DAD Method Research Articles

A Validated RP-HPLC–DAD Method for Analyzing Flavonoids in Caatinga Brazilian Green Propolis from Mimosa tenuiflora Produced by Apis mellifera

A Validated RP-HPLC–DAD Method for Analyzing Flavonoids in Caatinga Brazilian Green Propolis from Mimosa tenuiflora Produced by Apis mellifera

Phytochemical Analysis and Antioxidant, Antimicrobial, and Antibiofilm Effects of a New Himalayan Lichen Placidium deosaiense Usman and Khalid Growing in Pakistan

Phytochemical composition and antimicrobial, antibiofilm, and antioxidant effects of a newly described Himalayan lichen Placidium deosaiense Usman and Khalid growing in Pakistan were investigated. HPLC–DAD methods were used for identification of secondary metabolites in acetone and methanol extracts. The total phenolics content was measured using a spectrophotometric method. The study investigated the antioxidant (DPPH-scavenging activity assay and reducing-power assay), antibacterial (minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC)), and antibiofilm (inhibition of biofilm formation and reduction in mature biofilm) activities of extracts of the lichen P. deosaiense and isolated parietin. The chemical constituents olivetol, olivetolic acid, haematommic acid, fallacinol, and parietin were identified as major compounds in the tested extracts of the lichen. Parietin was isolated from the acetone extract on a separation column. The methanol extract had higher values of TPC (21.67 mg GAE/g) than the acetone extract. Isolated parietin showed the best antioxidant activity measures, according to the DPPH-scavenging activity assay (IC50 = 51.616 μg/mL) and reducing-power assay. Although the extracts showed the best antibacterial activity (especially against Proteus mirabilis ATCC 12453), parietin demonstrated superior antibiofilm activity (especially against Staphylococcus aureus ATCC 25923). This is the first report on the phytochemical composition of the lichen Placidium deosaiense and the first description of the chemical composition of some of the 45 species of the genus Placidium. This research will pave the way for further exploration of new activities of this lichen and its metabolites, which are important for medicine and pharmacy.

A sustainable HPLC method coupled with diode array detection for versatile quantification of telmisartan, chlorthalidone and amlodipine in a fixed-dose antihypertensive formulation and dissolution studies

Cardiovascular diseases, especially hypertension, stand as prominent contributors to global mortality. Hypertension, often referred to as a silent killer syndrome, necessitates the use of multiple medications for effective control and management. A new environmentally friendly HPLC–DAD method is introduced in this study for the concurrent analysis of telmisartan (TEL), chlorthalidone (CHT) and amlodipine besylate (AML), in both pure forms and combined pharmaceutical dosage form. An isocratic elution mode was employed to achieve chromatographic separation, utilizing an Inertsil C18 column (250 × 4.6 mm, 5.0 µm) and a mobile phase mixture of acetonitrile and phosphate buffer (pH 3.0 ± 0.1) with ratio of 35:65, v/v. The separation was achieved within 10 min at a flow rate of 1.0 mL/min. The proposed method's validation was carried out following the guidelines outlined by the International Council for Harmonisation (ICH). The achieved linearity range was 1.0–140.0 μg/mL for TEL and 1.0–100.0 μg/mL for CHT and AML with quantification limits of 0.061, 0.177, and 0.313 μg/mL for TEL, CHT, and AML, respectively. The fixed combination tablet dosage form demonstrated acceptable release profile, as indicated by the in-vitro dissolution studies. The studied dissolution media were phosphate buffer pH 7.5, 0.01 N HCl, and water, utilizing a USP type II apparatus at 37 ± 0.5 °C with a stirring rate of 75 rpm. The proposed method was applied successfully for the quality assessment of Telma-ACT® Tablets with good precision and accuracy. Various tools were used for evaluating the level of greenness, including Green Analytical Procedure Index (GAPI), Analytical Greenness Metric for Sample Preparation (AGREEprep), Analytical Eco-Scale (AES), and Analytical Method Greenness Score (AMGS). These tools had confirmed the eco-friendliness of the proposed method. Additionally, the newly introduced White Analytical Chemistry (WAC), and the Blue Applicability Grade Index (BAGI) have been specifically developed to evaluate the sustainability and the applicability of the method.

