HPLC Methodology Research Articles

Antinociceptive effect and identification of berberine alkaloid in Berberis ruscifolia extracts

Ethnopharmacological relevanceAerial parts (leaves and stems) of Berberis ruscifolia Lam. are a usual preparation as an analgesic, anti-inflammatory, antimalarial, antibacterial, and digestive in folk medicine. However, there were no previous studies of its chemical composition and biological activity related to analgesic effects. The objective of the studyThe evaluation of the anti-nociception of the infusion (I), the decoction (D), and the ethanolic extract (EE) obtained from aerial parts of B. ruscifolia and its main chemical constituent in them, in mouse models. Material and methodsThe chemical constituent of B. ruscifolia extracts was evaluated and quantified by LC-MS and HPLC methodology. The inhibition of nociception in mice was analyzed by formalin and acetic acid-induced contortions tests. Also, when the formalin test was performed to evaluate the antinociceptive activity, the inhibition of edema formation and the antipyretic effect of each extract were simultaneously evaluated in the same experiment. For the oral administration in the in vivo assays, doses ranging from 10 to 1000 mg/kg and 10–30 mg/kg were used for extract and the chemical compound, respectively. ResultsThe presence of berberine (Berb) was identified in the three evaluated extracts where the EE showed the highest content of this compound getting a yield of 2%, while in the I and D, Berb is present at 0.2%. The three extracts promoted a reduction of the contortions induced by acetic acid, being observed in EE the highest activity with 63 ± 6% of significant inhibition of the nociceptive behavior at a dose of 300 mg/kg, while D significantly inhibited 32 ± 12% at the same dose and for I at a dose of 1000 mg/kg an inhibition of 44 ± 8% was observed. Likewise, in the formalin trial, I and EE reduced nociception at a dose of 1000 (31 ± 5%) and 300 (35 ± 3%) mg/kg, respectively in the neurogenic phase, while in the second phase of the experiment, all the extracts evaluated showed an antinociceptive effect, with significant inhibition of I of 54 ± 6% and D of 44 ± 5% at a dose of 1000 mg/kg and for EE showed a 63 ± 2% inhibition at a dose of 300 mg/kg being the one with the highest antinociceptive activity. These extracts showed no inhibition in temperature and formalin-injected paw edema formation when compared to the control. As for Berb, at a 30 mg/kg dose, it showed significant inhibition of 70 ± 5% in the acetic acid-induced contortion test. ConclusionAltogether, the present results evidenced the analgesic properties of B. ruscifolia, scientific information presented for the first time, and also provided important knowledge not reported so far about the chemical composition of its extracts, by identifying the presence of Berb in them. Finally, we were able to conclude that the analgesic effect demonstrated by this medicinal plant is partly due to the presence of Berb.

A new HPLC methodology for the analysis of metronidazole and dexibuprofen: Application to pharmacokinetic study and comparative greenness assessment

For the simultaneous analysis of metronidazole and dexibuprofen in real human plasma samples, a new HPLC method with a 6-minute analysis time was developed. Furthermore, the method was used to investigate their pharmacokinetics and bioavailability. The internal standard was chosen to be guaifenesin (IS). Following acetonitrile precipitation of plasma proteins, the drugs were chromatographically separated using a Zorbax Eclipse® C 18 column with acetonitrile: water (pH 4 with aqueous phosphoric acid) gradient mixture. Metronidazole and dexibuprofen calibration curves were found to be linear over the ranges of 0.5–20 and 1–30 µg/mL, respectively, with good correlation coefficients. The analysis of quality control samples revealed outstanding accuracy and precision. Drug stability was also examined throughout storage and processing, and all coefficients of variation were under 8.23 %. After this optimized method was successfully applied to plasma samples and the pharmacokinetic parameters of each of the targeted medicines were examined separately and compared to their parameters when they were administered concurrently. The outcomes supported each drug's significant impact on the pharmacokinetic characteristics of the other. Using five metrics, including the Analytical Eco-scale, Analytical GREEnness Calculator (AGREE), RGB-12, Hexagon, and AGREE-prep, the environmental impact of the suggested approach and some reported methods utilized for the determination of either medicines in biological matrix were evaluated. The promising results demonstrated lower environmental and health risks associated with sample preparation and analysis.

