Method development and validation for Cabotegravir and Rilpivirine by using HPLC and its degradants are characterized by LCMS and FTIR

Background Using a Symmetry C18 (4.6 × 150 mm, 3.5) column, a high-performance liquid chromatographic method for quantification of Rilpivirine and Cabotegravir in active pharmaceutical ingredients was developed and validated. The mobile phase is made up of buffer, acetonitrile, and 0.1 percent formic acid in a 20:80v/v ratio. The flow rate was kept constant at 1.0 ml/min, and detection was accomplished through absorption at 231 nm with a photodiode array detector. Results The calibration curve was linear, with a regression coefficient (R2) value of 0.999 and concentrations ranging from 30 to 450 g/ml of Rilpivirine and 20–300 g/ml of Cabotegravir. The method's LOD and LOQ were 0.375 g/ml, 1.238 g/ml, and 0.25 g/ml, 0.825 g/ml for Rilpivirine and Cabotegravir, respectively. Conclusions In the forced degradation studies, the degradants were characterized by using LCMS and FTIR. The current application was found to be simple, economical, and suitable, and validated according to ICH guidelines.

Development of a Validated Stability Indicating Method for Quantification of Amoxicillin, Clarithromycin and Lansoprazole in Bulk and Pharmaceutical Dosage form by RP-HPLC

Objective: The main objective of the present work is to develop an efficient, unique, reliable Reverse Phase High Performance Liquid Chromatography (RP-HPLC) method for the simultaneous quantification of Amoxicillin (AMX), Clarithromycin (CTM) and Lansoprazole (LPZ) in bulk and pharmaceutical formulations. Methods: The chromatographic separation was achieved by using Kinetex column C18 (100 x 4.6 mm, 2.6 µm) with Buffer (2.5 g of hexane sulphonic acid and 1ml of Triethylamine which are added to 1000 ml of HPLC water and adjusted its pH at 5.0 with Ortho phosphoric acid) and acetonitrile in the ratio of 70: 30 (%v/v) as a mobile phase at flow rate of 1.0 ml/min. The column effluents were monitored by a photodiode array detector at wavelength predetermined at 240 nm. Results: The method produced reliable results at optimized chromatographic conditions. The method was linear at concentration range of 15-225 µg/ml of AMX, 15-225 µg/ml of CTM and 0.9-13.5 µg/ml of LPZ with regression coefficients of 0.9999, 0.9999, and 0.9999 respectively. The retention times of AMX, CTM, LPZ were obtained as 1.513, 3.124, 3.770 min respectively. Results obtained for system suitability, precision, LOD and LOQ were in acceptable range and were validated according to the guidelines of the International Council for Harmonization (ICH). Conclusion: The proposed method was validated in accordance with ICH and all the obtained results were found satisfactory and were successfully applicable to the analysis of the bulk and the pharmaceutical formulations.

Screening and Characterization of Phenolic Compounds from Australian Grown Bananas and Their Antioxidant Capacity

Bananas are an essential source of staple food and fruit worldwide and are widely regarded as the world’s largest fruit crop, with more than 100 million tons total annual production. Banana peel, a by-product that represents about 40% of the entire banana’s weight, and pulp are rich in bioactive compounds and have a high antioxidant capacity. As the production of polyphenols in fruit and vegetables is highly dependent on environmental conditions, genetic factors, and the level of maturity, this study aims to characterize six Australian banana cultivars in various stages of ripening for their phenolic compounds using the liquid chromatography-electrospray ionization quadrupole time of flight mass spectrometry (LC-ESI-QTOF-MS/MS), polyphenols quantification with the high-performance liquid chromatography coupled with photodiode array detector (HPLC-PDA), and their antioxidant capacity. All bananas were analysed for total polyphenols content (TPC), total flavonoids content (TFC), and total tannin content (TTC) and their antioxidant activities. Ripe Ducasse peel and pulp contained the highest amounts of total polyphenols content (1.32 and 1.28 mg gallic acid equivalent (GAE) per gram of sample), total tannin contents (3.34 mg catechin equivalent (CE) per gram of sample), and free radical scavenging capacity (106.67 mg ascorbic acid equivalent (AAE) per g of sample). In contrast, ripe Plantain peel had the greatest total flavonoids (0.03 mg quercetin equivalent (QE) per g of sample). On the other hand, unripe Ladyfinger pulp possessed the highest total antioxidant activity (1.03 mg AAE/g of sample). There was a positive correlation between flavonoids and antioxidant activities. By using LC-ESI-QTOF-MS/MS, a total of 24 phenolic compounds were tentatively characterized in this research, including six phenolic acids, 13 flavonoids, and five other polyphenols. Quantification of phenolic compounds by the high-performance liquid chromatography coupled with photodiode array detector (HPLC-PDA) revealed a higher content of phenolic acids. These findings confirmed that banana peel and pulp have considerable antioxidant activity and can be employed in human food and animal feed for variant health enhancement uses.

