An ultra-performance liquid chromatography (UPLC) technique with photodiode array-mass spectrometry (PDA-MS) detectors has been developed and validated for the determination of glycopyrronium bromide (GB), formoterol fumarate dihydrate (FFD) and their related substances in pressurized meter dose inhalers. This marks the first time the method has been implemented. Chromatographic analyses have been carried out on a UPLC system with a single quadrupole mass spectrometer (QDa) and a PDA detector. A C18 column has been used prior to the injection valve. The column temperature was set to 40°C. For each analysis, 1 μL of sample was injected at a flow rate of 0.1 mL/minute. The mobile phases used were A (0.5% formic acid in methanol) and B (0.5% formic acid in water). The total run time of the method was 30 minutes. The PDA detector was employed at 215 nm while the mass detector was set to extracted ion chromatography (XIC) mode. The range for detecting impurities of GB was found to be between 0.28 and 0.30 ng/mL, with a limits of 0.92-1.01 ng/mL for quantification. GB and FFD demonstrated linear accuracy. We studied the accuracy of related substances within the limit of quantification value range of corresponding impurities and up to 400% of the limit value. The method we used was linear, rapid, reproducible and accurate. Our proposed method is fast, linear, accurate and reproducible. The UPLC-MS method, which utilized QDa with XIC mode, was developed to ascertain the related substances and assay of GB and FFD.

Currently, many researchers strive to create eco-conscious methods for drug separation to avoid hazardous solvents. In line with this objective, this study focuses on developing two green chromatographic methods, TLC and UPLC, to determine the mixture of carvedilol (CAR) and ivabradine (IVA) used for controlling stable angina symptoms, along with their oxidative degradates. The TLC method employed toluene: methanol: ammonia (8:2:0.2, by volume) as a mobile phase and silica gel plates as the stationary phase, with detection at 270.0 nm using concentration ranges of 0.5-8.0 μg/band for each drug. The optimized UPLC method utilized a mobile phase of ethanol: 0.1% triethylamine adjusted by acetic acid to pH 6.4 (40:60 v/v) on an Agilent Poroshell C18 (150 mm × 3 mm) column, with a 0.3 mL/min flow rate at 30°C and fluorimetric detection at 285.0 nm excitation and 330.0 nm emission, with linearity of 0.2-12.0 μg/mL for both drugs. Green assessment of the proposed methods with 82 and 78 green Eco-scores, AGREE score values of 0.77 and 0.72 for UPLC and TLC methods, respectively were performed. Moreover, statistical comparison of the proposed methods with reported and official methods was conducted. Additionally, structure elucidation, molecular docking and ADMET assessment of the degradates were successfully performed.

In this study, ultra high-performance liquid chromatography and quadrupole time-of-flight tandem mass spectrometry was employed to characterize and identify the chemical constituents in the total extract of Tinosporae Radix. Chromatographic separation of the total extract was carried out on a Waters chromatographic column (100 mm × 2.1 mm, 1.7 μm) with a gradient elution program using a mobile phase system consisting of 0.1% aqueous formic acid (v/v, solvent A) and acetonitrile (solvent B). Mass spectrometric detection was conducted in both positive and negative ion modes. A total of 43 chemical components were successfully identified, including 29 alkaloids and 14 terpenoids. Furthermore, the mass fragmentation behaviors and characteristic fragment ions of protoberberine isoquinoline alkaloids, aporphine alkaloids and diterpenes were systematically analyzed. This study provides reliable reference data for the identification and compositional analysis of chemical constituents in Tinosporae Radix and its homologous medicinal herbs. The findings lay a solid foundation for exploring the pharmacodynamic material basis and chemical profiling of this traditional Chinese medicinal herb, supporting its quality evaluation and further pharmacological mechanism research.

