ICH Recommendations Research Articles

Identification and Structural Characterization of Forced Degradation Impurities of Sacubitril by HRMS – Development and Validation of a Stability indicating RP-HPLC method

The current work identifies and characterises the stress degradation products of the anti-hypertensive drug sacubitril. The stability profile of sacubitril was evaluated under several stress settings including hydrolytic, oxidative, thermal and photolytic conditions in accordance with ICH recommendations. The degradation products of sacubitril were separated using reverse phase liquid chromatography and structural characterization was carried out using HRMS. The chromatographic separation was carried out by Reverse Phase-High Performance Liquid Chromatography (RP-HPLC) using a Fortis C18 column (150 mm X 4.6 mm; 5μm particle size) with mobile phase A (consisting of 0.1% TFA in Milli-Q water) and mobile phase B (100% Acetonitrile) in gradient mode. With a sample injection volume of 10μL, the mobile phase flow rate was 0.7 mL/min. A quadrupole time-of-flight (Q-TOF) mass spectrometer was used for the LC-MS analysis. Sacubitril is relatively stable in oxidative, thermal and photolytic conditions; however, considerable degradation was observed in acid, base and neutral hydrolysis. Three degradation products were identified and the structural characterization of these impurities was performed using the HRMS data and mass fragmentation. The proposed RP-HPLC method was robust, precise and specific in determining the impurity profile of sacubitril under different stress conditions.

Sustainable UV approaches supported by greenness and whiteness assessments for estimating a recently FDA-approved combination for managing urologic disorders: Tukey's test

Erectile dysfunction caused by a urinary tract infection and an enlarged prostate is a common problem among men nowadays. A brand-new FDA-approved combination of finasteride (FNS) and tadalafil (TDF) is currently recommended, particularly for those with prostatic problems in addition to erectile dysfunction. TDF boosts FNS's short-term efficacy in addition to treating erectile dysfunction, one of the sexual adverse effects typically associated with FNS therapy. Accordingly, this research introduces three straightforward spectrophotometric techniques for determining FNS and TDF in their pure and pharmaceutical forms concurrently, in addition to providing a dissolution study for their formulation as per the FDA’s directions. FNS and TDF have superimposed UV spectra, making direct concurrent analysis of these medications impossible. To resolve this overlap, we utilized the following UV techniques: the dual-wavelength method, the deconvoluted Fourier method, and the ratio difference method, with linear ranges of (0.50–15) µg/mL for TDF and (5–100) µg/mL for FNS. The LODs ranged from (0.12–0.15) µg/mL and (0.88–1.14) µg/mL, while the LOQs ranged from (0.37–0.46) µg/mL and (2.66–3.45) µg/mL for TDF and FNS, respectively. The proposed techniques were verified as per ICH recommendations and compared statistically to the published approach using three statistical tests. A complete green profile was provided to assess the foregoing techniques’ greenness using four metrics, along with the whiteness assessment using the RGB algorithm. The foregoing UV methods have been demonstrated to be sustainable, highly sensitive, specific, and suitable for quality assurance testing.

Different methods for resolving overlapping UV spectra of combination medicinal dose forms of ciprofloxacin and metronidazole

Four simple, specific, easy, precise and accurate spectrophotometric methods were developed for the first time to examine ciprofloxacin and metronidazole in combination, without having been separated beforehand by the developed methods. Ciprofloxacin and metronidazole were determined by utilizing advanced absorbance subtraction (AAS), spectrum subtraction, bivariate and ratio difference methods. Precision, repeatability, robustness, and accuracy were all determined to be within acceptable levels after each of these procedures underwent validation in accordance with ICH recommendations. Each method’s benefits and drawbacks are illustrated, and the proposed and reported methodologies were statistically compared.

