HPLC Method For Simultaneous Determination Research Articles

Development of an High Performance Liquid Chromatography Method for Simultaneous Determination of Hesperidin and Sodium Benzoate in Kien Ty Syrup

The Kien ty syrup is formulated from six medicinal materials, including Pericarpium Citri Reticulatae, Fructus Aurantii immaturus, Radix Codonopsis, Rhizoma Atractylodis macrocephalae, Fructus Crataegi, Fructus Hordei germinatus and sodium benzoate as a preservative. Hesperidin, a bioflavonoid found in Pericarpium Citri Reticulatae, Fructus Aurantii immaturus, is used to assess the syrup's quality in terms of assay. In this study, an HPLC method for simultaneous determination of hesperidin and sodium benzoate in Kien ty syrup was developed. The method was performed on an Agilent C18 column (250 × 4.6 mm; 5 μm) with the mobile phase being a mixture of 0.05 M phosphate buffer solution pH 5.5: acetonitrile (70:30, v/v), running in a isocratic elution mode. The flow rate was set at 1.0 mL/min, and the injection volume was 10 μL. The analytes were detected at 225 nm. The method was validated and met the requirements according to ICH, AOAC guidelines. Applying the developed quantification method, the analysis of three sirup lots revealed that the content of hesperidin and sodium benzoate ranged from 0.480-0.504 mg/mL and 2.869-2.306 mg/mL, respectively. The study results indicate that this quantification method can be applied for simultaneous determination of hesperidin and sodium benzoate in Kien ty syrup and similar products.

Development of a rapid, accurate and efficient HPLC method for simultaneous determination of catechins and theaflavins in black tea

Catechins and theaflavins, as the primary bioactive constituents of black tea, have a significant impact on its color, aroma, and flavor profile.We developed a rapid and precise method using HPLC to detect catechins and theaflavins in black tea. The method employed 0.1% formic acid-acetonitrile (phase A) and 1% formic acid-water (phase B) as mobile phases. Separation occurred on an Agilent SB-C18 column (4.6 mm × 250 mm, 5 µm), with 20 μL injection volume. Detection took place at 30 °C with a flow rate of 1 mL/min for 55 min.Quantitative determination utilized the external standard method, exhibiting high sensitivity (limit of detection: 0.19 to 5.14 μg/mL) and a strong linear correlation (R2 ≥ 0.9993) between concentration range and peak area. Precision, repeatability, and stability were excellent, with relative standard deviations (RSDs) below 5% (n = 5). The average recovery rate of standard addition ranged from 98.02% to 102.57%, with an RSD of less than 3.17% (n = 5). In summary, this straightforward, efficient, and precise approach is well-suited for both qualitative and quantitative analysis of catechins and theaflavins in tea. It overcomes the limitations of previous methods, providing a valuable tool for evaluating the quality and composition of black tea.

Greenness Assessment and Validation of HPLC Method for Simultaneous Determination of Resveratrol and Vitamin E in Dietary Supplements.

There is an increasing interest in the use of a combination of trans-resveratrol and vitamin E in dietary supplements. Determination of the content of both components is essential for confirmation of the quality of the product. To establish the applicability and ensure the greenness of the previously developed high-throughput HPLC/UV method for the simultaneous determination of trans-resveratrol and alpha-tocopherol acetate (vitamin E) in dietary supplements. Separation was performed on RP C8 Select B chromatographic column, using acetonitrile and water in the mobile phase, with gradient elution. Full method validation was performed in accordance with ICH Q2(R1). The greenness of the method was assessed using the analytical eco-scale (AES) methodology and the analytical greenness metric (AGREE). The method is selective, linear, precise, and accurate over defined concentration ranges (185-369 µg/mL of trans-resveratrol and 37-75 µg/mL of alpha-tocopherol acetate), and it has a suitable sensitivity (limits of detection and quantification are 7.7 and 23.3 µg/mL for resveratrol and 2.6 and 7.8 µg/mL for tocopherol acetate, respectively). The obtained analytical eco-scale score of 77 and the pale green AGREE pictogram with an overall score of 0.61 confirm the method's greenness. The sensitivity and selectivity of the method, its short analysis time (7 min), the low negative environmental impact, and the simple sample preparation make the method readily applicable to inline quality control procedures. A method for simultaneously analyzing vitamin E and resveratrol in dietary supplements is presented. The method is rapid, includes a simple sample preparation procedure, and has a low environmental impact.

