HPLC.

A sensitive and rapid HPLC method was developed and validated for the determination of potential genotoxic impurities i.e (Bromomethyl)biphenyl methyl ester and (Dibromomethyl)biphenyl methylester at trace level in Telmisartan drug substance by applying the concept of threshold of toxicological concern (TTC). The HPLC method was developed and optimized on Symmetry Shield RP18, 3.5 m (150mm × 4.6mm) column with oven temperature maintaining at 40°C and 0.02M Phosphate buffer pH 2.5 was chosen as mobile phase A and mixture of acetonitrile and Phosphate buffer (55:45) was selected as mobile phase B in gradient reverse phase mode in isocratic mode of composition. Chromatographic parameters i.e flow rate: 1.0 ml/min, wavelength detection: 205 nm, injection volume: 20µl and run time: 25 min were applied in this methodology.  Based on validation data, the method is found to be specific, sensitive, accurate and precise. The established limits of Limit of detection and Limit of quantification for subjected impurities are found to be 2.4 µg/g and 4.7 µg/g respectively for each impurity. The recovery at LOQ level obtained was 98.2% for (Bromomethyl) biphenyl methyl ester and 99.2% for (Dibromomethyl) biphenyl methyl ester. This method can be used as good quality control tool for quantization of these impurities at low level. The experimental results are discussed in detail in this research paper.

This study examines the stability of paracetamol pediatric oral suspension (120mg/5ml) in simulated in-home storage conditions, temperature ranging from (300±5C/65±5RH) representing room condition. Samples from suspension were assayed for active pharmaceutical ingredient and tested for related substance (degradants) using B.P 2012 pharmacopeia HPLC method. The study was carried out in day zero, seven, fourteen, thirty and forty five.  The instrument utilize column ® C8, 100 x 4.6 mm, 3.5 μm particle size. The flow rate was 1.5 ml/minute. The mobile phase consisted of methanol, tetrabutylammonium hydroxide and sodium orthophosphate buffer. The results obtained showed that the drug assay content was out of the limits from day zero Furthermore, the half live was found to be 35.06 days

Andrographis paniculata is an important medicinal plant in Indian and Chinese systems of medicine used for a number of ailments. Andrographolide (AP), a diterpene lactone, has been reported to be responsible for many of the activities. A number of methods are available for the isolation of AP.  However, they are time consuming and not cost effective. We employed five relatively simpler methods for the isolation of AP. The identities and purity of the compounds isolated by these five methods were established by thin layer chromatography (TLC), high performance liquid chromatography (HPLC), melting point, ultraviolet (UV), infrared (FTIR), mass, and nuclear magnetic resonance (1H NMR) spectra. Treatment of the extracts with activated charcoal proved to be quite effective in removing all coloring matter and resulted in isolation of pure AP. Of the five methods, prior maceration with methanol for 16 h followed by refluxing for 1h yielded 0.6%w/w of AP and was found to be quite satisfactory. This method does not need prior extraction of the powder with toluene nor does it require washing of the concentrated extract with toluene.  In this method only methanol (MeOH) was employed. All the spectral data and melting point of andrographolide isolated by this method confirmed its purity. This method is simple as it involves only three steps and has the potential for commercial scale up.

The objective of the study is to compare the analytical performance in terms of precision and accuracy of three methods viz., Immunoturbidimetry (IT), Capillary Electrophoresis (CEP) and High Performance Liquid Chromotography (HPLC) for the measurement of glycosylated haemoglobin (HbA1c). A total of 40 routine patients samples were used for the estimation of HbA1c using the three methods and the results obtained were correlated between the three methods. The correlation coefficient ‘r’ obtained between IT & CEP, IT& HPLC and CEP & HPLC were 0.96, 0.95 & 0.99 respectively, indicating that an ‘r’ value of 0.99 obtained between CEP & HPLC is in agreement with the CLIA guidelines value of > 0.98,while poor ‘r’ values of 0.96 and 0.95 were obtained when IT method was compared with both CEP and HPLC methods. As per this study both CEP & HPLC methods gave good precision and accuracy for Bio-Rad controls. Further, CEP and HPLC methods have provisions to identify interference by Haemoglobin variants present in patient sample. The outcome this study recommends the use of one of the two methods viz., CEP and HPLC for routine measurement of HbA1c.

