Targeted Separation of COX-2 Inhibitor from Pterocephalus hookeri Using Preparative High-Performance Liquid Chromatography Directed by the Affinity Solid-Phase Extraction HPLC System

Pterocephalus hookeri, as a kind of popular traditional Tibetan medicine, is reputed to treat inflammatory related diseases. In the present work, a cyclooxygenase-2 functionalized affinity solid-phase extraction HPLC system was developed and combined with preparative-HPLC for rapidly screening and separating cyclooxygenase-2 ligand from P. hookeri extracts. Firstly, ligands of cyclooxygenase-2 were screened from extracts by affinity solid-phase extraction HPLC system. Then directed by the screening results, the recognized potential active compounds were targeted separated. As a result, the major cyclooxygenase-2 inhibitor of P. hookeri was obtained with a purity of >95%, which was identified as sylvestroside I. To test the accuracy of this method, the anti-inflammatory activity of sylvestroside I was inspected in lipopolysaccharide-induced RAW 264.7 cells. The results show that sylvestroside I significantly suppressed the release of prostaglandin E2 with dose-dependent, which was in good agreement with the screening result of the affinity solid-phase method. This method of integration of screening and targeted separation proved to be very efficient for the recognition and isolation of cyclooxygenase-2 inhibitors from natural products.

Qualitative Estimation of Seed of Butea monosperma Lam. by using Chromatography Technique

The objective of present studies deals with the Qualitative estimation of seed of Butea monosperma Lam. By using Chromatography Technique. The phytochemical study of different extract of seed of Butea monosperma Lam. Were observed various active chemical constituent like phytosterol, flavonoid, saponin and sterol etc. Qualitative estimation of Gallic acid, Rutin and Quarcetin was carried out by HPTLC and HPLC system.

Development and Validation of RP-HPLC Method for Analysis of Aclidinium Bromide and Formoterol Fumarate in Pharmaceuticals

A fast, sensitive, and reliable RP-HPLC method involving cyberlab HPLC System with PDA detection was developed and validated for the quantification of Aclidinium bromide and Formoterol fumarate in inhalation preparations. Chromatography was performed on the Inertsil -ODS C18 (250 x 4.6mm, 5μ) column using filtered and mixed degassed methanol: buffer (75:25 v/v) as a mobile phase with a flow rate of 1.0mL/min and the column effluent was monitored at 240nm. Retention times for Aclidinium bromide 4.713min and Formoterol fumarate 6.691min. The method obeyed linearity in the concentration range of 20-80µg/mL for the two drugs when validated according to standard procedures.

IDENTIFICATION OF A BACTERIAL STRAIN ISOLATED FROM PICKLED MELON SOLUTION AND DETERMINATION OF ITS ABILITY TO PRODUCE LACTIC ACID

Bacterial strain LB7 isolated from pickled melon solution was identified as Lactobacillus plantarum using molecular marker 16S rRNA gene (rDNA) sequences. The content of lactic acid in the isolate culture was relatively high (33.7 g/l) when quantified using HPLC system.

Development and validation of a short runtime method for separation of trace amounts of 4-aminophenol, phenol, 3-nitrosalicylic acid and mesalamine by using HPLC system

In a short runtime process (about 6 minutes), with isocratic method without need of reconditioning before the next run, a mixture of some molecules containing 4-aminophenol (4APh), phenol, 3-nitrosalicylic acid (3NSA), and mesalamine (5-aminosalicylic acid) (MLZ), were separated and detected by means of HPLC/UV-Vis system. The resolutions of the separated and sharp peaks referring to the mentioned compounds were at least more than 2, and the tailing factor (TF) was about 1, even in 1 µg/l (ppm) concentration. In order to optimize the method, the effects of the pH, eluent percentage, flow rate, and buffer concentration, and wavelength on the chromatogram were investigated. Also, the results of the validation processes showed that the method trustable to be used in laboratories.

Analysis on Bulk Pharmaceutical Formulation, Validation and Estimation Using Levetiracetam Methods

A new sensitive, specific, direct, exact and correct RP-HPLC technique is established and authenticated to estimate Levetiracetam in Majority and Pharmaceutical Tablet Formulations. An isocratic, turned around period HPLC system might have been created should differentiate the pill starting with the corruption products, Phenomenex Gemini 5µ C18 (2) 100A (250 x 4.60mm, 5 µ) section. Hamilton syringe (705 NR, 50 µL) might have been utilized to injecting example Furthermore standard result. The versatile stage comprises about mixture of Methanol: Acetonitrile in the proportion (90:10 v/v) toward A stream rate about 1.0 ml /min. UV identification might have been performed toward 210 nm. The linearity might have been made for Levetiracetam in the extent from claiming 5- 30µg/ml for relationship coefficient about 0.9997. LOD Also LOQ were found will make 0.076µg/ml Furthermore 0.23µg/ml individually. Maintenance duration of the time of Levetiracetam were found with make 2.281min and 2.274min. % recuperation might have been discovered on be 99.78-100.45 What's more %RSD might have been found for over ±2. Those system needs been approved as stated by ICH rules to linearity, precision, accuracy, robustness, ruggedness, LOD furthermore LOQ. Those produced approved system might have been effectively connected to dependable quantification about Levetiracetam to mass and pharmaceutical measurement type.

THE OPTIMIZATION OF RP-HPLC CONDITION USING RESPONSE SURFACE METHODOLOGY BOX-BEHNKEN DESIGN FOR SIMULTANEOUS DETERMINATION OF METFORMIN HCL AND GLIMEPIRIDE IN SPIKED PLASMA

Objective: Aim of this study was to develop and validate the RP-HPLC method using Box-Behnken Design (BBD) for simultaneous analysis metformin HCl and glimepiride in spiked plasma. Methods: The chromatographic system was comprised of acetonitrile-phosphate buffer 0.0125 M+Sodium Dodecyl Sulphate (SDS) 1 mmol as a mobile phase and Ascentis® Phenyl C18 (250 x 4.6 mm i.d.; 5 µm) column as a stationary phase with UV detector at 210 nm. Three independent variables included phosphate buffer (%), pH and flow rate were optimized using Box-Behnken Design. The observed responses were retention time, peak area and resolution. Results: The predicted optimum condition of the RP-HPLC system consisted of phosphate buffer solution of 72%, pH at 4.3 and flow rate at 0.8 ml/min. By using this condition, the duration of analysis was more than 18 min, so it was necessary to modify the flow rate to be 1.0 ml/min to get shorter analysis duration. This condition was then applied to analyze metformin and glimepiride in spiked plasma and validated according to the EMA guideline. AUC of interfering components at the IS retention time between 588-1092 mV, the linearity of metformin was 0.9993 and glimepiride was 0.9991, accuracy and precision were between-13.33% until 16.08%, dilution integrity and metformin stability studies were between-4.01% until 11.82%, and for glimepiride stability studies were between-37.48% until-4.76%. Conclusion: Box-Behnken Design can help optimize the HPLC system, and the optimum condition was valid to analyze metformin and glimepiride in spiked plasma by considering the storage time of plasma samples.