Formulation and Evaluation of Film-Coated Alalevonadifloxacin Mesylate Tablet using Opadry®

Purpose: In this work, film-coated tablets of the antibiotic alalevonadifloxacin mesylate (AFM) were made using Opadry® yellow as the film coating material, and their different parameters, including in-vitro drug release, were assessed. Method: Direct compression was used in the production of film-coated tablets of AFM. The film-coating substance was an aqueous coating dispersion that was opadry yellow. In 900 mL of 0.1N HCL solution, the film-coated tablets’ rate of dissolution was assessed. In-vitro drug release was also evaluated. The generated film-coated tablets’ stability was assessed in accordance with ICH recommendations. Result: The coated tablets were subjected to gastrointestinal pH testing to ascertain the effectiveness of the film coating, and high-performance liquid chromatography was used to assess drug release. The coated tablets were in perfect condition. The AFM release satisfied the study’s requirement of being at least after 30 minutes, 80% was dissolved in a 0.5% HCl solution. Conclusion: According to these results, opadry yellow aqueous film coating is a simple, repeatable, and affordable method for creating stable AFM film-coated tablets without changing the properties of the medication release.

Condition optimization of eco-friendly RP-HPLC and MCR methods via Box–Behnken design and six sigma approach for detecting antibiotic residues

A precise, Eco-friendly, and highly sensitive RP-HPLC method was employed using quality-by-design principles to concurrently identify cephalexin and cefixime residues in the manufacturing machines using a hypersil BDS C18 column (250 × 4.6 mm, 5 μm) at wavelength 254 nm. The Box–Behnken design was applied to obtain the best chromatographic conditions with the fewest possible trials. Three independent factors viz organic composition, flow rate, and pH were used to assess their effects on the responses' resolution and retention time. Overlay plot and desirability functions were implemented to predict responses of the high resolution and relatively short retention time using a mobile phase composed of acidic water: acetonitrile (85:15, v/v) at pH 4.5 adjusted by phosphoric acid with a flow rate of 2.0 mL/min. The spectral overlapping of the drugs was successfully resolved by the mean centering ratio (MCR) spectra approach at 261 nm and 298 nm for cephalexin and cefixime, respectively. Good linearity results were obtained for the suggested HPLC and MCR methods over the concentration range of (0.05–10 ppm) and (5–30 ppm) with a detection limit of 0.003, 0.004, 0.26, and 0.23 ppm, and quantitation limits of 0.008, 0.013, 0.79, and 0.68 ppm for cephalexin and cefixime, respectively, with a correlation coefficient of ≥ 0.9998 and good swab recovery results of 99–99.5%. A process capability index was accomplished for chemical and micro results, illustrating that both are extremely capable. The suggested method was effectively validated using ICH recommendations.

Green and fast microwave-driven synthesis of fluorescent N,S-carbon dots for determination of two 5-nitroimidazole antimicrobials

A green, fast, and economical technique for the manufacture of N,S-CQDS has been devised based on 3-min microwave irradiation of citric acid and thiosemicarbazide that furnished highly fluorescent quantum dots (λex/em of 360/430 nm) with high water solubility and stability. Transmission electron microscopy, X-ray photoelectron spectroscopy (XPS), energy-dispersive X-ray spectroscopy (EDX), Zeta potential analysis, Fourier transform infrared, UV-Visible spectroscopy, and fluorescence spectroscopy were used to fully characterize the N,S-CQDS. The obtained N,S-CQDS are spherical shaped with average size of 13.2 ± 4.3 nm and Zeta potential of -32.46 ± 1.55 mv. The effect of UV light and pH on the fluorescence intensity of the synthesized N,S-CQDS were also investigated. The synthesized N,S-CQDS were used as a fluorescent nanoprobe for the determination of two 5-nitroimidazole antimicrobials; secnidazole (SNZ) and tinidazole (TNZ) by fluorescence turn-off via inner filter effect and static quenching mechanisms. This approach demonstrated a linear dependency of the fluorescence signal on the concentration of both drugs over the range of 15-200 μM. The limit of detection and limit of quantification were 2.18 and 6.62 μM, respectively, for SNZ, and 2.29 and 6.96 μM, respectively, for TNZ. The developed method was used to determine both drugs in pharmaceutical dosage forms and human urine samples with % recovery of 100.65 ± 1.35 for SNZ, 98.66 ± 0.47 for TNZ in pharmaceutical dosage forms and 102.24 ± 2.93 for SNZ, 98.2 ± 1.78 for TNZ in case of spiked human urine samples. The approach's selectivity was confirmed for determining both analytes in the presence of various co-existing and co-administered species. The method validity was assessed in accordance with ICH recommendations. Various metrics were used to evaluate the greenness of the proposed probe. The developed probe is environmentally-friendly due to the quick synthesis of luminous N,S-CQDS using affordable and low-energy microwave radiation, and the absence of organic solvents.