Ecofriendly HPLC Method for Simultaneous Determination of the Co-Prescribed Drugs in Chemotherapy Omeprazole, Ondansetron and Deflazacort in Spiked Human Plasma.

Green, selective and accurate high-performance liquid chromatography (HPLC) chromatographic method is presented for simultaneous separation and quantitation of the co-prescribed drugs in chemotherapy omeprazole, ondansetron and deflazacort in spiked human plasma. An isocratic HPLC separation was performed on X Bridge C18 (4.6 × 250mm) column with 5μm particle size using mobile phase consisting of methanol: ammonium acetate buffer pH4 adjusted by acetic acid (60: 40, v/v). The injection volume was 20μL with UV detection wavelength at 237nm at room temperature. Flow rate of the mobile phase was adjusted to be 2.0ml/min. Dexamethasone was used as internal standard to correct the variation during sample pretreatment. FDA guidelines were followed to validate the developed method. Successful application of the developed method was revealed by simultaneous determination of omeprazole, ondansetron and deflazacort in spiked human plasma in ranges of 1-20, 0.1-8 and 0.2-8μgmL-1 for omeprazole, ondansetron and deflazacort, respectively. Four greenness assessment tools were used to evaluate the greenness of the developed method and the results were accepted. This method permitted the accurate simultaneous determination of the studied drugs, thus it can be used during therapeutic drug monitoring in daily clinical practice.

Efficient isolation of polyunsaturated amides from Zanthoxylum bungeanum and assessment of their anti-inflammatory potential

A targeted extraction, enrichment and separation method for polyunsaturated amides from Zanthoxylum bungeanum was established. Three main polyunsaturated amide monomers with high purity were isolated at the same time: hydroxyl-α-sanshool (HAS), hydroxyl-β-sanshool (HBS), hydroxyl-ε-sanshool (HES). This efficient method combined high-pressure supercritical fluid extraction (HPSFE) and industrial chromatography purification technology. By studying the extraction kinetics of target polyunsaturated amide monomers, the operation parameters of HPSFE were optimized. Meanwhile, a HPLC method for simultaneous determination of three polyunsaturated amide monomers was established in order to optimize process. Under the optimal conditions of high-pressure extraction, the extraction equilibration time was 45 min, the content of total polyunsaturated amides reached 27.53 %, and the yield was 2.89 %. At last, three high-purity polyunsaturated amide monomers were obtained, namely HES (95.16 %), HAS (98.43 %), and HBS (98.72 %). The in vitro cell assay showed that IC50 values of HES, HAS and HBS on the inflammatory factor NO were 14.5, 14.7 and 13.4 μg/mL, respectively. IC50 values on TNF-α and IL-6 were 21.2, 29.4, 23.9 μg/mL and 8.2, 7.0, 8.7 μg/mL, similarly. The Pharmacological studies showed that PA have obvious anti-inflammatory activity as one of the main effective components of ZB.

Transferrin functionalized nanostructured lipid carriers for targeting Rivastigmine and Resveratrol to Alzheimer's disease: Synthesis, in vitro characterization and brain uptake analysis

The management of Alzheimer's disease (AD) remains challenging owing to its complex pathophysiology and blood-brain barrier (BBB) restrictions. A rationally designed combination drug multi-target strategy has emerged as a promising approach for AD management. Considering the lipophilic nature of BBB and the need to embed dual drugs- Rivastigmine hydrogen tartrate and Resveratrol, nanostructured lipid carriers (NLCs) were used as drug delivery vehicle. NLCs were formulated by water/oil/water (w/o/w) double emulsion method and optimized using central composite design. HPLC method for simultaneous determination of drugs was developed and validated. To target the overexpressed Transferrin (TF) receptors, NLCs were surface functionalized with TF by covalent coupling employing carbodiimide chemistry. The success of conjugation was ascertained by Zeta potential, FTIR and TEM and the Bradford assay which estimated the conjugation efficiency of 47.52 ± 3.2%. A particle size of <100 nm was obtained which is favourable for brain delivery. Sustained release of drugs from nanoparticles was obtained, where TF-NLCs demonstrated a significantly slow release compared to NLCs. The functional stability and biocompatibility of the nanosystems was ensured by DPPH assay and hemolysis test, respectively. The confocal laser scanning microscopy (CLSM) revealed significantly higher (∼1.7 folds) brain uptake of TF-NLCs compared to NLCs in vivo following intraperitoneal administration. Dual drugs loaded TF functionalized NLCs demonstrated favourable in vitro and in vivo attributes for efficient brain targeting and hence AD management.