Development of salting-out extraction methodology for the determination of piroxicam from polymeric based nanocarriers and biological samples

The aim of the present study is to develop the polymeric nanoparticulate drug delivery systems of piroxicam and to evaluate the in-vitro characteristics such as entrapment efficiency, surface morphology, in-vitro drug release performance, etc. For this reason, a novel HPLC methodology was developed for the determination of piroxicam from its bulk form, pharmaceutical preparation, and nanoparticulate delivery systems. Furthermore, the developed formulation was applied to the rats and the biological samples (plasma, liver, heart, spleen, kidney, and lung homogenates) were analyzed by the developed HPLC method following a salting-out assisted liquid-liquid extraction strategy for the first time in the literature. A Kinetex C18 analytical column (150 mm × 4.6 mm i.d., 5 µm) was used as a stationary phase with a 0.8 mL/min flow rate of acetonitrile: phosphate buffer (40:60, v/v), the column oven was adjusted to 40 °C and detection wavelength is set to 360 nm. Developed method were validated as per selectivity, linearity, LOD, LOQ, precision, and accuracy specified in the International Council for Harmonisation guidelines. As a result of the present study, it has been shown that the analysis of piroxicam from the bulk form, pharmaceutical preparation, developed polymeric-based drug delivery system, and biological samples can be successfully performed and no interferences were observed in any matrix. The developed method was also successfully utilized to study the tissue distribution of piroxicam in rats.

Development of Method and Validation of Related Substances In NAPROXEN Injection by Applying Stability Indicating HPLC Methodology

A validated HPLC method was developed for the determination of Naproxen in pharma formulation. Isocratic elution at a flow rate of 1.0 ml/min was employed on NOVAPAK C18 4μm (3.9 x 150 mm) or equivalent, or similar is used for this chromatography analysis. Acetonitrile as 500v/v, water as 490 v/v furthermore Glacial Acetic acid as 10v/v is used a mobile phase. The UV detection wavelength was 254nm and 10.0µl sample was injected. The run time is About 120minutes for Sample, Unmarked, Placebo, System suitability, Sensitivity solution and 60minutes for diluted Regular. The Approximate retention time was founded for Naproxen is ± 3 minutes. The% R.S.D Naproxen was identified. The mean Percentage recovery for Naproxen is found within the specification limit. The method was validated as per the ICH guidelines. Thus, the proposed HPLC method can be successfully applied for routine quality control analysis of formulations. This method developed is simple and is better than the methods reported in the literature. Key words: RP-HPLC Refractive index detector, Naproxen, flow rate, column, ICH Guidelines, USP reference

Development of Method and Validation for Related Substances in Pemetrexed Injection formulation With HPLC

A validated HPLC method was developed for the determination of Pemetrexed (PMD) in pharmaceutical formulation. Isocratic elution at a flow rate of 1.00ml/min is employed on waters X bridge C18 50 mm × 4.6mm, 3.5μ Column, or equivalent. As a mobile phase 90 volumes of buffer solution and 10 volumes of Acetonitrile. Acetonitrile is utilized as mobile phase B. UV detection wavelength is set at 230nm. Injected sample is 5.0μl. Run time is 16 minutes to Sample, Blank, Placebo, System suitability along with 60minutes to diluted Regular. Approximate retention time is identified to PMD is ± 2.4 minutes. % R.S.D PMD is identified. Mean Percentage recovery to PMD is identified that within specification limit. This work is validated by using rules and regulations given by ICH. Therefore, proposed HPLC process should successfully applied to routine quality control analysis formulations. This process developed is simple also is better to that of different processes those are reported in earlier literature. Values permits application for proposed stability indicating HPLC methodology in syrup dose forms. Keywords: RP-HPLC Refractive index detector, PMD, flow rate, column, ICH Guidelines, USP reference.

