Column Oven Research Articles

A New Stability Indicating Reverse Phase HPLC Method for the Determination of Related Substances of DPP4 Inhibitor Drug: Linagliptin

A simple, sensitive, selective and reproducible stability indicating reverse phase HPLC method has been developed and validated for the determination of related substances of linagliptin drug substance. Separation of impurities was carried out on a YMC Pack ODS-AQ, 5 μm (250 × 4.6 mm) column. Column eluent consists of gradient mixing of phosphate buffer at pH-3.7 as mobile phase A and a mixture of acetonitrile, methanol and water in the ratio of 600:250:150 v/v/v as mobile phase-B. Gradient elution at a flow rate of 1.0 mL/min and injection volume is 10 μL. The UV/vis detector is set to a wavelength of 226 nm and the column oven is set at 30 ºC. The detector response was found to be linear over the concentration range of 0.07-1.1 μg/mL with correlation coefficients greater than 0.999 for linagliptin and its related substances. LOD values for specified impurities range from 0.004-0.005% w/w and LOQ values from 0.013-0.015% w/w. The accuracy obtained by the proposed method was found to be in the range of 93.2-101.3%. Degraded and process impurities are well separated and the method has been validated for specificity, precision, ruggedness, linearity, accuracy and robustness as per ICH guidelines.

Design and application of online derivatization device for polar organics on atmospheric particulate filter

设计制作了一套用于气相色谱-质谱(GC-MS)分析极性有机物的在线衍生装置,并将其应用于大气颗粒物样品中极性有机物的检测。将大气颗粒物滤膜样品置于GC-MS进样口,通过使用套针组件,匀速引入气态衍生试剂N-甲基-N-(三甲基硅烷)三氟乙酰胺(MSTFA),使其在衬管内于310 ℃下与待测物接触,10 min即可完成硅烷化在线反应。反应过程中,色谱柱箱保持低温,衍生产物得以在柱头保留,反应完成后色谱柱箱程序升温,使衍生产物直接进行后续分离检测。应用在线衍生装置建立有机酸分析方法,获得了一元酸、二元酸、芳香酸、醇等极性有机物的检测信息,涵盖了大气化学分析常见的大部分目标化合物。该方法检出限为0.02~0.53 mg/L,线性相关系数为0.976~0.996,日内、日间RSD为0.27%~7.28%,适用于批量大气环境样品检测。与传统离线衍生技术相比,本装置使衍生反应处于高温惰性气体氛围,排除空气中水分对衍生试剂的损耗和衍生产物降解风险,反应稳定、效率高;固体滤膜上有机物进行热解吸的同时完成在线衍生,样品需求量小,操作简单,零有机试剂污染,并有望应用于醇类、酚类等多种极性有机化合物的分析。此外, 该装置搭建简单,可模块化设计,适用于不同品牌气相色谱仪,具有商业化推广前景。

Gas chromatography method to quantify α-monobenzoate glycerol in methanolic solutions using methyl benzoate as internal standard and a Rxi-1ms column

The 2,3-dihydroxypropyl benzoate, also known as α-monobenzoate glycerol (α-MBG), is an important building block used in the organic chemistry industry and its synthesis can be performed by the direct esterification of glycerol and benzoic acid. On the other hand, gas chromatography is a powerful laboratory tool applied for the quantification of known and unknown molar concentrations of chemical compounds. Under those conditions, I developed a method and a calibration curve for the analysis of unknown concentrations of α-MBG in methanolic solutions using a gas chromatograph equipped with a Rxi-1ms column and methyl benzoate (MBe) as an internal standard. Thus, by a suitable chromatograph setup, such as injector temperature and column oven temperature program, it was achieved a good resolution of the chromatographic peaks of each individual compound (MBe and α-MBG). Additionally, it was possible to generate an appropriated calibration curve with a corresponding equation for the calculation of unknow molar concentrations of α-monobenzoate glycerol in methanolic solutions. The highlights of the method described are:•There were no derivatization steps in this method, only dilution of the α-monobenzoate glycerol in methanol.•The calibration curve was made using only four calibration mixtures.•This method also promotes the appropriate separation of glycerol, benzoic acid, and methyl benzoate, which can be useful for the analysis of the progress of an esterification reaction of glycerol and benzoic acid for the synthesis of α-MBG.

