Development of validated UHPLC–PDA with ESI–MS-MS method for concurrent estimation of magnoflorine, berbamine, columbamine, jatrorrhizine, palmatine and berberine in Berberis aristata

A validated UHPLC-PDA with an ESI-MS/MS method has been developed for simultaneous estimation of six bioactive alkaloids (magnoflorine, berbamine, columbamine, jatrorrhizine, palmatine and berberine) in the different extracts of the roots of Berberis aristata DC (Family:Berberdiaceae). It is an important medicinal herb native to Northern Himalaya and commonly known as ‘daruharidra’, ‘daruhaldi’, ‘Indian barberry’ or ‘tree turmeric’. An insight into the research literature uncovered reports on isoquinoline alkaloids like magnoflorine, berbamine, columbamine, jatrorrhizine, palmatine, and berberine as major bioactives in B. aristata roots, possessing different pharmacological and therapeutic effects. In the present study, these aforementioned alkaloids were separated on Phenomenex Luna®, 5 µm-C8 analytical column. The HPLC-MS analysis was performed at a flow rate of 0.90 mL min−1. Each alkaloid that is resolved was characterized by precursor ions and fragment ions with electrospray ionization (ESI) source in both positive and negative ionization using scan mode. The limit of detections (LODs) were 0.087, 0.727, 0.035, 0.124, 0.782 and 0.794 μg mL−1 for magnoflorine, berbamine, columbamine, jatrorrhizine, palmatine and berberine, respectively. The proposed UHPLC-PDA method was fully validated according to international (ICH) guidelines and was found to be selective, sensitive and highly accurate for the concomitant estimation of the aforementioned symbolic bio-markers of B. aristata roots.

A simple reverse phase UPLC validated method for concurrent estimation of Emtricitabine, Darunavir, Tenofovir and Cobicistat in drug substances and drug product

A fast, precise, accurate and steady isocratic liquid chromatographic technique was created for the synchronous assurance of the Emtricitabine, Darunavir, Tenofovir and Cobicistat in bulk and in formulation. To optimize a column HSS C18 100 x 2.1 mm, 1.8mm, mobile phase including Buffer 0.01N KH2PO4(5.4pH): Acetonitrile pick in the proportion 60:40v/v was pumped through column at a flow rate of 0.3 ml/min at 260nm. The retention times of Emtricitabine, Darunavir, Tenofovir and Cobicistat were initiated to be 1.066 min, 1.727 min, 2.574 min and 2.977 min. % recovery was 99.75%, 100.05%, 100.42% and 100.59% for Emtricitabine, Darunavir, Tenofovir and Cobicistat respectively. LOD and LOQ values got from relapse formula of Emtricitabine, Darunavir, Tenofovir and Cobicistat and were 0.18, 0.54, 0.71,2.14, 0.01,0.05 0.44 and 1.32 correspondingly. Relapse equation of Emtricitabine is y = 13375x + 2577.4, for Darunavir it is y = 7196.3x + 2981.2, for Tenofovir it is y = 66663x + 1338.9 and y = 22723x + 6978.7 for Cobicistat.

Valsartan in combination with metformin and gliclazide in diabetic rat model using developed RP-HPLC method

Background Oral administration of biguanides (metformin) and sulfonylureas (gliclazide) are the most common approach of management of type 2 diabetes in humans. Among these diabetic patients, approximately 40–60% suffers from hypertension. Hence, the need of the day is application of polytherapy. A major challenge in polytherapy is the drug-drug interactions that may arise. Hence, this study is focused to develop a reverse phase high-performance liquid chromatography (RP-HPLC) method for concurrent estimation of diabetic drug metformin and hypertension drug valsartan using C18 column and find any possible pharmacokinetic interactions between the two drug combinations strategies, i.e., metformin-valsartan and gliclazide-valsartan in streptozotocin-induced diabetic rats. Result The bioanalysis of drug-drug interaction pharmacokinetic result showed no significant difference in the tmax of single treatment of gliclazide and single treatment of metformin or upon co-administration with valsartan. Conclusion Our study has shown that polytherapy of valsartan, a drug administered for hypertension along with hypoglycemic drugs metformin and gliclazide, can be advantageous and safe in patients suffering from both diabetes and hypertension.

