validity Of Method Research Articles

QbD Assisted Development and Validation of UV Spectroscopic Method in Estimation of Silymarin

Background: Silymarin is a flavonoid utilised in liver dysfunction for years; new studies have emphasised its potential utility as a therapy for nanoparticulate targeting in many other disorders. In order to establish the product quality of such compounds, researchers have been trying to develop a robust method of analysis, but the methods developed till now are too expensive and time-consuming. Here we proposed quality by design-assisted development and validation of the UV spectroscopic method using Design-Expert® software in the estimation of Silymarin for fabrication of nanoparticulate formulations, which is simple, accurate, cost-effective and non-tedious. Aim: The proposed method is a simple, new, robust, accurate, and precise UV visible spectroscopic technique for estimating silymarin produced in nano-formulations utilising an analytical quality by design (AQbD) approach. Methods: A UV spectrophotometric technique was established, as maximum absorption (287.7) was measured using a Shimadzu UV-1800 double-beam UV visible spectrophotometer. The characterization of silymarin was done by melting point, DSC and FTIR techniques. The two critical method variables chosen were scanning speed and sample interval to be analysed by the design of experiment methodology utilizing the central composite design principle, which shows robustness and optimized technique involved in this work. Results: The spectroscopy technique was developed and validated as per International Conference of Harmonization recommendations. The Beer's-Lambert rule was followed in a series of 2-12 μg/ml dilution increments, with a correlation value of R2 = 0.999. The method's linearity was shown to be excellent across the concentration range. The percent recovery of the current method approach was determined to be within the confidential limitations, i.e., less than 2% expressed as % RSD, and the methodology was proved to be precise at inter and intraday variations (% RSD). The LOD and LOQ were found to be 0.264 μg/ml and 0.801 μg/ml which were also determined correctly. During specificity testing, no interfering peaks were found. Conclusion: This UV approach has been used successfully to determine the quantity of silymarin present in the nanoparticulate formulation, which can be used for testing its other pharmaceutical dosage forms.

Development and validation of a UV spectrophotometric method for biowaiver studies in pyrazinamide tablets

Development and validation of a UV spectrophotometric method for biowaiver studies in pyrazinamide tablets

Development and Validation of Novel HPLC Methods for Quantitative Determination of Vitamin D3 in Tablet Dosage Form

In the present work, an efficient isocratic HPLC method was developed for the precise and accurate estimation of vitamin D3 in tablet form. The chromatographic conditions comprised an L3 silica column (5 µm in particle size, 4.6 mm × 250 mm) with a mobile phase n-hexane/ethyl acetate (85:15 v/v) with a flow rate of 2.0 mL/min and a detection wavelength of 292 nm. The new methodology was validated for accuracy, precision, specificity, robustness, and quantification limits according to an official monograph of USP/BP and ICH guidelines. The peak areas of the six replicates of the homogeneous sample were recorded. The mean value obtained was 67,301, and the relative standard deviation (RSD) was 0.1741. The linearity and range were in the acceptable bounds, i.e., 0.999, which was calculated using regression line analysis. The results show that the method is truly acceptable as the RSD, as the flow rate was 0.81%, while for the mobile phase composition, it was 0.72%, which lies in the acceptable range. The limit of detection (LOD) and the limit of quantification (LOQ) values were 0.0539 µg/mL and 0.1633 µg/mL, respectively. The % RSD of the intra and inter-day precision of the method was deemed acceptable according to the international commission for harmonization guidelines. The developed method has potential to be used for the detection and quantification of vitamin D3 during routine analysis for tablets in dosage form.

UV Spectrophotometric Method Development and Validation of Butorphanol Tartrate in Bulk Drug and Pharmaceutical Formulation

The present research work discusses the development and validation of a UV spectrophotometric method for butorphanol tartrate. Simple, accurate and cost efficient spectrophotometric method has been developed for the estimation of butorphanol tartrate in tablet dosage form. The optimum conditions for the analysis of the drug were established. The maximum wavelength (λ max) was found to be 278nm. The percentage recovery of butorphanol tartrate was in the 100.016±0.68. Beers law was obeyed in the concentration range of 2-12μg/ml. Calibration curves shows a linear relationship between the absorbance and concentration. The line equation y = 0.0674x - 0.0057 with r2 of 0.9998was obtained. Validation was performed according to ICH guidelines for linearity, accuracy, precision, LOD and LOQ. The sample solution was stable up to 36 hours. The proposed method may be suitable for the analysis of butorphanol tartrate in tablet formulation for quality control purposes.
 Keywords: Butorphanol tartrate, Validation, Precision, Accuracy, LOQ, LOD, ICH guidelines.