GALLIC ACID ANALYSIS BY HIGH-RESOLUTION LIQUID CHROMATOGRAPHY ON REVERSE PHASES IN RASPBERRY FLOWER EXTRACT (Rubus idaeus L.)

Gallic acid is a phenolic acid widely spread in various plants, where it is present either in free form or as part of tannins, called gallotannins. It shows several positive effects on human health. It has different biological effects: antibacterial, antiviral, and anti-inflammatory. Many scientists are showing great interest in gallic acid precisely because of its antitumor effect. The content of gallic acid was determined in (Rubus idaeus L.) Polka raspberry by applying reverse-phase high-resolution liquid chromatography on reverse phases on Shimadzu Prominence Modular HPLC with UV/Vis detector, mobile phase degasser, pump, autosampler, and column oven. The analysis was performed in ethanolic extracts of Polka raspberry flower obtained by the Soxhlet extraction and the ultrasonic methods. Raspberry flowers (Rubus idaeus L.) Polka varieties were collected from two different localities in Bosnia and Herzegovina, namely: Starposle near Kakanj and Moševac near Maglaj. Gallic acid was determined in all analyzed samples of Polka raspberry. The highest gallic acid content was determined in the extract of Polka raspberry flower from the Maglaj-Moševac site, using the ultrasonic method (0.1789%), and the smallest in the Polka raspberry flower from the Kakanj Starposle site, using Soxhlet extraction (0.0995%). The proportion of gallic acid was higher in the extract of Polka raspberry flower from the Maglaj-Moševac site, which suggests that these flowers of Polka variety are recommended as better natural sources of this phenolic acid and that ultrasonic extraction has proven to be a more efficient method for extraction of gallic acid in ethanolic extracts, which is the goal of research.

Isolation and Structural Elucidation of Impurity in Sulfonamide Intermediate of Glyburide

A reproducible isolation method by Reverse Phase (RP) preparative HPLC technique for the isolation of one crucial impurity at 1.65 RRT (Relative Retention Time) in sulfonamide stage of Glyburide API (Active Pharmaceuticals Ingredient) was developed. Preparative chromatography was done on Luna C8, 10µm (250 mm x 21.2mm) preparative HPLC column with acetonitrile: water in 70:30 % v/v proportion as a mobile phase and 8 ml/min as a flow rate. This impurity was detected at 300 nm UV-wavelength maximum. This impurity was isolated from synthesized crude impurity of sulfonamide stage of Glyburide substance by preparative HPLC by injecting 50 mg/ml concentration over 5 ml fixed loop. Isolated impurity was elucidated as N-methyl impurity of sulfonamide intermediate of Glyburide API by means of chromatographic and spectral data. Structural elucidation carried out by spectral data was reviewed. This impurity was analyzed by reverse phase HPLC for purity analysis. A Inertsil C8 (250 x 4.6) mm, 5µ particle size was employed for separation. The mobile phase consisted of Water: Acetonitrile: Methanol in the ratio of 60:15:25 % v/v. The flow rate was set at 1 ml/min. Detection was carried out at 300 nm. 10µL of 2 mg/ml concentration of sample in methanol was injected. The column oven temperature was at 25°C.

A Stability Indicating Method Development and Validation for The Simultaneous Estimation of Emtricitabine, Tenofovir Alafenamide and Doultegravir in Pharmaceutical Formulation by Ultra Performance Liquid Chromatography