A simple and precise reversed-phase high performance liquid chromatography (HPLC) method for the determination of hydrochlorothiazide (HCT) in human plasma was developed and validated. Using hydroflumethiazide as an internal standard (IS), separation was achieved on Atlantis dC 18 column. The mobile phase, 10 mM monobasic potassium phosphate and acetonitrile (80:20, v: v), was delivered at a flow rate of 1.2 ml/min. The eluent was monitored by photodiode array detector, with the wavelength set at 272 nm. Plasma samples containing HCT and IS were extracted with methyl tert butyl ether and reconstituted in mobile phase. No interference in blank plasma or of commonly used drugs was observed. The relationship between the concentration of HCT in plasma and peak area ratio of HCT to the IS was linear over the range of 5-300 ng/ml. The intra-day and inter inter-day coefficient of variation and bias were

A novel, simple, precise, accurate and reproducible RP-HPLC method was developed and validated for simultaneous estimation of paracetamol, tramadol and dicyclomine in bulk and pharmaceutical formulations. Separation of paracetamol, tramadol and dicyclomine was successfully achieved on a Inertsil ODS C18 (250mm x 4.6mm x 5 µm). The mobile consisted of ammonium acetate buffer (pH 4.5): methanol (80:20 v/v) at a flow rate of 1.0 mL/min. The detection was performed at 271 nm. The method was validated according to ICH guidelines for linearity, sensitivity, accuracy, precision and robustness. The response was found to be linear in the concentration range of 500 µg/mL to1500 µg/mL for paracetamol; 50µg/mL to150 µg/mL for tramadol; 10 µg/mL to 30 µg/mL for dicyclomine. The LOD and LOQ for paracetamol were found to be 2.712 µg/mL and 9.042 µg/mL, respectively. The LOD and LOQ for tramadol was 0.9009 µg/mL and 3.003 µg/mL, respectively and for dicyclomine it was 0.380 µg/mL and 1.265 µg/mL, respectively.  The percentage recovery for paracetamol, tramadol, and dicyclomine were found to be 99.00%, 100.00% and 99.00%, respectively. The excellent percentage recovery values indicate the high accuracy of the proposed method. The method specifically determines the analytes in the sample without interference from excipients of tablet dosage forms. The method was extensively validated according to ICH guidelines for Linearity, Range, Accuracy, Precision, Specificity and Robustness.

The present report deals with the study of phytochemicals present in Padina tetrastromatica Hauck collected from the south east coast of Tamil Nadu, India. The phytochemicals of different extracts was analyzed by the standard procedure for UV-Vis spectroscopic and HPLC. The UV-Vis phytochemical profile of different extracts of Padina tetrastromatica Hauck was predicted. The qualitative HPLC fingerprint profile of methanol extract of Padina tetrastromatica Hauck was chosen at a wavelength of 660 nm due to sharpness of the peaks and suitable baseline. The profile showed one prominent peak at a retention time of 1.500 min and seven moderate peaks were also noted at the retention times of 2.220, 2.400, 2.500, 2.800, 3.000, 3.300 and 6.300 min respectively. The present study on Padina tetrastromatica Hauck fashioned a novel phytochemical marker in standardization of the quality of the drug used for medicine.

An accurate, precise and reproducible high performance liquid chromatographic method was developed for quantitative estimation of telmisartan in bulk drug samples and tablet dosage forms. Chromatographic separation of the drug was achieved on a Kromasil C18 column (150 x 4.6 mm; 5µ) using a mixture of phosphate buffer (pH 4.0) and acetonitrile (40:60 v/v) as the mobile phase at a flow rate of 1.0 mL/min. Under optimized conditions, the retention time of the drug was found to be 2.887 min. Good detecting sensitivity for the analyte was observed at 224 nm. The quantitation calibration curve for the drug was linear over the range of 20-60 µg/mL. The performance of the proposed method was validated as per ICH guidelines. The method was proved to be suitable for the estimation of telmisartan in tablet dosage forms. Key words: Telmisartan, Estimation, Tablets, HPLC.

The present study was aimed to explore the phytochemical constituents present in Turbinaria ornata (Turner) J.Ag. collected from the south east coast of Tamil Nadu, India. The phytochemical screening of different extracts was estimated using the standard procedure for UV-Vis spectroscopic and HPLC. The UV-Vis phytochemical profile of various extracts of Turbinaria ornata (Turner) J.Ag. was analyzed. The qualitative HPLC fingerprint profile of methanol extract of Turbinaria ornata (Turner) J.Ag. was selected at a wavelength of 254 nm due to sharpness of the peaks and proper baseline. The profile displayed one prominent peak at a retention time of 1.953 min and some moderate peaks were observed at a retention time of 3.000, 2.570 and 2.467 min respectively. The present study on Turbinaria ornata (Turner) J.Ag. produced novel phytochemical markers in standardization as useful analytical tools to check not only the quality of the powder but also the presence of adulterants in ayurvedic drugs.

  The present study investigates the beneficial role of anthocyanin extracted from two varieties of onion peel (Allium cepa L.,). And also to find out the quercetin and cyanidin present in the red onion peel and big onion peel by using HPLC. Key words; Allium cepa, red onion peel, big onion peel, anthocyanin, quercetin, cyanidin, HPLC.