Chromatography Research International
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Open Access journal
ISSN (Print) 2090-3502 - ISSN (Online) 2090-3510
Published by Hindawi [333 journals]
- Validated Stability Indicating Chromatographic Method for the Simultaneous
Estimation of Camylofin with NSAID Drugs and a New Approach of Method
Transfer from Classical HPLC to a Modern UPLC Instrument
Abstract: The presented work describes the method development of simultaneous determination of camylofin dihydrochloride (CMF), diclofenac potassium (DCF), and Paracetamol (PCM) using reversed phase high performance liquid chromatography (HPLC-UV) and the method was further transferred to a new generation instrument, ultraperformance liquid chromatography (UPLC-PDA). The detailed validation was carried out for the combination tablet formulation of CMF and DCF by UPLC-PDA. From the method development study, Acquity UPLC HSS C18 (2.1 × 50 mm, 1.8 μm) was finally selected for validation. The satisfactory results were observed for peak shape, retention time, and resolution with a mobile phase of 20 mM ammonium acetate buffer (pH 3.0 with dilute orthophosphoric acid) : methanol (33 : 67 v/v). The isocratic elution of mobile phase was carried out at a flow rate of 0.250 mL/min and detection at 220 nm. Both drugs were efficiently separated out in less than 3.5 min with 1.1 and 3.2 min of retention time of the CMF and DCF with 11.87 of resolution. The linearity was obtained in the 20.0–80.0 μg/mL range of concentration with 0.9998 of correlation coefficients for the substances. The method was analyzed for specificity with detailed force degradation study, which is a simple, precise, and accurate method, as per the International Conference on Harmonization (ICH) guidelines.
PubDate: Tue, 11 Oct 2016 14:19:43 +000
- Thermodynamic Study of Racemic Ibuprofen Separation by Liquid
Chromatography Using Cellulose-Based Stationary Phase
Abstract: Ibuprofen is a nonsteroidal anti-inflammatory drug (NSAID), also known for its significant antipyretic and analgesic properties. This chiral drug is commercialized in racemic form; however, only S-(+)-ibuprofen has clinical activities. In this paper the effect of temperature change (from 288.15 to 308.15 K) on the ibuprofen resolution was studied. A column ( mm) packed with tris(3,5-dimethylphenylcarbamate) was used to obtain the thermodynamic parameters, such as enthalpy change (), entropy change (), variation enthalpy change (), variation entropy change (), and isoenantioselective temperature (). The mobile phase was a combination of hexane (99%), isopropyl alcohol (1%), and TFA (0.1%), as an additive. The conditions led to a selectivity of 1.20 and resolution of 4.55. The first peak, R-(−)-ibuprofen, presented an enthalpy change of 7.21 kJ/mol and entropy change of 42.88 kJ/K·mol; the last peak, S-(+)-ibuprofen, has an enthalpy change of 8.76 kJ/mol and 49.40 kJ/K·mol of entropy change.
PubDate: Thu, 29 Sep 2016 13:59:16 +000
- Development and Validation of RP-HPLC Method for the Determination of
Hydrochlorothiazide in Bulk Drug and Pharmaceutical Dosage Form
Abstract: An HPLC-PDA method was developed and validated for the determination of hydrochlorothiazide in bulk and pharmaceutical formulation. The method was optimized selecting chromatographic conditions of 50 : 50 acetonitrile : water, Inertsil® column (ODS-3 250 mm × 4.6 mm 5 μm), 20 μL injection volume, flow rate of 1 mL/min at ambient temperature (30°C), and 272 nm. Another column of C18Zorbax® (Eclipse Plus, 4.6 × 250 mm, 5 μm) was tested showing no big difference in the method results. The method was validated giving good precision (RSD% < 1), acceptable linearity ( ≥ 0.9978), and low LOD and LOQ (0.5 and 1.7 μg/mL, resp.) on both columns. Successful application on pharmaceutical dosage tablet form gave high recovery of 99.93%. The method was compared with official BP and other reported methods. The proposed method is economic, simple, and rapid and hence can be employed for routine analysis in quality control laboratories.
PubDate: Thu, 25 Aug 2016 16:54:36 +000
- Development and Validation of New RP-HPLC Method for Simultaneous
Determination of Methyl and Propyl Parabens with Levetiracetam in Pure
Form and Pharmaceutical Formulation
Abstract: A simple and robust high-performance liquid chromatography (HPLC) method is described for the assay for levetiracetam (LTC), methyl paraben (MHB), and propyl paraben (PHB) either in their pure form or in commercial Levepsy® syrup. The method is selective and stability indicating and all chromatographic conditions were studied to obtain adequate separation of LTC, MHB, and PHB from their degradation products and from excipients. The HPLC separation was carried out on a RP C18 Hypersil BDS analytical column (150 mm × 4.6 mm ID) using gradient elution system. The mobile phase flow rate was 1.5 mLmin−1 and the column temperature was kept at 40°C. Complete separation of the studied components was obtained within a cycle time of 8 min. LTC, MHB, and PHB were eluted at 1.56, 5.86, and 7.85 min, respectively. Detection was carried out at 240 nm using a dual wavelength detector. The method has been validated for linearity, accuracy, precision, specificity, limit of detection, limit of quantitation, robustness, and ruggedness. The proposed method was successfully applied for the determination of LTC in the presence of parabens in Levepsy syrup.