Development and validation of HPLC method for simultaneous determination of Leflunomide and folic acid in the nanoparticulate system by reversed-phase HPLC

Objective The main objective of this study was to develop a highly sensitive, accurate, and reproducible analytical method for the simultaneous detection of LEF and FA in polymeric nanocarriers. Significance Leflunomide (LEF), is widely employed in the treatment of rheumatoid arthritis (RA). However, long-term delivery of the drug is associated with systemic side effects. Therefore, folate (FA) conjugated LEF nanocarriers were fabricated for targeting the nanocarriers toward activated macrophages. HPLC is considered one of the most sensitive and precise analytical techniques for the simultaneous detection and estimation of different components in a particular sample. Methods Analysis was performed on HPLC (Shimadzu 10 A), having a reversed-phase C-18 column (Beckmen, 250 X 4.6 mm, 5 µm) equipped with a photodiode detector set at a wavelength of 260 nm (LEF) and 285 nm (Folic acid). The isocratic mobile phase was composed of acetonitrile, water, and trimethylamine in a ratio of 65:35:0.5 at pH 4. Rapid analysis of both agents was performed, with a total run time of 10 min (FA = 2.1 ± 0.1 min, LEF = 5.9 ± 1 min) at a 1 mL/min flow rate. Results The assay demonstrated good linearity of 0.9989 of 0.9997 for LEF and FA respectively with a recovery in the range of 95–100%. The method also depicted good specificity, and intra and inter-day precision based on relative standard deviation (RSD) values. Conclusions The study concludes, that the developed method was helpful in the detection and quantitation of lower values of both agents from polymeric nanocarriers.

HPLC analysis of biomarkers of Toluene and Xylene in human urine samples

Introduction: Urinary Hippuric acid (HA) and Methylhippuric acid (MHA) are considered reliable biomarkers for monitoring exposure to toluene and xylenes. A simple and robust HPLC method for simultaneous determination of HA and MHA in urine samples was developed and validated as per Internation Conference on Harmonization (ICH) Q2 guidelines. Methods: Mobile phase consisting of two solvents A &amp; B, solvent A was 0.1% TFA in water, and solvent B was 0.1% TFA in acetonitrile. Separation was performed in gradient elution mode and the time program was as follows - Time (min)/%B: 0/20, 4/20, 11/80, 11.1/90, 16/90, 16.1/20, 25/20. The detection wavelength was 205 nm. Results: The limits of detection (LOD) and quantification (LOQ) of HA in urine were 0.3 and 1.0 mg/L, and for MHA were 0.25 mg/L and 1.0 mg/L respectively and the recovery for HA and MHA were 95% and 98.9% respectively. Conclusion: The developed method was successfully applied for the analysis of urine samples.

Development and validation of HPLC method for simultaneous determination of amlodipine besylate, telmisartan and hydrochlorothiazide in combined tablets

