International Council For Harmonisation Research Articles

Quality by Design Approach for the Stability Indicating Method Development and Validation of Selpercatinib Drug Formulation by using RP-HPLC

In method development quality design approach is useful for method goal identification evaluation, method selection and risk assessment. Selpercatinib is a drug used in the treatment of certain types of cancer. The method development includes selecting appropriate chromatographic conditions such as the mobile phase composition, column, and selection wave length. The aim is to achieve optimum separation and quantification of selpercatinib in the dosage form. Optimizing chromatographic conditions by using design expert software. The mobile phase methanol and 0.1% Acetic acid are used. The mobile phase range is 80:20 v/v. The range of this mobile phase is 80:20 .0.7 mL/min found flow rate. The retention time of selpercatinib is 3.20 minutes. The assay was found to be 99.01% for selpercatinib. Optimization was performed. Factorial design performed, i.e., flow rate and percentages of methanol. Analysis of variance confirmed that method parameters. UV detection at 220 nm. The International Council of Harmonization (ICH) guidelines ensure the accurate, precise and reliable results to validate the parameters is an acceptable range. Stress studies reveal that drugs were degraded acidic conditions, alkaline conditions, neutral conditions, oxidative conditions, and photolytic conditions. Hence the proposed method was stability, indicating using quality by design (QbD) approach all the method parameters were better understood. This systematic approach ensures a thorough understanding of the assay, facilitates process optimization, and enables effective control strategies to reduce variability and enhance product quality.

Simultaneous Quantification of Dutasteride and Silodosin Using a Stability-Indicating RP-HPLC Approach

An alternative stability-indicating reverse-phase high-performance liquid chromatography (RP-HPLC) approach remained established to quantify silodosin (SLD) and dutasteride (DTS) in the mixture, validated per International Council of Harmonization (ICH) recommendations. A Shimadzu Model LC-2030 PLUS (IND) HPLC system, 231 nm detection wavelength, and 10 minutes run duration were used to create the method. A 75:25 mobile phase of methanol and water with 0.05% OPA was used at 1-mL/min. The concentrations of SLD (8–40 μg/mL) and DTS (0.5–2.5 μg/mL) were shown to be linearly related, with R2 values of 0.9998 for SLD and 0.9993 for DTS. Both of the drugs had accuracy and precision within 2% RSD. SLD and DTS mean recoveries were 97.5 to 102%. According to ICH criteria, degradation trials with acid, alkali, oxidizing agent, and heat assessed the approach’s stability. The established approach is fast, accurate, sensitive, and precise for routine SLD and DTS analysis from bulk as well as pharmaceutical dosage forms.

Development and Validation of HPTLC for the Determination of Gefitinib HCl in Tablet Dosage Form

A simple, high-performance thin-layer chromatography (HPTLC) method of gefitinib HCl in pharmaceutical formulation was developed. Separation was done on TLC aluminum plates precoated by silica gel 60GF254 (20 × 20 cm) with 200 μm layer depth. With UV detection set to 254nm, chromatography was conceded out by the mobile phase of dichloromethane and methanol (9:1, v/v). Linearity was found to range of 50 to 300 ng/spot and, the correlation coefficient was found to be 0.99 and the Rf value was found to be 0.46. The system’s suitability was considered to ascertain the performance of ascertaining quality performance of the developed chromatographic method. The method’s accuracy, precision, and specificity were all verified against the International Council of Harmonization (ICH) standards. Application of the approach to the determination of gefitinib in tablet form was successful.

Simultaneous RP-HPLC Estimation, Validation and Stability Indicating Assay of Two-Component Tablet Formulation Containing Grazoprevir and Pibrentasvir

The determination of a two-component tablet formulation containing grazoprevir and pibrentasvir for method development and validation has been done using the reverse-phase high-performance liquid chromatography (RP-HPLC) method as per International Council of Harmonization (ICH) guidelines. Analytical grade acetonitrile and 0.01 N of potassium dihydrogen phosphate buffer were used as mobile phase while Kromasil C18 column was opted for separation. In order to elute the analyte a flow rate of 1-mL/min having 260 nm λ has been maintained. An RT of 2.909 and 2.358 minutes while linearity concentration range from 25 to 150 μg/mL and 10 to 60 μg/mL was obtained for grazoprevir and pibrentasvir, respectively, with 4.4 resolution. Both had a high correlation coefficient of 0.999. The limit of quantification was 0.24 and 0.09 μg/mL, while the detection limit at 0.72 and 0.27 μg/mL was noticed for grazoprevir and pibrentasvir, respectively. For grazoprevir and pibrentasvir, the regression equation was determined to be y = 16332x + 5313.1 and y = 17261x + 601.95, correspondingly

