Quality by design-assisted green uv-spectrophotometric method development and validation for the quantitative estimation of letrozole

Spectrophotometric methods were developed according to Quality by Design (QbD) approach as per ICH Q8(R2) guidelines for estimation of dolutegravir. QbD approach was carried out by varying various parameters, and these variable parameters were designed into Ishikawa diagram. The present work deals with the development of sensitive, simple, accurate, precise and cost-effective UV-spectrophotometric method for the determination of dolutegravir, an anti-retroviral drug, in bulk and pharmaceutical dosage form by UV spectrophotometric method as per International Conference on Harmonisation (ICH) guidelines. The critical parameters were determined by using principal component analysis as well as by observation. Estimated critical parameters in the spectrophotometric method were solvent methanol, wavelength: 260nm, slit width: 0.5, scan speed fast, sampling interval: 0.2nm and proposed method was validated for various parameters like system suitability, linearity, precision, accuracy, detection limits and quantification limits as per the International Conference on Harmonization guidelines ICH Q2(R1). The method's linearity was found to be excellent over the concentration range 5 to 25μg/ml with high correlation coefficient value of 0.999. Limits of detection and quantification were found to be 0.20μg/ml and 0.60μg/ml, respectively. The mean recovery was found to be 100.35% with low percentage relative standard deviation (% RSD) value. The precision study also has shown low % RSD value (less than 1). No interfering peaks were observed during specificity studies. A simple, rapid, sensitive, accurate, precise and inexpensive spectrophotometric method was developed for estimation of dolutegravirin bulk by using analytical quality by design (AQbD) approach. The same method is also applied for plasma samples study in bioanalytical work.

Objective: The present work is aimed to develop a simple, rapid, selective and economical UV spectrophotometric method for quantitative determination of Glipizideinbulk and pharmaceutical dosage form.
 Methods: In this method Dimethyl Form amide (DMF) was used as solvent, the absorption maxima was found to be275 nm in DMF. The developed method was validated for linearity, accuracy, precision, ruggedness, robustness, LOD and LOQ in accordance with the requirements of ICH guideline.
 Results: The linearity was found to be 10-60 µg/ml having linear equation y=0.017x-0.006 with correlation coefficient of 0.997. The% recovery was found to be in the range of 98.7-100%. The % RSD for intra-day and inter-day precision was found to be 0.569923 and 0.40169 respectively. The limit of detection (LOD) and limit of quantification (LOQ) was found to be3.06 µg/ml and 9.27 µg/ml respectively.
 Conclusion: The developed method was validated as per ICH Q2(R1) guidelines. The novel method is applicable for the analysis of bulk drug in its pharmaceutical dosage form.

BackgroundThe Analytical Quality by Design (AQbD) methodology extends the application of Quality by Design (QbD) principles to the management of the analytical procedure life cycle, encompassing method creation, optimization, validation, and continuous improvement. AQbD assists in creating analytical procedures that are robust, reliable, precise, and cost-efficient. Opdualag™ is a combination of Nivolumab and Relatlimab, which are antibodies that block programmed death receptor-1 (PD-1) and lymphocyte-activation gene 3 (LAG-3) receptors, used to treat advanced melanoma. This work aims to develop and validate a reversed-phase ultra-performance liquid chromatography (RP-UPLC) method using AQbD principles to determine NLB and RTB in pharmaceutical products.ResultsA central composite design (CCD) comprising three factors arranged in five distinct levels was implemented via Design-expert® software to optimize the chromatographic conditions. A mathematical model was constructed and the effects of three independent factors namely flow rate (X1), percentage of methanol in the mobile phase (X2), and temperature (X3) on responses including retention time (Y1–2), resolution factor (Y3), theoretical plates (Y4–5), and tailing factor (Y6–7) were investigated. The software determined the optimal chromatographic conditions for the separation of NLB and RTB, which were as follows: 32.80% methanol in the mobile phase, 0.272 mL/min flow rate, 29.42 °C column temperature, and 260 nm UV detection. The retention time for NLB and RTB were 1.46 and 1.88 min, respectively. The method exhibited linearity across the concentration ranges of 4–24 µg/mL for RTB and 12–72 µg/mL for NLB. The limits of detection (LOD) and limit of quantification (LOQ) for NLB and RTB, respectively, were 0.89 µg/mL, 2.69 µg/mL and 0.15 µg/mL and 0.46 µg/mL. The percentage relative standard deviation (%RSD) of intraday and interday precision for NLB and RTB was below 2. The recovery percentages for NLB and RTB were determined to be 99.57–100.43% and 99.59–100.61%, respectively. Both drugs were found to be susceptible to oxidative and photolytic degradation in forced degradation studies.ConclusionsEmploying the AQbD-based methodology, a straightforward, fast, accurate, precise, specific, and stability-indicating RP-UPLC method has been established for the quantitative analysis of NLB and its RTB in pharmaceutical formulations.