Effect of salt stress on growth and phenolic compounds production in callus suspension culture of the dioecious species thyrse sorrel (Rumex thyrsiflorus Fingerh.)

The sex-dependent differences in the response to salt stress of the dioecious Rumex thyrsiflorus and the influence of different concentrations of sodium chloride (NaCl) on the biosynthesis of phenolic compounds in callus suspension cultures were evaluated. The cultures originated from callus obtained on hypocotyls isolated from male and female seedlings were cultured on Murashige and Skoog medium supplemented with 0.4 mg/L 6-benzylaminopurine and 1 mg/L 2,4-dichlorophenoxyacetic acid. The results showed that the response to salt stress depends on both the sex of the plant and the NaCl concentration in the culture medium. The analysis of the main morphometric parameters showed that callus tissue derived from the hypocotyls of female seedlings was less sensitive to salt stress than that from male seedlings, which was correlated with higher concentrations of phenolic compounds. The optimal NaCl concentration to increase the production of phenolic compounds was 129 mM for females and 43 mM for males. In the methanolic tissue extracts 22 compounds were determined using the HPLC–DAD method. In general, higher amounts of compounds were detected in the extracts from the female tissue. The following dominated: catechin (max. 213.31), cryptochlorogenic acid (max. 76.35) and epicatechin (max. 54.84) (mg/100 g DW). This comprehensive phytochemical analysis of the sex-related aspects of the response to salt stress was performed for the first time in this dioecious model species. The results revealed potential application of NaCl as the ecological friendly and inexpensive elicitor to increase the production of pharmaceutically valuable compounds and highlight the importance of dioecy in phytochemistry.

Green HPLC–DAD method for the analysis of analgesic and antihistamine drugs adulterated in herbal mixtures

This study presents a HPLC-DAD method developed for the simultaneous detection of histamine H1 antagonist drugs and analgesic compounds, including paracetamol, cetirizine hydrochloride, cinnarizine, chlorpheniramine maleate, and loratadine, in herbal formulations. This method systematically investigates critical parameters for enhancing separation and quantification using HPLC-DAD. Chemometric techniques were employed to optimize the method for detecting adulterants, with sample preparation involving liquid-phase extraction and a mobile phase for separation using HPLC-DAD. The calibration curves were linear over concentrations ranging from 10 to 40 µg/mL, with correlation coefficients exceeding 0.99. The method exhibits good accuracy, with mean recoveries ranging from 90 to 105 %, and precision (% RSD) for intraday and interday precision: 0.71 to 2.94 %. The limits of detection (LODs) and limits of quantification (LOQs) for each compound are provided. The HPLC-DAD method ensures rigorous quality control, detecting potential adulteration in 32 traditional medicine samples and emphasizing its safety and efficacy in managing allergic conditions. The proposed approach is considered environmentally friendly based on evaluations from commonly used greenness assessment tools, Green analytical chemistry metrics (ComplexGAPI, AGREE, BAGI) confirm the method's sustainability and practicality.

Comparative analysis and validation of analytical techniques for quantification active component in pharmaceuticals: Green approach

Method validation is crucial for ensuring the reliability, accuracy, and reproducibility of analytical measurements. The primary goal is to demonstrate that an analytical method is suitable for its intended purpose. In this study, we developed and validated a method for extracting and quantifying active components from commercial β-blocker tablets. We employed Ultra-Fast Liquid Chromatography-Diode Array Detector (UFLC−DAD) and spectrophotometric techniques to validate the method, assessing key parameters such as specificity/selectivity, sensitivity, linearity, and dynamic range, limit of detection, limit of quantification, accuracy, precision, and robustness.Validation using UFLC−DAD successfully covered 50 mg and 100 mg tablets of metoprolol tartrate (MET), while the spectrophotometric method was explicitly applied to analyze the 50 mg tablets due to concentration limitations. Statistical analysis, including ANOVA and Student's t-test, indicated that both validated methods are suitable for routine analysis of MET in commercial tablets.Furthermore, we evaluated the methods' environmental impact using the analytical greenness metric approach. Findings suggest that UFLC−DAD and spectrophotometric methods contribute to an environmentally friendly overall analytical process.By systematically validating both methods and considering their environmental impact, we ensure the reliability and sustainability of observed analytical processes, thereby enhancing the quality of analytical results.

