HPLC Method For Simultaneous Analysis Research Articles

Quality characteristics and HPLC detection of phosphodiesterase inhibitors in some herbal capsules indicated for male sexual disorders in Ghana

BackgroundHerbal medicines used for male sexual disorders are widespread across the globe and have been noted as likely candidates for adulteration. To assure access to safe and quality herbal products, this study aimed at sampling some herbal capsules indicated for male vitality for quality analytical checks.MethodsHerbal capsules sampled from two regions were subjected to physicochemical assessment, using pharmacopeial and regulatory standards. Microbial quality analysis was also performed on the selected products. A validated HPLC method for simultaneous analysis of sildenafil citrate and tadalafil was used to detect and quantify the possible presence of phosphodiesterase inhibitors in the selected products.ResultsOut of a total of 57 herbal capsules, 19 were indicated for male sexual related illnesses of which only 8 fulfilled the inclusion criteria of being used exclusively for male vitality. The majority (62.5%) of the herbal products failed the weight uniformity test but all of them disintegrated within the acceptable limit of 30 min. The moisture and heavy metal contents were within limits of acceptability. Majority (87.5%) of the herbal products failed the test for the aerobic bacteria count; however, none of the isolated bacteria was pathogenic. Sildenafil citrate was detected in the majority (87.5%) of the herbal products in concentrations of 0.98–33.95 mg/g. Two products contained both sildenafil and tadalafil. High batch-to-batch variability was recorded in some physicochemical parameters and pharmaceutical adulterants.ConclusionHigh incidence of quality fails and adulteration of herbal capsules for sexual disorders observed means there is a real risk of unintended dosing with phosphodiesterase inhibitors. This represents a public health issue and necessitates tighter monitoring of production standards by the regulator.

Developed and validated HPLC method for simultaneous analysis of key flavonoids and phenolcarboxylic acids in the hawthorn-based cardiotonic Forticor®