Analytical Method Development by Using UV-Spectrophotometer for Estimation of Valsartan in Bulk

In order to accurately determine valsartan (API), this work aims to build a straightforward and exact UV spectroscopic approach. To improve sensitivity and accuracy, the procedure entails methodically optimizing experimental parameters, including solvent choice and wavelength selection. The produced stock solutions were tested for absorbance at 249 nm using a UV spectrophotometer. With %RSD values below 2% for both intra- and inter-day, the devised approach was determined to be quite accurate. At each additional concentration, the approach was shown to be accurate, yielding good drug recoveries ranging from 98 to 101%. The process was validated for linearity, specificity, accuracy, precision and limit of detection (LoD) and limit of quantification (LoQ) as per International Council of Harmonization (ICH) criteria. Its dependability for quantitative analysis of valsartan in pharmaceutical formulations was demonstrated. This UV spectroscopic approach offers a cost-effective and efficient alternative for routine quality control in pharmaceutical industries.

RP-HPLC Method for Estimation of Caspofungin in Bulk and in Parenteral Dosage Form

In order to more precisely and accurately estimate the antifungal medicine caspofungin and its parenteral dosage form, a new, straightforward, and cost-effective reverse-phase high-performance liquid chromatography (RP-HPLC) technique has been created. The accuracy, precision, linearity, specificity, and reproducibility of the method for the measurement of caspofungin in used equipment were determined during the present study’s validation according to the International Council of Harmonization (ICH) criteria. Elution was accomplished using a CHEMSIL ODS-C18 (5μm) column in isocratic mode with a mobile phase consisting of acetonitrile:ammonium acetate buffer at pH 4.8 (60:40%v/v). The mobile phase was pumped into the column at a flow rate of 1.0 mL/min, and the eluent was detected using a variable wavelength UV detector set at 210 nm. The column dimensions are 250 × 4.6 mm. The chromatogram for the optimized RP-HPLC method of caspofungin estimation had 28,99 theoretical plates (N) and a strong peak at a resolution time of 16,105 minutes. The thorough validation process led to the conclusion that the approach is stable and could be applied for the duration of the drug’s shelf life.

Estimation of Glyoxylic Acid in Emtricitabine by a New Validated RP-HPLC Method

To establish a simple, sensitive, as well as specified method for the separation and quantitative determination of glyoxylic acid in emtricitabine by using reversed-phase high-performance liquid chromatography (RP-HPLC). A superior separation of glyoxylic acid in emtricitabine was done by using a C8 column and a mobile system of (0.1% OPA and ACN: H2O) with 1-mL/min flow rate in gradient mode and a wavelength of 210 nm. The glyoxylic acid was eluted at 5.37 minutes with good efficiency and system suitability. The method has shown good linearity ranges from a limit of detection (LoQ) to 150% level of glyoxylic acid standard solution (0.1% or 0.001 mg/mL). ICH principles validated the suggested approach, and every parameter met the requirements for the International Council of Harmonization (ICH) Q2 recognition. %RSD of System, method precision, and precision at LoQ were in the range of 0.46 to 1.91. The developed method was sensitive, specific, and precise in estimating the trace levels of glyoxylic acid in emtricitabine.

Development of a Robust and Reliable RP-HPLC Method for the Estimation of Finerenone in Tablet Dosage Form

An advanced analytical method was explored, specifically tailored for estimating finerenone, a vital drug found in tablet formulations intended to reduce the risk of chronic kidney disease in patients with type-2 diabetes. This sophisticated technique employs an Inertsil ODS 3V C18 column (5 μm particle size x 250 x 4.6 mm) for precise separation of analyte. The mobile phase consists of an ammonium dihydrogen phosphate buffer (pH 4.5) and acetonitrile (60:40), which is a carefully balanced mixture that flows consistently at 1.0 mL/min. UV detection is achieved at a wavelength of 238 nm. Finerenone’s characteristics are unveiled with a brief retention time of 4.06 minutes, highlighting the efficiency of the method. The method demonstrates linearity within the concentration range of 2.5 to 15 μg/mL, offering a comprehensive scope for accurate analysis. This robust reversed-phase high-performance liquid chromatography (RP-HPLC) method undergoes thorough validation following International Council for Harmonisation (ICH) guidelines, ensuring its reliability. Notably, the accuracy is impressive, with a mean recovery falling comfortably within the acceptable range of 99.76 to 101.09%. In summary, our analytical approach stands out for its simplicity, precision, sensitivity, rapidity, and robustness in estimating finerenone in tablet formulations.