Transformation profiles of the isoflavones in germinated soybean based on UPLC–DAD quantification and LC–QTOF–MS/MS confirmation

Germinated soybean is one kind of food and a medicine. In the actual process of producing a large amount of naturally germinated soybean, it is difficult to strictly control the germination process conditions. However, sprout length may be more suitable as the terminal judgment indicator for naturally germinated soybean. An UPLC–DAD method was developed and validated to explore the transformation profiles of soybean isoflavones in germinated yellow or black soybean with different sprout lengths. Moreover, an LC − QTOF−MS/MS method was used to avoid false positive results. The contents of daidzein, glycitein, and genistein almost reached their corresponding maximum values when the sprout length ranged from 1.0 cm to 1.5 cm (P < 0.05). Therefore, yellow soybean is suggested to be the processing raw material with higher contents of those isoflavones, and the optimal sprout length for germinated soybean may be in the range of 1.0–1.5 cm.

Simultaneous determination of 14 analgesics in postoperative analgesic solution by HPLC–DAD and LC–MS/MS

A green, efficient, sensitive and accurate detection method by HPLC–DAD and LC–MS/MS was developed and validated for the quantification of morphine, hydromorphone, oxycodone, ketamine tramadol, dezocine, ropivacaine, remifentanil, butorphanol, bupivacaine, droperidol, fentanyl, lornoxicam and sufentanil. The 14 mixtures were chromatographed via HPLC–DAD method which employed 0.05 mol/L potassium dihydrogen phosphate solution-acetonitrile as the mobile phase, the analytes were gradient elution on a SinoChrom ODS-BP C18 column with a total separation time of 35 min, and 14 mixtures showed a good linear relationship in the linear range. The Limit of Quantitation (LOQ) ranged from 0.10 to 20.0 µg/mL, the inter-day and intra-day precision of each analyte is within 1.1–2.0% and 0.4–1.3%, and the average absolute recovery of all compounds was above 98%. The LC–MS/MS method was used to successfully separate the 14 mixtures within 10 min which employed 0.1% formic acid-acetonitrile as the mobile phase, the analytes were gradient elution on a ACQUITY UPLC-BEH C18 column with a total separation time of 13 min, and 14 mixtures showed a good linear relationship in the linear range. The LOQ ranged from 0.005 to 0.2 ng/mL, the inter-day and intra-day precision of each analyte is within 1.2–4.1% and 0.6–3.3%, and the average absolute recovery of all compounds was above 93%. The proposed method has been successfully applied in the clinic and provides a strong technical basis for the quantitative detection of these 14 mixtures for detecting drug abuse, and for studying the stability and compatibility of analgesic solutions. The proposed methods were validated against ICH guidelines.

Simultaneous Quantification Method of Flavonoids in Jeju Native Citrus from Different Harvest Times Using a High-Performance Liquid Chromatography–Diode Array Detector (HPLC–DAD)

In Jeju-native Citrus, flavonoids are the main contributors to the various types of biological activity, such as antioxidant, antitumor, and anti-inflammatory activity. Thus, we developed simultaneous quantification methods for the analysis of ten bioactive flavonoids in Jeju Citrus fruits (Dangyuja, Gamja, Jigak, Sadugam, and Soyuja) harvested at six different time points using a high-performance liquid chromatography–diode array detector (HPLC-DAD). Separation was performed using a flow rate of 0.8 mL/min, a column temperature of 40 °C, a mobile phase buffer of 0.5% acetic acid, and a detection wavelength of 278 nm. The established analytical method showed good linearity (R2 ≥ 0.9997), precision (inter-day &lt; 0.599%, intra-day &lt; 0.055%), and accuracy (recoveries 92.30–108.80%). The HPLC–DAD method was subsequently applied to analyze flavonoids in Citrus samples. Overall, the quantification results indicated that the compositions and content of flavonoids differed for each Citrus species. The harvesting period also influenced the changes in flavonoid content within each Citrus species. The analytical results with chemometrics revealed that higher flavonoid levels in early-harvested Citrus were derived from the improved fruit size and reduced flavonoid synthesis during maturation. This study provides a practical and reliable method for the analysis of ten flavonoids that can be further utilized in the quality assessment of Jeju Citrus.