A combination of three active ingredients amlodipine besylate, telmisartan and hydrochlorothiazide has been used and proven effective in the treatment of hypertension in patients with moderate to severe hypertension, while it contributes to a reduction in product costs and increases patient compliance. However, there are no monographs in pharmacopoeias for the simultaneous quantification of these three active ingredients. Therefore, in order to contribute to the establishment of an in-house specification for domestic generic drugs, shorten the time of quantifying each ingredient separately, the study was carried out with the aim of developing and validating procedure for the simultaneous quantification of amlodipine besylate, telmisartan and hydrochlorothiazide by high performance liquid chromatography. In this study, we conducted investigation into different chromatographic conditions, developed an analytical procedure for the simultaneous quantification of amlodipine besylate, telmisartan and hydrochlorothiazide in tablets preparation. The procedure was validated in compliance with ICH guidelines. Suitable chromatographic conditions for simultaneous quantification of amlodipine besylate, telmisartan and hydrochlorothiazide are: Xterra C18 column (250 x 4.6 mm, 5 μm), detection wavelength 230 nm, gradient program of mobile phase: acetonitrile and phosphate buffer pH 3.0, flow rate 1.0 ml/min, sample injection volume 10 μl, column temperature 30°C. The procedure was validated for systematic suitability, accuracy, specificity, linearity, precision, working range and robustness. A procedure for the simultaneous quantification of amlodipine besylate, telmisartan and hydrochlorothiazide in tablets by high performance liquid chromatography has been successfully developed and validated.

Determination of Acephate, Dinotefuran, and Emamectin Benzoate in a Pesticide Formulation: A Single-Laboratory Validation.

The advantage of simultaneous separation and quantification is the reduction of analysis time and consumption of solvents and reagents. The objective of the present investigation was to optimize and validate a novel, rapid, and simple reverse-phase high-performance liquid chromatographic method for the simultaneous determination of acephate, dinotefuran, and emamectin benzoate in a pesticide formulation. The chromatographic separation and quantification were accomplished by using Kromasil CN column (250 mm × 4.6 mm; 5 μm) with a mobile phase consisting of acetonitrile and water [0.1% (v/v) triethylamine, pH 2.7 with 10% (v/v) orthophosphoric acid] in the ratio of 50:50 (v/v) with a flow rate of 1.0 mL/min and diode array detection at the wavelength of (215 nm and 245 nm). The HPLC method was able to separate and quantify all the actives in the formulation by isocratic elution within 10 min. The method was fully validated in accordance with the SANCO and Collaborative International Pesticide Analytical Council guidelines concerning system suitability, specificity, linearity, precision, accuracy, and robustness. All the analytical parameters are within the range of acceptable limits in the guidelines. The validated method was successfully applied to a pesticide formulation. The novelty of the current research work lies in the development of the simple and rapid HPLC method for simultaneous determination of acephate, dinotefuran, and emamectin benzoate in wettable granular formulation.

Development and Validation of an HPLC Method for Simultaneous Determination of Miconazole Nitrate and Chlorhexidine Digluconate in Chitosan-Based Gel Formulations

Miconazole nitrate (MN) and chlorhexidine digluconate (CHX) are the commonly used antimicrobials for topical treatment of dermal infections. Combination of antimicrobials has been investigated to enhance the efficacy of the treatment. Gel formulations based on bioadhesive polymers are preferred for delivery of these drugs. Chitosan is a promising bioadhesive polymer due to its penetration enhancing, antimicrobial and tissue healing properties. Yet, most of the gel-based formulations present analytical challenges during testing the drug content. It was aimed to develop an HPLC method for simultaneous determination of MN and CHX in chitosan-based gel formulations. Different solvent combinations were investigated for extraction of drugs from the gels. HPLC conditions such as mobile phase, flow rate, run time, column temperature and wavelength were explored. The method was validated according to ICH guideline Q2(R1). MN and CHX were extracted in solvent composition same with the mobile phase. The method was employed on ACE-C8 column at 40°C by isocratic elution using the mobile phase consisting of methanol:phosphate (75:25 v/v) buffer (containing triethylamine). Flow rate was 1 mL/min. The drugs were detected at 254 nm (CHX) and 230 nm (MN). Linearity was obtained between 5 to 80 μg/mL for both drugs. LOD and LOQ obtained for CHX was 1.61 and 1.06, for MN: 4.87 and 3.21 µg/mL, respectively. A new validated HPLC method was developed for simultaneous determination of CHX and MN in chitosan-based gels, with 98 to 102% recovery, without any interference with the excipients.