LC-MS/MS CHARACTERIZATION OF FORCED DEGRADATION PRODUCTS OF TUCATINIB, A NOVEL TYROSINE KINASE INHIBITOR: DEVELOPMENT AND VALIDATION OF RP-HPLC METHOD

Objective: The current study focused on the development, validation, and characterization of forced degradation products using LC-MS/MS. Methods: A simple, selective, validated and well-defined isocratic HPLC methodology for the quantitative determination of Tucatinib at a wavelength of 239 nm. An isocratic elution of samples was performed on an Inertsil ODS (250x4.6 mm, 5m) column with a mobile phase of 70:30v/v Acetonitrile and formic acid (0.1%) delivered at a flow rate of 1.0 ml/min. MS/MS was used to characterize degradation products formed in the forced degradation study. The validation and characterization of forced degradation products were performed in accordance with ICH guidelines. Results: Over the concentration range of 5-100μg/ml, a good linear response was obtained. Tucatinib's LOD and LOQ were determined to be 0.05 and 0.5, respectively. According to standard guidelines, the method was quantitatively evaluated in terms of system suitability, linearity, precision, accuracy, and robustness, and the results were found to be within acceptable limits. The drug was degraded under acidic, alkaline, and reduction conditions in forced degradation studies. Conclusion: The method was found to be applicable for routine tucatinib analysis. Because no LC-MS/MS method for estimating tucatinib and its degradation products has been reported in the literature. There is a need to develop a method for studying the entire tucatinib degradation pathway.

A quick and convenient 1H quantitative NMR method for determination of bioactive pyranocoumarins from Clausena excavata

Clausena excavata, a shrub in the Rutaceae family, has been used in traditional medicine to alleviate cold, malaria, abdominal pain, bacterial infection, and HIV infection. The major biologically active constituents in the rhizomes and roots of C. excavata are the three pyranocoumarins; clausenidin, dentatin, and nordentatin. Due to the medicinal values of this plant, we have developed a quick and convenient qHNMR method to quantify clausenidin, dentatin, and nordentatin from the rhizomes and roots of C. excavata collected from six geographical locations throughout Thailand. The dried and powdered samples were extracted with deuterated methanol. The parameters for 1H NMR measurements were optimized according to the measured T1 relaxation time of the protons from the bioactive pyranocoumarins and of 2,3,5,6-tetrachloronitrobenzene (TCNB), the internal standard. HPLC assays were also conducted to verify the results obtained from the qHNMR method. The amount of clausenidin, dentatin, and nordentatin varied from different geographical locations, and the results from the qHNMR method were comparable to those obtained from the HPLC assays. The reported qHNMR method was quick, more convenient, and more applicable to large-scale quality control or screening than conventional HPLC methodology. The method developed herein is beneficial to the pharmaceutical and health industries.

Current Strategies for Studying the Natural and Synthetic Bioactive Compounds in Food by Chromatographic Separation Techniques

The present study summarizes the new strategies including advanced equipment and validation parameters of liquid and gas chromatography methods i.e., thin-layer chromatography (TLC), column liquid chromatography (CLC), and gas chromatography (GC) suitable for the identification and quantitative determination of different natural and synthetic bioactive compounds present in food and food products, which play an important role in human health, within the period of 2019–2021 (January). Full characteristic of some of these procedures with their validation parameters is discussed in this work. The present review confirms the vital role of HPLC methodology in combination with different detection modes i.e., HPLC-UV, HPLC-DAD, HPLC-MS, and HPLC-MS/MS for the determination of natural and synthetic bioactive molecules for different purposes i.e., to characterize the chemical composition of food as well as in the multi-residue analysis of pesticides, NSAIDs, antibiotics, steroids, and others in food and food products.

Intestinal/uterine antispasmodics, sedative effects of Fuchsia magellanica Lam. leaves’ and flowers’ extracts and their flavonolic components

