Development Of HPLC Method Research Articles

Determination of compounds with laxative action in aloe latex and aloe vera‐based products—Chemometric classification of aloe vera plants (Aloe barbadensis Miller) under different conditions

AbstractBackgroundThe bioactive compounds of latex in aloe vera are associated with several of its medicinal properties. Aloe emodin, aloins A and B, which are present in aloe latex, are known for their laxative action.ResultsThe developed HPLC method for the determination of compounds with purgative effect was performed with a C18 column, and the total analysis time was 22 min. Each leaf from the plants was studied separately for its content in aloins. It was observed that, in contrast with field cultivation plants, the concentrations of aloins in potted plants follow a specific pattern based on the position of the leaves. It was also observed that climatic and environmental conditions have a significant impact on the concentrations of aloins. Principal Component Analysis was performed to evaluate correlations among measurements. The model was able to separate the aloe plants based on their seasonal variation and growing conditions. Finally, it was observed that, in only two out of thirteen products, aloins were present, with the total content exceeding the maximum levels set by EU legislation.ConclusionThe present work provides useful information for the aloins variation among leaves in different positions and plants under various conditions.

Development and validation of the RP-HPLC method for quantification of tavaborole.

The stability-indicating approach for tavaborole quantification was developed and validated to establish a precise, linear, accurate, and robust HPLC method. The development section includes optimizing the detection wavelength, the mobile phase ratio, and the type of column used to achieve the best possible separation and sensitivity for analysis. The chromatographic conditions were established, considering peak symmetry, resolution, and retention time. The mobile phase composition, comprising a buffer: acetonitrile (75 : 25, %v/v), with an injection volume of 15 μL, showed suitable elution and recovery at 265 nm. A constant column oven temperature of 35 °C and a 1 mL min-1 flow rate were maintained. The pH of the buffer was changed to 3.0 by using orthophosphoric acid. Linearity was observed from 5 to 1000 ppm (r2 = 1.00000). The capacity (retention) factor (k) of 3.43 was observed, indicating significant interaction and good separation. Forced degradation (FD) or stress tests were performed for chemical and physical photolytic stress conditions, and the results observed were within the specified limits. The stability in the analytical solution was observed for up to 35 hours at 5 °C, confirming the stability of the solution. Validation of the developed HPLC method confirmed the system's suitability, precision, linearity, accuracy, FD, robustness, and results. All validation criteria for the technique were within acceptable limits.

DoE-assisted HPLC method development and validation of enzalutamide and repaglinide in rat plasma.

Aim: A simple and rapid HPLC technique was developed and validated to simultaneously estimate enzalutamide (ENZ) and repaglinide (REP) in rat plasma.Methods: In silico predictions using DDinter and DDI-Pred indicated possible drug-drug interactions between ENZ and REP. A central composite design was used to identify factors influencing the separation of the drugs. Interactions between chromatographic parameters were studied through 51 experiments, followed by illustration with three-dimensional response surface plots. The four factors optimized for the separation of the two drugs are column temperature (A), % organic strength (B), pH (C) and column type (D).Results: Plate count(R1), tailing factor (R2) and resolution (R3) responses in the experimental design were analyzed with the favorable chromatographic conditions predicted to be 0.1% formic acid and acetonitrile as mobile phases on a Phenomenex C18 LC column (250×4.6mm, 5μm). The method was applied to estimate the drugs in rat plasma using a simple protein-precipitation step and found to be linear, accurate and precise within the ranges of 0.5-16 and 5-50μg/ml for ENZ and REP, respectively.Conclusion: The optimized method can be used in future bioanalytical workflow for drug quantification and drug-drug compatible studies.