Development and Validation of an UPLC Method for Quality Control of Plantago major

Development and Validation of an UPLC Method for Quality Control of Plantago major

HPLC-MS/MS method development and validation for the determination of tetradecapeptide in human plasma

Introduction. The number of peptide drugs being developed and registered has increased in recent years. Therefore, modern analytical approaches and methods are required to determine these substances in biological matrices during pharmacokinetic studies. Peptides are structurally intermediate between small molecules and biopolymers, making it difficult to develop methods for determining them using High Performance Liquid Chromatography with Tandem Mass Spectrometry (HPLC-MS/MS). Peptide derivatization can help achieve optimal chromatographic separation and increase method sensitivity.Aim. To develop and validate a method for the determination of the tetradecapeptide (TDP) threonyl-glutamyl-lysyl-lysyl-arginyl-arginyl-glutamayl-threonyl-valyl-glutamyl-arginyl-glutamyl-lysyl-glutamate in human plasma by HPLC-MS/MS.Materials and methods. The determination of TDP in human plasma was performed by HPLC-MS/MS. Sample preparation included a combination of blood plasma protein precipitation with propionic acid solution in methanol, liquid-liquid extraction with chloroform, and peptide derivatization with propionic anhydride. Internal standard (IS) was threonyl-glutamyl-lysyl-lysyl-arginyl-arginyl-glutamayl-threonyl-leucyl-glutamyl-arginyl-glutamyl-lysyl-glutamate. Chromatographic separation was performed in gradient mode, eluent A was 0.1 % formic acid solution in water, eluent B was 0.1 % formic acid in acetonitrile. Column: Waters XBridge C18, 4.6 × 50 mm, 5 µm. Ionization source was electrospray in positive mode. Multiple reaction monitoring (MRM) transitions for 4-substituted TDP propionate were: 681.30 → 73.95 m/z, 681.30 → 84.00 m/z, 681.30 → 101.90 m/z, 681.30 → 140.10 m/z, and for 4-substituted IS propionate: 686.00 → 74.10 m/z, 686.00 → 84.05 m/z, 686.00 → 102.00 m/z, 686.00 → 140.00 m/z.Results and discussion. Validation of the developed method was carried out in accordance with the requirements of Eurasian Economic Union and the following parameters were determined: selectivity, matrix effect, calibration curve, accuracy and precision, recovery, lower limit of quantification, sample carryover, stability.Conclusion. The method for the determination of TDP in human blood plasma by HPLC-MS/MS was developed and validated. The analytical range was 5.00–1000.00 ng/mL, allowing the method to be used to study TDP pharmacokinetics.

Development and Validation of HPLC-UV Method for the Determination of Levothyroxine in Orodispersible Minitablets for Pediatric Application

Background: Thyroid hormones play an important role in cognition and brain development. The determination of the content of levothyroxine, as well as related substances and dissolution test analysis, should be carried out by methods that are selective and highly sensitive due to the low concentration used in low dose orodispersible minitablets. Objectives: This study aims to develop and validate an analytical method by HPLC-UV for the quantification of levothyroxine and its related substances in pharmaceutical formulations; looking forward to being this method suitable for a future dissolution test analysis using tandem mass spectrometry detector. Methods: A Hypersil GOLD C18 (100 x 2.1 mm, 3µm) column was used with 25°C column temperature, 5 µL injection volume, 0.3 mL/min flow rate and detection at 225 nm. The mobile phase consisted of methanol: 0.05% formic acid (55:45). The developed method was validated for specificity, linearity, precision, accuracy and robustness. Results: The method is linear within the range of 2-20 µg mL-1 (R2=0.9982), which makes the method suitable for the evaluation of levothyroxine in pharmaceuticals formulations. LOQ was 0.17 µg/mL (0.85 ng on column) and LOD 0.05 µg/mL (0.25 ng on column) of LT4. Therefore, in terms of efficiency (1671), retention factor, k (6.79), Tailing factor, T (1.09) and resolution, Rs (5.11) the chromatographic method was found to be suitable according to USP 43. Conclusion: The HPLC UV method was found to be linear, specific, precise, accurate and robust, therefore it is suitable for the quality control of levothyroxine in pharmaceutical ODMTs.