BackgroundFuchsia magellanica Lam. (Onagraceae) is a native plant of the Andean-Patagonian region, and it is also naturalized in Argentina and other parts of the world. Leaves are used for treating indigestion, stomachache, as sedative, for difficult delivery and as antiemmenagogue. However, up to now the pharmacological basis of such uses were not studied. PurposeThis work evaluates the gastrointestinal, uterine and sedative effects of leaves (L) and flowers (F) tinctures of F. magellanica from wild patagonian (T-Fm-P) and naturalized (T-Fm-BA) plants, as well as the mechanism of action and the flavonoids profile of both plants. MethodsPhytochemical studies were evaluated by using TLC and HPLC methodologies. The ex-vivo effects of T-Fm-BA and T-Fm-P were evaluated on contractile concentration-response curves (CRC) of carbachol (CCh) and calcium (Ca2+) in rat isolated intestinal and uterine tissues. In vivo tests for intestinal transit, elevated cross plus-maze and open-field tests were performed in mice. ResultsWild and cultivated leaves and flowers ethanolic extracts induced antispasmodic effect, as a non-competitive inhibitor of the CCh-CRC in intestine, with IC50 of 272.8 ± 64.3 µg/ml for T-L-Fm-P and 257.4 ± 36 µg/ml for T-L-Fm-BA. A similar effect was obtained in Ca2+-CRC with IC50 of 152.9 ± 29.1 µg/ml and 138.4 ± 48.6 µg/ml respectively. Mice intestinal transit was reduced at oral doses of 73.5 mg/kg of T-L-Fm-P. In uterine tissues, both T-L-Fm reduced the contractions of CCh CRC´s as non-competitive antagonists, with IC50 near those in intestine. The T-L-Fm-BA also inhibited the serotonin CRC´s and contribution of other relaxing mechanisms was investigated. Although extracts didn't show anxiolytic effect in the elevated cross plus-maze test, they reduced the spontaneous activity of mice in the open-field at 32.7 mg/kg T-L-Fm-P and 41.7 mg/kg T-L-Fm-BA. ConclusionsLeaves and flowers ethanolic extracts from wild and cultivated plant F. magellanica showed intestinal and uterine antispasmodic effects, mainly by interfering with Ca2+ influx. In mice, the ethanolic extracts reduced the intestinal transit and showed sedative effect. The effects agree with the presence of flavonoids, such as quercetin and kaempferol. This is the first study which gives experimental support for some of the traditional uses of F. magellanica.

Synthesis of Triphenylethylene-Naphthalimide Conjugates as topoisomerase-IIα inhibitor and HSA binder.

A series of triphenylethylene-naphthalimide (TPE-naph) conjugates was synthesized by a molecular hybridization technique, and their anticancer activity was evaluated in vitro on 60 human cancer cell lines through their cytotoxicity. The ratios of E and Z isomers were determined on the basis of HPLC methodology and NMR spectroscopy. The structure-activity relationship for anticancer activity was deduced on the basis of the nature and bulkiness of the amine attached to the C-4 position of the naphthalene ring. Experimental and molecular modeling studies of the most active TPE-naph conjugate bearing a morpholinyl group showed that it was able to inhibit topoisomerase-II (TOPO-II) as a possible intracellular target. Moreover, the transportation behavior of TPE-naph conjugate towards human serum albumin (HSA) indicated efficient binding affinity. The steady-state and time-dependent fluorescent results suggested that this conjugate quenched HSA significantly through static as well as dynamic quenching. Thus, this report discloses the scope of triphenylethylene-naphthalimide (TPE-naph) conjugates as efficient anticancer agents.

Negative Pharmacokinetics Evaluation of Liquorice and Pomegranate Juices on Potential of Salbutamol in Rats

Objectives: Beverages or food may alter the pharmacokinetic parameters and pharmacodynamics of drugs-substances’ active pharmaceutical ingredients which may cause adverse circumstances. Salbutamol is widely indicated to prevent asthma and acute/chronic obstructive pulmonary disorders. Liquorice and/or Pomegranate are prescribed to support cough and asthma management. Nonetheless, an interaction thereafter with a Salbutamol pharmacokinetics was not investigated yet. Thus, the aim of this research is to evaluate the effect of Liquorice and/or Pomegranate on Salbutamol pharmacokinetic-parameters in animals (rats). Methods and Materials: A Chromatography method of (HPLC) was carried out. Partially validated after a development profitably conformed to the European Medical Agency’s guidelines to measure Salbutamol in rat serum. Liquorice and/or Pomegranate juices were given to rats for three days and, an hour sooner than Salbutamol (5.0 mg/kg), it was administered as an oral solution. Blood samples were collected at scheduled intervals, then handled and investigated by HPLC methodology. Results: The linearity of the calibration curve was over 98%. The intra-day and the inter-day accuracy values of Salbutamol were (97.93%-105.18%), and (98.67%-105.91%), respectively. While the intra- and inter-day precision values were equal to or less than 3.178% and 3.653%, respectively. In addition, no significant change was seen in the pharmacokinetic parameters of Salbutamol when administered to rats that were given either pomegranate or Liquorice juice. Conclusions: It was established that the given Liquorice or pomegranate juices with the above-mentioned orally administrated Salbutamol solution, did not result in a significant parameter in the pharmacokinetic interaction.

