The convergence of statistical design and computational intelligence: a systematic review of DOE-driven AI/ML in analytical method development and validation

Analytical method development, validation, and transfer are essential elements of any pharmaceutical development program. Effective method development confirms that laboratory resources are optimized, while methods meet the purposes required at each stage of drug development. High performance liquid chromatography is most accurate methods extensively used for the qualitative and quantitative analysis of drug product. Analytical method development and validation play vital role in the drug discovery, Drug development and manufacture of pharmaceuticals. It includes detection of the purity and toxicity of a drug substance. A number of chromatographic parameters have been evaluated in order to optimize the methods in the analysis of method development in HPLC. Ansuitable mobile phase, column, column temperature, wavelength, and gradient are developed. Keywords: Analytical method validation, ICH, HPLC, Method Validation, Regulatory Requirements.

Analytical methods plays vital role in the process of identification, separation and then quantification of chemical components in natural materials or synthetic materials based on their chemistry. The main purpose of the analytical method development and validation is to prove that proposed analytical method is accurate, specific, precise and robust in the pharmaceutical industry for analysis of a drug moiety. Analytical method development gives important information in the pharmaceutical industry, on the potency of a drug, the drug’s bioavailability, the drug’s stability and also its effects. The analytical method validation is essential for analytical method development and tested for specificity, linearity, accuracy, precision, range, detection limit, quantitation limit and robustness. In summary, analytical method development and validation confirms that an accurate, precise and reliable potency measurement of a pharmaceutical preparation can be performed.
 Keywords: HPLC, HPTLC, UPLC, GC, MS, SOP

Evogliptin is an anti-diabetic drug, which comes under the class of gliptin derivatives for the inhibition of selective dipeptidyl peptidase-4 inhibitor. The developed ultraviolet spectrophotometric method was simple, sensitive, accurate, precise and economic for the development and validation of evogliptin in bulk and tablet dosage form. In this present study, the analytical method validation and development of evogliptin was done using the different parameters of method validation as per International Council for Harmonisation Q2(R1) guidelines. Water using as a solvent and it shows the maximum wavelength at 266 nm and then performed all the parameters of analytical method validation like accuracy, precision, linearity, range, robustness, ruggedness, limit of detection and limit of quantitation. Evogliptin showed linearity over the range of 2-48 µg/ml. The correlation coefficient value obtained was 0.996 with the regression equation y=0.0032x+0.0005. The accuracy studies was done in spiking method and the recoveries ranging from 97.07 %-106.13 %. The percentage relative standard deviation for intra-day precision was 0.44 and inter-day precision was 0.59. The limit of detection was 1.1 µg/ml and limit of quantitation was 3.33 µg/ml respectively. The method has shown good and consistent recoveries and is validated as per International Council for Harmonisation guidelines and can be used for routine quality control analysis of evogliptin in dosage form.

Alfuzosin hydrochloride, an alpha-adrenergic blocker, is extensively used in the treatment of benign prostatic hyperplasia. Robust and reliable analytical methods are indispensable for quantifying alfuzosin hydrochloride in various matrices, ensuring the quality, efficacy, and safety of the drug. Recent advancements in analytical method development and validation for alfuzosin hydrochloride encompass a spectrum of techniques, including UV spectrometry, RP-HPLC, HPTLC, UPLC, and voltammetry. Optimization of chromatographic conditions, such as mobile phase composition, column selection, and detection wavelength, is crucial for achieving optimal separation and quantification of the drug. Method validation parameters, including accuracy, precision, linearity, and robustness, are assessed in accordance with ICH guidelines. These analytical methods find application in the determination of alfuzosin hydrochloride in bulk drugs, pharmaceutical formulations, and biological matrices. Stability-indicating methods are developed to evaluate drug stability under various stress conditions. The current state of analytical method development and validation for alfuzosin hydrochloride facilitates the selection of suitable analytical techniques for routine quality control and research purposes

Method validation is an important part of analytical chemistry to confirm that the method employed for a specific test is suitable for its intended use. As such, it is an essential requirement for any package of information submitted to regulatory agencies in support of new product marketing or clinical trial applications. Currently, there is no single source or final guideline on analytical method validation that helps analysts to perform validation in a systematic manner. Therefore, industry depends on the analyst's knowledge and experience to develop simple and efficient methods of analysis. The intention of this paper is to review regulatory requirements and role of the pharmacopeias and to study how analytical method development and validation are typically carried out at present, and to formulate this into a simple step by step approach. Such a template was not only used as the foundation of this research programme, but could also serve as a simple systematic guide for other practitioners and those new to the field. Furthermore, it was recognized that this protocol should satisfy the requirements of the most strategically important regulatory bodies.