A Validated HPTLC Method for Quantification of Linoleic Acid and Beta-Sitosterol in <i>Solanum nigrum</i> Extract

Solanum nigrum, a member of the Solanaceae family, has long been used as both food and medicine. This plant has been recognized for its immunomodulatory, anti-ulcerogenic, hepatoprotective, and anti-hyperlipidemic effects. It contains various chemical components such as alkaloids, coumarins, flavonoids, tannins, saponins, proteins, carbohydrates, glycosides, and phytosterols. The objective of the current work was to create a repeatable and consistent HPTLC-validated technique for concurrently detecting Beta-Sitosterol (BTS) and Linoleic Acid (LIA) in S. nigrum in accordance with ICH recommendations. The HPTLC process involved using silica gel 60 F254 TLC plates as the stationary phase and a mobile phase consisting of n-hexane, ethyl acetate, and formic acid in a ratio of 6:4:0.2 (v/v/v). The resulting bands were uniformly visualized after derivatization with anisaldehyde sulfuric acid. The calibration curves of standard LIA and BTS exhibited satisfactory linearity within concentration ranges of 400-1150 ng/spot and 350-1200 ng/spot, respectively, with correlation coefficients (r2) of 0.99153 and 0.99389. LOD and LOQ were determined as 91.96 and 118.14 ng/spot for LIA, and 278.66 and 314.62 ng/spot for BTS, respectively. In conclusion, this HPTLC method proved to be efficient, simple, precise, and reproducible for measuring LIA and BTS in S. nigrum.

Application of Box-Behnken Design and Response Surface Methodology for Selecting the Optimum RP-HPLC Conditions for the Simultaneous Determination of Paracetamol and Diclofenac Sodium Along With Three Skeletal Muscle Relaxants in Three Different Pharmaceutical Dosage Forms.

An isocratic HPLC method has been developed and validated for estimating paracetamol and diclofenac sodium simultaneously with three skeletal muscle relaxants, namely, methocarbamol, tizanidine hydrochloride and chlorzoxazone in their pure standard mixtures and in different multi-component dosage forms in a single chromatographic run. HPLC separation was achieved on a C18 Inertsil ODS-3V 5μm column (250 × 4.6mm) using a mobile phase mixture containing acetonitrile and 25mM phosphate buffer (pH7.4 adjusted with NaOH) in the proportion of (39.7:60.3, v/v) pumped at 1.2mL/min flow rate with UV detection at 220nm. An experimental design was used by applying Plackett-Burman design for screening the most critical predictors affecting the chromatographic separation and Box-Behnken design for optimizing the selected predictors and creating the response surface between the selected predictors and the interested responses. ICH recommendations were applied for validating the proposed method with regard to linearity, precision, accuracy, selectivity, limits of detection and quantitation, and robustness. There are many applications for the optimized method that can be applied for routine estimation of the cited drugs in laboratories of quality control and pharmaceutical industries to save money and time and to reduce material waste and effort.