Enantioselective Green HPLC Method for Simultaneous Determination of Enantiomer, and Potential Impurities in Apremilast Drug Substance

A single reversed-phase HPLC method was developed for the quantification of seven impurities of Apremilast and its enantiomer in the drug substance. An immobilized chiral stationary phase with a chiral selector “tris (3,5-dimethyl phenyl carbamate) derivative of amylose-Chiralpak IA-3 (250 mm × 4.6 mm, 3 μm) was employed to achieve the desired separation. A mobile phase consists of buffer (0.01M NH4HCO3, PH 8.0) and acetonitrile in the ratio 50:50 (v/v) and is pumped at a flow rate of 0.4 mL/min with an isocratic elution mode. The column oven temperature is set at 25°C, and the chromatographic output is monitored at 225 nm with a total run time of 45 min. A test concentration of 500 µg/mL of Apremilast in the mobile phase is used with an injection volume of 10 µL. Induced degradation studies were carried out to study the intrinsic chemical behavior of the drug. The degradation sample solutions were utilized to demonstrate the stability-indicating nature of the developed analytical method. The obtained mass number [M+H]+ for the primary degradation product formed in acid hydrolysis was identified as 418.36 and in base hydrolysis 478.12. The two impurities were identified as impurity-5(Deacetylated impurity), and impurity-2(open ring acid impurity), respectively, and the degradation pathways were established. Following ICH Q2 and USP<1225>guidelines, complete method validation was carried out. The RSD for the drug and impurities in interday and intraday studies are less than 4.0%, and the recoveries for the impurities are between 96.1-102.1% and linearity r ≥ 0.9997. LOQ results for the drug and impurities are between 0.052 µg/mL and 0.107 µg/mL, and LOD results are between 0.016 µg/mL and 0.032 µg/mL. The greenness of the method was evaluated by using an analytical eco scale, GAPI, and AGREE, and it was found that the method is green.

The Uncertainty Profile Used for Full Validation of the HPLC Method to Determine 22 Azo Amines in Fabrics.

Azo dyes are synthetic dyes that can degrade to carcinogenic amines, therefore determining their exact concentration is crucial. The purpose of this study is to demonstrate that an HPLC method may be used to quantify 22 azo dye amines in a textile matrix. The HPLC-DAD (Diode Array Detector) technique for measuring 22 amines in a textile matrix was tested using a new graphical statistical strategy based on the building of β-content, γ-confidence tolerance intervals, which allows for good qualimetry of analytical procedures. In this way, we established the approach's ability to perform a full validation of the analytical technique as well as its ability to estimate measurement uncertainty in a simple and uncomplicated way with minimum effort, utilizing only the data collected during the analytical validation. The findings suggest that this technique can properly assess the validity of the HPLC method for simultaneous determination of 22 amines, with 67-90% of compliance results falling within acceptable limits. Using the uncertainty profile methodology, the full validation of an HPLC method for the determination of 22 azo amines in a textile matrix was successfully established. The development and evaluation of an efficient HPLC technique for the simultaneous determination of 22 azo amines in a textile matrix have been completed. The total reliability of the analytical procedure was established by evaluating its measurement uncertainty at each concentration level using the suggested uncertainty profile approach and confirming proportions 67 to 90% of compliance.

Concepts for a New Rapid and Simple HPLC Method for Simultaneous Determination of Metoprolol and Meldonium in Pharmaceutical Dosage Forms

Simultaneous determination of the tandem of drugs, like meldonium and metoprolol, with enormous polarity differences between them, requires thorough research and careful selection of chromatographic conditions. The three different CN-cyano groups with link-based particle columns, LiChrospher CN, Waters Spherisorb CNRP, Zorbax CN SB stationary phases, were tested, in an isocratic elution system, with a running mobile phase containing various concepts of composition contents. They were first with buffering salts which included acetonitrile and ammonium phosphate in one group, and then without buffering salts but with diluted acids, composed of acetonitrile and diluted acids as the second group. We can conclude that the most optimal concepts, in terms of expressiveness and environmental friendliness, were concepts using of column Zorbax CN SB (4.6 mm i.d. × 250 mm, 5 μm) and mobile phase ACN—0.15% NH4H2PO4 (50:50 and 60:40, v/v). There are very poor available data about ideas and usable information about the development of methods for simultaneous determination of these two active substances with polarity differences between them. We suggest that our work offered detailed and successful solutions for the mentioned aim using less sophisticated equipment for quality control and a lab for routine manufacturing control.