Stability Indicating Force Degradation Study of Nintedanib in Bulk and Pharmaceutical Dosage Form

Assay of the nintedanib drug substance can be determined using a newly designed and verified reverse-phase high-performance liquid chromatography (RP-HPLC) method, even when degradation products from forced degradation experiments are present. By using gradient elution, the mobile phase was a mixture of 70% acetonitrile and 30% water by volume. Acid, base, peroxide, thermal, and photolytic degradation were among the stress conditions that the product was subjected to. During the thermal and photolytic breakdown processes, no extra contaminants were detected. Using validation criteria such as specificity, linearity, limit of quantitation (LoQ), accuracy, precision, robustness, and ruggedness, the developed technique was validated in accordance with International Council for Harmonization (ICH) recommendations. At a concentration of 12.4 ng/mL, the LoQ value was attained. The results showed good linearity (r2 > 1.00) across doses ranging from 2 to 10 μg/mL. Verification of recovery was done by adding concentrated solutions of 5, 10, and 15 μg/mL. So, newly developed RP-HPLC technology can separate Nintedanib from its main degradation products, and it can also estimate the drug substance’s concentration.

Validated Area Under Curve Quantitative UV Spectrophotometric Analysis of Rebamipide in Drug Formulations

A very simple, accurately perfect, precisely repeatable procedure has been proposed for the estimate of rebamipide in medicines. Evaluation of previous works indicates the fact that, so far no UV spectrophotometric scheme for quantifiable estimate by area under curve technique for the drug rebamipide is not described. Therefore, there was a need to plan a different methodology to analyze the medicine employing dimethyl formamide in the form of solvent. This medication monitors Beer’s law in the range of concentration 20 to 200 ìg/mL in the ultraviolet range, particularly areas between 320 and 340 nm for are under curve because the absorption maximum was found to be 330 nm in the selected solvent. The recovery readings proved the offered technique’s accuracy and outcomes were authenticated par accordance with the International Council for Harmonisation (ICH) references. The discoveries were reasoned to be consistent as well as agreeable. As a consequence, this optional technique was successfully used to estimate rebamipide quantitatively in conventional analytical applications.

Application of estimand framework to the design and analysis of multi-regional clinical trials

ABSTRACT With the increasing globalization of drug development and the publication of the International Council for Harmonisation (ICH) E17 guideline (ICH International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use 2017), multi-regional clinical trials (MRCTs) have become a preferred option to accelerate the availability of new medical products by design, execution and simultaneous submission under one protocol. MRCTs, with the participation of all major regions including countries from both developed and emerging markets, surely make new drug development more efficient. Even though the proposed estimand framework (ICH E9 (R1) (2019), came later in 2019 and was not mentioned in ICH E17, the application of the estimand framework has the potential to enhance the design, execution, and analysis in MRCTs. Defining an estimand within the regional context in MRCTs is an important issue that requires careful consideration. Given that consistency evaluation of treatment effects across regions is critical in MRCTs, the utilization of the estimand framework for regional consistency evaluation is also worth discussion. This paper aims to address these two questions. The five attributes of the estimand definition are discussed within a multi-regional context. It is imperative to thoroughly consider regional intrinsic/extrinsic factors when planning the estimand and estimation of MRCTs. A holistic approach is summarized to conduct consistency evaluation. When a regional inconsistency is observed, the possible reasons need to be further explored under five attributes of the estimand framework. Two real case studies are discussed to illustrate the application of the estimand framework in the consistency evaluation.