Composition of Polygonatum zanlanscianense Pamp. Steam and Leaf Phenolic Extract and Its Protective Mechanism on t-BHP-Induced Oxidative Damage of HepG2 Cells.

Plant phenolic compounds have attracted considerable attention because of their health benefits. This study aimed to investigate the composition and antioxidant activity of phenol extracts from Polygonatum zanlanscianense Pamp. steam and leaf (PPP). The FTIR, UPLC-Q-Obtrip-MS, and HPLC-DAD methods were used to analyze the composition of PPP, and 20 phenolic compounds were preliminarily identified. Among them, the contents of hyperin, astragalin, and diosmetin levels were the highest. Treatment with PPP can significantly reduce t-BHP-induced cell damage in HepG2 cells, reactive oxygen species (ROS) accumulation, and malondialdehyde (MDA) content. Meanwhile, the superoxide dismutase (SOD), catalase (CAT), glutathione (GSH), and glutathione peroxidase (GSH-Px) activities can be increased. Moreover, PPP enhanced Nrf2 expression, which was consistent with that of heme oxygenase-1 (HO-1), glutamate-cysteine ligase catalytic subunit (GCLC), and NAD(P)H quinone oxidoreductase-1 (NQO1), whereas the expression of Keap1, the Nrf2 inhibitor, was decreased. All findings indicate that PPP can serve as a natural bioactive substance for preventing oxidative stress.

White targeted chromatographic screening method of Molnupiravir and its metabolite with degradation kinetics characterization and in-silico toxicity

SARS-CoV-2 virus triggered a worldwide crisis, with world nations putting up massive efforts to halt its spread. Molnupiravir (MLN) was the first oral, direct-acting antiviral drug approved for nasopharyngeal SARS-CoV-2 infection with favorable safety and tolerability profile. This study aims at determination of MLN and N4-hydroxycytidine (NHC), its main degradation product and its main metabolite, using sensitive, simple, and green HPLC–DAD method. Moreover, under different stress conditions using NaOH, HCl, neutral, H2O2, dry heat and sun light, the method was applied for MLN assay along with kinetics degradation investigation. The linearity range for MLN and NHC were both 0.1–100 µg/mL with LOD and LOQ of 0.013 & 0.043 and 0.003 & 0.011 µg/mL, for MLN and NHC, respectively. MLN was found to be extremely vulnerable to alkali hydrolysis compared with acid and dry heat degradation. In contrast, MLN was stable under conditions of oxidative, neutral, and sunlight-induced deterioration. Acid and alkali-induced degradation followed pseudo first-order kinetics model. In addition, LC–MS-UV was used to suggest the mechanism of the stress-induced degradation route and to characterize the eluted degradation products. Toxicities of both MLN and its degradation products were evaluated using ProTox-II and they were found to be negligibly harmful. The proposed HPLC–DAD was effectively used for the analysis of MLN in commercial pharmaceutical formulations. The proposed method for MLN determination after greenness and whiteness appraisal was found to be superior compared to the reported methods for MLN analysis.