A Precise, specific, linear, accurate and robust technique used to be developed for the simultaneous estimation of the, Emtricitabine, Tenofovir alafenamide and Doultegravir in pill dosage structure and Validated as per ICH Validation guidelines. Method was once optimised with the useful resource of Acquity BEH C18 125A° (100 × 2.1 mm, 1.7μ) column at a flow rate of 0.6ml/min, Mobile part was as soon as pH 3.8 Buffer, Methanol and Acetonitrile (50:40:10 respectively). The Column Oven temperature used to be maintained at 35°C and working wave dimension used to be selected at 274nm. The retention times of Emtricitabine, Tenofovir Alafenamide and Doultegravir have been determined to be 3.77, 6.34 and 7.67minutes respectively. The % RSD of the Emtricitabine, Tenofovir Alafenamide and Doultegravir had been and located to be 0.27%, 1.24% and 0.11% respectivelyIn Method precision Parameter, % Assay had been determined ninety five to 105.0% and p.c Recovery had been acquired as 98.9%, 100.4% and 100.0% for Emtricitabine, Tenofovir Alafenamide and Doultegravir respectively. Linearity was as soon as acquired as 0.999, 0.999 and 0.999 for Emtricitabine, Tenofovir Alafenamide and Doultegravir respectively. Analytical Range used to be determined from the linearity and accuracy for Emitricitabine used to be 50µg/mL to 150µg/mL, Tenofovir Alafenamide used to be as soon as 6.25µg/mL to 18.75µg/mL and Doultegravir was as soon as 12.5µg/mL to 37.5µg/mL.

Stability indicating Analytical Method Development using Quality by Design (QbD) approach for simultaneous estimation of Ivabradine and Metoprolol

The applicability of a quality by design (QbD) approach for the development of a sensitive and selective stability indicating reversed-phase high performance liquid chromatography (RP-HPLC) method for the estimation of Ivabradine and Metoprolol was investigated. Design of experiments using a fractional factorial design approach was used for method development. Fifteen experimental runs were performed to optimize the chromatographic conditions like mobile phase, flow rate and column oven temperature. Mobile phase composition was optimized by changing Acetonitrile composition ranging between 13 and 17% v/v, Flow rate from 0.6 to 1.0 ml/min and temperature between 30 to 50 0C. The optimized method produced sharp peaks with good resolution (>2) for Metoprolol and Ivabradine with retention times of 3.3 and 9.2 min, respectively. The experimental data revealed that volume of Acetonitrile in mobile phase was prominently affecting the Retention time, Resolution & tailing factor of both the drugs. Normal probability plots revealed that the residual and predicted data fall approximately on a straight line, indicating that the experimental error for these studies was evenly distributed suggesting that the model could be used to navigate the design space. This approach is useful to expedite method development and optimization activities in analytical laboratories.

Hydrogel Coating with Temperature Response Retention Behavior and Its Application in Selective Separation of Liquid Chromatography

We reporte the double-layer hydrogel-coated mesoporous silica material as a new stationary phase for liquid chromatography. The method of combining physical coating and chemical coating was to apply hydrogel coating on the surface of silica, and finally, a new type of liquid chromatography stationary phase with in situ coating of the functional hydrogel on silica was obtained. This hydrogel-functionalized liquid chromatography stationary phase also exhibits a certain temperature responsiveness. Experimental results show that this temperature response is mainly due to changes in the hydrogen bonding between the stationary phase and the analyte at different temperatures in the column oven, which leads to changes in retention behavior. The hydrogel-coated mesoporous silica microspheres showed excellent selectivity for many polar analytes. An excellent column efficiency was obtained (139 000 plates/m for terephthalic acid) after optimization of chromatographic conditions. In addition to rapid separation of some analytes, this new hydrogel stationary phase also has certain superiority in chromatographic performance compared with other new excellent liquid chromatography stationary phases functioned by three-dimensional cross-linking systems. The important thing is that this strategy is relatively easy to prepare a new stationary phase with different properties.

Development and validation of HPLC method for enantioseparation of Ibrutinib on immobilized chiral stationary phase

Novel HPLC method was developed for enantioresolution of tyrosine kinase inhibitor drug Ibrutinib (IBT). Separation and identification of IBT enantiomers was investigated using cellulose tris (3,5-dichlorophenylcarbamate) immobilized on silica gel (Chiral-pack-IC). Good baseline enantioseparation was achieved on Chiral-pack-IC under normal-phase condition composed of n-Hexane, ethanol in a ratio of 55 and 45 (%v/v) with 0.1% Diethyl amine and 0.3% Trifluoro acetic acid. Resolution between IBT and its enantiomer was above 4. The flow rate and column oven temperature were 0.9 mL/min and 30 °C respectively. Detection was carried out at 260 nm. Injection volume optimized was 10 µL. Developed method was validated in accordance with ICH guide lines for specificity, limit of detection, limit of quantification, linearity, accuracy, precision, ruggedness and robustness.