HPLC-DAD method for concurrent estimation of netarsudil, latanoprost and benzalkonium chloride: Stability indicating study, impurity separation, assay of ophthalmic preparations and greenness/whiteness valuation

A validated, stability indicating, multiuse and sustainable HPLC-DAD method was developed for simultaneous determination and separation of the ophthalmic ternary mixture of netarsudil (NTS), latanoprost (LTP) and benzalkonium chloride (BZC) in presence of four related compounds and suspected impurities. Effectual liquid chromatographic separation was accomplished by using a Zorbax-SB Phenyl column with gradient elution of a mobile phase consisting of buffer mixture solution of phosphate and sodium pentane sulfonate (pH 3.0) and acetonitrile at flow rate 1.5 mL/min. DAD was tuned at 254 nm for measuring peak area of NTS and 210 nm for LTP and BZC. The proposed procedure showed to be effective in the stability investigation of the cited drugs. The suggested HPLC procedure was validated based on the International Council for Harmonization (ICH) guiding principles. The three analytes were determined within linearity ranges of 1–100 µg/mL, 5–200 µg/mL and 10–200 µg/mL for NTS, LTP and BZC respectively with excellent analytical performance. Assay of several ophthalmic preparations was successfully carried out using the validated method, which was further appraised together with some relevant reported methods in regard to green and white analytical chemistry aspects using AGREE and RGB-12 tools. Very few published analytical records were found for NTS/LTP mixture. After surveying the scientific literature, no documentation was retrieved for a similar inclusive stability-indicating impurity-profiling separative method for the synchronized quantitation of NTS, LTP together with co-formulated preservative BZC.

Green analytical chemistry-based spectrophotometric techniques for ternary component analysis of pain relievers

BackgroundThe management of pain presents a significant challenge in healthcare, particularly in cases where conventional therapies prove inadequate. In response to this need, this study aims to devise two innovative UV spectrophotometric techniques rooted in the principles of green analytical chemistry for the analysis of Aceclofenac (ACE), Paracetamol (PAR), and Tramadol (TRM) in both bulk and tablet forms.ResultsUtilizing advanced mathematical methodologies such as the double divisor ratio spectra method and area under the curve, the concentrations of these drugs were accurately determined. Validation of the developed methods adhered to the guidelines outlined by the International Council for Harmonisation in the Q2 (R1), revealing linear calibration curves for ACE (8–12 µg/mL), PAR (22.75–35.75 µg/mL), and TRM (2.62–4.12 µg/mL). Furthermore, statistical analyses employing Student’s t test and F test were conducted to ensure the robustness of the proposed method. The evaluation of environmental impact through green metric tools confirmed the eco-friendliness of the proposed methodologies.ConclusionThe assessment performed utilizing green metric tools has substantiated the environmental sustainability of the proposed approach. Thus, this methodology offers accurate and reliable outcomes for the determination of three drugs, as indicated by the complete overlap observed in the zero-order spectra.

Green and fast quantification of Bisphenol A in instant noodles cup using iridium nanoparticle-modified glassy carbon electrode

This study presents a method for quantifying the endocrine-disrupting compound Bisphenol A (BPA) in food packages. Carbon-supported iridium nanoparticles were synthesized via formic acid reduction and used to modify a glassy carbon electrode, creating the Ir/C/GCE sensor. Employing this sensor in conjunction with square wave voltammetry, a fast and green electroanalytical methodology was developed and subsequently validated for quantifying BPA in instant noodle cup packages. The optimized and validated analytical method exhibited linearity, with low limits of detection and quantification recorded at 0.011 μmol L−1 and 0.037 μmol L−1, respectively. Besides this, it demonstrated good selectivity and accuracy, complying with major validation guidelines, including those from the International Council for Harmonisation, Eurachem, the European Union, and AOAC International. Furthermore, the environmental sustainability of the method was confirmed using the AGREE metric.

Eco‐Friendly Analytical Approaches for the Quantification of Molnupiravir and Favipiravir Using Capillary Zone Electrophoresis and Derivative Ratio Spectrophotometry

AbstractDesigning new validated analytical procedures with a focus on many objectives, such as simplicity, environmental friendliness, analytical effectiveness, and sustainability, has grown to be a top priority in quality control departments. Applying green and white analytical chemistry concepts is no longer a luxury. In this report, two new approaches, Capillary Zone Electrophoresis (CZE) with Photo Diode Array (PDA) detection and derivative ratio spectrophotometric methods, are established and validated for the simultaneous analysis of molnupiravir (MLP) and favipiravir (FAV) in bulk, single dosage forms, and their expected combined dosage forms. For the two drugs, linearity ranges of 5–50 μg/mL and 1–30 μg/mL were achieved using the CZE and spectrophotometric methods, respectively. Both techniques have been validated and used for the estimation of the two medications in lab‐made capsules in accordance with the International Council for Harmonization's (ICH) recommendations. Seven assessment models were used to evaluate the greenness and sustainability of the introduced approaches compared to three previously published HPLC, chemometric, and HPTLC techniques. ChlorTox and BAGI tools are applied for the first time for the evaluation of an analytical procedure. Higher green and sustainable qualities were declared by suggested approaches than by earlier ones.