<p>The article entitled “Analytical Method Development and Validation for Determination of Assay of Antibacterial Drugs Besifloxacin Hydrochloride and Phenoxyethanol in Gel Formulation”, by Pradeep Kundu, Neelam Pawar, Neha Minocha and Asha Poonia, published in Anti-Infective Agents 2023; 21(4): e310323215342. https://dx.doi.org/10.2174/2211352521666230331111623. <p> The corresponding author of this article submitted it simultaneously to the journal “Biochemistry & Analytical Biochemistry of Walsh Medical Media publishers and to the journal of “Anti-Infective Agents” of Bentham Science Publishers. Therefore, the article has been retracted due to the unethical practice of the author and during plagiarism checking with a previously published article titled: “Analytical Method Development and Validation for the Determination of Assay of Antibacterial Drugs Besifloxacin Hydrochloride and Phenoxyethanol in Gel Formulation. Biochem Anal Biochem. 11:426. doi: 10.35248/2161-1009.22.11.426 <p> The authors were informed of this submission and were suggested to give justification on the matter in their defense. Instead of sending justification, the corresponding author Dr. Neelam Pawar suggested Bentham Science Publishers to retract this article. <p> Bentham Science apologizes to the readers of the journal for any inconvenience this may have caused. <p> The Editorial Policy of Bentham Science Publishers on Article Retraction can be found at the link https://benthamscience.com/editorial-policies-main.php. <p> BENTHAM SCIENCE DISCLAIMER: <p> It is a condition of publication that manuscripts submitted to this journal have not been published and will not be simultaneously submitted or published elsewhere. Furthermore, any data, illustration, structure or table that has been published elsewhere must be reported, and copyright permission for reproduction must be obtained. Plagiarism is strictly forbidden, and by submitting the article for publication, the authors agree that the publishers have the legal right to take appropriate action against the authors if plagiarism or fabricated information is discovered. By submitting a manuscript, the authors agree that the copyright of their article is transferred to the publishers if and when the article is accepted for publication.

Aim: The present work aims to develop an analytical method and validate it to determine the assay of an antibacterial dug-in gel formulation. background: Background: Analytical Method Validation is a process involving confirmation studies that procedure/ method/ system/ analyst provides precise and reproducible outcome recognized by research laboratory studies that the performance features of the technique follows the necessities required for the analytical applications. Introduction: Analytical Method Validation is a process involving confirmation studies that procedure/ method/ system/ analyst provides precise and reproducible outcomes recognized by research laboratory studies that the performance features of the technique follow the necessities required for the analytical applications. Objective: Objective: To improve the conditions and parameters which should be followed in the development and validation by developing a new sensitive and accurate RP-HPLC method. Validating the proposed newly developed methods per the analytical parameters mentioned in the IP, USP, BP and ICH guidelines. Methods: HPLC method was validated to indicate that the analytical procedure used is suitable for intended use by using various parameters like specificity, linearity, LOD, LOQ, precision, accuracy, range, robustness, stability in analytical solution and system suitability. Results: The standard retention times for the Drug Besifloxacin were 7.781 min, and the sample was 7.731, respectively. The area of standard besifloxacin was 1828547, and the sample area was 1825315. The assay of the sample was 98%. The retention times for the drug Phenoxyethanol standard were found to be 2.010 min, and the sample was at 2.004, respectively. The Area of standard Phenoxyethanol was 438025, and the sample area was 438103. The assay of the sample was 97.04%. The RSD for 5 replicate injections for each peak is 0.33% in system suitability. In specificity, peaks of Diluent, Placebo & Impurities are not interfering with the Besifloxacin peaks. Peaks of Besifloxacin were found to be pure. Degradation products were found to be well separated from the besifloxacin peak. The peak purity factor was NLT 0.9995. In the precision study, the System Precision RSD of the Retention time for Besifloxacin obtained from six replicate injections was 0.33%. The RSD of the Area of Besifloxacin obtained from six replicate injections is 0.46%. Method precision RSD was calculated on 6 determinations assay value of Drug besifloxacin is 0.56%. The RSD calculated on 6 determinations for the assay value of the Drug besifloxacin is 0.50%. In Intermediate precision, RSD was calculated on 6 determinations for the assay value of the Drug besifloxacin is 0.50%. The RSD calculated on 12 determinations (Method precision & Intermediate precision) for assay value is 0.50%. Stability in the analytical solution for the standard and sample, the area difference of besifloxacin peak was within ±2.0% from initial Linearity. The correlation coefficient & regression coefficient (R square) should be not less than 0.995 for Besifloxacin Correlation Coefficient is 0.998 Regression coefficient is 1.000. The % intercept should be within ±5.0% of the response at 100% level Precision at 50% and 150% levels; the RSD is 0.01%. Precision at 50% & 200% level: the RSD was found to be NMT 2.0%. Accuracy means % recovery at each level found to be between 98 to 101 % of the drug besifloxacin. RSD on 9 (3 levels X 3) determinations is 1.2, following the NMT 2% range criteria. A correlation was 1.0% for the accuracy and linearity parameters. Conclusion: The % recovery is between 98% to 101%, and the % RSD for all recovery values is 1.41% which is within limits. The HPLC method optimized the conditions to obtain an adequate separation of eluted compounds.

