Response Surface Methodology Driven Systematic Development of a Stability- indicating RP-UPLC Method for the Quantification of Aliskiren: A Renin Inhibitor

Aims: The current study envisages experimental design enabled rapid, sensitive, and stability-indicating RP-UPLC method to quantify Aliskiren in its pharmaceutical formulations. Study Design: Box-Benkhen experimental Design using Response surface methodology. Place and Duration of Study: Department of Analytical Research and Development, Brawn laboratories ltd., Gurugram, India, and Department of Pharmacy KL College Pharmacy, KL Deemed to be University, Vaddesearam, Guntur, Andhra Pradesh, between May 2021 and September 2021. Methodology: The chromatographic partitioning was achieved on a Waters Acuity H class UPLC system, with BEH 130οA, C18 column (100 x 2.1 mm,1.7 μm) having isocratic elution containing (50:50 %v/v) of 0.2% Glacial acetic acid (GAA) : acetonitrile, at constant flow rate using PDA detection. The optimum conditions were delineated, selecting three influential factors (CMPs), i.e., mobile phase composition, flow rate, and injection volume. Systematic optimization was accomplished by 32 Box-Benkhen design using response surface methodology (RSM). Results: The selected variables are evaluated for obtained responses (CAAs), i.e., peak area, retention time (Rt), USP Plate count. The final optimized method employed, organic phase composition 0.2 % GAA (pH 3.0) and acetonitrile 50:50 (% v/v) with 0.3 mL min-1 flow rate. The injection volume was maintained as 2μL with 2 minutes run time and λmax 280 nm. Conclusion: The method was linear for 5-300 ppm, with regression co-efficient (R2) 0.9995. As per ICH guidelines, forced degradation studies were carried out to analyse the stability profile of drug. The short Rt 1.214, minute implies superior robustness, sensitivity, and cost-effectiveness for routine analysis. The results exhibited that RSM approach of QbD will be competently used to optimize the RP-UPLC method with fewer experimental trials and error-free investigation.

Validation of a simple HPLC method to quantify methotrexate concentrations in human plasma

Methotrexate (MTX) is a chemotherapy and immunosuppressive agent widely used to treat cancer, autoimmune diseases in children and adult patients, and ectopic pregnancy. However, MTX is highly toxic to the liver, kidney, and nervous system. This study aimed to quantify the concentration of MTX in human plasma using high-performance liquid chromatography (HPLC). MTX and its internal standard (para aminoacetophenone-PAPA) in plasma samples were extracted simultaneously with methanol. Sample purity was performed using the 1 cc OASIS HLB cartridges. Sample injection volume of 10 µL was analyzed on a Lichrocart Supersil 125-4 column C18 maintained at 40 °C on a Waters 2695 XE equipped with a PDA detector set at 303 nm. The mobile phase contained phosphate buffer (pH 6.0) and methanol at a ratio of 80:20 (v/v) and was maintained at a flow rate of 1 ml/min. The results showed that the total time of chromatographic analysis was 15 min. MTX and PAAP were found in the chromatograms at retention times of 2.3 and 5.2 min, respectively. The linear range of the MTX from 0.5 to 25 µg/mL. Intra-day and inter-day imprecision for MTX ranged from 3.42 to 8.128%. LLOQ of MTX was 0.5 µg/mL and the extraction effects were above 77%. In conclusion, we developed and validated a simple HPLC method to determine the MTX concentrations in human plasma.

Experimental Research on the Measurement of High-Pressure Microflow Based on Momentum Principle

The fuel injector is an important component of the diesel engine. It has a great influence on the atomization of diesel fuel injection, the formation of mixed gas, and combustion emissions. Due to the current nozzle structure, processing level, and the internal hydraulic conditions of each nozzle, there are certain differences between the injection rules of each hole, and there are few methods to quantify the quality of the injector using mathematical methods in engineering. Based on the principle of spray momentum, this paper measures the injection characteristics of each hole of four five-hole pressureless chamber injectors of the same model and analyzes the circulating fuel injection volume and flow coefficient of each injector and each hole under different working conditions. It is proposed to evaluate the quality of the injector with the average circulating fuel injection volume, average flow coefficient, and nonuniformity as indicators. The test results are as follows: there are differences in the circulating fuel injection volume and flow coefficient between each hole of the same fuel injector. With the increase of the fuel injection pump speed, the average circulating fuel injection volume of each hole differs by 2.8%–47.5%, and the average flow coefficient differs by 3.7%–30%; as the fuel injection volume increases, the average circulating fuel injection volume of each injector differs 1.8%–36%, and the average flow coefficient difference is 2.5%–28.7%. The circulating fuel injection volume and flow coefficient of different fuel injectors of the same model are different. With the increase of the fuel injection pump speed, the average circulating fuel injection volume of each injector differs by 3.5%–9.6%, and the average flow coefficient differs by 1.4%–5.7%; as the fuel injection volume increases, the average circulating fuel injection volume of each injector differs 0.3%–5.5%, and the average flow coefficient difference is 2.8–4.2%. The relative flow coefficient of each hole differs from 0 to 0.02, and the nonuniformity differs from 1.8% to 16.9%. The relative circulating fuel injection amount of each hole differs from 0.02 to 0.1, and the nonuniformity differs from 1.1% to 6.9%. The relative flow coefficient of each hole and its nonuniformity is smaller than the relative circulating fuel injection volume of each hole and its nonuniformity.

