Method Development and Validation of Degradation Studies of Favipiravir by RP-HPLC

RP HPLC method was developed by this study estimation of favipiravir. This method is developed by Shimadzu LC -2010 HT by using a C18 (250 X 4.6 X mm X 5µ) column in solvents Water(OPA)+ACN (60:40)v/v as mobile phase and the temperature was maintained at 25°C. The mobile phase flow rate of 1ml/min was pumped and sample wavelength was detected at 324 nm by ultraviolet -visible spectrophotometer. The Rt was found 4.453 min. The number of theoretical plates and tailing factor for favipiravir was observed 82651(NLT 2000) and 1.265 (NMT 2). The method was validated for analytical standards such as linearity, accuracy, precision, LOD, LOQ and robustness. LOD and LOQ values were calculated from regression of favipiravir 1.26 and 3.83 µg/ml.The regression equation of validated method for favipiravir is Y=253.5x+1881.In a wide range of 4 to 20 (µg/ml) the linearity was observed. Degradation methods were also performed.

RP-HPLC Method for Determination of Gemfibrozil using Central Composite Design (CCD)

The high-Performance Analytical Liquid Chromatography (HPLC) method (AQbD) for routine analysis of Gemfibrozil in dosage form was developed in column C18 using an experimental design. The central composite design (CCD) was adopted in evaluating the responses and robustness of the method. In the project, the combined effect of buffer pH, % organic phase and flow rate, each at five levels, was selected for responses such as retention time and number of theoretical plates, then interpreted and optimized statistically with the help of the surface methodology of response and therefore of the analysis of the constructed models and of the outline graphs was obtained. Acetonitrile: phosphate buffer (pH-4) (59: 41% v / v) as eluent at a flow rate of 1.0ml/min was found to be the optimal condition for obtaining the desired answers.

METHOD DEVELOPMENT AND VALIDATION OF IXAZOMIB DRUG BY RP-HPLC IN BULK AND PHARMACEUTICAL DOSAGE FORM

A novel approach was used for develop and validate a rapid, accurate, and an isocratic RP-HPLC method with PDA detector for the estimation of ixazomib drug in pharmaceutical dosage forms. Ixazomib was seperated using Agilent 4.6*150 mm, 5μm analytical column, a Waters HPLC system (USA) and a mobile phase consisting of water and acetonitrile in the ratio of 40:60 V/V. The flow rate was set to 0.7 mL/min with 10µL injection volume. The column was maintained at ambient temperature, detector was set at wavelength of 274 nm. The retention time of ixazomib was found to be 2.17 min. The system suitability parameters for ixazomib such as theoretical plates and tailing factor were found to be 4146. Linearity was established for ixazomib such as theoretical plates and telling factor were found to be 4146. Linearilty was established for ixazomib in the range of 50-250 µg/ml concentration levels with correlation coefficients (r2) of 0.999. The intra-and inter-day precision % RSD values were found to be 0.47 and 0.31, respectively. The limit of detection (LOD) and limit of quantification (LOQ) were found to be 2.03 and 6.17 µg/mL respectively. The method was validated for all of the above parameters according to the International Conference on Harmonization (ICH) guidelines. This method can be used for estimation and analysis of ixazomib drug in active pharmaceutical ingredients and pharmaceuticals.

Beta-Alanine and Tris-(hydroxyl methyl) Aminomethane as Peak Modifiers in the Development of RP–HPLC Methods Using Aceclofenac and Haloperidol Hydrochloride as Exemplar Drugs

In the present analytical approach, beta-alanine (ALA) and tris-(hydroxyl methyl) aminomethane (TRIS) were investigated as peak modifiers due to their water solubility and their possible peak modifying a property. These reagents were tested for their efficacy on the elution of aceclofenac (ACF) and haloperidol hydrochloride (HLC) from C18 column (250 mm × 4.6 mm, 5 μ) equipped with a photodiode array detector. The test reagents were investigated at 0.25 ± 0.05% concentration with a varying % aqueous composition on elution efficacy of HLC and ACF. The added ALA/TRIS in the mobile phase significantly (P < 0.05) improvised the symmetrical elution of HLC with 3-fold theoretical plates increase (P < 0.05) and 10-fold reduced capacity factor as compared to the control run. For ACF, the shoulder effect observed for ACF peak was eliminated. The optimized mobile phase was a combination of acetonitrile and water containing 0.25% beta-alanine/TRIS (pH 3.5 with ortho-phosphoric acid) at the ratio of 70:30 and 60:40% v/v, respectively, for ACF and HLC. The method was validated as per ICHQ2 guidelines. The column performance was tested for reproducibility in non-peak modifier applications and revealed a null effect on the column, thus these agents are relatively less toxic to HPLC columns.