Cannabinoid Modulation of Monoamine Levels in Mouse Brain: Unveiling Neurochemical Dynamics through an Innovative High-performance Liquid Chromatography-Fluorescence Detection Bioanalysis

Homeostasis of monoaminergic pathways is compromised in aging and neurodegenerative/neurological diseases such as Alzheimer’s disease and depression. On the other hand, their modulation has also been linked to the mechanism of action of several drugs. Therefore, monitoring the levels of noradrenaline (NA), adrenaline (AD), dopamine (DA), and serotonin (5-HT), as well as those of tryptophan (TRYP), the precursor of 5-HT, and DA metabolites, namely 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA), is fundamental for assessing disease severity and progression. This work aimed to develop and validate the first High-Performance Liquid Chromatography (HPLC) coupled with fluorescence detector (FLD) method that simultaneously and accurately quantifies NA, AD, DA, DOPAC, HVA, 5-HT and TRYP in mouse brain and prefrontal cortex (PFC) matrices. Previous sample preparation by protein precipitation was required to extract the compounds. Calibration curves were plotted using the background subtraction approach to reduce the interference of the endogenous analytes. Intra and inter-day accuracy and precision were within the ranges defined by ICH (The International Council for Harmonisation) guideline for bioanalytical method validation. Following validation, the impact of cannabidiol (CBD), cannabigerol (CBG), and cannabidivarin (CBDV) was explored in mice brains post-administration, revealing significant alterations in specific neurotransmitter levels upon cannabinoid exposure and shedding light on the complex modulation of neurochemical dynamics by cannabinoids. This research highlights the fit-for-purpose of the HPLC-FLD method and provides insights into potential mechanisms underlying phytocannabinoid actions in the central nervous system (CNS).

IDENTIFICATION, SEPARATION, AND CHARACTERIZATION OF DEGRADATION PRODUCTS OF TRIAMCINOLONE HEXACETONIDE USING LC AND LC-MS/MS

Objective: The study aimed to separate the forced degradation products of Triamcinolone hexacetonide using HPLC and characterize the degradation product by LC-MS/MS fragmentation pattern. Methods: Triamcinolone hexacetonide (THA) and its primary degradation products were identified using a liquid chromatography-mass spectrometry/Mass spectrometry (LC-MS/MS) approach. The degradation study was based on in-depth stress testing with acid, base, peroxide, heat, and light. A Zorbax SB C18 column and a greener mobile phase composed of methanol and 10 mmol ammonium acetate buffer in water at pH 3 were employed to accomplish separation and quantitation at a flow rate of 0.7 ml/min in an isocratic mode with a 239 nm detection wavelength. Results: A major degradation product of the drug was obtained in acidic and alkaline stress conditions. The drug was found to be stable for all other stress conditions. The LC-MS/MS analysis results of the active pharmaceutical ingredient and resulting product after degradation were interpreted to identify the novel degradation product and fragments. The developed method was validated as per International Council for Harmonization (ICH) guidelines. The square root of the correlation coefficients, which indicated linearity for THA in 50 to 150 % of the workload, was 0.99. Method Precision assay was performed on six different preparations, percentage relative standard deviation (% RSD) of assay value is 0.17 % and system precision is 0.30 %. In accuracy, overall % RSD of 50 %, 100 %, and 150 % in triplicate is 0.95. Conclusion: It is concluded that the drug is stable to all other stress conditions except for acidic and alkaline stress conditions and generates a novel degradation product. The developed LC (Liquid chromatography) method separates and identifies the degradation product.

Development and Validation of Fast and Sensitive RP-HPLC Stability-Indicating Method for Quantification of Piroxicam in Bulk Drug.

