RESUMEN
A new high-performance liquid chromatography (HPLC) method was applied for the quantification of the active substance of tofacitinib. Analysis was performed on a Chromasil 100 C18 (100.0 × 4.0 mm, 3.5 µm) stationary phase. The mobile phase consisted of acetonitrile:0.2% phosphoric acid in water (12:88, v/v). The prepared sample (20.0 µL) was injected into the system. A detection wavelength of 285.0 nm was chosen for the compound, and the flow rate was 0.8 mL/min. The experiment was completed in 5.0 min. The analysis temperature was set to 40.0°C. The method was evaluated using green chemistry. The method was validated according to the International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use guidelines. For linearity studies calibration curves were constructed in the range of 10.0-200.0 µg/mL. The recovery values were calculated at 97.66% and 105.68%. The method developed for the analysis of the active substance had a short analysis time and was cost-effective. It is an environmentally friendly method due to the mobile phase content used. The technique can be used in laboratory analysis and bioequivalence experiments.
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Estabilidad de Medicamentos , Tecnología Química Verde , Piperidinas , Pirimidinas , Cromatografía Líquida de Alta Presión/métodos , Piperidinas/análisis , Piperidinas/química , Pirimidinas/análisis , Pirimidinas/química , Tecnología Química Verde/métodos , Reproducibilidad de los Resultados , Modelos Lineales , Límite de Detección , Pirroles/análisis , Pirroles/químicaRESUMEN
Two spectrophotometric techniques and a novel HPLC method were consecutively applied for the simultaneous quantification of the active ingredients of emtricitabine (EMC), tenofovir (TNF), and bictegravir (BIC). The first spectrophotometric method is the dual amplitude difference method coupled with the ratio difference method. TNF was determined using the dual amplitude difference method, while BIC and EMC were determined using the ratio difference method. The second spectrophotometric method was the constant multiplication with absorbance extraction method, and was applied for the determination of active substances used in the treatment of human immunodeficiency virus (HIV) infection. BIC was determined by the constant multiplication method, whereas EMC and TNF were determined using the absorbance extraction method. For the HPLC method, the XBridge C18 column was used. The solvent system comprised acetonitrile:phosphate buffer (pH 6.8; 30:70 v/v). All active ingredients were detected at 260.0 nm, and the flow rate was 0.5 mL/min. The experiment was completed within 5.5 min. The experiments carried out enabled the simultaneous analysis of the three active substances and they were economical, fast, environmentally friendly, and simple. The methods have been successfully applied to prepare mixtures and tablets without matrix interference. The methods were evaluated in terms of green chemistry. The methods have been validated according to International Council for Harmonisation (ICH) guidelines.
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Compuestos Heterocíclicos de 4 o más Anillos , Humanos , Tenofovir , Emtricitabina , Cromatografía Líquida de Alta Presión , Preparaciones FarmacéuticasRESUMEN
The present study aims to develop an electroanalytical method to determine one of the most significant antineoplastic agents, topotecan (TPT), using a novel and selective molecular imprinted polymer (MIP) method for the first time. The MIP was synthesized using the electropolymerization method using TPT as a template molecule and pyrrole (Pyr) as the functional monomer on a metal-organic framework decorated with chitosan-stabilized gold nanoparticles (Au-CH@MOF-5). The materials' morphological and physical characteristics were characterized using various physical techniques. The analytical characteristics of the obtained sensors were examined by cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), and differential pulse voltammetry (DPV). After all characterizations and optimizing the experimental conditions, MIP-Au-CH@MOF-5 and NIP-Au-CH@MOF-5 were evaluated on the glassy carbon electrode (GCE). MIP-Au-CH@MOF-5/GCE indicated a wide linear response of 0.4-70.0 nM and a low detection limit (LOD) of 0.298 nM. The developed sensor also showed excellent recovery in human plasma and nasal samples with recoveries of 94.41-106.16 % and 95.1-107.0 %, respectively, confirming its potential for future on-site monitoring of TPT in real samples. This methodology offers a different approach to electroanalytical procedures using MIP methods. Moreover, the high sensitivity and selectivity of the developed sensor were illustrated by the ability to recognize TPT over potentially interfering agents. Hence, it can be speculated that the fabricated MIP-Au-CH@MOF-5/GCE may be utilized in a multitude of areas, including public health and food quality.