A stability-indicating liquid chromatographic method was developed employing the principles of quality by design (QbD) to quantify vilazodone hydrochloride (VLN) in pharmaceutical dosage form. A Box-Behnken experimental design was employed to establish optimum conditions including method robustness by selecting organic phase proportion (%), mobile phase flow rate (mL/min) and pH of buffer as the factors, to study their effect on plate number as the response variable. Chromatography was performed on a C-18 column using methanol:phosphate buffer of pH 7.0 (85:15, v/v) as the mobile phase at a flow rate of 1.2 mL/min with photo diode array (PDA) detection at 285 nm. Calibration curve was linear over 5-80 µg/mL with values of accuracy within 99.4-100.8%. The limit of detection and quantification were found to be 1.5 and 5.0 µg/mL, respectively. The developed method revealed high specificity for VLN and its degradation products formed during forced degradation conditions. Furthermore, control strategies were developed based on system suitability test result. The QbD-based developed liquid chromatographic method was found suitable for routine analysis of VLN in bulk drug and pharmaceutical dosage form.

Objective: To develop a simple, rapid, accurate, robust and inexpensive spectrophotometric method for the estimation of benidipine hydrochloride by using quality by design (QbD)” approach.Methods: A UV spectrophotometric method was developed on Shimadzu UV-1800 double beam spectrophotometer using methanol as solvent and wavelength of 236 nm was selected as absorbance maxima (𝜆max). Effect of input variables on spectrum characteristics were studied for the selection of critical parameters and proposed method was validated for various parameters like system suitability, linearity, precision, accuracy, detection limits and quantification limits as per the International Conference on Harmonization guidelines ICH Q2(R1).Results: Linearity of the method was found to be excellent over the concentration range 3 to 18 µg/ml with high correlation coefficient value of 0.9999. Limits of detection and quantification were found to be 0.20 µg/ml and 0.60 µg/ml respectively. The mean recovery was found to be 100.35 % with low percentage relative standard deviation (% RSD) value. The precision study also has shown low % RSD value (<1). No interfering peaks were observed during specificity studies.Conclusion: Obtained result indicated that the developed spectrophotometric method is robust and efficient for the determination of benidipine hydrochloride.

This study emphasizes the crucial role of Quality by Design (QbD) in developing pharmaceutical procedures, particularly in risk assessment. It demonstrates how QbD principles were applied to create a precise and effective HPLC method for Silymarin Tablets, ensuring consistent quality within specified criteria. The optimized method, developed using a Design of Experiment approach, employs a C18 column (150 mm x 4.6 mm, 5μm) with isocratic elution using a 95:25 ratio of acetonitrile to orthophosphoric acid buffer (pH 3). Peaks were detected using a PDA detector calibrated at 287 nm, with a flow rate of 1.0 mL/min. The column oven temperature was maintained at 25°C, and a 10 μL injection volume was used. Thorough validation, adhering to USP <1225> and ICH Q2 (R1) standards, ensures the method's reliability. Key factors such as accuracy, precision, robustness, limit of detection (LOD), and limit of quantification (LOQ) were comprehensively assessed. The method exhibits exceptional sensitivity, selectivity, efficiency, precision, accuracy, and cost-effectiveness, making it ideal for pharmaceutical analysis of Silymarin tablets. It has been validated to effectively differentiate between marketed products, including those closely resembling the original. This method is intended for routine quality control analysis in the pharmaceutical industry, highlighting its suitability and reliability for ongoing use.