New ecological, sustainability and practicality analytical methodologies became constant in the modern pharmaceutical analysis. Likewise, a simple, rapid, stability-indicating high-performance liquid chromatography (HPLC) method was developed and validated for simultaneous determination of lamivudine (3TC), zidovudine (ZDV) and nevirapine (NVP) in pharmaceutical formulations. The HPLC method was carried out on a monolithic C18 column (100 mm × 4.6 mm i.d.), maintained at 30°C. The mobile-phase consisted of ammonium acetate (pH 4.5; 10 mM)-methanol-acetonitrile (65:28:7, v/v/v), run at a flow rate of 2.0 mL/min, using photodiode array (PDA) detection at 270 nm. Validation parameters such as the specificity, linearity, precision, accuracy and robustness were evaluated according to the International Council for Harmonisation (ICH) guidelines. From the results obtained, the developed HPLC method was successfully applied for the simultaneous quantitative analysis of 3TC, ZDV and NVP in tablet dosage forms. The chromatographic separation was obtained in less than 3 min and it was suitable for high-throughput analysis. In addition, the greenness assessment, the holistic overall evaluation and the practicality check using the novel Analytical GREEnness Metric Approach and Software (AGREE) calculator, the RGB-12 model and Blue Applicability Grade Index (BAGI), respectively, were performed. The results demonstrated the environmental friendliness (AGREE), practicality (BAGI) and holistic sustainability profile (RGB-12 model) of the proposed method.

The present study aimed to develop and validate a simple, rapid and stability-indicating high-performance liquid chromatographic (HPLC) method for the simultaneous quantification of nystatin (Nys), neomycin sulfate (NS), triamcinolone acetonide (TAC) and gramicidin D (Gr) in bulk drug samples and transferosomal gel formulations. Chromatographic separation was achieved on an Agilent Eclipse C18 (250 × 4.6 mm, 5 μm) using a mobile consisting of methanol and 0.1% orthophosphoric acid (30:70, v/v). The flow rate was maintained at 1.0 mL/min, and detection was carried out at 260 nm using a Photo Diode Analyzer (PDA) detector. Under the optimized chromatographic conditions, all four analytes were eluted with symmetrical peaks, satisfactory resolution and acceptable theoretical plate counts within a run time of 12 min. The method exhibited excellent linearity over concentration ranges of 25-150 μg/mL for TAC, 62.5-375 μg/mL for Nys, 250-1,500 μg/mL for NS and 6.25-37.5 μg/mL for Gr, with correlation coefficients greater than 0.999 for all analytes. The developed method was validated according to International Council for Harmonization guidelines and demonstrated acceptable accuracy, with percentage recoveries ranging from 98 to 102% and precision, with percentage relative standard deviation values below 2%. Forced degradation studies were performed under acidic, alkaline, oxidative, reductive, thermal, photolytic, peroxide and hydrolytic conditions. The analytes exhibited degradation ranging from ~11 to 12.1%, confirming the stability-indicating capability of the method. The proposed method is simple, accurate, precise, robust and suitable for the routine quantification of Nys, NS, TAC and Gr in bulk drug substances, transferosomal gel formulations and marketed pharmaceutical products.

Erectile dysfunction (ED) is defined as the persistent inability to achieve or maintain an erection sufficient for satisfactory performance for a duration of >3 months. However, chewable tadalafil formulations for treating ED and corresponding dissolution analysis methods for their quantification are lacking. Therefore, this study aims to investigate Quality by Design considerations for developing a reversed-phase high-performance liquid chromatography method. The test method was designed to perform a quantitative dissolution analysis of tadalafil chewable tablets to enhance medication compliance. This method required an Agilent C8 column (4.6 × 5.0 cm, 5 μm) as the stationary phase and a mobile phase comprising a 1:1 methanol-water mixture. The column temperature and detection wavelength were maintained at 40°C and 225 nm, respectively. The development and validation of the dissolution method resulted in a relative standard deviation of 0.03% for system suitability, assessed in a test solution containing a solubilizer. The test method demonstrated specificity by clearly distinguishing the active pharmaceutical ingredient from excipients. Tadalafil accuracy was confirmed with an average recovery rate of 100.22%, and the precision showed a relative standard deviation of 0.35%. Excellent linearity (R2 = 1.000) was observed over the concentration range of 1.5 μg/mL to 9.0 μg/mL. Additionally, the detection and quantification limits for tadalafil were 0.003 μg/mL and 0.008 μg/mL, respectively. The developed method facilitated rapid quantification of tadalafil in chewable tablets, completing each run within 5 min and reducing analysis time by ~60% compared to conventional methods. Additionally, reagent consumption and operational costs were reduced by ~45%, improving overall efficiency. These findings validate the suitability of the method for reliable industrial quality control of tadalafil chewable tablets and new formulations.