Development and Validation by UV Spectrophotometric Method for Simultaneous Estimation of Gliclazide and Quercetin in Formulation

Abstract: A simple, quick, accurate, precise, and cost-effective spectrophotometric approach for simultaneous estimation of Gliclazide and Quercetin has been developed by Simultaneous equation method. Gliclazide and Quercetin shows absorbance maximum at 226 and 255 nm respectively, as a result absorbance was measured at these wavelengths for the estimation of Gliclazide and Quercetin. In concentration ranges from 2-18 g/ml, and 2-22 g/ml for Gliclazide and Quercetin respectively followed Beer-Lambert's law. The method has been developed and validated in accordance with ICH recommendations and can be used for estimation of Gliclazide and Quercetin in formulations.

Selective Separation and Mass Spectral Characterization of Degradants in Viloxazine by LC-MS/MS

This research describes a novel technique for the selective separation of degradants from API employing HPLC and online coupling of a triple quadrupole mass analyzer and PDA detector with a SCIEX QTRAP 5500 mass spectrometer. Chromatography was used to separate all degradants on the column Agilent eclipse XDB (150 mm x 4.6 mm, 3.5 μ) with mobile phase ACN: 0.1% TEA (40:60) %v/v. The highest absorption was found to occur at 220 nm, which allows for simultaneous detection without being impacted by the placebo matrix. According to the general ICH recommendations, the suggested RP-HPLC method was accepted. All of the metrics- specificity, linearity, LoD, LoQ, accuracy, precision and robustness of validation were deemed sufficient. The proposed method exhibits strong correlation and great linearity over the range of (12.5–75 μg/mL). The accuracy trials produced consistent recoveries (95–105%), while the precision experiments' percent RSD was less than 2%. The intrinsic stability of the drug molecules in the current formulation could be ascertained by conducting forced degradation studies to assess the degradation products produced under various stress settings. The degradants produced were well separated and further characterized by MS/MS studies. The newly devised approach was demonstrated to be stable and sensitive to all degradants during validation tests. Validation trials demonstrate that the newly developed method was also accurate, precise, resilient, selective, and linear within the necessary operating range.

UV Spectrophotometric and UPLC assay methods for the determination of Palbociclib in bulk and Tablet Dosage Form

The goal of this study was to create a sensitive and reliable UV-spectrophotometric and reverse phase liquid chromatographic method for determining Palbociclib in bulk and pharmaceutical dosage forms. The current UV approach uses methanol and phosphate buffer as a solvent to evaluate absorption at a maximum wavelength of 220nm. Waters Acquity BEH C18 (2.1 x 50mm, 1.7µm) UPLC column with Empower 2 was used in the UPLC procedure. The mobile phase was made up of 50:50 (v/v) methanol and phosphate buffer.The drug obeyed Beer Lambert's law in the UV technique in the concentration range of 1-70µg/ml with a regression coefficient of 0.999. The linearity level in the UPLC technique was 1-100µg/ml. In the validation investigation, the developed UV and UPLC procedures were successful. For the UPLC and UV methods, the percentage purity of the marketed dosage form of Palbociclib was found to be 98.76 percent and 98.84 percent, respectively. The percent RSD of all validation parameters investigated, as suggested by the ICH recommendations, was determined to be less than 2 in both techniques. The two methods established for determining Palbociclib in bulk and tablet doses were found to be accurate, simple, and validated.

A Validated Sensitive Stability Indicating HPLC Method for the Determination of Etoricoxib in Bulk and Formulation

Aim: The current research aims to create a high-resolution and validated liquid chromatographic method used for quantitative detection of etoricoxib within dosage forms that is both easy to use and reliable in terms of accuracy, sensitivity, and reproducibility. Materials and Methods: Analysis was carried out at isocratic conditions at flow rate of 0.7 mL/min using a mobile phase consisting of 40 parts methanol to 60 parts water (0.1% OPA) at pH 3.2. The eluents were tested using UV detection at 236 nm. Results: Etoricoxib had clearly separated peaks with a retention duration of 4.347 minutes in the optimized settings. The concentration range used to generate the calibration curve was 10 to 60 μg/mL. LoD was 0.0779 μg/mL, and upper LoQ was 0.23 μg/mL. The approach has been effective in separating a known quantity of etoricoxib, and the percentage of degradation was shown to be very low across all stress settings. Conclusion: Etoricoxib in bulk drug and commercial formulations can be identified and quantified using the proposed method, which has been validated in accordance with ICH recommendations.