Bovine clinical mastitis has significant repercussions for farmers across the globe. Meloxicam, a COX-2 inhibitor, attenuates mastitis symptoms and is also approved for veterinary use. An RP-HPLC (Reverse Phase-High Performance Liquid Chromatography) method development and validation is essential in the pharmaceutical industry to assess API (Active Pharmaceutical Ingredient) quantity present in the pharmaceutical dosage forms. RP-HPLC method of meloxicam was developed and optimized with the aid of QbD (Quality by Design) using Box-Behnken design (BBD). The pH of the aqueous mobile phase, acetonitrile (ACN) percentage, and column temperature were chosen as influence variables, and retention time (RT) and tailing factor (Tf) were selected as response variables. The optimum experimental conditions were selected as pH ~ 3 of the aqueous mobile phase, 65% v/v ACN, and 30˚C as column temperature. The drug was eluted at 6.02min RT with 1.18 as Tf. The method was subjected to validation for accuracy, linearity, precision, range, sensitivity, and robustness and was found to comply with ICH Q2 (R1) guidelines. The in vitro bioequivalent studies were performed in hydrochloric acid, pH ~ 1.2; acetate buffer, pH ~ 4.5; and phosphate buffer, pH ~ 6.8 for two veterinary brands of meloxicam tablets, and their release profiles were compared by mathematical models. Both the brands, brand 1 and 2 exhibited significant (Unpaired t-test, P < 0.05) differences in dissolution efficiency (DE) and mean dissolution time (MDT) except DE at pH 1.2. However, brands 1 and 2 showed similarity (f2 > 50) in terms of release of meloxicam except at pH 6.8 (f2 = 47.01). The in vitro release of meloxicam followed Peppas kinetics except for brand 2 at pH 6.8, where it followed the Higuchi model. Moreover, the recovery of meloxicam extracted with ACN in the milk sample was estimated to be 99.67 ± 0.58% significantly (Unpaired t-test, P < 0.05) higher than 90.34 ± 6.77% extracted with methanol. In conclusion, the optimized and validated RP-HPLC method of meloxicam may further be used for the analysis of drug content in pharmaceutical dosage forms in addition to biological fluids.

Xanthohumol (XH) is a prenylated chalcone available naturally and has diverse pharmacological activities. It has some limitations in the physiological environment such as biotransformation and less gastrointestinal tract absorption. To overcome the limitations, we prepared nanoformulations [solid lipid nanoparticles (SLNs)] of XH. Therefore, an analytical method is required for the estimation of XH in the bulk nanoformulations, so we developed and validated a quality by design (QbD)-based ultraviolet (UV)-spectrophotometric method as per the International Conference of Harmonization (ICH) Q2 (R1) guidelines. The new analytical Qbd based UV-visible spectrophotometric technique is developed and validated for estimation of XH in bulk and SLNs as per ICH guidelines Q2 (R1). Critical method variables are selected on the basis of risk assessment studies. Optimization of method variables was performed using the a central composite design (CCD) model. Multiregression ANOVA analysis showed an R2 value of 0.8698, which is nearer to 1, indicating that the model was best fitted. The optimized method by CCD was validated for its linearity, precision, accuracy, repeatability, limit of detection (LOD), limit of quantification (LOQ), and specificity. All validated parameters were found to be within the acceptable limits [% relative standard deviation (RSD) <2]. The method was linear between 2-12 g/mL concentration with R2 value 0.9981. Method was accurate with percent recovery 99.3-100.1%. LOD and LOQ were found to be 0.77 and 2.36 μg/mL, respectively. The precision investigation confirmed that the method was precise with %RSD <2. The developed and validated method was applied to estimate XH in bulk and SLNs. The developed method was specific to XH, which was confined by the specificity study.