Formulation, optimization, qualitative and quantitative analysis of new dosage form of corticosteroid

BackgroundThe objective of the present research work is to develop the entirely new drug product formulation (Ophthalmic Suspension) which belongs to the therapeutic class of steroids drugs (Corticosteroid). There are different dosage forms of Triamcinolone acetonide to treat different systemic as well as topical diseases as per patient symptomatic conditions. Due to non-availability of this active drug (Triamcinolone acetonide) ophthalmic dosage form in the national and international market research work is carried out to enter in this dosage form.ResultsIn the formulation design and development, different inactive like polysorbate 30 mg/mL, PEG-12 glyceryl dimyristate 70 mg/mL, ethyl alcohol 10 µL/mL, citric acid 1 mg/mL, sodium citrate 3.5–4.0 mg/mL, BKC 0.15 mg/mL and water for injection have been used at different stages in different proportions during the manufacturing procedure. Gradient HPLC, C18 column, 5 µm, 100 A, 3.9 mm × 300 mm, detector UV/Vis @ 254 nm, column oven, auto-sampler, degasser, is used and this analytical testing method is validated to obtain the accurate results. The excipients play different roles including solvent, antioxidant, solubilizer, emulsifier, antibacterial, preservative, anticoagulant, antimicrobial agent, surfactant and buffers in the stability of active drug for its excellent therapeutic output results. Correlation coefficient factor is greater than 0.999 which indicates that the method has good linearity at proposed concentration against triamcinolone acetonide.ConclusionsExcellent physical and chemical properties like physical appearance, pH, specific gravity, viscosity, re-suspendability, sedimentation volume is accomplished to meet the requirements of the eye treatment products. On the basis of experimental results achieved during validation process, it can be assumed that the current analysis method is more accurate, precise, specific, linear and consistency indicating in the range of 70 to 130% of the analyte concentration. From the physical, chemical behaviors and chemical assay of the newly developed drug formulation of corticosteroid (triamcinolone acetonide) it is concluded that it can be probably put for further clinical trial in different phases for human use.

Analysis of short-chain bioactive peptides by unified chromatography-electrospray ionization mass spectrometry. Part I. Method development

A method to analyse short-chain bioactive peptides (MW < 800 Da) and their impurities was developed with a unified chromatography (UC) analysis, including a wide mobile phase gradient ranging from supercritical fluid to near-liquid conditions, with UV and electrospray ionization mass spectrometry detection (ESI-MS). Four stationary phases and three mobile phase compositions were examined. Ten model peptides were first selected to identify the best operating conditions, including five linear tripeptides and five cyclic pentapeptides, with log P values ranging from -5.9 to 3.6, and including isomeric species. Derringer desirability functions were designed to identify optimal operating conditions based on 7 criteria, namely the number of peaks detected (including all impurities resolved), the proportion of the chromatogram occupied by target peaks, the least favourable resolution observed between the main peptide and impurities, peak shape features (asymmetry and peak width at half height), and finally the signal-to-noise ratio observed both with UV (210 nm) and ESI-MS in positive ionization mode.The optimum conditions were obtained on Ascentis Express OH5 stationary phase, with a mobile phase composed of carbon dioxide and methanol, comprising 2% water and 20 mM ammonium hydroxide. The final gradient program ranged from 5 to 80% co-solvent in CO2, with a reversed flow rate gradient ranging from 3.0 to 1.5 mL/min. Back-pressure was set at 120 bar and the column oven temperature at 60°C. Optimal conditions were applied to a large set of 76 peptides (34 linear tripeptides and 42 cyclic pentapeptides) and provided adequate scattering of the peaks in the retention space, together with some separation of isomeric species, particularly for the cyclic peptides.

A NOVEL VALIDATED HIGH-PERFORMANCE LIQUID CHROMATOGRAPHIC STABILITY-INDICATING ASSAY METHOD IN REVERSE PHASE FOR ESTIMATION OF HYDROXYPROGESTERONE CAPROATE IN AN INJECTABLE FORMULATION.