PubDate: Mon, 25 Jul 2016 12:13:50 +000
- A Simple Analytical Method for High-Throughput Screening of Major Sugars
from Soybean by Normal-Phase HPLC with Evaporative Light Scattering
Abstract: This paper presents a simple analytical method for determining sugars in soybean (Glycine max (L.) Merr.) tissues. Sample preparation was modified from several early published methods. High-performance liquid chromatography (HPLC) equipped with an evaporative light scattering detector (ELSD) was used to separate, identify, and quantify seven sugars, including glucose, galactose, fructose, sucrose, melibiose, raffinose, and stachyose. Two mobile phases were programed into a gradient elution. Mobile phase A is pure water and mobile phase B is a mixture of acetonitrile and acetone 75 : 25 (v/v). Total chromatographic retention time is less than 20 minutes. This method has been validated for detection limit, calibration range, and intraday and interday repeatability. This method has been used analyzing more than 5000 soybean samples in the experiments studying natural genetic variation of sugar contents and components in soybean seeds and other tissues.
PubDate: Wed, 01 Jul 2015 06:51:47 +000
- Development and Validation of an HPLC Method for the Analysis of
Chlorpropham and 3-Chloroaniline in Potato Extracts
Abstract: Chlorpropham (CIPC) is the main sprout inhibitor used by potato industry. There is concern about the residues of CIPC and its degradation product 3-chloroaniline, 3-CA; hence, analytical methods are required to analyse their residues in potato samples. An HPLC-UV method was developed and validated for the separation and quantification of these compounds using propham (IPC) as an internal standard. The chromatographic conditions required to achieve good separation were 60% mobile phase of methanol, 15-minute run time at a flow rate of 1.5 mL/min, and a detection wavelength of 210 nm using Phenomenex (ODS-2 250 mm 4.60 mm 5 µm Sphereclone) column at an ambient temperature. The method was validated for precision, linearity, the limit of detection (LOD) and the limit of quantification (LOQ), producing high precision through RSD ≤ 0.03%, and acceptable criteria of the coefficient of determination () of the calibration curves (0.990). LOD values of CIPC and 3-CA were approximately 0.01 µg/mL whereas the LOQ values were approximately 0.04 µg/mL using repeated injection approach. The proposed HPLC method was compared with the standard GC method of the CIPC residues extracted showing good agreement . Despite using the same extract, the recovery results for the proposed HPLC method were 13% higher than GC analysis.
PubDate: Mon, 29 Dec 2014 00:10:15 +000
- Practical Implication of Chromatographic Method for Estimation of
Aceclofenac and Pregabalin in Bulk and Pharmaceutical Dosage Forms
Abstract: Background. Aceclofenac and Pregabalin in combination significantly reduce pain as compared to individual drug in chronic low back pain. Literature reveals that all the reported spectrophotometric methods either need tedious extraction procedures, do not offer high sensitivity, use nonspecific reagent, or recommend the measurement of absorbance in the near UV region where interference most probably occurs that does not offer suitable linearity range. Result. A selective, sensitive, accurate, and precise, high performance liquid chromatographic method with UV detector analysis of Aceclofenac and Pregabalin was investigated. Good chromatographic separation was achieved using an ODS-BP hypersil C18 column (250 mm × 4.6 mm, i.d., 5 μm) and a mobile phase consisting of 0.05 M phosphate buffer (KH2PO4) (pH 6.0) : methanol (60 : 40, v/v) at a flow rate 1 mL/min. The ultraviolet detector was set at wavelength 218 nm. Retention time for Aceclofenac and Pregabalin was found to be 3.220 and 5.910 min, respectively. Rectilinear relationship with good regression coefficients 0.999 and 0.999 was found over the concentration ranges of 5–25 μg/mL and 3.75–18.75 μg/mL for ACF and PGB, respectively, with detection limits 0.64 and 0.35 μg/mL and quantitation limits 1.95 and 1.06 μg/mL. Conclusion. The mean percentage recoveries were in the range of 98.45–100.08 and 99.69–100.48 for ACF and PGB, respectively. The developed method was successfully applied to the analysis of the drugs in their commercial tablets.