A quick, accurate, and cost-effective UV spectroscopy method was developed to estimate the Phenytoin sodium in bulk, tablet dosage formulation using a solvent 0.1 N NaOH. Due to the high cost of antiepileptic drugs there is a need to develop a simple, accurate and cost effective method. UV-Spectrophotometric method was developed using 0.1 N NaOH as a solvent. According to ICH Q2(R1) requirements, the created technique was standardized in terms of validation parameters linearity, precision, accuracy, specificity, Limit of Detection (LOD), and Limit of Quantification (LOQ), like parameters like parameters were validated by using UV/visible spectroscopy technique and developed method was so simple, sensitive, precise, linear, accurate, robust, reproducible, and cost-effective. This newly established approach was effectively used to estimate phenytoin sodium in marketed formulation Eptoin. Phenytoin sodium API exhibits λmax at 216nm in the Beer’s range 2 to 10µg/ml So, Beer’s law was obeyed. The limit of detection (LOD) was found to be 0.564µg/ml and limit of quantification (LOQ) was found to be 1.71µg/ml. Recovery of phenytoin sodium in marketed formulation as Eptoin was observed in the range of 100 – 104%. The entire precision range was achieved at under 2% in accordance with ICH Q2(R1) standards. In both inter and intra-day examinations, it was found that the created UV method was precise, with % relative standard deviation varying from 0.27 to 0.46 and 0.26 to 0.46, respectively. The overall recovery percentage of Vinpocetine was found to range from 98.42 to 99.82%. LOQ and LOD were determined to assess the sensitivity of the method, yielding values of 0.4565 µg/ml and 0.1506 µg/ml, respectively. The estimation of Phenytoin sodium in its bulk form and commercial formulation was accomplished using the established method: A rapid, precise, and cost-effective UV spectrophotometric technique has been created to measure the concentration of Phenytoin sodium in bulk and tablet dosage forms.

Lobeglitazone sulfate is a novel thiazolidinedione drug used for the therapy of type 2 diabetes. Recent pre-clinical studies show that the drug is also effective in attenuating mucus hypersecretion and airway inflammation caused due to ovalbumininduced asthma. This study aimed to develop an accurate, simple, and cost-effective analytical method for the quantification of lobeglitazone sulfate in bulk and pharmaceutical dosage forms by UV spectrophotometric technique. The drug was identified by FTIR (ATR) instrument and the method development was performed with methanol as the solvent. In compliance with the ICH Q2(R2) guidelines, all the important validation parameters, such as linearity, accuracy, Limit of Detection (LOD), precision, specificity, Limit of Quantification (LOQ), and robustness were determined. The maximum absorption wavelength of lobeglitazone sulfate was found to be 248 nm. The linearity range was established from 2-14 μg/mL with a linearity equation of y = 0.0361x + 0.0024 and r² = 0.9987. The accuracy of the analytical method was found to be in the range of 99.37-100.41%. The precision studies were performed under three subsets of repeatability, intra-day, and inter-day, and the results recorded were within the acceptance limits, i.e., %RSD (<2%). The developed analytical method's Limit of Detection (LOD) and Limit of Quantification (LOQ) values were 0.07 μg/mL and 0.22 μg/mL, respectively. The developed analytical method has been found to be valuable in regular quality control analysis of lobeglitazone sulfate in pharmaceutical industries. The adopted approach of the developed method has been found to be facile, accurate, precise, and economical.

A simple, specific, precise, and accurate UV spectrophotometric method has been created for the simultaneous measurement of Nirmatrelvir and Ritonavir in pharmaceutical dosage forms. The absorption maxima of the drugs were found to be at 240 nm and 258 nm for Nirmatrelvir and Ritonavir respectively. Nirmatrelvir and Ritonavir obeyed Beer’s law in the concentration range of 12-18 µg/ml and 8-12 µg/ml respectively. Different analytical parameters such as linearity, precision, accuracy, limit of detection (LOD) and limit of quantification (LOQ) were determined as per ICH guidelines. Limit of detection and quantification values for Nirmatrelvir 1.25 and 3.85 μg/ml and for Ritonavir 0.34 and 1.12 μg/ml respectively. The accuracy of the methods was assessed by recovery studies and recovery values between prescribed limit of 99-101 % shows that method is free from interference of excipients present in formulation. The developed method was free from interferences due to excipients present in formulation and it can be used for routine quality control analysis. The results were validated statistically as per ICH Q2 R1 guideline and were found to be satisfactory. The proposed methods were successfully applied for the determination of for Nirmatrelvir and Ritonavir in commercial pharmaceutical dosage form