The relationship between heavy metals and metabolic syndrome using machine learning

BackgroundExposure to high levels of heavy metals has been widely recognized as an important risk factor for metabolic syndrome (MetS). The main purpose of this study is to assess the associations between the level of heavy metal exposure and Mets using machine learning (ML) method.MethodsThe data used in this study are from the national health and nutrition examination survey 2003–2018. According to the demographic information and heavy metal exposure level of participants, a total of 22 variables were included. Lasso was used to screen out the key variables, and 9 commonly used ML models were selected to establish the associations with the 5-fold cross validation method. Finally, we choose the SHapley Additive exPlanations (SHAP) method to explain the prediction results of Adaboost model.Results11,667 eligible individuals were randomly divided into two groups to train and verify the prediction model. Through lasso, characteristic variables were selected from 24 variables as predictors. The AUC (area under curve) of the models selected in this study were all greater than 0.7, and AdaBoost was the best model. The AUC value of AdaBoost was 0.807, the accuracy was 0.720, and the sensitivity was 0.792. It is noteworthy that higher levels of cadmium, body mass index, cesium, being female, and increasing age were associated with an increased probability of MetS. Conversely, lower levels of cobalt and molybdenum were linked to a decrease in the estimated probability of MetS.ConclusionOur study highlights the AdaBoost model proved to be highly effective, precise, and resilient in detecting a correlation between exposure to heavy metals and MetS. Through the use of interpretable methods, we identified cadmium, molybdenum, cobalt, cesium, uranium, and barium as prominent contributors within the predictive model.

The Health Monitoring of Bonded Composite Joints Using Both Thickness and Radial Impedance Resonance Responses

With the advent of bonded composites in today’s aircraft, there is a need to verify the structural integrity of the bonded joints that comprise their structure. To produce adequate joint integrity, strict process control is required during bonding operations. The latest non-destructive joint inspection techniques cannot quantify the strength of the bond and only indicate the presence of disbonds or delaminations. Expensive and timely proof-load testing of the joints is required to demonstrate structural performance. This work focuses on experimentally evaluating joint-health monitoring with piezoelectric sensors exposed to repeated loadings until failure. Single-lap-shear composite joints are structurally tested while using sensor electromechanical impedance response as a health indicator. Based on these experiments, validation of this novel method is achieved through detailed evaluation of the sensor impedance response characteristics during loading, which enable initial and prognostic joint health assessments. The experimental results indicate that the embedded piezoelectric sensors are able to measure the sensor impedance radial and thickness resonance response changes prior to joint failure, without sacrificing the joints’ structural performance.

Development and Validation of UV Spectroscopic method for the estimation of favipiravir in bulk and tablet dosage form

Simple, precise and accurate area under curve spectroscopic method has been developed and validated for the estimation of Favipiravir in bulk and pharmaceutical dosage form. The drug shows maximum absorption (λ max) at 361nm in 0.1N NaOH solution and Area under Curve [AUC] in absorption spectra were measured between the wavelength range 356 to 366nm which obeys Beer’s law in the concentration range of 2-10µg/ml. The linearity study carried and regression coefficient was found to be 0.9982 and it has showed good linearity, precision during this concentration range. The % recovery was found to be 98.6-101.1. The LOD and LOQ were found to be 0.055 and 0.168µg/ml. The % relative standard deviation were found less than 2. According to ICH guidelines the method has been validated for linearity, precision, accuracy, robustness, ruggedness, LOD and LOQ. The developed and validated method can be successfully applied for reliable quantification of Favipiravir in bulk form and pharmaceutical dosage form.

Validation and recalibration of sex estimation methods using pubic nonmetric traits for the Chilean population.