Effect of Wuzi Yanzong prescription on oligoasthenozoospermia rats based on UPLC-Q-TOF-MS metabolomics

Context Wuzi Yanzong Prescription (WYP) has long been used to treat male infertility, with convincing clinical evidence. However, its mechanism of action is not clear. Objective WYP chemical components were quantified by HPLC, and the effect on oligoasthenozoospermia rats was explored based on UPLC-Q-TOF-MS metabonomics technology. Materials and methods The solution was extracted by alcohol and water; the content of eight components in the extracting solution was determined by HPLC. Twenty-four Sprague-Dawley rats were randomly divided into control (CG), model (MG) and administration (PG) groups. Oligoasthenozoospermia was induced for 30 days in all, but the CG with daily oral doses of 20 mg/kg Tripterysium glycosides (TG). PG was given daily oral doses of WYP solution (1.08 g/kg), CG and MG received the same volume of distilled water, all administered for 30 days. After the last administration, the serum samples were collected and detected by UPLC-Q-TOF-MS. Results An HPLC method for simultaneous determination of chlorogenic acid (0.081%), ellagic acid (0.021%), hyperoside (0.032%), verbascoside (0.011%), isoquercitrin (0.041%), astragalin (0.026%), kaempferol (0.009%) and schisandrin (0.014%) was established. Moreover, the 74 potential biomarkers and eight metabolic pathways related to oligoasthenozoospermia were identified by multivariate analysis and metabolite profiling. WYP can regulate 36 markers, mainly involving amino acid metabolism, lipid metabolism and nucleotide metabolism. Discussion and conclusions This is a simple and accurate method for quality control of WYP. It further enriches the potential mechanism of WYP in the treatment of oligoasthenozoospermia. It can provide a theoretical basis for the rational application of WYP.

Development and validation of RP HPLC method for simultaneous determination of picroside I, picroside II, phyllanthin and boeravinone B in a polyherbal hepatoprotective tablet formulation

Purpose: To develop and validate RP-HPLC method for simultaneous determination of four active ingredients: picroside I (PSI), picroside II (PSII), phyllanthin (PHY) and boeravinone-B (BVB) in a polyherbal hepatoprotective tablet formulation.Methods: The study was carried out using Waters X-Bridge, C18 (250 mm x 4.6 mm, 5 μm) column with mobile phase consisting of 5 mM ammonium acetate in 10 % methanol and acetonitrile, with gradient programme at dual wavelengths of 220 nm and 274 nm and flow speed of 1 mL.min-1. The procedure was validated with respect to specificity, linearity, precision, accuracy, system suitability, limit of detection (LOD) and of quantification (LOQ), and robustness in line with ICH specifications.Results: The method was linear within the concentration range of 25 to 200 %, and the values of correlation coefficients (R2) were &gt; 0.999. Intra-day and inter-day RSDs of PAs and RTs were &lt; 5.0 %, with recovery in the range of 100.0 - 106.0 %.Conclusion: The four active ingredients have with good resolution with regard to the method used. The method is rapid, simple, highly selective, sensitive and cost effective, which make it an efficient method for quality assurance.

Development and validation of HPLC method for simultaneous determination of Ketotifen Fumarate and Salbutamol Sulfate in bulk and tablets dosage forms

The aim of the study was to develop and validate a new, rapid, sensitive, simple, accurate and reproducible HPLC method for simultaneous determination of ketotifen fumarate and salbutamol sulfate. Simultaneous HPLC method was developed using RP-C18 stainless steel analytical column 4.6×150 mm C18.Acetonitrile and phosphate buffer pH 4 (30 : 70) were used as mobile phase and wavelength was adjusted to 276 nm for detection of drugs. Developed method was validated for its specificity, accuracy, precision, linearity and robustness. Method was also applied to quantify drugs in commercial tablets. Chromatogram obtained by newly developed method for simultaneous determination of two anti-asthmatic drugs, having well distinguished peaks for both drugs. Retention time of ketotifen fumarate and salbutamol sulfate were 2.69 minutes and 9.47 minutes respectively. Total run time for both anti-asthmatic drugs was 12 minutes. Limit of quantification for ketotifen fumarate and salbutamol sulfate was 1 ng/ml and 1.50 ng/ml respectively. Limit of detection of ketotifen fumarate and salbutamol sulfate was 3.03 and 4.54 respectively. A simple, easy, precise and new method was developed for simultaneous quantification of frequently used anti-asthmatic drugs. Developed method may prove effective and beneficial in determination of ketotifen fumarate and salbutamol sulfate in bulk and other pharmaceutical dosage forms.