<p>Relatively simple, but very reliable, one-step qualitative/quantitative HPLC analysis of four key flavonoids, and a phenolcarboxylic acid, has been introduced. For substrate/sample preparations, either triple percolations, using 70% ethanol-water (V/V), or/and ultrasonic bath methanol extractions, were carried out to obtain the desired isolates yielding quercetin, isoquercitrin, hyperoside, vitexin and chlorogenic acid. The method is linear, over the studied range of 1.05 <span style="background-color: var(--bs-body-bg); font-weight: var(--bs-body-font-weight); text-align: var(--bs-body-text-align);">– </span><span style="background-color: var(--bs-body-bg); font-weight: var(--bs-body-font-weight); text-align: var(--bs-body-text-align);">210.00, 6.25 </span><span style="font-weight: var(--bs-body-font-weight); text-align: var(--bs-body-text-align); background-color: var(--bs-body-bg);">–</span><span style="font-weight: var(--bs-body-font-weight); text-align: var(--bs-body-text-align); background-color: var(--bs-body-bg);"> </span><span style="font-weight: var(--bs-body-font-weight); text-align: var(--bs-body-text-align); background-color: var(--bs-body-bg);"> 250.00, 25.00 – </span><span style="font-weight: var(--bs-body-font-weight); text-align: var(--bs-body-text-align); background-color: var(--bs-body-bg);">250.00, 7.50 – 150.00 and 1.04 - 10.40 µg/mL for chlorogenic acid, vitexin, hyperoside, isoquercitrin and </span><span style="font-weight: var(--bs-body-font-weight); text-align: var(--bs-body-text-align); background-color: var(--bs-body-bg);">quercetin, respectively. The correlation coefficient for each of the analytes was greater than 0.999. The </span><span style="font-weight: var(--bs-body-font-weight); text-align: var(--bs-body-text-align); background-color: var(--bs-body-bg);">intra-day and inter-day precision of the analysis was below 2.00 and 3.00 %, respectively. The accuracy of the analysis is verified by the standard addition method, using three different concentrations of each component in the tested materials, with recovery values obtained in the range of 98.04 </span><span style="font-weight: var(--bs-body-font-weight); text-align: var(--bs-body-text-align); background-color: var(--bs-body-bg);">–</span><span style="font-weight: var(--bs-body-font-weight); text-align: var(--bs-body-text-align); background-color: var(--bs-body-bg);"> </span><span style="font-weight: var(--bs-body-font-weight); text-align: var(--bs-body-text-align); background-color: var(--bs-body-bg);">102.47% </span><span style="font-weight: var(--bs-body-font-weight); text-align: var(--bs-body-text-align); background-color: var(--bs-body-bg);">(RSD ≤ 1.85%). The detection limits were 0.6, 0.5, 0.5, 0.8 and 0.3 µg/mL chlorogenic acid, vitexin, </span><span style="font-weight: var(--bs-body-font-weight); text-align: var(--bs-body-text-align); background-color: var(--bs-body-bg);">hyperoside, isoquercitrin and quercetine, respectively. The developed method is convenient in the </span><span style="font-weight: var(--bs-body-font-weight); text-align: var(--bs-body-text-align); background-color: var(--bs-body-bg);">routine control of hawthorn raw materials and pharmaceutical dosage forms made from hawthorn </span><span style="font-weight: var(--bs-body-font-weight); text-align: var(--bs-body-text-align); background-color: var(--bs-body-bg);">berries or flowerbearing branches. The new HPLC method may be used for the quality control of the </span><span style="font-weight: var(--bs-body-font-weight); text-align: var(--bs-body-text-align); background-color: var(--bs-body-bg);">commercially used cardiotonic Forticor<sup>®</sup> intended for increasing/strengthening cardiac muscles, in </span><span style="font-weight: var(--bs-body-font-weight); text-align: var(--bs-body-text-align); background-color: var(--bs-body-bg);">order to lower the risks of atherosclerosis, hypertension and congestive heart failure.</span></p>

Green HPLC Method for Simultaneous Analysis of Three Natural Antioxidants by Analytical Quality by Design.

Glutathione, silybin, and curcumin are well-known potential antioxidants that are recommended as adjuvant therapy in cancer treatment. Based on the principles of Analytical Quality by Design (AQbD) and green analytical chemistry, a simple, robust, and environmentally benign HPLC method for the simultaneous estimation of glutathione, silybin, and curcumin in bulk and formulation was performed. Elution was achieved by an Agilent Eclipse XDB C18 (150 mm × 4.6 mm id, 3.5 μm) column using a gradient mobile phase composed of ethanol-water pH 6.7 (with 0.1%, v/v orthophosphoric acid) and 1.07 mL/min flow rate with PDA detection at 215 nm. Critical method variables were identified by risk assessment using an Ishikawa diagram, and multivariate optimization of the experimental conditions for the HPLC technique was accomplished by central composite design using design of experiments (DoE) software. The separation was achieved within 15 min, where the retention time of glutathione, silybin, and curcumin were 3.3, 4.9, and 7.3 min, respectively. The standard curve was linear in the range of 3.75-26.25 µg/mL for glutathione, 62.50-437.50 µg/mL for silybin, and 12.5-87.50 µg/mL for curcumin. The developed method was validated as per ICH guidelines Q2 (R1), and all the parameters are within specified limits. The proposed method is simple, precise, and robust, which can be employed for routine analysis and also concluded to be a greener approach according to AGREE, Green Analytical Procedure Index, and analytical eco-scale tools. The chosen antioxidants were evaluated for the very first time simultaneously using the chromatographic technique in bulk and dosage forms employing green solvents. The peak purity of all three compounds was studied using a PDA detector. Wastage was reduced in terms of time, cost, and solvents by employing AQbD elements and tools. Complete application of environmentally sustainable safe solvents were employed.