Chile had a violent military coup (1973-1990) that resulted in 3,000 victims declared detained, missing or killed; many are still missing and unidentified. Currently, the Human Rights Unit of the Forensic Medical Service in Chile applies globally recognised forensic anthropological approaches, but many of these methods have not been validated in a Chilean sample. As current research has demonstrated population-specificity with extant methods, the present study aims to validate sex estimation methods in a Chilean population and thereafter establish population-specific equations. A sample of 265 os coxae of known age and sex of adult Chileans from the Santiago Subactual Osteology Collection were analysed. Visual assessment and scoring of the pelvic traits were performed in accordance with the Phenice (1969) and Klales et al. (2012) methods. The accuracy of Phenice (1969) in the Chilean sample was 96.98%, with a sex bias of 7.68%. Klales et al. (2012) achieved 87.17% accuracy with a sex bias of -15.39%. Although both methods showed acceptable classification accuracy, the associated sex bias values are unacceptable in forensic practice. Therefore, six univariate and eight multivariate predictive models were formulated for the Chilean population. The most accurate univariate model was the ventral arc at 96.6%, with a sex bias of 5.2%. Classification accuracy using all traits was 97.0%, with a sex bias of 7.7%. This study provides Chilean practitioners a population-specific morphoscopic standard with associated classification probabilities acceptable to accomplish legal admissibility requirements in human rights and criminal cases specific to the second half of the 20th century.

Joint Implementation Method for Clutter Suppression and Coherent Maneuvering Target Detection Based on Sub-Aperture Processing with Airborne Bistatic Radar

An airborne bistatic radar working in downward-looking mode confronts two major challenges for low-altitude target detection. One is range cell migration (RCM) and Doppler migration (DM) resulting from the relative motion of the radar and target. The other is the non-stationarity characteristic of clutter due to the radar configuration. To solve these problems, this paper proposes a joint implementation method based on sub-aperture processing to achieve clutter suppression and coherent maneuvering target detection. Specifically, clutter Doppler compensation and sliding window processing are carried out to realize sub-aperture space–time processing, removing the clutter non-stationarity resulting from the bistatic geometric configuration. Thus, the output matrix of clutter suppression in the sub-aperture could be obtained. Then, the elements with the same phase of this matrix are superimposed and rearranged to achieve the reconstructed 2-D range-pluse echo matrix. Next, the aperture division with respect to slow time is conducted and the RCM correction based on modified location rotation transform (MLRT) and coherent integration (CI) are realized within each sub-aperture. Finally, the matched filtering process (MFP) is applied to compensate for the RCM/DM among different sub-apertures to coherently integrate the maneuvering target energy of all sub-apertures. The simulation and measured data processing results prove the validity of the proposed method.

Sensitivity analysis for matched observational studies with continuous exposures and binary outcomes

Summary Matching is one of the most widely used study designs for adjusting for measured confounders in observational studies. However, unmeasured confounding may exist and cannot be removed by matching. Therefore, a sensitivity analysis is typically needed to assess a causal conclusion’s sensitivity to unmeasured confounding. Sensitivity analysis frameworks for binary exposures have been well-established for various matching designs and are commonly used in various studies. However, unlike the binary exposure case, there still lacks valid and general sensitivity analysis methods for continuous exposures, except in some special cases such as pair matching. To fill this gap in the binary outcome case, we develop a sensitivity analysis framework for general matching designs with continuous exposures and binary outcomes. First, we use probabilistic lattice theory to show our sensitivity analysis approach is finite-population- exact under Fisher’s sharp null. Second, we prove a novel design sensitivity formula as a powerful tool for asymptotically evaluating the performance of our sensitivity analysis approach. Third, to allow effect heterogeneity with binary outcomes, we introduce a framework for conducting asymptotically exact inference and sensitivity analysis on generalized attributable effects with binary outcomes via mixed- integer programming. Fourth, for the continuous outcomes case, we show that conducting an asymptotically exact sensitivity analysis in matched observational studies when both the exposures and outcomes are continuous is generally NP-hard, except in some special cases such as pair matching. As a real data application, we apply our new methods to study the effect of early-life lead exposure on juvenile delinquency. An implementation of the methods in this work is available in the R package doseSens.

Development, Validation, Identification and Estimation of Quinoline Yellow in the Levodopa and Carbidopa Pharmaceutical Combined dosage form by Ultra violet Visible Spectrophotometer

A simple, cost-effective, précised, accurate and robust Ultra Violet spectrophotometric method [1, 2 ] has been developed for the Identification and estimation of Quinoline Yellow in the Levodopa and Carbidopa tablet dosage forms. It is used as a colourant in the pharmaceutical tablet’s dosage form. UV scan of Quinoline yellow was taken from the entire UV range of 200 to 800 nm. From the spectrum, maximum absorption was observed at 414nm (λmax) for the Quinoline Yellow. Method was found to be specific with no interference due to blank and placebo, The method was linear with a correlation coefficient of more than 0.999, and Accuracy was in the range of 98.3 to 101.3, In the robustness study employed for standard and sample preparation showed no impact on the results, by deliberate changes proves method is robust and can be utilized for regular analysis. Method validation was performed with reference to ICH guidelines Q2R1.