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Quitosano , Nanopartículas del Metal , Estructuras Metalorgánicas , Impresión Molecular , Humanos , Polímeros Impresos Molecularmente , Quitosano/química , Topotecan , Oro/química , Nanopartículas del Metal/química , Técnicas Electroquímicas/métodos , Impresión Molecular/métodos , Límite de Detección , Polímeros/química , Carbono/químicaRESUMEN
An HPLC method with UV detection was developed for the determination of carnosic acid in human plasma and applied to a pharmacokinetic study after oral administration of Rosemary extract to a healthy volunteer. Sample preparation depends on liquid-liquid extraction with hexane. Chromatographic separation was achieved with C18 column (150 mm × 4.6 mm × 5 µm), at 25°C with isocratic elution, mobile phase composed of solution A (methanol), and solution B (2% o-phosphoric acid in water) (90:10, v/v) at flow rate of 1.0 mL/min. The analyte was detected at 230 nm. The retention time is 4.20 ± 0.03 min. The method was validated in terms of accuracy, precision, specificity, robustness and detection and quantification limits, in accordance with European Medicines Agency guidelines. LOD and LOQ were found to be 0.075 and 0.25 ng/mL, respectively. The method was applied to the analysis of carnosic acid in human plasma with good recovery as 91.7%. The plasma concentration-time profile and pharmacokinetic parameters: AUC0-t, AUC0-∞, Cmax, tmax, t1/2 were calculated according to the assays. The method can certainly be used for routine analysis of carnosic acid in human plasma after oral administration of Rosemary extract, and for phase I clinical studies and bioavailability-bioequivalance studies as well.
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Abietanos , Humanos , Cromatografía Líquida de Alta Presión/métodos , Administración Oral , Disponibilidad Biológica , Reproducibilidad de los ResultadosRESUMEN
A fast procedure obtained by the combination of fabric phase extraction (FPSE) with high performance liquid chromatography (HPLC) has been developed and validated for the quantification of favipiravir (FVP) in human plasma and breast milk. A sol-gel polycaprolactone-block-polydimethylsiloxane-block-polycaprolactone (sol-gel PCAP-PDMS-PCAP) coated on 100% cellose cotton fabric was selected as the most efficient membrane for FPSE in human plasma and breast milk samples. HPLC-UV analysis were performed using a RP C18 column under isocratic conditions. Under these optimezed settings, the overall chromatographic analysis time was limited to only 5 min without encountering any observable matrix interferences. Following the method validation procedure, the herein assay shows a linear calibration curve over the range of 0.2-50 µg/mL and 0.5-25 µg/mL for plasma and breast milk, respectively. The method sensitivities in terms of limit of detection (LOD) and limit of quantification (LOQ), validated in both the matrices, have been found to be 0.06 and 0.2 µg/mL for plasma and 0.15 and 0.5 µg/mL for milk, respectively. Intraday and interday precision and trueness, accordingly to the International Guidelines, were validated and were below 3.61% for both the matrices. The herein method was further tested on real samples in order to highlight the applicability and the advantage for therapeutic drug monitoring (TDM) applications. To the best of our knowledge, this is the first validated FPSE-HPLC-UV method in human plasma and breast milk for TDM purposes applied on real samples. The validated method provides fast, simple, cost reduced, and sensitive assay for the direct quantification of favipiravir in real biological matrices, also appliyng a well-known rugged and cheap instrument configuration.
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Leche Humana , Pirazinas , Femenino , Humanos , Cromatografía Líquida de Alta Presión/métodos , Límite de DetecciónRESUMEN
Biogenic amines (BAs) are group of substances that are formed from amino acids by decarboxylation or amination and transamination of aldehydes and ketones. They may have either aliphatic, aromatic or heterocyclic structure. Their quantity determines their effects, optimum amounts are essential for physiological functions, but excess of BAs causes various toxic effects through out human body. BAs are presented in a wide variety of fermented foods such as fish, meat, milk products and some kinds of beverages like wine, beer and some fruit juices. In order to quantify their intake by food products are important, the methods that provide determination of BAs in food products are a matter of priority. Mostly, liquid chromatographic (LC) methods are preffered. Their amine groups are able to be derivatized by so many fluorogenic reagents. It is possible to combine LC systems with UV-vis. absorption spectrometric, fluorimetric and mass spectrometric detectors. Due to the fact that BAs are important markers for food quality and important for health, in this article LC methods for the determination of BAs in foods were reviewed from 2012 to present.