A high-performance liquid chromatographic stability-indicating assay method in reverse-phase has been developed and validated for the quantitative measurement of Hydroxyprogesterone corporate in injectable formulation prepared in castor oil base containing excipient benzyl benzoate as preservative. The reverse-phase method was developed by using Sunfire C18 column having dimensions (150 x 4.6 mm x 5μm) with a mobile phase[A] comprised of water, acetonitrile, and Methanol mixed in 40:30:30 v/v ratio and mobile phase [B] comprised of water, acetonitrile, and Methanol mixed in 20:40:40 v/v ratio pumped gradient program at a flow rate of 1.0 ml/min where the thermostat of the column oven controlled at 30°C and sample compartment at 5°C. The injection volume is set at 20μL. Hydroxyprogesterone corporate has a retention time of 10 minutes with UV detection at 240 nm. The detector was showing linear response in a range of 0.25 μg/ml to 150 μg/ml for Hydroxyprogesterone corporate. The guidelines of the International Conference on Harmonization on guidance for industry (2005) validation of analytical procedures, text, and methodology Q2(R1) were followed for the validation of this novel method. For quantification of Hydroxyprogesterone corporate the developed method was found to be genuinely exact precise, accurate, linear, robust, rugged, stable. and cost-effective.

Development and Validation of a Diethyl Phosphite Content in Foscarnet Sodium USP by GC MS Technique

A simple, rapid, selective, and reproducible Gas chromatographic mass spectrometry (GC-MS) method has been developed and validated for the estimation of Diethyl Phosphite content in Foscarnet Sodium USP Drug substance. The drugs were estimated using HP-5, Length-30 M, Internal diameter 0.32 mm; Film thickness 1 μ at a total flow rate of 11.9 ml/min, and column flow of 1.49 ml/min was used for the separation. Flow control mode was pressure. Column oven temperature 70°C and injector temperature 220°C. Oven program modified for proper elution of peak. The linearity range used was 0.025-0.120µg/ml and (Rt) was 6.7 min. The correlation coefficient values were found to be 0.997. Precession studies showed % RSD values less than 15.0% for all the selected concentrations. The percentage recovery of Diethyl phosphite from LOQ to 150% was found in range of 100.7 -116.7%. The content results of Phophite content were within the limits of less than 0.12 ppm. The method was validated as per the International Conference on Harmonization (ICH) guidelines. The developed method was successfully used for the quantitative analysis of commercially available dosage forms.

Innovative Method Development Comprehensive Separation of Impurities and Validation for a novel Antipsychotic Drug Blonanserin

Blonanserin an antipsychotic novel drug used for the treatment of schizophrenia has antagonist properties for dopamine D2 and serotonin 5-HT2. On the other hand, it lacks adrenergic-α1, muscarinic M1, and histamine H1 antagonist activities. Clinical studies demonstrated in Japan had shown to be more effective for treating negative as well as positive schizophrenic symptoms. This drug was accepted and approved worldwide in the treatment of schizophrenia. A new HPLC method was developed and validated for the estimation of Impurities of Blonanserin (BNS) to ensure that the methodology meets the requirements of the target analysis application. Active and efficient chromatographic separation was achieved on a Zorbax Bonus RP EP C18 column having a particle size of 5μm, with dimensions of 250mm × 4.6 mm, mobile phase containing pH 2.4 buffer and Organic, with 1.0 ml /min flow rate, column oven temperature at 30oC and the eluent detection at 245 nm. The method shows well-separated impurities, is specific without interference from blank solution with resolution more than 1.2 between any of the impurity, correlation coefficient more than 0.99 showing good linearity; mean recovery ranging from 97% to 105% and is very sensitive at lower detection and quantification limits. This method was well developed and has been applied successfully to monitor and estimate impurities in Blonanserin.