Histological analysis of the dermal and hypodermal layers of the face and correlation with high-frequency 24 MHz ultrasonography and elastosonography.

Knowledge of the structure of the face is of fundamental importance. In fact, the face is treated in many areas of medicine, from dermatology, to maxillofacial surgery, to otorhinolaryngology, to ophthalmology, etc. and anti-aging aesthetic treatments, and those for the resolution of blemishes are on the increase. For ethical reasons it is not possible to take biopsy samples for facial analysis in the aesthetic field. The main aim of this study was to demonstrate that a high-resolution bimodal ultrasound examination, combined with elastosonography, could be a valid tool for pre-treatment morphological evaluation. To achieve this goal, skin samples were taken from the forehead, zygomatic area, nasolabial fold, upper and lower lip from cadavers to histologically characterize their structure. Subsequently, these same areas were evaluated in vivo using conventional B-mode ultrasound with a 24 MHz high-frequency probe, and elastosonography. The data obtained with the different techniques were compared, in order to state that modern ultrasound techniques can provide similar histological information. The analysis showed that the superficial hypodermis presented a different shape and structure in the different areas, with the exception of the areas of the upper and lower lip, which appeared similar. With aging, the forehead and zygomatic area showed a volumetric increase in the superficial hypodermic layer, while the lip showed non-structural changes. The morphology of the nasolabial fold remained unchanged. When it is not possible to perform histological investigations on the face, to understand its characteristics and dynamics, ultrasound with a 24 MHz probe would seem to be the most suitable method, while elastosonography could be a valid method for evaluating the stiffness of the structural components.

Optimization of the QuEChERS Extraction of Mycotoxins in Maize Using Response Surface Methodology (RSM)

AbstractSeveral methods have been developed for the analysis and detection of mycotoxins in food; however, most do not make use of critical statistics and mathematical tools for precise optimization. This study developed, optimized, and validated a modified quick, easy, cheap, effective, rugged, and safe (QuEChERS) extraction procedure for the extraction of multiple mycotoxins in maize and subsequent validation using liquid chromatography tandem mass spectrometry (LC–MS/MS). Central composite design (CCD) was used to optimize extraction conditions. Data analysis of full factorial screening experiments revealed that MeCN (%), FA (%), and extraction time significantly affected the mycotoxins recovery. Assessment of the statistical significance of the generated model using analysis of variance (ANOVA), coefficient tables, and surface plots showed the relative interactions of factors and the adequacy of the model. Thus, P values from the lack of fit (LOF) test ranged from 0.137– 0.467 and a composite desirability function of 0.91 was obtained. Using the optimum extraction conditions of 0.1% (v/v) FA in 80.2% MeCN for 74 min, 10 mycotoxins were effectively extracted with satisfactory recoveries (85–114%), coefficients of regression (R2 > 0.98), coefficients of variation (CVs < 15%), limit of quantifications (LOQs) (0.33–60.45 µg/kg), and other associated method validation parameters. The method validation was carried out according to Commission Implementing Regulation 2021/808 and Commission Regulation (EC) No 401/2006 of 23 February 2006. Application of this method to 20 maize samples collected from markets in Botswana showed detectable mycotoxins in 13 samples, with 2 exceeding the European Union (EU) maximum aflatoxin B1 (AFB1) limit, suggesting potential exposure to high levels of toxic mycotoxins in Botswana.

Automatic detection of skate strokes in short-track speed skating using one single IMU: validation of a new method

ABSTRACT Greater impulse is a key performance indicator of success in short track speed skating. The main objective of this study was to develop a method to measure skating strokes using a single IMU. Eight elite or world-class speed skaters had one IMU placed against their skin on the lower back, and a camera setup was positioned to capture the test. A maximal speed trial was then executed by each participant, and the data were analysed to estimate agreement between the camera and IMU estimates of skate stroke events. Inter-evaluator reliability was assessed on a dataset of 22 athletes performing speed trials as well. The algorithm detected 100% of the strokes identified on the video capture system with a root mean square error of 0.06s. Bland-Altman analysis showed a bias of 0.03s between the two methods, which corresponds to the frame rate of the camera. The inter-evaluator reliability yielded an intra-class correlation of 1.00 (ICC3,1) from a dataset of 7089 strokes. This study provides an example of on-ice evaluation of speed skating strokes using a single IMU. This equipment is less expensive than that employed by previous authors and can be implemented in training situations with low invasiveness.