Greening of the method for simultaneous determining the enisamium iodide and tilorone dihydrochloride using GC-FID assay

Pharmaceutical companies in Ukraine aspire to develop their innovative medicinal products and successfully introduce them to the global market. However, along with the prospects of increased usage of these pharmaceuticals, there arises a challenge of heightened waste production, making them a part of the over twenty million tons of PPCPs produced annually. Consequently, one of the tasks in producing new pharmaceuticals is the development of methodologies and approaches not only for quality control but also for their determination in the environment matrices.
 The aim. Develop and validate GC-FID chromatographic method for the simultaneous determination of Enisamium iodide and Tilorone dihydrochloride, evaluate their applicability, and compare their "greenness" with the previously developed HPLC method.
 Materials and methods. The determination of the Tilorone dihydrochloride and Enisamium iodide was carried out by gas chromatography with a flame ionization detector using the Rxi-5 ms (30 m long, 0.25 mm outer diameter and 0.25 μm liquid stationary phase thickness)
 Results. Chromatographic GC-FID methods have been developed for the simultaneous determination of Enisamium iodide and Tilorone dihydrochloride. Optimal sample preparation conditions were established, and a validation process was conducted. A comparison with the previously developed HPLC method was made regarding "greenness."
 Conclusions. The developed GC-FID methodology is accurate and more environmentally friendly compared to the previously established methods. It can be recommended to determine Enisamium iodide and Tilorone dihydrochloride in the environmental matrices. It is considered environmentally friendly based on the overall GREENness (AGREE) scale, scoring 0.73 (>0.70), which demonstrates the environmentally favourable nature of the proposed analytical approach

Development and Validation of HPLC Method for Simultaneous Estimation of Minoxidil and Finasteride in Topical Solution

A simple, precise, rapid, accurate HPLC method has been developed and validated for the simultaneous determination of Minoxidil and Finasteride in pharmaceutical dosage form. The chromatographic separation was achieved on ODS C18 column (250mm*4.6mm,5 micrometer particle size) using a mobile phase comprising Buffer(7.0PH); ACN 80:20% v/v. The flow rate was 1ml/min and eluents were detected by UV detector at 210 nm. Retention times were found to be 2.967 min and 5.750 min Finasteride and Minoxidil respectively. The calibration curve was linear over the range of 20-80 microgram/ml of Minoxidil and 0.5 -1.6 microgram/ml of Finasteride. The developed method was successfully applied for determination of the two drugs from its pharmaceutical formulation. The excipients in the formulation do not pose any hindrance in determination of two drugs. The proposed method is suitable for routine quality control analysis

Development and validation of an HPLC method for the simultaneous estimation of salbutamol, theophylline and ambroxol in tablet dosage form

A simple, accurate and precise reverse phase high performance liquid chromatographic (RP-HPLC) method has been developed and validated for the simultaneous estimation of Salbutamol, Theophylline and Ambroxol in pharmaceutical formulations. The chromatographic separation was achieved on a Inertsil ODS-3V (250 × 4.6 mm, 5μ) column using a mobile phase consisting of phosphate buffer (pH 3.0): acetonitrile in the ratio of 55:45 v/v at a flow rate of 1 mL/min. Detection was carried out at 225 nm. The retention times for Salbutamol, Theophylline and Ambroxol were found to be 2.317, 3.808 and 5.863 min respectively. The proposed method was validated as per the ICH guidelines and was found to be linear in the concentration range of 0.5-3.0 μg/mL, 25-150 μg/mL and 7.5-45 μg/mL for Salbutamol, Theophylline and Ambroxol respectively with correlation coefficients greater than 0.999. The developed method was found to be precise, accurate, robust and specific. Forced degradation studies proved the stability-indicating capability of the developed HPLC method. The developed method can be successfully applied for the routine analysis of Salbutamol, Theophylline and Ambroxol in pharmaceutical dosage forms.