The primary objective of this study was to develop and validate a liquid chromatography-atmospheric pressure chemical ionization-tandem mass spectrometric (LC-APCI-MS/MS) method for the determination of trace levels of N-nitroso vonoprazan (NVNP) in vonoprazan active pharmaceutical ingredient (API) and pharmaceutical products. Chromatographic separation was achieved on an Acquity® Premier HSS T3 column under isocratic conditions using 0.1% formic acid in water and acetonitrile as the mobile phase. Detection was performed in positive ion mode using multiple reaction monitoring. The method was validated according to ICH Q2(R2) guidelines for selectivity, linearity, accuracy, precision, robustness, matrix effect, limit of detection (LOD), and limit of quantification (LOQ). The method was linear over the concentration range of 0.24-50.0 ng/mL (R2 = 0.9971), with recoveries of 85.6%-99.2% and relative standard deviation values below 6.1%. The validated LOD and LOQ were 0.10 and 0.24 ng/mL, respectively; the latter corresponds to 0.24 μg/g (240 ppb) in the pharmaceutical product. Matrix effect evaluation showed 91.6% response (8.4% ion suppression), indicating minimal matrix interference. The validated LC-APCI-MS/MS method was successfully applied to both API and finished products and is suitable for routine quality control, stability studies, nitrosamine risk assessment, and regulatory monitoring of NVNP.

Scutellariae barbatae Herba, the dried whole plant of S.barbatae D. Don, has been documented in the Chinese Pharmacopoeia since 1985. The chemical constituents of S. barbatae Herba are complex. And comprehensive analysis of the chemical components in S. barbatae Herba is still scarcely studied. In this study, ultra-high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry was used in both negative and positive ion modes to comprehensively analyze chemical constituents of S. barbatae Herba. Moreover, the fragmentation pathways of compounds were explained in brief. According to tandem mass spectrometry fragmentation data and previous reports, a total of 39 compounds were identified, including 19 flavonoids, 4 neo-clerodane diterpenoids, 1 neo-clerodane diterpenoid alkaloid, 3 triterpenes, 5 fatty acids, 1 lignin, 1 monoterpene phenol, and 4 other constituents. More fragments of information are formed in the compounds, and the fragmentation pathways were explained simply. The developed analytical method is simple, reliable, and effective, providing comprehensive information on the metabolite profile of S. barbatae Herba, which may enhance its quality control and further utilization.

The simultaneous determination of Abemaciclib and Olaparib in in-houseprepared tablets was accomplished by the development and validation of a novel and rapid and sensitive reverse-phase ultra-performance liquid chromatography (RP-UPLC) technique. Using a Waters X-Bridge C18 (4.6 × 100 mm, 3.5 μm) column and a mobile phase made up of 0.1% orthophosphoric acid in water, acetonitrile and methanol (70:20:10% v/v/v) in an isocratic elution mode at a flow rate of 1 mL/min, the chromatographic separation was accomplished. High sensitivity and selectivity were made possible by performing the detection at 219 nm. Following ICH Q2(R1) criteria, the technique was validated and showed high linearity (R2 > 0.999) for both drugs over the concentration range of 10-60 μg/mL. Within acceptable boundaries (<2% Relative Standard deviation (RSD)), both intra-day and inter-day precision and accuracy (recovery percentage) were proven. Forced degradation studies confirmed the method's stability-indicating capability, Stability studies indicated significant degradation under acid, alkali, peroxide and thermal degradation conditions. The degradation products were characterized by LC-MS technique, and possible degradation pathways were proposed. Additionally, in vitro pharmacokinetic evaluation, including dissolution and drug release kinetics, was performed to assess formulation performance. The developed method proved simple, precise and robust, making it suitable for routine quality control, stability study and pharmacokinetic studies of Abemaciclib and Olaparib co-formulated tablets.

A new cooled side arm tube headspace single drop microextraction (CSAT-HS-SDME) method was successfully applied to the extraction of Carum copticum volatile components. The aim of this research is to design and fabricate a new device for simultaneous cooling of the drop and evaporated sample. The innovated technique was evaluated by the extraction and Gas chromatography-mass spectrometer (GC-MS) determination of p-cymene, γ-terpinene and thymol in C. copticum samples. Optimization of the effective extraction parameters was carried out by the simplex method. Under the optimized conditions (i.e. cold and heat temperature, -10°C and 70°C; water addition, 20 μL and extraction time, 20 min), the relative standard deviations of peak areas obtained from five replications were between 3.62% and 4.69%. Compared to other similar equipment for cooling the extraction phase and simultaneously heating the sample, this device has advantages such as simplicity, very low cost, ease of use, portability and no need for additional equipment such as a liquid carbon dioxide cylinder or heater-stirrer. The proposed method was successfully applied to the extraction and identification of volatile components of some C. copticum samples.