Navigating the future of diabetes: innovative nomogram models for predicting all-cause mortality risk in diabetic nephropathy.

This study aims to establish and validate a nomogram model for the all-cause mortality rate in patients with diabetic nephropathy (DN). We analyzed data from the National Health and Nutrition Examination Survey (NHANES) spanning from 2007 to 2016. A random split of 7:3 was performed between the training and validation sets. Utilizing follow-up data until December 31, 2019, we examined the all-cause mortality rate. Cox regression models and Least Absolute Shrinkage and Selection Operator (LASSO) regression models were employed in the training cohort to develop a nomogram for predicting all-cause mortality in the studied population. Finally, various validation methods were employed to assess the predictive performance of the nomogram, and Decision Curve Analysis (DCA) was conducted to evaluate the clinical utility of the nomogram. After the results of LASSO regression models and Cox multivariate analyses, a total of 8 variables were selected, gender, age, poverty income ratio, heart failure, body mass index, albumin, blood urea nitrogen and serum uric acid. A nomogram model was built based on these predictors. The C-index values in training cohort of 3-year, 5-year, 10-year mortality rates were 0.820, 0.807, and 0.798. In the validation cohort, the C-index values of 3-year, 5-year, 10-year mortality rates were 0.773, 0.788, and 0.817, respectively. The calibration curve demonstrates satisfactory consistency between the two cohorts. The newly developed nomogram proves to be effective in predicting the all-cause mortality risk in patients with diabetic nephropathy, and it has undergone robust internal validation.

Highly sensitive liquid chromatography-mass spectrometry method for the quantitative analysis of mometasone furoate in human plasma: Method validation and application to clinical pharmacokinetic studies.

We report the development and the validation of a sensitive liquid chromatography-mass spectrometry (LC-MS/MS) method for mometasone furoate (MF) analysis in human plasma. Plasma samples were processed through liquid-liquid extraction and analyzed using LC-MS/MS operating in positive mode using multiple reaction monitoring of transitions m/z 520.9→ 355.0 and m/z 525.8→ 355.0 for MF and the internal standard (IS), respectively. Separation was achieved at 1.0mL/min on a C18 column using a gradient elution of mobile phase of 0.05% ammonia in water (phase A) and acetonitrile (phase B). The assay range was 0.250-100 pg/mL and proved to be accurate and precise MF. Normalized recoveries were consistent and reproducible with a coefficient of variation (CV%) value of 6.0. The CV (%) of the IS normalized matrix factor was not observed in normal, lipemic, and hemolyzed plasmas. Dilutions of 1:10 were accurately quantified. A cycle of three freeze and thaw and stabilities at room temperature and on the autosampler were demonstrated. In addition, MF in the presence of indacaterol and glycopyrronium was proven to be stable at -70°C for at least 157 days. The present method was successfully applied to quantify MF in patients receiving MF, indacaterol, and glycopyrronium as a fixed-dose combination.

Method development and validation for the estimation of gedatolisib in mouse plasma by tandem mass spectrometry and its application to pharmacokinetics studies.

A simple and a sensitive liquid chromatography-tandem mass spectrometry method was developed and validated for the quantification of gedatolisib in mouse plasma. The extraction technique involved a simple precipitation method to extract gedatolisib and idelalisib (internal standard) from mouse plasma. A clean chromatographic separation of gedatolisib and the internal standard was achieved on an Atlantis dC18 column using an isocratic mobile phase (10mm ammonium formate and acetonitrile; 30:70% v/v, both supplemented with 0.1% formic acid) delivered at a flow rate of 0.7ml/min. The total run time was 2.0min, and gedatolisib and idelalisib were eluted at 0.80 and 0.95 min, respectively. Gedatolisib was monitored at m/z 616.40 → 488.20 and idelalisib at 416.05 → 176.10. All the required parameters for the method validation were performed as per US Food and Drug Administration guidelines, and the results were within the acceptance criteria. The method was accurate and proved to be precise at a linearity range of 1.33-2667 ng/ml. The accuracy for gedatolisib in mouse plasma was in the ranges 0.99-1.06% (intra-day) and 0.96-1.04% (inter-day). Gedatolisib appeared to be stable in a series of stability conditions. Gedatolisib showed a good intravenous profile when administered through a solution formulation.