HPLC Method Development and Validation for Residue Analysis of Steroid

A High-Performance Liquid Chromatography technique was developed for Residue Analysis of Megestrol Acetate collected from pharmaceutical manufacturing equipment’s. The samples were collected by Texwipe swab, separated by using Primesil C18 150mm x 4.6 mm, 5µm (Part No.: PC181504650) stationary phase and 65:35 of Acetonitrile: water as mobile phase in isocratic mode of elution. The residues were extracted from the swab in ultrasonic bath and later determined at a wavelength of 280nm. The detection was done using PDA detector for improved sensitivity. The developed method was then validated depending on the parameters of Cleaning Validation suggested by FDA. The method was found to be linear in a concentration range of 0.03-15 μg/ml. RSD of six replicates (Precision) were found to be 1% and the Recovery was found to be >90%. This method can be used to determine trace residues by Megestrol Acetate API on the manufacturing equipment’s in order to avoid any cross contamination during batch manufacturing.

Development and Validation of a New HPLC Method for the Estimation of Sulfamoxole in Bulk and Tablet Formulations

To quantify the sulfamoxole content in bulk as well as tablet formulation, a new isocratic RP-HPLC method was developed and then validated. Kromasil C18 column with a dimension of 250 X 4.6mm was used which was filled with a particle size of 5μ. A mixture containing CH3OH, CH3CN and H2O in the volume ratio of 55, 30 and 15 was used as a mobile phase at room temperature with an optimized flow rate of 1ml/min. UV detector was set at 241nm for sulfamoxole determination. The run time of the current method is ten minutes. Accomplished the forced degradation studies to understand the stability indicating nature of the current method. As per the current ICH guides (Q2R1), validation of the method was conducted.

Quantitative Estimation of Variability in Content of Asiaticoside in Centella asiatica Linn Collected from Different Geographical Regions of India by HPLC

Centella asiatica Linn is a significant and highly valued medicinal plant in various indigenous system of medicine Ayurveda, Siddha and Unani Indian system of medicines and Chinese medicines. It is one of the chief herbs for treating skin disorder, wounds healing, gastrointestinal disorders, asthma, ulcers, leprosy, vein diseases and memory improvement. In the present study an attempt is made to develop HPLC method for qualitative and quantitative estimation of an important bio active marker Asiaticoside in Centella asiatica collected from different geographical regions of India. Three samples were collected from different geographical regions of India and analyzed by HPLC. The compound separation was achieved on C18 Eclipse, XDB, 4.6mm × 150mm, 5μm particle size column using mobile phase consisted of Acetonitrile: Buffer (25:75v/v) in isocratic elution with flow rate 1.8ml/min. The detection of analytes at 210nm was carried out by using Diode Array Detector (DAD). The comparative study was carried out for variations of Asiaticoside content in Centella Asiatica Linn Collected from different geographical regions of India i.e. Chennai (Tamilnadu), Jhansi (Uttar Pradesh) and Raniketh (Uttarakhand). The studies reveal that the maximum amount of the marker compounds Asiaticoside (0.6157%) is present in the sample collected from Chennai compare to the other regions samples.

Accurate determination of Biotinidase activity in serum by HPLC and its utilization as second tier test for the confirmation of initial positive newborn screening results

Diagnosis of Biotinidase deficiency (BTD) is extremely important to avoid several neurodevelopmental problems in early childhood. Colorimetric and fluorometric methods lack specificity and selectivity due to several interferences resulting in a high number of false positive results. We developed an HPLC method for BTD activity in serum with fluorescent detection. In colorimetric assays, biotinidase attacks the amide linkage of the artificial substrate biotinidyl-4-aminobenzoic acid (B-PABA) and releases p-aminobenzoic acid (PABA), which is converted to a purple dye by diazotization reaction. The newly developed method injects the reaction mixture directly into the HPLC column and quantifies using a six-point calibration curve without coupling and diazotization reaction. The method is linear over the 5–1000 μmol/L range. The detection and quantitation limits were 2.5 μmol/L and 5.0 μmol/L, respectively. When compared with colorimetric assay, the correlation coefficient (R2) was 0.9963. The within-assay and between-assay precision was <10.0% for four levels of quality control samples. No significant variation in BTD activity was detected due to hemolysis, icteric, and lipemic samples. The newly developed method eliminates the potential interference due to the presence of aromatic amines and significantly reduces the false positive results observed with the colorimetric method. It is simple, specific, sensitive, faster in sample preparation, and requires a small sample volume. The newly developed HPLC method was used in our laboratory as a secondary tier test for initial positive BTD samples from newborn screening programs. To our knowledge, no similar HPLC method has been reported to date.