Optimization of injection parameters for temperature time response gel plugging agent

Aiming at the temperature time response gel plugging agent developed by Daqing exploration and development institute. In this paper, the injection volume, injection speed and injection concentration are optimized. The experimental results show that: the temperature time response gel can achieve the best plugging effect under the injection volume 0.1PV, injection concentration 1000mg/L and injection speed 0.6ml/min. In the subsequent water injection stage, the conductivity of high permeability layer decreases from 72.7% to 0.7%, that of low permeability layer increases from 4.1% to 14.3%, and that of medium permeability layer increases from 23.2% to 85%.

Evaluation of computed tomography settings in the context of visualization and discrimination of low dose injections of a novel liquid soft tissue fiducial marker in head and neck imaging

Background Intraoperative incorporation of radiopaque fiducial markers at the tumor resection surface can provide useful assistance in identifying the tumor bed in postoperative imaging for RT planning and radiological follow-up. Besides titanium clips, iodine containing injectable liquid fiducial markers represent an option that has emerged more recently for this purpose. In this study, marking oral soft tissue resection surfaces, applying low dose injections of a novel Conformité Européenne (CE)-marked liquid fiducial marker based on sucrose acetoisobutyrate (SAIB) and iodinated SAIB (x-SAIB) was investigated. Methods Visibility and discriminability of low dose injections of SAIB/x-SAIB (10 µl, 20 µl, 30 µl) were systematically studied at different kV settings used in clinical routine in an ex-vivo porcine mandible model. Transferability of the preclinical results into the clinical setting and applicability of DE-CT were investigated in initial patients. Results Markers created by injection volumes as low as 10 µl were visible in CT imaging at all kV settings applied in clinical routine (70–120 kV). An injection volume of 30 µl allowed differentiation from an injection volume of 10 µl. In a total of 118 injections performed in two head and neck cancer patients, markers were clearly visible in 83% and 86% of injections. DE-CT allowed for differentiation between SAIB/x-SAIB markers and other hyperdense structures. Conclusions Injection of low doses of SAIB/x-SAIB was found to be a feasible approach to mark oral soft tissue resection surfaces, with injection volumes as low as 10 µl found to be visible at all kV settings applied in clinical routine. With the application of SAIB/x-SAIB reported for tumors of different organs already, mostly applying relatively large volumes for IGRT, this study adds information on the applicability of low dose injections to facilitate identification of the tumor bed in postoperative CT and on performance of the marker at different kV settings used in clinical routine.

Establishment of the Quantitative Analysis of Multiindex in Euphorbia lathyris by the Single Marker Method for Euphorbia lathyris Based on the Quality by Design Concept

Methods. The influences of methanol proportion, flow rate, column temperature, and injection volume in the mobile phase on the chromatographic resolution of chromatographic peak of euphorbia factor L1 were experimentally studied via Plackett–Burman design, and the key analysis parameters were screened out; the key analysis parameters were optimized through the central composite design, and the chromatographic analysis conditions were established. Euphorbia factor L1 was taken as the internal reference to construct the relative correction factors for L3 and L4 relative to L1, and their contents were calculated, thus realizing the QAMS. Meanwhile, the euphorbia factor L3 and euphorbia factor L4 were determined using the external standard method, and the differences of values measured by the external standard method from the values predicted by the QAMS method were compared, in an effort to verify the accuracy and feasibility of the QAMS method. Results. The methanol proportion and column temperature in the mobile phase were the key analysis parameters P < 0.05 , and the chromatographic conditions were determined as follows. The methanol/water ratio, column temperature, detection wavelength, flow rate, and injection volume were 60 : 40, 30°C, 275 nm, 1.0 mL/min, and 10 μL, respectively. A total of 20 batches of samples were determined by the QAMS method and external standard method; the relative standard deviations (RSDs) of L3 and L4 determination results were less than 2.0%, without any significant difference. Conclusion. The QbD-based QAMS method can be used to determine the contents of euphorbia factor L3 and euphorbia factor L4 in Euphorbia lathyris L., and it is accurate and feasible.

A Simple HPLC/DAD Method Validation for the Quantification of Malondialdehyde in Rodent’s Brain

In the present study, a HPLC/DAD method was set up to allow for the determination and quantification of malondialdehyde (MDA) in the brain of rodents (rats). Chromatographic separation was achieved on Supelcosil LC-18 (3 μm) SUPELCO Column 3.3 cm × 4.6 mm and Supelco Column Saver 0.5 μm filter by using a mobile phase acetonitrile (A) and phosphate buffer (20 mM, pH = 6) (B). Isocratic elution was 14% for (A) and 86% for (B). The injection volume (loop mode) was 100 μL with an analysis time of 1.5 min. Flow rate was set at 1 mL/min. The eluted compound was detected at 532 nm by a DAD detector by keeping the column oven at room temperature. The results indicated that the method has good linearity in the range of 0.2–20 μg/g. Both intra- and inter-day precision, expressed as RSD, were ≤15% and the accuracies ranged between ±15%. The lower limit of quantification (LLOQ), stability, and robustness were evaluated and satisfied the validation criteria. The method was successfully applied in a study of chronic toxicology following different treatment regimens with haloperidol and metformin.