Development and Validation of Estimation of Genotoxic Impurity (Benzimidamidecontent) in Leflunomide by Using RP-HPLC Technique

The measurement of Genotoxic contaminant, a simple, selective, linear, accurate, and specific reverse phase high-performance liquid chromatographic (RP-HPLC) process was proposed. A Benzimidamide impurity in the medication Leflunomide has been discovered. Separation and analysis were carried out on Zorbax SB phenyl (4.6 mm x 250 mm) with a particle size of 5.0 μm. with 0.1 % Triethylamine in purified water with a pH of 7.0 and a buffer of phosphoric acid (20% in water). The mobile phase is a 40:60 mixture of buffer and Acetonitrile with degassing. Isocratic program mode was used. The elution was carried out at a rate of 1.0 mL/min with UV detection at a wavelength of 289 nm. The temperature of the selected column oven is 25°C. The linearity and accuracy of Benzimidamide are covered in this approach, with a LOQ limit of 150 percent (i.e.0.03 to 0.45 ppm). The observed correlation coefficient is 0.99994, with a range of 100.01 to 104.8 for recovery. The measured percent RSD of six spiked test preparation is below 5.0 percent in procedure precision (i.e. repeatability) and intermediate precision (IP). When maintained at room temperature, the standard and sample remained stable for three days. System appropriateness characteristics such as tailing factor and percent RSD do not exhibit significant changes in robustness experiments. For the detection of Benzimidamide the present RP-HPLC method is act as selective, robust, linear and precise.

Ultra-short ion-exchange columns for fast charge variants analysis of therapeutic proteins

The purpose of this work was to study the potential of recently developed ultra-short column hardware for ion exchange chromatography (IEX). Various prototype and commercial columns having lengths of 5, 10, 15, 20 and 50 mm and packed with non-porous 3 µm particles were systematically compared. Both pH and salt gradient modes of elution were evaluated. Similarly, what has been previously reported for reversed phase liquid chromatography (RPLC) mode, an “on-off” retention mechanism was observed in IEX for therapeutic proteins and their fragments (25–150 kDa range).Because of the non-porous nature of the IEX packing material, the column porosity was relatively low (ε = 0.42) and therefore the volumes of ultra-short columns were very small. Based on this observation, it was important to reduce as much as possible all the sources of extra-column volumes (i.e. injection volume, extra-bed volume, detector cell volume and connector tubing volume), to limit peak broadening. With a fully optimized UHPLC system, very fast separations of intact and IdeS digested mAb products were successfully performed in about 1 min using an IEX column with dimensions of 15 × 2.1 mm. This column was selected for high-throughput separations, since it probably offers the best compromise between efficiency and analysis time. For such ultra-fast separations, PEEK tubing was applied to bypass the column oven (column directly connected) to the optical detector via a zero dead volume connection.

A Development and Validation of RP-HPLC Method for the Determination of Degradation Impurities in Anagrelide Dosage Form

A simple and selective method for liquid chromatography has been developed to determine the degradation impurities in solid oral dosage forms of Anagrelide (Antithrombocythemic agent). An excellent resolution between Anagrelide (ANG) and its related substances was achieved with Symmetry C8, 250 mm × 4.6 mm, 3.0 μm column. and mobile phase A (phosphate buffer, pH 4.1) and mobile phase B (the mixture of acetonitrile, methanol, and buffer in the ratio of 40:40:20 v/v/v). The column oven temperature was maintained at 40°C. The flow rate was fixed at 0.8 mL/min, and elution was monitored at 254 nm using a UV detector. The method was validated for precision, accuracy, specificity, linearity, and sensitivity. The validation studies demonstrated that the current HPLC method is reproducible, specific and accurate for its intended purpose. Linearity was observed for ANG, and its impurities are more than (R2 > 0.995). The limit of detection and limit of quantitation (LOQ) was found to be adequate (< 0.05 %) for the estimation of impurities. The RSD for intra- day and inter-day precision was found to be between 0.5 % to 2.5 %. The percentage of recovery was found between 95 % to 105 %. Forced degradation studies were performed on ANG under different conditions such as acidic, basic, oxidation, photostability, and thermal degradation. Based on the results, this is found to be simple, specific, precise, accurate, linear, and stability-indicating method. Hence it can be used to determine the Anagrelide-related impurities in the capsule dosage form. It is economically feasible and user-friendly method. The proposed method was successfully applied in quality control labs for stability analysis.