The genus Tabebuia is widely used in traditional medicine of tropical and subtropical countries. We recently reported the antitumor activity toward several human cell lines of the ethyl acetate extract of the leaves of Tabebuia hypoleuca, a species endemic to Cuba, together with the high content of triterpenic acids. This work describes the development and the validation of a reversed-phase high-performance liquid chromatography with diode array detection method for the simultaneous quantitation of the contents of oleanolic acid (OA) and ursolic acid (UA). The method was carried out on a C18 column, using methanol and water as mobile phase (ratio 95:5 v/v), at the flow rate of 0.4 mL/min, recorded at λ = 210 nm. The retention times were 16.371 ± 0.045 and 16.943 ± 0.052 min for OA and UA, respectively, with a resolution between peaks of 1.29. The average contents of OA and UA within three sets of the ethyl acetate extract, considered as an active pharmaceutical ingredient (API) were 102.93 and 244.28 mg/g, respectively. The linearity of the response was found to be 10-700 μg/mL for OA and its equation was y = 0.2946x - 2.0432 (r2 = 0.9924). The linearity range for UA was 5-1000 μg/mL and its equation was y = 0.3284x - 17.508 (r2 = 0.9865). The limit of detection and limit of quantification of these constituents were 6.50 and 19.81 μg/mL for OA, 0.75 and 1.98 μg/mL for UA. OA and UA may be used as markers for the quality assessment of formulations containing this API. The developed method is accurate, specific, precise, reproducible and may be applied as a simple and rapid quality-control procedure for medicinal plant extracts with similar triterpene patterns.

In recent years, the government of India has banned single-use plastic, as it is more customer-friendly but poses major disadvantages, such as leaching. As impurities can be present in any source, from the raw material to the end product, and so on, these are scientifically termed extractables and leachables. To identify these types of impurities from the pharmaceutical formulation, predominantly parenteral and ophthalmic or any other products available in plastic packaging, a rapid, selective, and simultaneous method for four different analytes has been developed with the help of gas chromatography-mass spectrometry and validated completely according to International Council for Harmonization Q2(R1) guidelines. Selected extractables and leachables, namely cyclohexanone, 2,4,7,9-tetramethyl-5-decyne-4,7-diol, bis(2-ethylhexyl) sulfosuccinate sodium salt, and benzophenone, were detected by mass spectrometry and identified using the NIST17 library of the instrument. The developed method was within the range.

Atrazine (ATZ) is a persistent herbicide found in aquatic environments worldwide, and the evaluation and monitoring of potential contamination by this pesticide are of utmost importance. A properly validated and well-established analytical method is important to detect ATZ in the environment. In this work, a dispersive liquid-liquid microsyringe extraction method was developed for the pre-concentration of ATZ in lake water samples for subsequent determination in a gas chromatograph with an electron capture detection (GC-ECD) system. The method showed excellent linearity in the range of 2 μg L-1 to 30 μg L-1, with correlation coefficient (R) values ≥0.995. In addition, the method is selective, and the sample matrix does not interfere with the determinations (≤20%). The limits of detection and limits of quantification were 0.6 μg L-1 and 2 μg L-1, respectively. The method is accurate, with recoveries ranging from 100% to 114%, and precise (13%). The extraction method associated with GC-ECD is reliable, sensitive, simple, fast, reproducible, environmentally friendly and economical. The method was applied to water samples from the Municipal Lake of the city of Cascavel (PR, Brazil) and no ATZ was detected in any of the samples.

There are nine different varieties of Angelica sylvestris plants found in Asia, America, and Europe, and all of them, especially their roots, have medicinal properties. It is important to determine the surface properties of medicinal A. sylvestris that can be easily found in nature. In this study, it was aimed to investigate the surface characteristics of the root, stem, and flower parts of the A. sylvestris. For this purpose, gas chromatography columns were prepared by separating the root, stem, and flower parts of the plant, and the surface properties were determined by the inverse gas chromatography (IGC) method. The IGC study was performed at infinite dilution with polar and non-polar solvents. Surface characteristics of different parts of the plant were determined through analysis of retention diagrams. From IGC experiments, the changes in the acidic and basic character of the parts of the plant were determined and compared with each other. As a result of this comparison, it was found that all KD/KA values were less than 1 and changed as flower > root > stem.