Development of Reverse Phase HPLC Method and Validation: Sofosbuvir and Velpatasvir Quantification in Bulk and Tablets

A new reverse-phase high-performance liquid chromatography (RP-HPLC) method for measuring the amount of sofosbuvir and velpatasvir in active pharmaceutical ingredients (API) and tablet forms has been developed and the method was also validated. Discovery column C18 150 mm x 4.6 mm, pore size of 5 μm was used for the analysis of drugs. Elution of drugs was achieved with 0.1% OPA Buffer: Acetonitrile mixed in the proportion of 40:60 v/v was used as mobile phase, maintaining 1.0 mL/min as flow rate, column temperature maintained at 30°C and uv detector wavelength was fixed at 272 nm. Sofosbuvir and velpatasvir were eluted at retention time (RT) of 2.183 and 3.038 minutes, respectively. Linearity parameter was performed in the levels from 25 to 150% and the square of correlation coefficient (r2) was found to be 0.9992. The %recovery obtained for sofosbuvir was 99.6 and for velpatasvir was 99.4. In precision %RSD for sofosbuvir was found to be 0.4 and for velpatasvir was 0.9. Validation was performed for the developed method and all of the validated parameter results obtained was within the ICH Q2 guidelines acceptance range. So, the developed RP-HPLC method can be used in pharmaceutical industry for quantification of sofosbuvir as well as velpatasvir at once in API and tablet dosage form.

The Development and Validation of Simultaneous Multi-Component Quantitative Analysis via HPLC–PDA Detection of 12 Secondary Metabolites Isolated from Drynariae Rhizoma

Drynariae Rhizoma (DR) is a functional food and traditional medicine that has been widely used for bone and joint disorders for thousands of years. In this study, 14 compounds were isolated from DR, and their structures were identified using UPLC/QTOF–MS, UPLC–ESI/LTQ–Orbitrap–HRMS, and 2D NMR and compared with those obtained in previous studies. An HPLC–PDA multi-component simultaneous quantitative determination method was developed for 12 of the 14 DR-derived compounds, excluding compounds with a content &lt;1.5 mg. The developed HPLC method was validated based on linearity (r2 ≥ 0.999), limit of detection (0.01–0.65 μg/mL), limit of quantification (0.04–1.97 μg/mL), intra-day precision and accuracy ranges (0.06–2.85% and 95.03–104.75%, respectively), and inter-day precision and accuracy ranges (0.24–2.83% and 95.75–105.75%, respectively). The developed analysis method improved the resolution of compounds 4 and 5. In addition, this is the first quantitative analysis of compounds 7, 8, and 11 and the first simultaneous quantitative analysis of 12 compounds, including compounds 4, 7, 8, 10, 11, and 14. This study developed a rapid, accurate, and economical HPLC method for performing the simultaneous quantitative analysis of 12 secondary metabolites isolated from DR.