Development of a simple, eco-friendly HPLC-DAD method for tulathromycin analysis: Ensuring stability and assessing greenness

The core objective of this analytical study was to develop a simple, green, and efficient high-performance liquid chromatography-diode array detector (HPLC-DAD) method for analyzing Tulathromycin (TUL) in pure powder and injection solution without the need for complex derivatization procedures. A simple liquid mobile solution of 15 mL of phosphate buffer (pH = 7.5) and 85 mL of methyl alcohol was passed through a Hypersil BDS C18 (4.6 mm × 25 cm, 5µ) stationary phase in an isocratic mode. The flow speed was controlled at 1.0 mL/min, and the DAD detector was adjusted at 210 nm because of the extremely weak UV absorption characteristic of TUL. TUL was well separated and quantified in 10 min. The HPLC-DAD forced degradation assay was carried out in dissimilar stressful environments concerning international conference for harmonization (ICH) protocols. TUL was linearly quantified from 1000.0 to 3000.0 µg·mL−1. The moderate sensitivity of the HPLC-DAD is attributed to the aliphatic, less conjugated chemical structure of TUL. The maximum degradation ratios occurred in acidic and oxidative environments with proportions of 20.11 % and 18.87 %, respectively. Furthermore, the eco-friendly merits of the HPLC-DAD method were confirmed via the application of two greenness approaches, specifically the analytical greenness tool (AGREE) and the green analytical procedure index (GAPI). The seven green subdivisions in the GAPI pictogram and the total score of 0.72 in the AGREE pictogram illustrate the greenness appraisal of the RP-HPLC-DAD stability method. The innovative RP-HPLC–DAD method is the first sustainable stability-demonstrating assay for TUL in the co-existence of its degradation products in variable stressful environments.

Development of green micellar HPLC–DAD method for simultaneous determination of some sulbactam combinations used in COVID-19 regimen

In a new attempt to separate some sulbactam combinations by green chemical method we came up with this research in which an ecofriendly, green, sustainable and selective method was established for separation of four antibiotics, namely, cefoperazone (CFP), cefixime (CFX), ampicillin (AMP) and sulbactam (SLB). No organic solvents were used in the composition of the mobile phase as it was replaced by mixing two surfactants together, sodium dodecyl sulfate (SDS) and polyoxyethylene-23-lauryl ether (Brij-35). Effect of varying the concentrations of the two surfactants on chromatographic separation was studied. Optimum separation was maintained using a mobile phase consisting of 0.01 mol/L SDS, 0.03 mol/L Brij-35, 0.4% Tri-ethylamine (TEA) and pH of 2.8 adjusted by using 1 M ortho-phosphoric acid on reversed phase Isère C18 BDS column with temperature of 40 °C at flow rate 1 mL/min, wavelength 215 nm, and the total run time was 6 min. Validation of the proposed method has been made according International Conference of Harmonization (ICH) guidelines at linearity range of 10–200 µg mL−1 for all drugs under study, high accuracy results (recovery range 98.39–100.35%). and the variation coefficient (RSD) of the points on the calibration curve was ranged from (0.1–1.7%) indicating precise method. The LOQ was (6.09 μg mL−1) for CFP, (6.07 μg mL−1) for CFX, (3.85 μg mL−1) for AMP and (7.20 μg mL−1) for SLB. Successful applications were made on marketed dosage forms with recovery range of (100.16–102.25%) and RSD of (0.03–1.88%). The method was verified on the Green Analytical Procedure Index (GAPI) and Analytical Greenness metric approach (AGREE) and it was found to be an excellent green alternative method.

Sistem Informasi Penjualan Handpone Pada Toko Delta Cellular

Conventional trading, where sellers and buyers meet at certain locations to carry out transactions, is increasingly being abandoned with technological developments. One of the innovations that has emerged is e-commerce. E-commerce allows buying and selling transactions online, freeing from geographical limitations. In this research, attention is limited to the Design of a Mobile Phone Sales Information System at the Delta Cellular Store. The purpose of this research is to understand the information system for selling cellphones at the shop and design a web-based information system for selling cellphones. The research results show that this Mobile Sales Application makes it easier to process transaction data and stock of goods. The development methods used include old system analysis, user needs analysis, system feasibility evaluation, process design using the DAD method, data design by designing tables, and user interface design using appropriate designs. This application was built using a web-based programming language with MySQL as the database. The level of accuracy of this application in recording transactions and cellphone stock can be adjusted based on certain time periods, which helps reduce the level of human error. This application can also handle price changes and item additions by allowing users to change this information through the sales transaction form. This application is designed to be simple and uses language that is easy for users to understand, so training in its operation does not require a long time.