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Alimentos Fermentados , Vino , Animales , Cerveza/análisis , Aminas Biogénicas/análisis , Humanos , Carne/análisis , Vino/análisisRESUMEN
For the quantifiable amounts of Telmisartan (TLM) and Hydrochlorothiazide (HYD) in the presence of Amlodipine (AML) in a ternary mixture of synthetic laboratory mixture, a novel, sensitive, quick, and practical reversed-phase high-performance liquid chromatography (RP-HPLC) method was given. In order to separate, a Waters Spherisorb ODS-2 C18 column was used. For HYD, TLM, and AML, these techniques were viable over linearity ranges of 4-12 µg/mL, 4-25 µg/mL, and 5-40 µg/mL, respectively. The mobile phase system was acetonitrile:methanol: phosphate buffer at pH 2.5 (65:5:30 v/v/v), and the flow rate was 1.5 mL/min. Novel spectrophotometric methods were applied for active substances to determine simultaneously. The first method is absorptivity centering using factorized spectrum, and the second method is dual amplitude difference coupled with absorbance subtraction. These approaches have been effectively applied to bulk, laboratory synthetic mixtures to employ active components quantitatively. Correlation coefficients were found to be higher than 0.99 and the limit of detection values lower than 0.49 µg/mL in both spectrophotometric methods. The methodologies were validated following ICH recommendations. In the developed HPLC method, the limit of detection values was found to be 0.01 µg/mL for HYD and 0.02 µg/mL for AML and TLM. The correlation coefficients for the HPLC method were found to be 0.9971 for HYD, 0.9990 for AML, and 0.9983 for TLM. The suggested HPLC technique is a simple, effective, sensitive, environmentally friendly, and time-saving approach for determining TLM and HYD in the presence of AML.
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Amlodipino , Leucemia Mieloide Aguda , Amlodipino/análisis , Cromatografía Líquida de Alta Presión/métodos , Humanos , Hidroclorotiazida/análisis , TelmisartánRESUMEN
A novel, sensitive, fast, and pratic RP-HPLC methods were presented for the quantitative amounts of Telmisartan (TEL) and Olmesartan (OLM) in the presence of Amlodipin (AML) in a binary mixture of pharmaceutical preparation. Waters Spherisorb ODS-2 C18 column was used for separation. These methods were valid over linearity ranges of 2.5-30 µµg/mlL, 2-85 µµg/mlL, and 2-35 µµg/mlL for OLM, TEL, and AML, respectively. The mobile phase system consisted of acetonitrile:methanol: phosphate buffer at pH 3.0 (65:5:30 v/v/v), and the flow rate was 1,5 mlL/min for OLM and AML. The mobile system's other mixture (TEL and AML) was acetonitrile:methanol: phosphate buffer at pH 2.5 (65:5:30 v/v/v), and the flow rate was 1,5 mlL/min. These procedures were successfully applied to bulk, laboratory synthetic mixture, and medicinal dosage forms to use active ingredients quantitatively. The studied methods were validated according to ICH guidelines. In the developed HPLC method, the limit of detection values was found to be 0.020 µµg/mlL for TEL, 0.025 µµg/mlL for OML, and 0.070 µµg/mlL for AML. The correlation coefficients for the HPLC method were found to be 0.9938 for TEL, 0.9996 for OML, and 0.9982 for AML. The calibration range is between 2.5 and -30, 5-35, and 2-85 µµg/mlL for OLM, AML, and TEL, respectively. The proposed HPLC method is a convenient, effective, sensitive, green, and time-saving method for the rapid determination of TEL and OLM in the presence of AML.