A New Method Development and Validation of 5α Reductase Inhibitor (Dutasteride) Using R-HPLC In Bulk and Pharmaceutical Dosage Form (Dexamethasone Use as An Internal Standard)

The proposed HPLC Method was found to be simple,precise,accurate and economicalfor the estimation of dutasteride in Bulk and tablet dosage form. The present work, developed HPLC method was found suitable and validated. Luna – C-18 (250 mm x 4.6 mm, 20 µL) column was used for the assay. Mobile phase was Acetonitrile and Water in the ratio 80:20. Linearity was established in the range of 5-100 µg/ml with a coefficient correlation of 0.9998 and eluents were monitored at 242 nm and at 1 ml/min. flow rate. The Retention time was found to be 5.9 min. The efficiency of the column, expressed as the number of theoretical plates for six replicate injections was 11566 ± 119.695 (%CV1.03%) and USP tailing factor was 1.25 ± 0.012 (%CV 0.79). The regression equation for the calibration plot was Y = 0.0618 X – 0.008. Intra-day precision from 0.166 to 1.449 and Inter-day precision from 0.186 to 1.449. The% recovery of dutasteride from API and dosage form were 100.70% and 99.66%. The drug was subjected to stress conditions such as Oxidative and Photodegradation, Considerable degradation was found to occure in acidic and basic medium on heating at 100°C but no degradation was observed at room temperature. The drug was exposed to 10% H2O2{heating at 100°C forming degradation product at RT: 2.557}. The LOD and LOQ was 0.257µg and 0.779 µg respectively.When the flow rate was reduced, the retention time of dutasteride increased.The assay result with analyst #1 and Analyst #2 were % Assay = 98.40% (%RSD = 0.13%) and % Assay = 99.07% (%RSD =0.53) respectively. Robustness and ruggednesswasobserved that results were well within acceptance limits of 98–102%, with %RSD ±2.0%, indicating the method is rugged and provides consistent and reliable results which are not affected by small changes in experimental conditions. This method can be used for routine analysis of dutasteride. Running Title: A New Method Development and Validation of 5α-Reductase inhibitor using R-HPLC.

HPLC method development for the quantification of a mixture of reacting species: The case of lactose

Preparative and analytical chromatography are impaired by analytes that undergo a chemical reaction during the chromatographic separation, leading to peak distortion and systematic errors during the subsequent quantification phase. The pitfalls are highlighted through a combination of analytical results and numerical simulations. Two different quantification strategies for partially overlapping and reacting peaks are compared. A novel method development strategy based on the valley-to-peak ratio instead of the more common resolution is proposed. The method has been used to experimentally investigate the chromatographic behavior of a mutarotating sugar, lactose. The separation of the unprotected lactose isomers, α and β, has been optimized using a C18 column and pure water as the mobile phase. Phase dewetting phenomena during method development have also been studied and discussed.

A Comparative Analysis of Phytochemicals, Metal Ions, Volatile Metabolites in Heart Wood, Stem Bark and Leaves of Salix alba L. along with in Vitro Antioxidant, Antacid, Antimicrobial Activities for Sake of Environment Conservation by Substitution of Stem Bark With Leaf.

The genus of Salix is used in food, medicine and nutraceuticals, and standardized by using the single marker compound Salicin only. Stem bark is the official part used for the preparation of various drugs, nutraceuticals and food products, which may lead to overexploitation and damage of tree. There is need to search substitution of the stem bark with leaf of Salix alba L. (SA), which is yet not reported. Comparative phytochemicals viz. Salicin, Procyanidin B1 and Catechin were quantified in the various parts of SA viz. heart wood (SA-HW), stem bark (SA-SB) and leaves (SA-L) of Salix alba L.by using newly developed HPLC method. It was observed that SA-HW and SA-L contained far better amount of Salicin, Procyanidin B and Catechin as compared to SA-SB (SA-HW~SA-L≫SA-SB). Essential and toxic metal ions of all three parts were analysed using newly developed ICP-OES method, where SA-L were founded as a rich source of micronutrients and essential metal ions as compared to SA-SB and SA-HW. GC-MS analysis has shown the presence of fatty acids and volatile compounds. The observed TPC and TFC values for all three parts were ranged from 2.69 to 32.30 mg GAE/g of wt. and 37.57 to 220.76 mg QCE/g of wt. respectively. In DPPH assay the IC50 values of SA-SB, SA-HW, and SA-L were 1.09 (±0.02), 5.42 (±0.08), and 8.82 (±0.10) mg/mL, respectively. The order of antibacterial activities against E. coli, S. aureus, P. aeruginosa, and B. subtilis strains was SA-L>SA-HW>SA-SB with strong antibacterial activities against S. aureus, and B. subtilis strains. The antacid activities order was SA-L>SA-SB>SA-HW. The leaves of SA have shown significant source of nutrients, phytochemicals and medicinal properties than SA-HW and SA-SB. The leaves of SA may be considered as substitute of stem bark to save the environment or to avoid over exploitation, but after the complete pharmacological and toxicological studies.