Optimization and validation of HPLC–DAD method for simultaneous analysis of sweeteners, preservatives, and caffeine in sugar-free beverages

In this study, the optimization and performance evaluation of a high-performance liquid chromatography with diode array detection (HPLC–DAD) method for the simultaneous determination of four sweeteners (acesulfame-potassium, saccharin, aspartame, and rebaudioside A), two preservatives (sodium benzoate and potassium sorbate), and caffeine in sugar-free drinks are presented. The separation was carried out using a gradient elution of acetonitrile and phosphate buffer (12.5 mM, pH = 3.3) on a reversed-phase column. Under the optimized conditions, the method allowed for the specific and selective simultaneous separation of all target analytes in less than 9 min. The developed HPLC–DAD method was validated and demonstrated excellent linearity (all analytical curves showed R2 ≥ 0.9995), satisfactory accuracy (recovery values ranging between 94.1 and 99.2% in real samples), and repeatability (intra- and inter-day relative standard deviations were ≤ 2.49%). Overall, 69 products available in the Hungarian market were successfully tested with the applied method.

Comparison of Bioactive Secondary Metabolites and Cytotoxicity of Extracts from Inonotus obliquus Isolates from Different Host Species

Inonotus obliquus, a wood-decaying mushroom, has been used as a health-promoting supplement and nutraceutical for centuries. It is a source of bioactive compounds accumulated in both the conks (pseudosclerotia/sclerotia) and the biomass obtained in vitro. This study aimed to qualitatively and quantitatively analyze the bioelements and selected metabolites produced in mycelial cultures obtained from different host species. The mycochemical potential of mycelial cultures isolated from pseudosclerotia grown in Betula pendula, Alnus glutinosa, and Carpinus betulus was compared. Parent cultures were obtained in two types of medium (malt extract agar substrates without and with birch wood). Experimental cultures were developed in 2 L bioreactors for 10 days. The content of bioelements was determined using FAAS and FAES methods. Organic compounds were estimated using the RP–HPLC–DAD method. The cytotoxicity of the extracts was evaluated in human keratinocytes HaCaT, human skin fibroblasts BJ, human liver cancer HepG2, human melanoma A375, and mouse melanoma B16-F10. The extracts showed the presence of bioelements: sodium, potassium, magnesium, calcium, zinc, manganese, iron, and copper; phenolic acids: p-hydroxybenzoic, caffeic, p-coumaric, and protocatechuic; sterols: lanosterol, ergosterol, ergosterol peroxide; triterpene compounds: betulin, betulinic acid, inotodiol; indole compounds: L-tryptophan, tryptamine, 5-methyltryptamine, melatonin. The content of bioactive substances in the biomass was dependent on both the origin of the host species of the fungus isolate and the type of culture medium. Based on the results of this study, mycelial cultures can be proposed as a potential source of bioactive compounds and are promising naturally derived cytotoxic agents.

A new validated RP-HPLC-UV DAD method for assay of bisoprolol fumarate and related substances in tablets

The research focus of this work was development and validation of an efficient analytical method that can be used for separation and determination of related substances of bisoprolol in finished drug product using reversed-phase HPLC with UV-DAD. In a previous systematic study of various stationary phases and elution conditions it was found that several octadecylsilane phases can be successfully employed for separation of the critical pairs of bisoprolol and its related degradation products. Namely, method development led to conclusions that satisfactory resolution and peak shapes were obtained with gradient elution with water with 0.2% perchloric acid and acetonitrile and the reversed-phase columns: Hypersil 3 BDS C18 (100 x 4 mm, 3 m); Zorbax SB C18 (150 x 4.6 mm, 3.5 m); Acquity UPLC BEH C18 (50 x 2.1 mm, 1.7 m), and Xterra MS C18 (100 x 4.6 mm, 3.5 m). Since regular quality control includes the parameter related substances, which is of great importance for the overall quality of dosage forms, this study was oriented towards widening the available validated analytical methods for determination of this parameter. In this work, the results from the validation of the method for determination of related substances of bisoprolol using Zorbax SB C18 150 x 4.6 mm, 3.5 m, are presented. Validation parameters that were tested (linearity, accuracy, precision, selectivity) confirmed that the method is suitable for its intendance and it was used for characterization of the samples from a forced degradation study of bisoprolol tablets. Keywords: Bisoprolol, RP-HPLC-UV DAD, validation, related substances