Estimation of Cyproheptadine Hydrochloride and Tricholine Citrate Simultaneously in Syrup Dose by Applying Stability Indicating HPLC Methodology

An stability indicating HPLC methodology for the concurrent estimation of Tricholine citrate (TRC) and Cyproheptadine hydrochloride (CYH) in syrup dose and bulk using Waters column reverse phase C18 (5 µm, 250 mm and 4.6 mm) as stationary phase and 0.1M Na2HPO4 of pH 4.5 and acetonitrile in proportion of 60:40 (v/v) at flow of 1.0 ml/min rate as mobile phase was reported. The linear scales were 275-825 μg/ml for TRC and 2-6 μg/ml for CYH with correlation coefficients of 0.9999 for TRC and 0.9997 for CYH. Followed ICH Q2(R1) strategies for validating the suggested method for precision, sensitivity, robustness, specificity, selectivity and accuracy. The measures of LOD and LOQ are 0.023 µg/ml and 0.079 µg/ml for CYH, while for TRC it was 0.565 µg/ml and 1.885 µg/ml, respectively. The precision measures for CYH and TRC were 0.073 and 0.212 relative measured deviation percentage, respectively. The accuracy measures for CYH and TRC were 99.40% and 99.09% mean assay percentiles, respectively. Recovery percentiles measures of CYH and TRC were ranged between 99.48% to 100.35% and 100.38% and 100.41%, respectively. While in degradation investigation, peaks of degraded products are very well differentiated from TRC and CYH peaks suggesting the specificity and stability of suggested methodology. The results permit the application of the proposed stability indicating HPLC methodology in syrup dose forms.

A novel RP – HPLC methodology for method development and validation of aceclofenac and tizanidine pharmaceutical dosage forms

A simple and selective LC technique is chosen for the determination of Aceclofenac and Tizanidine in pill indefinite quantity forms. Chromatographic process separation was achieved on a c18 column victimization mobile part consisting of a combination of fifty volumes of Triethylamine buffer, fifty volumes of acetonitrile with detection of 230nm. Dimensionality was discovered within the vary 5-15 µg/ml for aceclofenac (r2 =0.999) and 1-3 µg /ml for tizanidine (r2 =0.998) for the number of medicine calculable by the planned strategies was in smart agreement with the label claim. The planned strategies have a sound procedure. At three completely different levels the accuracy of the strategies was assessed by recovery studies. The recovery experiments indicated the absence of interference from unremarkably encountered pharmaceutical additives showing %RSD below a pair of this technique was found to be precise as indicated by the repeatability analysis. All applied mathematics information proves all ways have valid procedure and might be used for routine analysis of pharmaceutical dose kind

Back Cover: Development and validation of HPLC methodology for quantitative estimation of Efinaconazole in topical pharmaceutical formulation prepared in‐house for the treatment of onychomycosis

DOI: 10.1002/sscp.202000050 The cover picture shows an orthogonal approach for method development and validation of three potential halo alkyl alcohol genotoxic impurities in miglitol drug substance by fast gas chromatography–mass spectrometry. The chloro ethanol/bromo ethanol/iodo ethanol is a key starting material (KSM) in miglitol drug substance synthesis. As per ICH M7, it is a potential genotoxic impurity (PGI), it should show cut-off in miglitol drug substance. The developed and validated FGC-MS quantitative method for chloro ethanol, bromo ethanol and iodo ethanol in miglitol drug substance are specific, accurate, precise, linear, robust and rugged at 2.0 ppm limit quantitation level and they can be detected up to 0.7 ppm.

Development and Validation of RP-HPLC Method using Photodiode Array or Diode-Array Detection Detector for simultaneous Estimation of the Amlodipine Besylate and Lisinopril in Fixed-Dose Formulation