Stability Indicating Photodiode Array Detector Based Estimation of Dapagliflozin Propanediol Monohydrate in API and Tablet Dosage Form by RP-UPLC

Stability indicating RP-UPLC photo diode array detector based method for determination of Dapagliflozin propanediol monohydrate (DPM) in active pharmaceutical ingredient (API) and in tablet dosage form (5mg dapagliflozin) has been developed and validated on Bridge Ethylene Hybride (BEH) C18 column (50mm × 2.1 mm, 1.7µm). Mobile phase composition was water: acetonitrile (60:40 v/v), flow rate 0.5ml/min and detection carried out at 223nm at column temperature 30ºC. Chromatographic separation achieved within 2 min with retention time 0.77 min. Linearity of the method was found over the concentration range of 25-75µg/ml (R2 = 0.9977). The degradation was carried out in five different stress conditions. The developed method was able to resolve peak of API from all generated peaks. Sufficient degradation was achieved in the range of 5.25 to 12.31%. The peak purity is acceptable, Method validation was performed as per ICH guideline Q2(R1).

MITRAGYNINE PERCENTAGES OF VARIOUS KRATOM VARIANTS SEIZED IN INDONESIA: A QUANTITATIVE ANALYSIS USING LIQUID CHROMATOGRAPHY-PHOTO DIODE ARRAY DETECTOR

Objective: This study details the determination of mitragynine in various kratom samples using the thin-layer chromatography (TLC) technique and validation of analytical methods for quantifying the concentration of mitragynine in various kratom samples using liquid chromatography with photodiode array detector (LC-PDA). Methods: TLC technique using n-hexane: ethyl acetate: ammonia 25% (30: 15: 1 v/v/v) was applied to isolate mitragynine from kratom samples. Several interesting spots obtained were visualized under UV light at 254 nm. Samples were also prepared with organic solvent extraction directly prior to LC analysis (non-isolation method of preparation) to quantify the concentration of mitragynine. Mobile phases used were acetonitrile (MP A) and 0.1% formic acid in water (MP B). Samples and standards were run by gradient elution with a flow rate of 0.3 ml/min, detection using PDA detector at 254 nm. Results: Mitragynine was successfully isolated from kratom samples in Rf 0.50 by TLC system applied. The validation of analytical methods for mitragynine passed the acceptance criteria as described by UNODC Guidance. The concentration of mitragynine in various kratom samples seized in Indonesia ranged from 0.37%-1.70% (%w/w). Conclusion: Both TLC and LC analytical methods could be applied to determine and quantify the concentration of mitragynine in each examined sample, respectively.

Method Development and Validation for the Determination of Linezolid Drug in Human Plasma by Reversed-Phase High-Performance Liquid Chromatography

Objective: Linezolid is a significant antibiotic used against severe infections initiated by multi-resistant bacterial pathogens. Method: Linezolid extraction from plasma is obtained using methanol. Chromatographic separation is achieved isocratically on a C18 column [Zorbax C18, 5 µm particle size, 150 mm ˟ 4.6 mm] making use of a mobile phase of acetonitrile / 0.05 M phosphate buffer, pH = 4.5 (30 : 70 v/v) at a flow rate of 1.2 mL/min with photodiode array detector DAD, at a wavelength of 256 nm. Method: Linezolid extraction from plasma is obtained using methanol. Chromatographic separation is achieved isocratically on a C18 column [Zorbax C18, 5 µm particle size, 150 mm ˟ 4.6 mm] making use of a mobile phase of acetonitrile / 0.05 M phosphate buffer, pH = 4.5 (30 : 70 v/v) at a flow rate of 1.2 mL/min with photodiode array detector DAD, at a wavelength of 256 nm. Results : The retention time of linezolid was 2.5 min. The analytical method was linear (r2 > 0.998) over the calibration range of 0.30 to 50.0 µg/mL. The extraction recoveries of linezolid range from 71.03 to 91.93 %. The limit of quantification and the limit of detection were 0.112 µg and 0.037 µg, respectively. The RSDs for intraday and interday assays were < 7.77 and 4.32 %, respectively. The intraday and interday accuracies were in the range 80.6-112 % and 77.44-104.85 %, respectively. Conclusion: The applied method is precise, accurate and appropriate for pharmacokinetic studies and therapeutic drug monitoring of linezolid in routine clinical practice.