Concurrent estimation of lamivudine, tenofovir disoproxil fumarate, and efavirenz in blended mixture and triple combination tablet formulation by a new stability indicating RP-HPLC method

Background An easy, defined, rapid, and accurate reverse phase high-performance liquid chromatography method was developed and subsequently validated for the concurrent estimation of lamivudine, efavirenz, and tenofovir disoproxil fumarate in their pure blend and combined tablet formulation. An efficient and appropriate separation of the three analytes was attained with Zorbax eclipse XDB-Phenyl column, with a mobile phase of methanol: buffer (0.1% v/v formic acid in water) (73:27 v/v) at a flow rate of 1mL/min and isocratic elution by using 260nm as detection wavelength. Equal ratio of acetonitrile and water was used as diluent. Results The retention times of lamivudine, tenofovir disoproxil fumarate, and efavirenz were found at 2.6, 4.4, and 5.9 min respectively. The linear response for lamivudine, tenofovir disoproxil fumarate, and efavirenz was in the range of 15.0–45.0μg/mL, 15.0–45.0μg/mL, and 20.0–60.0 μg/mL respectively. The method validation was done in accordance to ICH guidelines and all validation parameters in compliance with ICH standards. The degradants produced by stress testing were well resolved from the peaks of active analytes, which stipulates the stability-indicating property of the method. Conclusion The method has the ability to separate lamivudine, efavirenz, and tenofovir disoproxil fumarate concurrently in blended powder and their combined tablet. All degradants produced by application of stress conditions were separated with high resolution and determined with good sensitivity that ensures the stability-indicating property of the method. Thus, the projected method has high probability to adopt in the pharmaceutical industrial sector.

Estimation of Cyproheptadine Hydrochloride and Tricholine Citrate Simultaneously in Syrup Dose by Applying Stability Indicating HPLC Methodology

An stability indicating HPLC methodology for the concurrent estimation of Tricholine citrate (TRC) and Cyproheptadine hydrochloride (CYH) in syrup dose and bulk using Waters column reverse phase C18 (5 µm, 250 mm and 4.6 mm) as stationary phase and 0.1M Na2HPO4 of pH 4.5 and acetonitrile in proportion of 60:40 (v/v) at flow of 1.0 ml/min rate as mobile phase was reported. The linear scales were 275-825 μg/ml for TRC and 2-6 μg/ml for CYH with correlation coefficients of 0.9999 for TRC and 0.9997 for CYH. Followed ICH Q2(R1) strategies for validating the suggested method for precision, sensitivity, robustness, specificity, selectivity and accuracy. The measures of LOD and LOQ are 0.023 µg/ml and 0.079 µg/ml for CYH, while for TRC it was 0.565 µg/ml and 1.885 µg/ml, respectively. The precision measures for CYH and TRC were 0.073 and 0.212 relative measured deviation percentage, respectively. The accuracy measures for CYH and TRC were 99.40% and 99.09% mean assay percentiles, respectively. Recovery percentiles measures of CYH and TRC were ranged between 99.48% to 100.35% and 100.38% and 100.41%, respectively. While in degradation investigation, peaks of degraded products are very well differentiated from TRC and CYH peaks suggesting the specificity and stability of suggested methodology. The results permit the application of the proposed stability indicating HPLC methodology in syrup dose forms.

A Simple Reverse Phase Ultra Performance Liquid Chromatography Validated Method for Concurrent Estimation of Daunorubicin and Cytarabine in Drug Substances and Drug Product

A fast, precise, accurate, and steadiness indicating isocratic liquid chromatographic technique was created for the synchronous assurance of the daunorubicin and cytarabine in bulk and formulation. To optimize a column CHS C18 100 × 2.1 mm, 1.8 μm, mobile phase, including buffer 0.1% orthophosphoric acid, acetonitrile pick in the proportion 70:30 v/v, was pumped through the column at a flow rate of 0.3 mL/min at 240 nm, initiate to be an efficient method for elution of drug with good peak shapes, as well as, retention times. The retention time of daunorubicin and cytarabine were initiated to be 0.556 and 0.743 minutes. The % recovery was got at 100.07 and 99.88% for daunorubicin and cytarabine separately. The limit of detection (LoD) and limit of quantitation (LoQ) values got from the relapse formula of daunorubicin and cytarabine were 0.16, 0.5, and 0.64, 1.93, correspondingly. The relapse equation of daunorubicin is y = 2974.3x + 648.32, and y = 4896.5x + 4851.5 of cytarabine.