Separation of Sb (V) and Sb (III) antimony compounds using anion exchange chromatography technique

Separation of Sb (V) and Sb (III) antimony compounds using anion exchange chromatography technique have been done. To obtain the separation of Sb (V) and Sb (III) antimony compounds which is good in this study have been studied several parameters of separation in anion exchange chromatography technique. Parameters that influence the process of separation of Sb (V) and Sb (III) antimony compounds is the concentration and pH of the mobile phase (eluent) have been evaluated. The separation of Sb (V) and Sb (III) antimony compounds is good and optimum obtained using an eluent 200 mM phosphate buffer at pH 7 with a flow rate of 1 mL/min. Based on the optimum conditions for the separation of Sb (V) and Sb (III) antimony compounds with anion exchange chromatography method has generated value the capacity factor (k ') Sb (V) and Sb (III) obtained are respectively 1.77 and 3.01. While the value of selectivity (α), Number of theoretical plates (N) and Resolution (Rs) obtained are respectively 1.70; 369.48; and 1.48.

Study on chemical constituents of different species of Dendrobium

The objective of this study is to determine the erianin of 10 species of Dendrobium by high performance liquid chromatography (HPLC). Processing techniques were washed, dried and ground. The research method for determining the content of erianin was adopted from Chinese Pharmacopoeia 2015 Edition. Acetonitrile-0.05% phosphoric acid (37:63) was used as mobile phase. The samples were separated on Sharpsil-TC18 column(4.6*150mm;5μm) at a flow rate of 1.2mL/min and detected at 230nm, and the column temperature was kept at 30℃. The injection volume was 20μL. The number of theoretical plates was not less than 6000 according to the chromatographic peak of erianin. The result of the study showed erianin was detected only in Dendrobium chrysotoxum among the 10 species of Dendrobium. and the content was 0.098%. Conclusion: The content of erianin in Dendrobium chrysotoxum met the specification of no less than 0.03% in the 2015 edition of Chinese Pharmacopoeia.

Two novel UPLC methods utilizing two different analytical columns and different detection approaches for the simultaneous analysis of velpatasvir and sofosbuvir: application to their co-formulated tablet

In the present study two different RSLC columns, Acclaim RSLC 120 C18, 5.0 µm, 4.6 × 150 mm (column A) and Acclaim RSLC 120 C18, 2.2 µm, 2.1 × 100 mm (Column B) were utilized for the analysis of velpatasvir (VPS) in presence of sofosbuvir (SFV), where due to the encountered fluorescent properties of VPS fluorescent detection at 405 nm after excitation at 340 nm (Method 1) was used for its detection where the non-fluorescent SFV did not interfere. The same columns were further utilized for the simultaneous determination of SFV and VPS either in bulk form or in their combined tablet, where UV- spectrophotometric detection at 260 nm was selected for the simultaneous analysis of both drugs (Method 2). A mobile phase consisting of NaH2PO4, pH 2.5 (with phosphoric acid) and acetonitrile in a ratio of 60:40 v/v was used for both methods. The mobile phase was pumped at a flow rate of 1.0 mL/min when using column, A and 0.5 mL/min when using column B. The methods showed good linearity over the concentration ranges of 1.0–5.0 and 2.5–10.0 ng/mL for VPS when utilizing Method 1 A and B respectively. Where the linearity concentration range was from 30.0–150.0 to 120–600.0 ng/mL for VPS and SFV respectively when applying Method 2. Both methods 1 and 2 were performed by utilizing the two analytical columns. The different chromatographic parameters as retention time, resolution, number of theoretical plates (N), capacity factor, tailing factor and selectivity were carefully optimized. The results show that comparing the performance of the two utilized columns revealed that shorter column (2.1 mm × 100 mm) with small particle packing was superior to the longer column (4.6 × 150 mm) for the analysis of the studied drugs allowing a reduction of the analysis time by 70% without any detrimental effect on performance. This prompts the decrease of the investigation costs by saving money on organic solvents and expanding the overall number of analyses per day.

Performance Comparison Between Monolithic, Core-Shell, and Totally Porous Particulate Columns for Application in Greener and Faster Chromatography

Background: The introduction of monolithic rods and core-shell particles as new morphologies of packing materials different from the conventional totally porous particles resulted in a leap forward for performance in LC. Meanwhile, environmental safety has become increasingly important in many areas, especially in industry and research laboratories. Objective: This study compared the efficiencies of commercially available columns of different lengths and diameters when greener chromatographic conditions were utilized. The main purpose of this study is to help practitioners select the most appropriate stationary phase for faster and greener analysis. Methods: The three types of stationary phases were compared in terms of separation efficiency, number of theoretical plates, peak shape, selectivity, resolution, analysis time, mobile phase consideration, and permeability using six drug molecules. Results: Results indicated that core-shell and monolithic stationary phases had superiority over the conventional totally porous particles in terms of efficiency and speed of analysis. Monolithic rods had lower column backpressure and higher permeability, so they are more suitable for higher mobile phase flow rates and viscosities. However, core-shell particles provided enhanced peak shapes and number of theoretical plates. Conclusions: The choice will depend on the main purpose of analysis and the composition of the mobile phase. Compromise must be made to obtain the best trade-off between separation efficiency and analysis speed. Highlights: This study is the first to consider green chromatography concepts for the selection of the best stationary phase of new morphologies.