A basic and specific HPLC-PDA (high-performance liquid chromatography-photodiode array) detector showing approach was created for the simultaneous estimation of anti-hypertensive medications—amlodipine besylate (AMD) and lisinopril (LIS). A successful chromatography method was developed using the SunFire C8 column (4.6 × 150 mm, 5 μm) with gradient elution of the mobile phase composed of 85:25 potassium dihydrogen phosphate in water and methanol at a flow rate of 0.7 mL/min. The wavelength was set at 212 nm for the simultaneous estimation of AMD and LIS. The retention time obtained was 5.1 and 10.5 minutes for AMD and LIS, respectively. Descriptive performance of the proposed HPLC methodology was measurably approved as for framework accuracy, linearity, robustness, specificity, precision, and forced degradation for identification and evaluation limits. The linearity range for AMD and LIS correlation coefficient value obtained was R2 andgt; 0.999. The drugs were exposed to forced degradation conditions—acidic, alkali, oxidation, etc. The newly developed reverse phase high performance liquid chromatography (RP-HPLC) method was applied for the detection of the referred to anti-hypertensive medications in their combination pharmaceutical tablets.

Back Cover: Development and validation of HPLC methodology for quantitative estimation of Efinaconazole in topical pharmaceutical formulation prepared in‐house for the treatment of onychomycosis

Back Cover: Development and validation of HPLC methodology for quantitative estimation of Efinaconazole in topical pharmaceutical formulation prepared in‐house for the treatment of onychomycosis

Effects of Several Physicochemical Factors on the Biodegradation of 1,4-Dioxane by Microbial Consortium CDIK-3 Enriched from Industrial Wastewater

Objective : With the ultimate aim of application for effluent treatment, the relationships between 1,4-dioxane degradation by microbial consortium CDIK-3 and several relevant physicochemical environmental parameters were investigated.Methods : Microbial consortium CDIK-3 enriched from an industrial wastewater sample developed to grow aerobically with 1,4-dioxane as the sole carbon substrate was used to evaluate the degradation of 1,4-dioxane depending on the various environmental parameters and HPLC methodology was used to measure residual 1,4-dioxane in the test consortium.Results and Discussion : The microbial consortium, designated CDIK-3, was originally enriched from industrial wastewater sample. Complete depletion of 1,000 mg/L 1,4-dioxane was achieved in this experiment within 168 h. Effective 1,4-dioxane degradation was observed in slightly alkaline conditions (pH 8) and 30℃. The effect of supplemented carbon and nitrogen (<i>e.g</i>., glucose, yeast extract, ethylene glycol, tetrahydrofuran) on the 1,4-dioxane degradation by the CDIK-3 test culture was evaluated. The results indicated that the addition of glucose, yeast extract, or tetrahydrofuran accelerated bacterial growth compared to growth in the absence of supplemented carbons. In contrast, the addition of low concentrations (≤100 mg/L) of ethylene glycol accelerated 1,4-dioxane degradation, but the degradation of 1,4-dioxane was delayed in the culture to which high concentrations (≥200 mg/L) of ethylene glycol were added.Conclusions : 1,4-Dioxane as a wastewater contaminant can be removed by the microbial consortium CDIK-3 enriched from industrial wastewater. Effective 1,4-dioxane removal was obtained by the control of various physicochemical parameters. These results provide clues for understanding improved 1,4-dioxane degradation by microbial consortium CDIK-3 with relevant environmental parameters and possible applications in the processing of industrial wastewater containing relatively high 1,4-dioxane concentrations.

UPLC-PDA: A greener miniaturized tool for the analysis and purity assessment of ebastine and phenylephrine hydrochloride

Recent improvements in miniaturization of the existing technologies, nevertheless analytical instruments have acquired worldwide acceptance and encouragement. The replacement of traditional analytical procedures by miniaturized alternatives has become inevitable to promote more ecologically benign techniques. The dominance of HPLC technique during the past decades is noticeable in the fields that deal with drugs’ analysis and quality control despite the provoked environmental concerns about it. Thus, the development of a miniaturized HPLC that retains the practicality and versatility of the conventional HPLC technique, whereas it is greener, has been a long thought after goal for the analysts. Lately, UPLC has been developed as a unique analytical approach that utilizes the chemistry of small particles to meet the need for a miniaturized HPLC methodology. Employing a smaller sized stationary phase that can withstand high pressure, UPLC leads to a substantial reduction in time, labor, waste, and cost of materials required to get results compared to traditional HPLC columns. In this contribution, a UPLC method was adopted for the first time for the analysis and purity assessment of a binary mixture of ebastine (EBS) and phenylephrine hydrochloride (PHE) formulated in Ebast DC® tablets. The proposed UPLC method was applied successfully to the determination of the investigated drugs over a linearity range of 1–50 µg/mL, with LODs equal to 0.31 µg/mL and 0.33 µg/mL for EBS and PHE, respectively. To evaluate the greenness of the developed UPLC methodology; the Eco-Scale semi-quantitative green metric tool was employed in a side-by-side comparison with the reported HPLC method to conclude whether UPLC provides a greener and more efficient technique or not.