Determination of Nifedipine by Validated RP-HPLC Method in Bulk and Pharmaceutical Dosage Form

The present paper deals with the development and validation of reverse phase HPLC method for the determination of Nifedipine on Nucleosil 100, 5 μm, C8, 250 x 4.0 mm column. A mobile phase consisting of 40 ml 2-propanol: 60 ml phosphoric acid 0.85% was employed in this study. The flow rate was kept at 0.8 ml/min and the injection volume was 10 µl. The separation was performed at 40°C. Eluents were monitored by UV detector set at 237 nm. The developed method was statistically validated for the linearity, precision, robustness, specificity and solution stability. The specificity of the method was ascertained by force degradation studies by acid and alkali hydrolysis, oxidation, heat and photo degradation. The degraded products were well resolved from the analyte peak with significant differences in their retention time values.

Application of high-performance liquid chromatography in determining saturates and aromatic hydrocarbons along with polars/additives in automotive finished and used lubricating oils

Lubricating oils help an internal combustion engine function effectively by reducing friction and wear on the engine's moving parts. They typically consist of petroleum-derived base oil and various additives to achieve the desired characteristics in automotive engine oils. Determination of aromatics and polar additives in the finished and used lubricating oils is not possible with existing methods hence their development is significant from the perspectives of environment and reuse/re-refining of used lubricating oils. This study reports the development of a new HPLC method to determine additives in the finished lubricating oils and/or polars in the used engine oils. The proposed method is simple, fast (runtime of 13 min), does not require sample pre-treatment, and exhibits high precision and superior limits of detection and quantification. The method demonstrated good linear response ranging from 0.1 to 30 mass for total aromatics and 0.1 to 20 % for additives. The method validation was carried out by analyzing brand-new commercial two and four-wheeler lubricants with used automotive lubricants. Based on the proposed method, the aromatics and additives concentration ranges in the studied finished lubricants were estimated between 0.20–1.70 % (mass) and 0.20–3.50 % (mass), respectively. Similarly, for used lubricants, the aromatics and additives were estimated to be 1.00–6.10 % and 0.60–2.40 % (mass), respectively.

Development of stability-indicating HPLC method for quantification of pharmacopeia impurities of Zuclopenthixol and characterization of its stress degradation products by LCMS/MS

BackgroundThe present study focused to develop a simple and sensitive HPLC method for resolution and estimation process-related impurities of zuclopenthixol and further assessment of forced degradation behavior of zuclopenthixol.ResultsThe chromatographic separation of drug substance, process-related impurities and its degradation products (DPs) was achieved on KNAUER C18 (250 mm × 4.6 mm, 5µ id) column at that was maintained at 35 °C temperature using 0.1 M sodium acetate buffer at pH 4.3 and methanol in 20:80 (v/v) as mobile phase A, 0.1% formic acid and acetonitrile in 75:25 (v/v) as mobile phase B. Equal volume of mobile phase A and B was pumped in isocratic elution at 0.8 mL/min. Detection wavelength was selected as 257 nm. In the proposed conditions, zuclopenthixol is identified at 6.91 and 1.91 min and 2.89 min, respectively, for impurity B and A min with acceptable system suitability and specificity. The method produces LOD at 0.009 for impurities with calibration range of 30–180 µg/mL for zuclopenthixol and 0.03–0.18 µg/mL for impurities. The other validation parameters were notices to be with in the acceptable levels for zuclopenthixol and its impurities. The drug was exposed to different stressed conditions (acid, base, peroxide, thermal and UV light) according to ICH Q1A (R2) guidelines. The DPs formed during the stress study were identified and characterized by LCMS/MS in ESI positive mode.ConclusionThe analysis involved a comparison of collision-induced dissociation mass spectrometry data between the degradation products and zuclopenthixol. As a result, potential structures for six degradation compounds were suggested. The results from additional validation studies were similarly pleasing and demonstrated their suitability for the routine analysis of zuclopenthixol and its associated impurities in both bulk drug and pharmaceutical dosage forms. Additionally, these findings can be extended to assess the mechanism of stress degradation in zuclopenthixol.