Formulation and Evaluation of Punica granatum L Seed Oil-loaded Cosmetic Cream: Its In-vitro Antioxidant Activity

Cosmetic creams for the antioxidant activity of Punica granutum L seed oil (PSO) were formulated in different concentrations of oil in F1 to F6 formulations of cosmetic creams and their physicochemical properties were evaluated. There are several different pharmacological actions attributed to P. granutum L seed oil. The prepared cream was evaluated its properties like pH, resistance to flow, spreadability size, and surface charge value, and ex-vivo release study were performed. All results are found within the limit and it complies with the specification as per requirements. Good in-vitro DPPH scavenging activity was observed from the optimized batches throughout the preparation. To determine the stability profile, cosmetic creams containing PSO (Batches F1 through F6) were exposed to a three-month accelerated stability study in accordance with ICH recommendations. From this study it was concluded that prepared batches used for various treatments of skin disorders.

A sustainable approach for the degradation kinetics study and stability assay of the SARS-CoV-2 oral antiviral drug Molnupiravir

Molnupiravir (MPV) is the first direct-acting oral antiviral drug that effectively decreases nasopharyngeal infections with SARS-CoV-2 virus. The stability of MPV was tested by subjecting the drug to various stress conditions. The drug is liable to oxidative, acidic, and alkaline degradation and showed significant stability against thermal degradation. Mass spectrometry identified the degradation products and guided suggestion of the degradation patterns. Interestingly, while inspecting the UV-absorption spectra, we observed no absorbance at 270 nm for the products of the three degradation pathways (c.f. intact MPV). Direct spectrophotometry seemed a solution that perfectly fit the purpose of the stability assay method in our case. It avoids sophisticated instrumentation and complex mathematical data manipulation. The method determined MPV accurately (100.32% ± 1.62) and selectively (99.49% ± 1.63) within the linear range of 1.50 × 10–5–4.0 × 10–4 M and down to a detection limit of 0.48 × 10–5 M. The proposed method is simple and does not require any preliminary separation or derivatization steps. The procedure proved its validity as per the ICH recommendations. The specificity was assessed in the presence of up to 90% degradation products. The study evaluated the greenness profile of the proposed analytical procedure using the National Environmental Methods Index (NEMI), the Analytical Eco-Scale, and the Green Analytical Procedure Index (GAPI). The three metrics unanimously agreed that the developed procedure results in a greener profile than the reported method. The method investigated the degradation reactions' kinetics and evaluated the reaction order, rate constant, and half-life time for each degradation process.

Determination of Irinotecan enantiomer impurity in Irinotecan Hydrochloride API by using reverse-phase liquid chromatography

To evaluate and quantify Irinotecan (R-enantiomer) / Irinotecan related compound D in Irinotecan hydrochloride trihydrate API, a high stereo-specific liquid chromatography technique was developed and validated. The partition was accomplished on ChiralpakIC-3 (150 x 4.6 mm 3µm) through a mobile fragment comprising 0.1 % v/v Formic acid in water and acetonitrile with 1mL/min, 250C, 20µL, 50C and 370 nmas flow rate, column temparture, injection volume, sample cooler temperature and detection wavelength. At 8.903 and 9.75 min, the retention time of Irinotecan (R-enantiomer) and Irinotecan (S-enantiomer) was determined. The resolution between Irinotecan (R-enantiomer) and Irinotecan (S-enantiomer) was found to be 2.4. The impurity acceptance limit is 0.2 %. The established method's precision, accuracy, sensitivity, linearity, specificity, and ruggedness were all verified in accordance with ICH recommendations. The qualifying sample LOQ was found to be 0.4 g/ml, while the minimal amount of sample needed for LOD detection was found to be 0.12 g/ml. The proposed reversed-phase method has been sophisticated and authenticated in accordance with ICH criteria and is capable of quantifying irinotecan enantiomer in irinotecan hydrochloride trihydrate API at trace level concentration. The specificity, linearity, and accuracy of the approach were used to guarantee its efficacy; as a result, it is appropriate for the task at hand, may be used successfully for routine laboratory analysis, and can be utilised for quality control.
 Keywords: Irinotecan, liquid chromatography technique, enantiomer impurity