Experimental Design Approach for Development of HPLC Method for Simultaneous Analysis of Triamcinolone, Nystatin, and Gramicidin in Industrial Wastewater

This study used an experimental design approach to optimize an HPLC method for the simultaneous determination of three pharmaceutical residues (triamcinolone, nystatin, and gramicidin) in industrial wastewater samples. The goal of using an experimental design approach was to maximize the method performance through separation enhancement and shortening the time of analysis and/or minimizing the environmental effects through the reduction in wastes and sample treatment. To achieve this goal, two steps were performed: a full factorial screening design for the three chromatographic variables, and optimization design using central composite design to select the optimum conditions that accomplished the highest resolution between adjacent peaks within a minimum run time of less than 5 min. The optimal chromatographic conditions derived from Minitab software using the desirability function were applied. Separation was carried out on a Zorbax C18 column (250 mm × 4.6, 5 μm) with gradient elution of a mobile phase composed of methanol and 0.25 M potassium dihydrogen phosphate buffer (pH 3.6) at different UV detections. For the validation of the developed HPLC method, ICH guidelines were followed, and the obtained results were found to be in compliance with the acceptance criteria. Linearity was over the concentration range of 1.00–25.00 μg/mL for triamcinilone and nystatin and 10.00–50.00 µg/mL for gramicidin. The proposed method was successfully applied to quantify the three studied pharmaceutical compounds in rinsing wastewater samples.

Micellar eco-friendly HPLC method for simultaneous analysis of ternary combination of aspirin, atorvastatin and ramipril: application to content uniformity testing

BackgroundCardiovascular disease medications such as aspirin (ASP), statins like atorvastatin (ATR), and blood pressure-lowering drugs including ACE inhibitors like ramipril (RAM) have been included in the World Health Organization (WHO) Essential Medicines List (EML) for many years. Therefore, there is a strong demand to develop a simple, rapid, and sensitive analytical method that can detect and quantitate the ternary mixture of these analytes in pharmaceutical preparations in a short run time. Lately, the analytical community focused on eliminating or reducing hazardous chemicals and solvents usage.ResultsA green, fast, selective, and cost-effective micellar HPLC method was established and validated for the concurrent determination of ternary combination of ASP, ATR, and RAM in the pure form and pharmaceutical preparations. Resolution of the three drugs was achieved by using a monolithic column and a micellar mobile phase consists of 0.3% triethylamine (TEA) in 90: 10 an aqueous solution of 0.12 M sodium dodecyl sulfate (SDS): n-propanol, (v/v). The pH was adjusted to 2.5 using orthophosphoric acid and a flow rate of 1.5 mL/min. was applied. To ensure method reproducibility, Valsartan (VAL) was utilized as an internal standard (IS). The UV detection of the studied drugs was performed at 210 nm. Good linearity for the three drugs was obtained over the concentration ranges of 1.0-200.0 mg/mL, 0.5-200.0 mg/mL, and 5.0-100.0 mg/mL with correlation coefficients of 0.9998,0.9999 and 0.9999 for ASP, ATR, and RAM respectively. The method sensitivity was revealed by the relatively small values of limits of detection (LOD) (0.19, 0.13 and 0.30 mg/mL) and limits of quantitation (LOQ) (0.63, 0.44 and 0.99 mg/mL) for ASP, ATR, and RAM, respectively. The retention times of ASP, ATR and RAM were 1.50, 2.3 and 4.3 min., respectively.ConclusionsThe suggested technique was employed for the analysis of the three drugs in their prepared tablets maintaining the recommended pharmaceutical ratio without any interference from excipients. The method was further extended to content uniformity testing of RAM. The results were validated according to international council for harmonisation (ICH) guidelines.

A new HPLC method for simultaneous analysis of sterols, tocopherols, tocotrienols, and squalene in olive oil deodorizer distillates using a monolithic column with chemometric techniques

Simultaneous determination: sterols, tocopherols, tocotrienols, and squalene in an olive oil deodorizer distillate have been separated using a monolithic column.