Development and Validation of an HPLC Method for Simultaneous Determination of Capsaicinoids and Camphor in Over-the-Counter Medication for Topical Use.

A reverse-phase high-performance liquid chromatography method was developed to determine and quantify capsaicin (trans-8-methyl-N-vanillyl-6- nonenamid), dihydrocapsaicin (8-methyl-N-vanillylnonanamide), and camphor (trimethylbicyclo[2.2.1]heptan-2-one). It is applicable in analyses of over-the-counter (OTC) medications for topical use and raw materials such as chili pepper oleoresin. Chromatographic separation was carried out on a C18 column using an isocratic flow of the mobile phase containing acetonitrile and ultrapure water in a ratio of 2:3, with pH adjusted to 3.2 using glacial acetic acid, and a flow rate of 1.5 mL/min. The concentration of the eluting compounds was monitored by a diode-array detector at a wavelength of 281 nm. The method was evaluated for several validation parameters, including selectivity, accuracy (confidence intervals < 0.05%), repeatability, and intermediate precision. The limit of detection (LOD) was determined to be 0.070 µg/mL for capsaicin, 0.211 µg/mL for dihydrocapsaicin, and 0.060 µg/mL for camphor. The limit of quantification (LOQ) was determined to be 0.212 µg/mL for capsaicin, 0.640 µg/mL for dihydrocapsaicin, and 0.320 µg/mL for camphor. Linearity was set in the range of 2.5–200 µg/mL for capsaicin and dihydrocapsaicin and 25–2000 µg/mL for camphor. The suggested analytical method can be used for quality control of formulated pharmaceutical products containing capsaicinoids, camphor, and propolis.

Development and validation of an HPLC method for simultaneous determination of imatinib mesylate and rifampicin

Development and validation of an HPLC method for simultaneous determination of imatinib mesylate and rifampicin

Quality by design approach for green HPLC method development for simultaneous analysis of two thalassemia drugs in biological fluid with pharmacokinetic study.

This work implements a combined experimental approach of analytical quality-by-design (AQbD) and green analytical chemistry (GAC) to develop an HPLC method for simultaneous determination of the two thalassemia drugs, deferasirox (DFX) and deferiprone (DFP), in biological fluid for the first time. This integration was designed to maximize efficiency and minimize environmental impacts, as well as energy and solvent consumption. To accomplish this goal, an analytical quality-by-design approach was performed, beginning with quality risk assessment and scouting analysis, followed by Placket–Burman design screening for five chromatographic parameters. Critical method parameters were thoroughly recognized and then optimized by using a two levels-three factors custom experimental design to evaluate the optimum conditions that achieved the highest resolution with acceptable peak symmetry within the shortest run time. The desirability function was used to define the optimal chromatographic conditions, and the optimal separation was achieved using an XBridge® HPLC RP-C18 (4.6 × 250 mm, 5 μm) column with ethanol : acidic water at pH 3.0 adjusted by phosphoric acid in the ratio of (70 : 30, v/v) as the mobile phase at a flow rate of 1 mL min−1 with UV detection at 225 nm at a temperature of 25 °C. Linearity was obtained over the concentration range of 0.30–20.00 μg mL−1 and 0.20–20.00 μg mL−1 for DFX and DFP, respectively, using 20.00 μg mL−1 ibuprofen (IBF) as an internal standard. The established method's greenness profile was evaluated and measured using various assessment tools, and the developed method was green. For the validation of the developed method, FDA recommendations were followed, and all the results obtained met the acceptance criteria. The suggested method was successfully used to study the pharmacokinetic parameters of DFX and DFP in rat plasma. Due to the substantial increase in bioavailability of the two iron chelating drugs, the results from this study strongly recommend their co-administration.