Univariate versus multivariate spectrophotometric data analysis of triamterene and xipamide; a quantitative and qualitative greenly profiled comparative study

Triamterene (TRI) and xipamide (XIP) mixture is used as a binary medication of antihypertension which is considered as a major cause of premature death worldwide. The purpose of this research is the quantitative and qualitative analysis of this binary mixture by green univariate and multivariate spectrophotometric methods. Univariate methods were zero order absorption spectra method (D0) and Fourier self-deconvolution (FSD), as TRI was directly determined by D0 at 367.0 nm in the range (2.00–10.00 µg/mL), where XIP show no interference. While XIP was determined by FSD at 261.0 nm in the range (2.00–8.00 µg/mL), where TRI show zero crossing. Multivariate methods were Partial Least Squares, Principal Component Regression, Artificial Neural Networks, and Multivariate Curve Resolution-Alternating Least Squares. A training set of 25 mixtures with different quantities of the tested components was used to construct and evaluate them, 3 latent variables were displayed using an experimental design. A set of 18 synthetic mixtures with concentrations ranging from (3.00–7.00 µg/mL) for TRI and (2.00–6.00 µg/mL) for XIP, were used to construct the calibration models. A collection of seven synthetic mixtures with various quantities was applied to build the validation models. All the proposed approaches quantitative analyses were evaluated using recoveries as a percentage, root mean square error of prediction, and standard error of prediction. Strong multivariate statistical tools were presented by these models, and they were used to analyze the combined dosage form available on the Egyptian market. The proposed techniques were evaluated in accordance with ICH recommendations, where they are capable of overcoming challenges including spectral overlaps and collinearity. When the suggested approaches and the published one were statistically compared, there was no discernible difference between them. The green analytical method index and eco-scale tools were applied for assessment of the established models greenness. The suggested techniques can be used in product testing laboratories for standard pharmaceutical analysis of the substances being studied.

Development of Analytical Technique for Dichlofenac Sodium and Ibuprofen Utilizing UV Visible Spectroscopy and AUC Method

The purpose of this work was to create an analytical method combining UV visible spectroscopy and the AUC method for the simultaneous determination of diclofenac sodium and ibuprofen in their combination dosage form. The medications’ UV spectra were captured between 200 and 400 nm, and the calibration curves were built using AUC values for diclofenac sodium and ibuprofen at 274 nm and 222 nm, respectively. The developed method’s linearity, precision, accuracy, limit of detection, and limit of quantification were all validated in accordance with ICH recommendations. The findings demonstrated that the approach was linear, exact, accurate, and sensitive for the measurement of ibuprofen and diclofenac sodium. Without any excipient interference, the suggested approach was effectively used to determine the two medicines in their combination dose form. The created approach is straightforward, accurate, and cost-effective, and it may be utilised for regular analysis of ibuprofen and diclofenac sodium in their combination dose form.