Development of a validated stability-indicating HPLC- DAD method for dasabuvir and the characterization of its degradation products using LC-QToF-MS/MS

A stability-indicating HPLC-DAD method was developed and validated for the simultaneous determination of dasabuvir and its degradation products in the pharmaceutical formulation. The proposed method utilized a Symmetry® C18 (4.6 x 75 mm, 3.5 µm) column, and the mobile phase consisted of an isocratic elution of formic acid (0.1%) and acetonitrile (55:45, v/v), at a flow of 1 mL min-1; analytes were detected at 244 nm. Dasabuvir was submitted to different stress degradation conditions, such as acidic, alkaline, neutral, thermal, oxidative and photolytic, and the structural elucidation of degradation products was performed using LC-QToF-MS/MS. The HPLC-DAD stability-indicating method was validated for selectivity, linearity, limit of detection and quantification, accuracy, precision and robustness, according to ICH guidelines. Dasabuvir produced two degradation products (DP1 and DP2) from the alkaline stress conditions, which were characterized in negative ion mode. Dasabuvir was linear in the range 9.78 to 136.92 µg mL-1, and DP and DP were linear in the range 2.9 to 20.2 µg mL-1 and 1.3 to 14.9 µg mL-1, respectively. The 1 2 recovery ranged between 99.16 and 100.86%, while precision ranged from 1.02 to 2.89%. As the method can effectively separate the dasabuvir from its degradation products and quantitate them, it may be employed as a stability-indicating method for the pharmaceutical formulation.

Fast Quantitative Determination of Principal Phenolic Anti-oxidants in Rosemary Using Ultrasound-Assisted Extraction and Chemometrics-Enhanced HPLC–DAD Method

Fast Quantitative Determination of Principal Phenolic Anti-oxidants in Rosemary Using Ultrasound-Assisted Extraction and Chemometrics-Enhanced HPLC–DAD Method

A Validated HPLC Multichannel DAD Method for the Simultaneous Determination of Amoxicillin and Doxycycline in Pharmaceutical Formulations and Wastewater Samples

The quality of marketed pharmaceutical formulations must be guaranteed to attain better remedial effects and lower toxicity. The wide exploitation of antibiotics may lead to their presence as residues in body fluids and wastewaters, potentially toxic to human health. Consequently, determining antibiotics in pharmaceutical formulations and water samples is of significant importance. This paper aims to explore the possibilities of a high-performance liquid chromatography coupled with diode array detection (HPLC-DAD) method to obtain a simple, fast, and efficient analytical tool for the simultaneous determination of antibiotics in pharmaceutical formulations and environmental samples. The method was completely validated with regard to specificity, linearity, detection and quantification limits, precision, accuracy, and robustness according to the requirements of existing guidelines, and was proven to be reliable and suitable for the envisioned application. The linearity study was conducted for the calibration curves in the range of 10–100 μg/mL. The limits of detection and quantification were found to be 0.2 and 0.7 μg/mL for amoxicillin and 0.3 and 1.0 μg/mL for doxycycline, respectively. The high recovery of drugs from their commercial pharmaceutical formulations (93%) and from wastewater samples (98%) indicated good accuracy and precision. The method is robust for small or deliberate changes to the chromatographic parameters, and it was successfully applied for the quantitative determination of amoxicillin and doxycycline in wastewater and commercial tablets. The obtained results proved that the validated method is appropriate for its intended use in the routine quality control and assay of both antibiotics studied.