Application of central composite design and structural analysis to the optimization of HPLC method for simultaneous analysis of hydrochlorothiazide, amlodipine besylate and valsartan

Application of central composite design and structural analysis to the optimization of HPLC method for simultaneous analysis of hydrochlorothiazide, amlodipine besylate and valsartan

Development and Validation of Stability Indicating HPLC Method for the Simultaneous Estimation of LevocloperastineFendizoate and Chlorpheniramine Maleate in Pharmaceutical Dosage Form

The aim of the present study was to develop and validate stability indicating HPLC method for simultaneous estimation of LevocloperastineFendizoate (LCP) and Chlorpheniramine Maleate (CPM). HPLC method for simultaneous analysis of both drugs was developed and validated according to ICH guideline. Efficient chromatographic separation was achieved on ODS column C18 (250 mm × 4.6 mm, 5 μm) using the optimized mobile phase. Stability indicating assay method was carried out by different stress degradation conditions. In HPLC method, the Retention time for LCP and CPM was 3.173 min and 5.060 min using optimized mobile phase Phosphate buffer (pH-3.5): Methanol (60:40 %v/v) and UV detection at 273 nm. The degradation of LCP, CPM and Formulation was shown to be highest in alkaline condition. Linearity was observed in concentration range of 20-80 μg/ml for LCP and 4-16 μg/ml for CPM. The correlation coefficient of LCP and CPM were respectively 0.9992 and 0.9994. All validation parameters were within the acceptable range. The LOD and LOQ values for HPLC method were found to be 0.146 μg/ml and 0.444 μg/ml for LCP and 0.0113 μg/ml and 0.0344 μg/ml for CPM respectively. The Method validation parameters showed %RSD value less than 2.

Stability indicating simplified HPLC method for simultaneous analysis of resveratrol and quercetin in nanoparticles and human plasma

Resveratrol and quercetin are well-known polyphenolic compounds present in common foods, which have demonstrated enormous potential in the treatment of a wide variety of diseases. Owing to their exciting synergistic potential and combination delivery applications, we developed a simple and rapid RP-HPLC method based on isosbestic point detection. The separation was carried out on phenomenex Synergi 4μ Hydro-RP 80A column using methanol: acetonitrile (ACN): 0.1% phosphoric acid (60:10:30) as mobile phase. The method was able to quantify nanograms of analytes simultaneously on a single wavelength (269nm), making it highly sensitive, rapid as well as economical. Additionally, forced degradation studies of resveratrol and quercetin were established and the method’s applicability was evaluated on PLGA nanoparticles and human plasma. The analytes peaks were found to be well resolved in the presence of degradation products and excipients. The simplicity of the developed method potentializes its suitability for routine in vitro and in vivo analysis of resveratrol and quercetin.

Development and validation of a reversed-phase HPLC method for simultaneous analysis of butylhydroxyanisol, simvastatin and its impurities in tablet dosage forms

A simple, rapid, and sensitive RP-HPLC method using photodiode array detection was developed and validated for the simultaneous determination of butylhydroxyanisol and simvastatin with its impurities in tablet forms. Chromatographic separation was achieved on a Phenomenex Hypersil (250×4.6 mm, 5 μm) column using the mobile phase acetonitrile-sodium acetate (12 mM) buffered to 4.2 with glacial acetic acid. The flow rate was 1.7 mL/min, and the UV detection were made at 238 nm for simvastatin and its impurities and at 290 nm for butylhydroxyanisol. The system suitability solution used for peak impurity identification was generated in-situ without use of any impurity reference standard. The method was validated according to ICH Q2(R1) guidelines, and the acceptance criteria for accuracy, precision, linearity, specificity, robustness, LOD, and LOQ, were met in all cases. Moreover, the reproducibility results obtained by 22 Official Medicines Control Laboratories (OMCL) of European Directorate were satisfactory. The compounds selected for impurity validation were based on those found during long term and accelerate stability studies carried out on several formulation tablets from Moroccan and other markets. The described method was robust and successfully applied in quality control laboratories for routine analysis to determine the butylhydroxyanisol and simvastatin with its impurities content in tablet dosage forms.