Development and validation of HPLC methodology for quantitative estimation of Efinaconazole in topical pharmaceutical formulation prepared in‐house for the treatment of onychomycosis

AbstractThe purpose of this research work was to design, develop, and optimize new high‐performance liquid chromatography method for quantitative estimation of Efinaconazole in topical pharmaceutical formulation developed in‐house for the treatment of onychomycosis. Based on the outcomes of preliminary investigation, optimized high‐performance liquid chromatography conditions were reverse phase column (Luna C18) as stationary material, with simple isocratic mobile phase comprising mixture of water and acetonitrile (20:80 v/v), flow rate 1 mL/min, column oven temperature 30°C, injection volume 10 µL, detection wavelength of 230 nm, and total run time of 10 min. The optimized method was successfully validated as per International Council for Harmonization guideline Q2(R1), and statistical analysis proved that the method is specific (peak purity ≥ 0.999), accurate (99.21–99.63% w/w), precise (% relative standard deviation < 2.0), and sensitive (limit of detection = 9.26 µg and limit of quantitation = 28.05 µg). As a result, this research work provides a new and simple high‐performance liquid chromatography methodology with optimized chromatographic parameters for quantitation of Efinaconazole as a routine assay purpose, contributing to the technological and scientific domain and guaranteeing a safe and effective use of Efinaconazole for the treatment of onychomycosis.

Optimization of glycine and thiotriazoline compound analysis by high-performance liquid chromatography

Украина занимает одно из первых мест в Европе по показателям цереброваскулярной заболеваемости и смертности от инсульта. Поэтому создание новых лекарственных средств для лечения этих патологий – актуальная задача современной фармации, а создание нового комбинированного препарата на основе глицина (нейротрасмиттерной аминокислоты) и тиотриазолина (антиоксиданта) целесообразно и актуально. Для нового комбинированного лекарственного средства предложена рациональная лекарственная форма – таблетки. Для созданных комбинированных таблеток необходимо разработать методы стандартизации. Чаще всего для стандартизации готовых лекарственных форм и аптечного, и заводского изготовления используют физико-химические методы исследования. Наше внимание привлек метод высокоэффективной жидкостной хроматографии, который позволяет одновременно провести стандартизацию действующих веществ в одной навеске. Для разработки методики ВЭЖХ прежде всего необходимо подобрать оптимальные условия проведения анализа действующих веществ. Цель работы – подбор оптимальных условий одновременного определения глицина с тиотриазолином в модельной смеси методом ВЭЖХ. Материалы и методы . В ходе исследований использовали глицин, тиотриазолин. Исследования проводили с использованием модульной системы ВЭЖХ BISCHOFF со спектрофотометрическим детектором Lambda 1010. Использовали колонки Prontosil 120-5-CN, Hypersil ODS-C18-5u. В качестве элюента использовали воду, 0,05 % водный раствор трифторуксусной кислоты Bu 4 NHSO 4 3,4 г/л в воде, Bu 4 NHSO 4 3,4 г/л, 0,02 М Na 2 HPO 4 в воде, Bu 4 NHSO 4 3,4 г/л, 0,05 % раствор трифторуксусной кислоты в воде. Результаты. Согласно полученным результатам, среди различных использованных элюентов и фаз в дальнейшем для совместного определения глицина с тиотриазолином и в модельной смеси, и в комбинированных лекарственных формах целесообразно использовать в качестве элюента тетрабутиламмоний в условиях ион парного хроматографирования на обратной фазе с одновременным использованием кислого буфера – 0,05 % раствора трифторуксусной кислоты. Выводы. В ходе исследований подобраны оптимальные условия одновременного определения глицина с тиотриазолином в одной навеске. Установили, что определение действующих веществ необходимо проводить в условиях ион парного хроматографирования на обратной фазе при использовании элюента тетрабутиламмония с одновременным использованием кислого буфера – 0,05 % раствора трифторуксусной кислоты.