This write-up provides a review on the stability indicating analytical method development and validation. This review provides a basic detail about analytical method development, stability indicating analytical methods and validation as per regulatory guidelines and current scenario. What kind of content required for analytical procedure, steps involve in development of stability indicating analytical procedure also described. Details of Validation and its parameter like Specificity, Linearity, Range, Accuracy, Precision, Detection limit, Quantitation limit, Robustness, System suitability testing given.

A specific and sensitive analytical method development and validation plays an important role for building drug discovery, product development and manufacturing of pharmaceuticals. The main purpose of this review article is to check the report of specific and sensitive analytical method development and validation parameters involved in different pharmaceuticals manufacturing. Development of sensitive analytical method is extremely much significant because it ensures product efficacy and product quality. The sensitive analytical method development and validation is the process of proving that the analytical method meets the analytical acceptance criteria and it can use to measure the concentration of target molecule at its specification limit. The aim of method development is to produce a reproducible specific and sensitive analytical method in a cost effective manner. The parameters of validation involved are introduced as specificity, method precision, accuracy, linearity, limit of detection (LOD), limit of quantitation (LOQ), ruggedness, robustness and system precision of the target molecule.

Navigating ICH Q2(R2) compliance in analytical method validation: A gap analysis toolkit to streamline risk assessment and change management.

Analytical method development and validation are the continuous and inter-dependent task associated with the research and development, quality control and quality assurance departments. Analytical procedures play a critical role in equivalence and risk assessment, management. It helps in establishment of product-specific acceptance criteria and stability of results.Validation should demonstrate that the analytical procedure is suitable for its intented purpose. Design of experiment is a powerful tool for the method characterization and validation. Analytical professionals should be comfortable to use it to characterize and optimize the analytical method. An effective analytical method development and its validation can provide significant improvements in precision and a reduction in bias errors. It can further help to avoid costly and time consuming exercises.

The importance of the quality control unit to gatekeep pharmaceutical products to satisfy CGMP regulation is discussed. Scientific results and data are needed in most cases to answer PDA's questions. Laboratory's work functions, i.e. quality control (QC) , analytical method development and validation (AM) and stability (STB) have been included. The role of quality assurance (QA) has been discussed briefly. Analytical method validation, stability indicating method development and validation, specification and standard testing method preparations are critical tasks that may affect PDA's approval of the product and decision on whether a quality control unit of a comapny can ensure the products' good quality. Also mentioned are the "retest", "training/qualification" and "calibration of instruments" issues. A discussion is also made on the most recent trend in the US for considering using the first party or the third party to audit or inspect facilities to lessen the PDA's burden on frequent inspection.