HPLC Method for Simultaneous Analysis of Ranitidine and Metronidazole in Dosage Forms

HPLC Method for Simultaneous Analysis of Ranitidine and Metronidazole in Dosage Forms

A simple HPLC method for simultaneous analysis of 7 bioactive constituents for Liuwei Dihuang Pill and its application in quality consistency evaluation

A combinative method using HPLC fingerprinting and quantitative analysis was developed and validated for manufacturer-to-manufacturer quality consistency evaluation of Liuwei Dihuang pills (LDP). Seven bioactive constituents of LDP, including gallic acid, 5-hydroxymethyl furfural, morroniside, paeoniflorin, sweroside, loganin and paeonal, were selected as markers to evaluate the similarities of 20 samples from 16 manufacturers. The similarity values for the 20 LDP samples were all greater than 0.92, indicating that there is a general quality consistency for different manufacturers. Additionally, the 7 selected bioactive compounds were in accordance with the compatibility principle of “the monarch drug, the ministerial drug, the adjunctive drug and the messenger drug” of Traditional Chinese Medicine (TCM) prescription and the multi components quality evaluation requirements of TCM as well. The detection and quantification limits for these 7 bioactive components were all less than 0.11 μg mL−1 and 0.38 μg mL−1, respectively. The mean recovery for all the investigated constituents was acceptable with an accuracy of 98–102%. The intra- and inter-day precisions of this method were less than 4.73%, and stability and repeatability were less than 3.23%. This method is simple and reliable, and also shows that the combination of the chromatographic fingerprint and quantitative analysis offers an efficient solution to quality consistency evaluation of herbal preparations.

An Optimized Ion-Pair HPLC Method for Simultaneous Analysis of Nucleoside Triphosphate Levels in Hepatoma Cell Line

An improved ion-pair HPLC method was developed for the simultaneous determination of ribonucleoside triphosphates and their corresponding deoxyribonucleoside triphosphates in HepG2 cell extracts. HPLC conditions, flow rate and column temperature were optimized and good linearity (r 2 > 0.9993) was obtained over the investigated concentration ranges. Reproducibility was evaluated by intra- and inter-day assays and RSD values were below 5.39%. Recoveries ranged from 98.2 ± 3.49% to 103.1 ± 1.75%, respectively. Finally, the method was successfully applied to the analysis of eight compounds present in HepG2 cell extracts.

A Simple and Sensitive HPLC Method for Simultaneous Analysis of Nabumetone and Paracetamol in Pharmaceutical Formulations

This paper describes a high‐performance liquid chromatographic method for simultaneous estimation of nabumetone and paracetamol in binary mixture. The method was based on RP‐HPLC separation and quantitation of the two drugs on hypersil C‐18 column (250 mm × 4.6 mm) using a mobile phase consisting of acetonitrile and 0.05% aqueous acetic acid (70:30v/v) at flow rate of 1 mL min-1. Quantitation was achieved with PDA detector at 238 nm based on peak area with linear calibration curves at concentration ranges 5‐25 µg mL-1 for both the drugs. Naproxen sodium was used as internal standard. The method has been successively applied to pharmaceutical formulation. No chromatographic interference from the tablet excipients was found. The method was validated in terms of precision, robustness, recovery and limits of detection and quantitation. The intra and inter‐day precision and accuracy values were in the acceptance range as per ICH guidelines.