A simple, rapid, sensitive, and cost-effective green solvent-assisted reverse-phase high-performance liquid chromatographic technique, coupled with a photodiode array detector, was developed and validated for the estimation of piroxicam (PRXM). The chromatographic separation was achieved by using a C-18 (250 × 4.6) mm, 5-μm stationary phase and a mobile phase consisting of methanol and 0.1% ortho-phosphoric acid in water in a ratio of (80:20) v/v at a flow rate of 1ml/min. The detection was carried out at a wavelength of 254nm with a constant injection volume of 10μL throughout the analysis. The calibration curve was observed to be linear over the optimum concentration range of 50-300μgmL-1, with an R2 value of 0.9995. The developed method was validated as per the International Council for Harmonisation (ICH) Q2 (R1) guideline. Various parameters like selectivity/specificity, accuracy/recovery, linearity, precision, detection limit, quantitation limit, robustness and stability of analyte in solution were performed for the method validation. The PRXM was evaluated under stressed conditions, including acidic, basic, oxidative, thermal and photolytic, as per ICH Q1 (R2) guidelines. Significant degradation was observed in acidic and basic degradation conditions. Conversely, the drug substance showed stability when exposed to oxidative, photolytic and thermal degradation conditions.

Simultaneous measurement of duloxetine hydrochloride and avanafil at dual-wavelength using novel ecologically friendly TLC-densitometric method: application to synthetic mixture and spiked human plasma with evaluation of greenness and blueness

The simultaneous assay of duloxetine hydrochloride (DLX) and avanafil (AVN) in their pure forms, synthetic mixtures, and spiked human plasma was achieved using a novel, eco-friendly, sensitive, and specific HPTLC methodology that have been established and validated. Measuring the levels of co-administered antidepressants and sexual stimulants in biological fluids is an important step for individuals with depression and sexual problems. Separation was performed successfully using pre-coated silica gel 60-F254 as a stationary phase and a mobile phase composed of methanol, acetone, and 33% ammonia (8:2:0.05, v/v/v). Compact bands were produced by the optimized mobile phase that was chosen for development (Rf values were 0.23 and 0.75 for DLX and AVN, individually) after dual-wavelength detection for DLX and AVN at 232 and 253 nm, respectively. The results of polynomial regression analysis were exceptional (r = 0.9999 for both medicines) over concentration ranges of 5-800 and 10-800ng/spot for DLX and AVN, respectively. The quantitation limits were 4.69 and 9.53 ng/spot (0.31 and 0.94 µg/mL), whereas the detection limits were 1.55 and 3.15 ng/spot (0.63 and 1.91 µg/mL), for DLX and AVN, respectively. The International Council for Harmonization (ICH) criteria served as the basis for validating the established approach. Moreover, the proposed technique was evaluated in terms of greenness using four contemporary ecological metrics: The Analytical Greenness software (AGREE), the Green Analytical Procedure Index (GAPI), Eco-Scale, and the National Environmental Method Index (NEMI). Additionally, the Blue Applicability Grade Index (BAGI), a newly developed tool for evaluating the practicality (blueness) of procedures, was taken into consideration when evaluating the sustainability levels of the established approach.

Should You Run a Dedicated TQT Study? Sponsor and Regulatory Considerations on Substitution Pathways to Assess QT Liability.

Cardiac safety regulatory guidance for drug development has undergone several monumental shifts over the past decade as technological advancements, analysis models and study best practices have transformed this landscape. Once, clinical proarrhythmic risk assessment of a new chemical entity (NCE) was nearly exclusively evaluated in a dedicated thorough QT (TQT) study. However, since the introduction of the International Council for Harmonisation (ICH) E14/S7B Q&A 5.1 and 6.1 TQT substitutions, drug developers are offered an alternative pathway to evaluate proarrhythmic risk during an ascending dose study in healthy volunteers or during a powered patient study, respectively. In addition, the findings as well as the manner in which nonclinical studies are conducted (i.e., utilizing best practices) can dictate the need for a positive control in the clinical study and/or affect the labeling outcome. Drug sponsors are now faced with the option of pursuing a dedicated TQT study or requesting a TQT substitution. Potential factors influencing the choice of pathway include the NCE mechanism of action, pharmacokinetic properties, and safety profile, as well as business considerations. This tutorial will highlight the regulatory framework for integrated arrhythmia risk prediction models to outline drug safety, delineate potential reasons why a TQT substitution request may be rejected and discuss when a standalone TQT is recommended.