Ecofriendly stability-indicating UHPLC-PDA method for determination of the new influenza antiviral prodrug Baloxavir Marboxil; application to degradation kinetic studies and structure elucidation of the major degradation products using LC-MS

Implementation of green chemistry principles in analytical procedures is of great importance. Green analytical chemistry minimizes the dangerous impact on human beings and the environment that is the primary objective nowadays. In the proposed work, a well-developed, fully validated and ecofriendly stability indicating UHPLC coupled with photodiode array detector (PDA) method was established for the determination of Baloxavir Marboxil (BXM), a novel antiviral drug for the treatment of influenza A and B, in raw materials and pharmaceutical formulations. The method employed Inertsil ODS-3 C18 column to separate BXM from its forced degradation products using a mobile phase composed of 0.1% citric acid (pH 2.5 ± 0.05)-methanol (50:50, v/v), at a flow rate of 0.8 mL/min in an isocratic separation mode. BXM was monitored using diode array detector at 260.0 nm. BXM stability was inspected using various stress conditions as per ICH recommendations. The impact of acid, base hydrolysis, and oxidative stress conditions was greater on BXM in comparison to photo and thermal hydrolysis. The formed degradation products were injected into the mass spectrometer (MS) to elucidate their structures. The suggested pathways for degradation mechanisms were presented. Moreover, the proposed method was effectively utilized to study the degradation kinetics of acid, base and oxidative degradation of BXM. Additionally, four assessment tools were employed to evaluate the greenness of the developed method, namely; the National Environmental Method Index, the Analytical Eco-scale, Green Analytical Procedure tool, and the Analytical Greenness metric. Moreover, a comparative study of the method's greenness was conducted with the reported HPLC method and the results confirming that the suggested method is superior regarding the guidelines and principles of green chemistry. In addition to, having higher qualitative and quantitative scores in comparison to the reported method, and providing reliable and precise results of the method greenness.

Utility of Quality by Design approach in RP-HPLC method development for quantification of lamivudine and effavirenz in combination formulation

Objective: For the measurement of lamivudine(LAM) and effavirenz (EVZ) in combination formulation, a uncomplicated and reliable liquid chromatographic approach has been proposed.
 Material and Method: Multivariate optimization of the RP-HPLC method, experimental conditions were accomplished using the design of experiments (DoE). The crucial method parameters were determined by a risk assessment. The mathematical models were created using three independent variables: percentage of acetonitrile, percentage of methanol, and buffer pH. The impacts of these independent elements were thoroughly investigated using the central composite design (CCD), which was utilized to analyze the response surface methodology. 
 Result and Discussion: The LAM and EVZ retention time and resolution were both concurrently optimized using the desirability function. Acetonitrile, methanol, and phosphate buffer (pH 7.0) in the proportions of 40:20:40 v/v each were used in the optimized and anticipated data from the contour diagram, with detection occurring at a wavelength of 215 nm. Baseline separation of both pharmaceuticals with high resolution and a run time of under 6 minutes was accomplished under these ideal conditions. The validated test parameters followed ICH recommendations. As a consequence, the findings demonstrated that the Quality by Design methodology could be successfully used to optimize the RP-HPLC technique for the concurrent quantification of LAM and EVZ.

Development and validation of ultraviolet spectrophotometric method for Saquinavir estimation in bulk and dosage form

We developed and validated a simple, rapid, sensitive, precise, and reproducible specific UV spectrophotometric technique for determining Saquinavir in bulk medication and pharmaceutical dosage form. Methods: As per ICH recommendations, a simple double-beam UV spectrophotometric technique has been designed and validated with several criteria, such as linearity, precision, repeatability, the limit of detection (LOD), limit of quantification (LOQ), and accuracy. Results: UV-visible spectrophotometric approach, measurement of absorption at the maximum wavelength in 10 mL methanol, and volume made with water solvent system as reference Saquinavir was discovered at 239 nm. The medicine followed Beer's rule and showed a good correlation. Beer's law was followed for Saquinavir in the 1-5µg/ml concentration range, with a correlation coefficient of 0.999. Saquinavir's LOD and LOQ were determined to be 0.56 μg/ml and 2.78 μg/ml, respectively. The proposed method is simple, precise, accurate, and reproducible, and it may be utilized for routine analysis of Saquinavir in bulk and tablet dosage form.