Analysis and Identification of Astaxanthin and its Carotenoid Precursors from Xanthophyllomyces dendrorhous by High-Performance Liquid Chromatography

This study presents an HPLC method for simultaneous analysis of astaxanthin and its carotenoid precursors from Xanthophyllomyces dendrorhous. The HPLC method is accomplished by employing a C18 column and the mobile phase methanol/water/acetonitrile/ dichloromethane (70:4:13:13, v/v/v/v). Astaxanthin is quantified by detection at 480 nm. The carotenoid precursors are identified by LC-APCI-MS and UV-vis absorption spectra. Peaks showed in the HPLC chromatogram are identified as carotenoids in the monocyclic biosynthetic pathway or their derivatives. In the monocyclic carotenoid pathway, 3,3'-dihydroxy-beta,psi-carotene-4,4'-dione (DCD) is produced through gamma-carotene and torulene.

ABSORPTION OF INTRAVESICALLY APPLIED DRUGS: COMPARISON OF NORMAL AND ILEAL AUGMENTED RABBIT BLADDER

Drug absorption within urinary diversions has been reported to cause prolonged and higher grade toxicity. Therefore, continuous urine drainage has been recommended during chemotherapy in patients with continent urinary diversion. We developed an animal model in which to examine the significance of drug absorption in normal rabbit bladders compared with ileal augmented bladders. Ten rabbits with ileal bladder augmentation and 5 control animals were used for absorption studies with methotrexate and ofloxacin. One, 4 and 12 months after surgery the rabbits received an intravesical instillation of either drug. During 2 hours blood samples were drawn. To avoid overfilling the bladder by urine it was emptied after 30, 60, 90 and 120 minutes, and refilled with fresh solution to yield a relatively constant drug amount. After 12 months the animals were sacrificed. Area of the native bladder wall and of the intestinal segment was measured to allow the correction of absorbed drug amounts per surface area. Moreover, the median villous heights of native and augmented ileum were compared histologically. Serum levels of methotrexate and ofloxacin were determined by high performance liquid chromatography. The maximum serum concentration of the 2 drugs was typically seen after 60 to 120 minutes. During the 1-year period peak serum concentrations of ofloxacin remained consistently higher in ileal augmented than in control rabbits, although this did not achieve statistical significance at all instillation time points. For methotrexate a statistically significant difference was not shown for either time point. When absorption was corrected for total bladder surface area, the enhancement of ofloxacin absorption by ileal augmentation weakened and attained statistical significance only at the 1-month time point. Histological examinations after 12 months showed that augmented intestinal mucosa had a significantly smaller villous height than native ileum. Our data demonstrate that bladder surface is the most important factor for increased absorption but time dependent histological changes of the integrated intestinal mucosa also influence absorption. There is a broad interindividual discrepancy. Therefore, general rules in patients with urinary diversion may not be justified.

Improved HPLC method for simultaneous analysis of cortisol, 11-deoxycortisol, prednisolone, methylprednisolone, and dexamethasone in serum and urine

Improved HPLC method for simultaneous analysis of cortisol, 11-deoxycortisol, prednisolone, methylprednisolone, and dexamethasone in serum and urine

A HPLC method for simultaneous analysis of NAD(P)+ and NAD(P)H. Its application to the study of spinach ferredoxin: NADP+ reductase-catalyzed transhydrogenation.

A new HPLC system for simultaneous analysis of NAD+, NADH, NADP+, and NADPH was developed and used to measure the transhydrogenase activity of spinach ferredoxin-N ADP + reductase (EC 1.18.1.2, FNR). The system is based on a reverse-phase HPLC with isocratic elution on an ODS column (4.6 × 50 mm). The four nucleotides were completely separated by developing the column with 0.15 m sodium phosphate/citrate buffer (pH 6.8) containing 1 mm EDTA at 40°C with a flow rate of 1 ml/min. The four nucleotides can be simultaneously assayed within 13 min by monitoring the effluents with a UV detector. It was also indicated that the transhydrogenase activity of spinach FNR can be advantageously assayed by measuring the nucleotides by this method.