Background: Olaparib, Abiraterone acetate, and Pazopanib are critical anticancer agents used in the treatment of breast/ovarian, prostate, and renal cancers, respectively. Ensuring their quality through precise, cost-effective analytical techniques is vital for routine quality control (QC). Given their clinical importance, a robust method capable of quantifying these drugs individually and indicating their stability under stress is highly desirable. Methodology: High-performance thin-layer chromatography (HPTLC) methods were developed for each drug, guided by a Quality by Design (QbD) approach. A 2³ factorial design was employed to optimize three critical method parameters: mobile phase composition, chamber saturation time, and detection wavelength. Chromatographic analysis was performed on aluminum-backed silica gel plates with detection wavelengths set at 278 nm (Olaparib), 255 nm (Abiraterone acetate), and 254 nm (Pazopanib). Method validation followed ICH Q2(R2) guidelines, assessing linearity, accuracy, precision, LOD/LOQ, specificity, and robustness. Forced degradation under acidic, basic, oxidative, thermal, and photolytic conditions was evaluated to assess the stability-indicating capability. Results and Discussion: All methods exhibited strong linearity (r² &gt; 0.997), high accuracy (98–102% recovery), and precision (%RSD &lt; 2). Sensitivity was excellent, with LOD as low as 28 ng/spot for Olaparib. Under stress, degradation ranged from 5% to 20%, with distinct degradant peaks and peak purity indices of&gt; 0.995, confirming no co-elution and indicating stability. The validated methods were successfully applied to stressed marketed formulations. Conclusion: The QbD-optimized HPTLC methods are accurate, economical, and stability-indicating, making them suitable for routine QC of Olaparib, Abiraterone acetate, and Pazopanib in pharmaceutical dosage forms.

Stability indicating RP-HPLC-UV method for quantitative estimation of isomer and other organic impurities of atopic dermatitis drug abrocitinib in drug substance and pharmaceutical dosage form using quality by design (QbD) approach

Quality by design (QbD) approach thrives to achieve an assured and predicted quality product. A stability-indicating reversed phase ultrafast liquid chromatographic method was developed using the principles of QbD to quantify telaprevir (TEL) in pharmaceutical dosage form. A Box-Behnken experimental design was employed for identifying optimum chromatographic conditions by assessing the method robustness by selecting organic phase composition (%), mobile phase flow rate (mL/min) and pH of the borate buffer as the factors, to study their effect on the responses like retention time, theoretical plate count and tailing factor. Chromatographic separation was achieved on Enable-C18G (250 × 4.6 mm i.d., 5 µm) column using methanol: borate buffer of pH 9.0 (90 : 10, v/v) as mobile phase at a flow rate of 1.2 mL/min and PDA detection at 270 nm. Establishment of calibration curve yielded linearity in the range of 5-70 µg/mL along with values of accuracy and precision within the acceptance limit of mean percent recoveries between 98.9 and 100.7%. Limit of detection and limit of quantitation were found to be 1.60 and 4.75 µg/mL. Analysis of system suitability yielded high degree of method reproducibility and robustness. The developed method showed high specificity for TEL and its degradation products formed during forced degradation conditions. The developed method also demonstrated suitability for routine analysis of TEL in bulk drug and pharmaceutical dosage forms.

A UV spectrophotometric and RP-HPLC method for the Naringenin quantification in the developed solid dispersions is described. Both the methods were validated according to ICH Q2 R (1) guidelines for linearity, accuracy, precision, limit of detection, limit of quantification, specificity and robustness. HPLC was run in isocratic mode on a reversed-phase C18 column (250 × 4.6 mm internal diameter and particle size of 5 μm) with methanol:water (70:30, v/v) as the mobile phase maintaining a flow rate of 1.0 ml/min. Naringenin showed an absorbance maximum (λmax) at 288 nm which was used for the UV spectrophotometric determinations. The calibration curve of Naringenin showed linearity in the required concentration range (R2 > 0.999) by both the UV and HPLC methods. Both the methods were found to be precise and accurate with recovery range of 98–101% and relative standard deviation < 2%. Most importantly, the accuracy and precision achieved by the HPLC method, correlated closely with the UV method. Current study also involves detection and quantification of Naringenin released from its formulations by both the developed methods. This paper demonstrates the high correlation (R2 ≥ 0.98) between the UV and HPLC methods when determining the release of Naringenin from various formulations. With this established correlation, we hereby suggest that, for routine analysis, UV spectroscopy can be an economic, simple, reliable and less time-consuming alternative for expensive and time-consuming chromatographic analysis.