Introduction: Analytical method development and validation are crucial for ensuring drug quality and efficacy. As combination therapy is common in type 2 diabetes, a reliable method is needed for the simultaneous estimation of Metformin, Sitagliptin, and Imeglimin in pharmaceutical formulations. This study aims to develop and validate a simple, precise, and robust RP-HPLC-UV method for the simultaneous estimation of Metformin, Sitagliptin and Imeglimin in single combined dosage forms. Materials & Methods: Metformin, Sitagliptin, and Imeglimin were analyzed using a Shimadzu RP-HPLC system with a C18 column and UV detection at 260 nm. Stock and working solutions were prepared in aqueous media, and calibration curves were constructed over relevant concentration ranges. The optimized mobile phase (Methanol: Cyanomethane: 15 mM Ammonium Formate, 10:40:50 v/v, pH 6.5) under isocratic flow (0.9 mL/min) provided well-resolved peaks with retention times of 3.7, 4.7, and 8.3 min. The method was validated as per ICH guidelines for specificity, linearity, accuracy, precision, LOD, LOQ, robustness, and ruggedness. Results: The developed RP-HPLC-UV method enabled efficient and simultaneous estimation of Sitagliptin, Metformin, and Imeglimin with well-resolved peaks at retention times. The method exhibited high specificity, with no interference from diluents or other components. The LOD and LOQ were 85/265 ng/mL for Sitagliptin, 25/80 ng/mL for Metformin, and 55/175 ng/mL for Imeglimin. Accuracy, assessed via recovery studies, ranged from 98.6–99.9%, and precision studies showed low %RSD, confirming repeatability and reproducibility. System suitability parameters, including theoretical plates, tailing factor, and resolution, were within acceptable limits, indicating the method’s reliability for routine analysis of these antidiabetic drugs. Conclusions: A validated RP-HPLC-UV method was developed for simultaneous estimation of Sitagliptin, Metformin, and Imeglimin. The method showed high specificity, accuracy, and precision, suitable for routine pharmaceutical analysis. Keyword :Analytical method, Imeglimin, Metformin, RP-HPLC, Sitagliptin and Validation.

Analytical Method Development and Validation for Spironolactone and Hydrochlorothiazide in bulk and Combined Dosage Form by RP-HPLC. New method was established for simultaneous estimation of Spironolactone and Hydrochlorothiazideby RP-HPLC method. The chromatographic conditions were successfully developed for the separation of Spironolactone and Hydrochlorothiazideby using Inertsil C18 (4.6mm ×250mm, 5µm particle size), flow rate was 1.0ml/min, mobile phase ratio was (55:45% v/v) Methanol: Phosphate buffer pH 4.8 (pH was adjusted with ortho phosphoricacid), detection wavelength was 282nm. The instrument used was WATERS Alliance 2695 separation module, Software: Empower 2, 996 PDA detector. The retention times were found to be 1.688mins and 3.282mins. The %purity of Spironolactone and Hydrochlorothiazidewas found to be 99.86%. The system suitability parameters for Spironolactone and Hydrochlorothiazidesuch as theoretical plates and tailing factor were found to be 7586, 1.69 and 6235 and 1.58, the resolution were found to be 10.85. The analytical method was validated according to ICH guidelines (ICH, Q2 (R1)). The linearity study ofSpironolactone and Hydrochlorothiazidewas found in concentration range of 100µg-500µg and 30µg - 70µg and correlation coefficient (r2) was found to be 0.999 and 0.999, % recovery was found to be 100.112% and 100.16%, %RSD for repeatability was 0.1702 and 0.043 respectively. The precision study was precise, robust, and repeatable. The LOD value was found to be 2.1µg/ml and 1.28µg/ml, and LOQ value was 6.3µg/ml and 3.84µg/ml for Spironolactone and Hydrochlorothiaziderespectively. Hence the suggested RP-HPLC method can be used for routine analysis of Spironolactone and Hydrochlorothiazide in API and Pharmaceutical dosage form.

Analytical Method Development and Validation for Azithromycin and Cefpodoxime in bulk and Combine Dosage Form by RP-HPLC, New method was established for simultaneous estimation of Azithromycin and Cefpodoxime by RP-HPLC method. The chromatographic conditions were successfully developed for the separation of Azithromycin and Cefpodoxime by using Symmetry ODS C18 (4.6mm×250mm, 5µm) particle size, flow rate was 1.0 ml/min, mobile phase ratio was (70:30 v/v). The system suitability parameters for Azithromycin and Cefpodoxime such as theoretical plates and tailing factor were found to be 5387, 0.97 and 5398 and 1.26, the resolution was found to be 2.97. The analytical method was validated according to ICH guidelines (ICH, Q2 (R1)). The linearity study n Azithromycin and Cefpodoxime was found in concentration range of 30µg-70µg and 60µg-140µg and correlation coefficient (r2) was found to be 0.999 and 0.999, % recovery was found to be 100.14% and 100.56%, %RSD for repeatability was 0.1 and 0.5, % RSD for intermediate precision was 0.1 and 0.1 respectively. The precision study was precise, robust, and repeatable. LOD value was 0.56 and 1.2, and LOQ value was 1.7 and 3.6 respectively. Hence the suggested RP-HPLC method can be used for routine analysis of Azithromycin and Cefpodoxime in API and Pharmaceutical dosage form.