Investigation of Drug-Excipient Compatibility Studies Using Validated RP-HPLC Method for Azelnidipine and Telmisartan Tablets

Aims: The Drug-Excipient compatibility testing was conducted at an early product development stage to determined that Excipients were compatible with drugs used in formulation and to distinguish as many degradation products as possible using validated gradient RP-HPLC method.&#x0D; Study Design: Drug-Excipient Compatibility study was conducted in glass vials at different stability conditions namely, at 300C + 20C/75% + 5% RH, 400C + 20C/ 75% + 5% RH for 04 weeks and another set of closed vials were stored in stability chamber at temperature 600C + 20C for 02 weeks.&#x0D; Methodology: Samples were analyzed by validated RP-HPLC method using Inertsil C-18 Column 150 × 4.6 mm ×5 µm, column oven temperature 40°C, flow rate 1.5 mL/min, Injection volume 10 µL with run time 12.0 minutes at 254 nm using Acetonitrile and buffer as mobile phase in gradient mode.&#x0D; Results: The developed method meets all system suitability parameters and found specific to determine the drug in the presence of Excipient as no interference was observed at the Retention time (Rt) of analyte.&#x0D; Conclusion: There was no physical and chemical incompatibility observed with Drug-Excipient and did not observe significant increase in the related substances.

Determination of Chromatographic Assay of Deferasirox in Bulk and Pharmaceutical Dosage Form

Deferasirox belongs to the class antidote, which is not official in any pharmacopoeia. But only a few reported studies were availableemploying RP-HPLC technique. To validate the analytical process, we chose bulk powder and tablets of different intensity deferasirox tablets.The objective of the research or analysis is to develop a sensitive reverse phase chromatography method and validate quantitative perseverance of deferasirox in bulk and marketed formulation. A Simple, precise, accurate, economical and reliable UVspectrometric method has been developed toestimate deferasirox bulk and its tablet dosage form. The method was validated as per the international conference on harmonization (ICHQ2AR1) the validated dosage type was tested for device suitability, precision, linearity, specificity, robustness, and ruggedness.Statistical analysis proved that the method is repeatable and specific for the determination of the said drug. The established procedure or technique was done using Inertsil ODS 3V (C18 250mm X 4.6mm, 5µm) column oven temperature 40°C and sample temperature is 8°C with a flow rate of 1.5mL/min. By using HPLC waters PU-2487 pump and photodiode array detector at 245nm. The separation was carried out using a mobile phase consisting of amixture of sodium di hydrogen phosphate monohydrate buffer (pH3.0) and acetonitrile in the ratio of 45:55. The retention time of Deferasirox peak was considered to be about 7.5 min and the total run time was found to be 20 minutes.The calibration plot gave linear relationship over the concentration range of 6.28- 811.480 µg/ml The average percent recovery was proved to be 96.1,97.6&103.The correlation coefficient was found to be 0.999. The relative standard deviation (R.S.D) was found <2.0% for the UV-spectroscopic RP-HPLC method. The percent assessment of the drug was around 100% indicating the accuracy of the procedure. The suggested techniques were further confirmed and employed for the testing of the drug in tablet preparation.

Method Development and Validation of Simultaneous Estimation of Emtricitabine and Tenofovir Alafenamide in Bulk and tablet Dosage form using LC-MS/MS

A selective, sensitive, rugged and high throughput high performance liquid chromatography tandem mass spectrometric method was developed for the simultaneous estimation of Emtricitabine and Tenofovir Alafenamide in Bulk and Tablet Dosage form. Liquid chromatography system has been coupled with tandem quadrupole mass spectroscopy with LC-20AD pump, electrospray ionization (ESI) interface, PDA detector (SPD-M20), column oven (CTO-20AC), CEM-20 alite controller and SIL-20AC auto sampler. Lab solution data station software was used. The separation of both Tenofovir and Emtricitabine was achieved by Gemini C18 column (4.6×100mm, 5µm) as a stationary phase with 0.1 % formic acid: methanol at the ratio of 20:80, v/v at a flow rate of 0.5ml/min and injection volume of 10 µl. The percentage Recovery of the Drug Tenofovir Alafenamide and Emtricitabine was 96.59 and 96.84% respectively. The linearity range was 20-120 ng/ml for both the drugs and the LOD was found to be 5 and 10 ng/ml and LOQ were 15 and 20 ng/ml for Tenofovir and Emtricitabine Respectively. The developed method was precise, accurate, sensitive and can be used for the routine quality control of the drug formulation.