Development of an eco-friendly HPLC method for the stability indicating assay of binary mixture of ibuprofen and phenylephrine

The development and validation of the stability indicating HPLC technique has contributed to the understanding of the stability profile of ibuprofen (IBU) and phenylephrine (PHE). Stability profile was achieved for PHE; the drug was found to be liable to be influenced by stress oxidative conditions; two oxidative degradants (Deg1 & Deg2) were formed and their structures were confirmed using IR and mass spectrometry. The drugs and degradation products were successfully separated using a gradient elution method on YMC-C8 column with 0.1% hexanesulfonic acid and acetonitrile as a mobile phase at pH 6.6. The flow rate was 1.0 mL/min, and a diode array detector operating at 220 nm was used for UV detection. The retention times of degradants Deg1, Deg2, ibuprofen (IBU), and phenylephrine hydrochloride (PHE) were 2.0, 2.2, 3.2 and 7.0 min, respectively. The proposed method was validated with respect to linearity, accuracy, precision, specificity, and robustness using ICH guidelines. The linearities of ibuprofen and phenylephrine hydrochloride were in the range of 10–100 μg/mL and 0.3–10 μg/mL, respectively. The % recoveries of the two drugs were found to be 100.75 ± 1.44%, 99.67% ± 1.67, and the LOD was found to be 2.75/mL and 0.09/mL for IBU, and PHE, respectively. The method was successfully applied to the estimation of ibuprofen and phenylephrine hydrochloride combination in pharmaceutical dosage form. The proposed technique was validated using ICH guidelines and its greenness was assessed according to Analytical Eco Scale metric (AES). Molecular docking was used to assess the two drugs and PHE oxidative degradants interaction with the stationary phase and to confirm the outcomes of the proposed method with regard to the order of elution of the two drugs and PHE degradation products. Eco-friendly and environmental safety were assessed through the application of one of the most applicable greenness assessment tool; Analytical Eco Scale metric (AES).

Development and Validation of RP-HPLC Method for Simultaneous Determination of Aceclofenac and Piperine in Rat Plasma to Study Pharmacokinetic Parameters

The BCS class II medication aceclofenac is poorly soluble in water and undergoes significant metabolism in human hepatocytes and microsomes. It has a 15% oral bioavailability, which is low. Piperine is a naturally occurring bioenhancer that is used to increase the oral bioavailability of many different medications. In this study, the pharmacokinetics of aceclofenac (20 mg/kg) alone and in combination with piperine (10 mg/kg) in rats were tested. Despite their distinct chemistry and physical properties, in this study, piperine was used to enhance the bioavailability of aceclofenac, followed by the development and validation of HPLC method to determine their simultaneous measurement in rat plasma. Plasma samples were processed by solid phase extraction and subjected to HPLC analysis. The analysis identified retention time for aceclofenac and piperine to be 5.3 and 14.4 minutes respectively. We found the linear concentration range of aceclofenac in rat plasma between 0.1 to 20 μg/mL. The correlation coefficient (R2) determined was 0.9995. Cmax was increased from 23.59 to 48.52 μg/mL. Including piperine with aceclofenac in formulation can improve its oral bioavailability.