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Antihipertensivos , Leucemia Mieloide Aguda , Acetonitrilos , Cromatografía Líquida de Alta Presión/métodos , Humanos , Metanol , FosfatosRESUMEN
An ultrasensitive and selective voltammetric sensor with ultra-trace level detection limit is introduced for idarubicin (IDA) determination in real samples. The as-synthesized nanocomposite was characterized by several techniques, including Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD), Raman spectroscopy, Energy-dispersive X-ray spectroscopy (EDX), and Field emission scanning electron microscopy (FE-SEM). The electrocatalytic performance of the developed electrode was observed by cyclic voltammetry (CV), differential pulse voltammetry (DPV), electrochemical impedance spectroscopy (EIS), and chronoamperometry. The limit of detection (LOD) of the developed sensor for idarubicin is 1.0 nM, and the response is found to be in the dynamic concentration range of 0.01-1.9 µmol/L in a Britton-Robinson buffer (B-R, pH = 6.0). Moreover, the fabricated electrode illustrated high selectivity with good repeatability and reproducibility for diagnosing idarubicin as an anthracycline antileukemic drug. Furthermore, to evaluate the validity of the recommended method, three real samples, including human plasma, urine, and water samples, were analyzed with satisfactory recovery and compared with high-performance liquid chromatography (HPLC). The minor groove-binding mode of interaction was also supported by docking simulation studies, emphasizing that IDA can bind to ds-DNA preferably and confirmed experimental results. The reduced assay time and the possibility of measuring a single sample with another anticancer drug without any interference are significant advantages compared to the HPLC. The developed and validated sensor could be a valuable point-of-care diagnostic tool for IDA quantification in patients.
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Grafito , Nanosferas , Puntos Cuánticos , Técnicas Electroquímicas/métodos , Grafito/química , Humanos , Idarrubicina , Límite de Detección , Simulación del Acoplamiento Molecular , Reproducibilidad de los Resultados , Espectroscopía Infrarroja por Transformada de FourierRESUMEN
HIAM technique allows the extraction of the original constant signal of each single component out of interference signals of a mixture and further transformed into basic spectrum (D0). It includes the methods: ratio subtraction coupled with unified constant subtraction (RS-UCS), constant center (CC) and constant extraction (CE). The technique was introduced for the analysis of two pharmaceutical formulations used to treat cardiovascular diseases. The formulations are binary combinations of Amlodipine (AML) with either Atorvastatin (ATR) or Candesartan (CND) which shows interefernce absorbance signals. The technique was valid over the linearity range of (5.0-35.0 µg/ml) for AML, ATR and CND with recovery percentage 100.40 ± 1.88 , 100.00 ± 0.86 and 99.83 ± 1.07, respectively . The extracted signals were tested for its purity by spectral contrast angle (cos θ) to illustrate the efficency of the HIAM technique where cos θ values ranges from (0.9902 to 0.9986). The presented technique was fully validated regarding ICH guidelines and were statistically compared using one-way ANOVA at 95% confidence.
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Amlodipino , Amlodipino/análisis , Análisis de Varianza , Atorvastatina , Combinación de Medicamentos , Espectrofotometría/métodosRESUMEN
Versatile, extraction-free univariate spectrophotometric methods have been modified to get effective spectral resolution of mixtures in accordance with their feature challenges. The proposed methods have been applied and validated for analyzing some antihypertensive medicines in their co-formulated medicinal products. Two mixtures have been used: the first one [Mix I (OLM/ADB)] is composed of Olmesartan medoxomil (OLM) and Amlodipine besylate (ADB) with partly-overlapped spectra while the second [Mix II (TEL/HCT)] is made up Telmisartan (TEL) and Hydrochlorothiazide (HCT) with total-overlapped spectra. Induced dual wavelength, absorbance correction and ratio subtraction coupled with constant multiplication methods were applied to Mix I (OLM/ADB), while dual wavelength, advanced absorbance subtraction and constant center coupled with spectrum subtraction methods were applied to Mix II (TEL/HCT). Calibration graphs were established with good correlation coefficients. The methods exhibit significant advantages as simplicity, sensitivity, minimal data manipulation besides optimum robustness. Selectivity was inspected using lab-mixtures with diverse ratios. Accuracy, precision and repeatability were found to be within the acceptable limits. The proposed methods are good enough to be used for co-assay of analytes in combined forms without any interfering from excipients. Moreover, all results were estimated in accordance with ICH criteria and successfully compared with those of the reported methods applying t-test and F-test at 95% confidence level. Generally, these methods can be used efficiently for routine quality control testing.