PubDate: Tue, 16 Dec 2014 00:10:06 +000
- A Validated Stability Indicating RP-HPLC Method Development and Validation
for Simultaneous Estimation of Aliskiren Hemifumarate and Amlodipine
Besylate in Pharmaceutical Dosage Form
Abstract: The present study describes the stability indicating RP-HPLC method for simultaneous estimation of aliskiren hemifumarate and amlodipine besylate in pharmaceutical dosage forms. The proposed RP-HPLC method was developed by using waters 2695 separation module equipped with PDA detector and chromatographic separation was carried on C-8 Inertsil ODS (150 × 4.6 mm, 5 µ) column at a flow rate of 1 mL/min and the run time is 10 min. The mobile phase consisted of phosphate buffer and acetonitrile in the ratio of 40 : 60% v/v and pH was adjusted to 3 with orthophosphoric acid and eluents were scanned using PDA detector at 237 nm. The retention time of aliskiren and amlodipine was found to be 3.98 and 5.14 min, respectively. A linearity response was observed in the concentration range of 30–225 µg/mL for aliskiren and 2–15 µg/mL for amlodipine, respectively. Limit of detection and limit of quantification for aliskiren are 0.161 µg/mL and 0.489 µg/mL and for amlodipine are 0.133 µg/mL and 0.404 µg/mL, respectively. The stability indicating method was developed by subjecting the drugs to stress conditions such as acid and base hydrolysis, oxidation, and photo- and thermal degradation and the degraded products formed were resolved successfully from the samples.
PubDate: Sun, 07 Dec 2014 09:47:17 +000
- Method Development and Validation for Determination of Febuxostat
Spiked Human Plasma Using RP-HPLC with UV Detection
Abstract: A rapid, simple, selective, and specific reverse phase high performance liquid chromatography (RP-HPLC) method with UV detection (315 nm) was developed and validated for estimation of febuxostat from spiked human plasma. The analyte and internal standard (diclofenac) were extracted using LLE with diethyl ether. The chromatographic separation was performed on Shodex C-18-4E (5 μm; mm) with a mobile phase comprised of methanol : acetate buffer pH 4, 20 mM (90 : 10 v/v), at a flow rate of 1 mL/min. Febuxostat was well resolved from plasma constituents and internal standard. The calibration curve was linear in the range of 250–8000 ng/mL. The heteroscedasticity was minimized by using weighted least square regression with weighing factor of . The intraday and interday %RSD was less than 15. Results of recovery studies prove the extraction efficiency. Stability data indicated that febuxostat was stable in plasma after three freeze thaw cycles and upon storage at −20°C for 30 days.
PubDate: Thu, 27 Nov 2014 07:11:19 +000
- Identification and Quantification of Aldose Reductase Inhibitory
Flavonoids in Herbal Formulation and Extract of Gymnema sylvestre Using
HPLC-PDA and LC-MS/MS
Abstract: Adulteration of herbal supplements is a major issue for many countries. A simple and reliable HPLC-PDA method was developed for quantification of aldose reductase inhibitory flavonoids rutin, quercetin, and kaempferol. The chromatographic separation was performed on a Fortis C18 column in gradient mode with detection at 267 nm. The presence of these markers was confirmed through the accurate m/z values and MS/MS data obtained using quadruple time of flight mass spectrometry (QTOF-MS). The proposed method was successfully applied to examine the amount of these active constituents in antiobese polyherbal formulation and plant extract of Gymnema sylvestre.
PubDate: Wed, 26 Nov 2014 12:49:15 +000
- High-Performance Liquid Chromatography Determination of Meloxicam and
Piroxicam with Ultraviolet Detection
Abstract: A simple accurate and sensitive high-performance liquid chromatographic method for the determination of meloxicam and piroxicam concentrations in small volume plasma samples has been developed. Following a liquid extraction using chloroform, samples were separated by reversed-phase high-performance liquid chromatography on an XBridge C18 column (4.6 × 250 mm) and quantified using ultraviolet detection at 360 nm. The mobile phase was a mixture of water with glacial acetic acid (pH 3.0) and acetonitrile (50 : 50), with a flow rate of 1.0 mL/min. The standard curve ranged from 5 to 10,000 ng/mL for meloxicam in bearded dragon (Pogona vitticeps) plasma and piroxicam in crane (Grus rubicunda) plasma. Intra- and interassay variability for meloxicam and piroxicam were less than 10% and the average recovery was greater than 90% for both drugs. This method was developed in bearded dragon and crane plasma and should be applicable to any species, making it useful for those investigators dealing with small sample volumes, particularly when conducting pharmacokinetics studies which require multiple sampling from the same animal.
PubDate: Mon, 27 Oct 2014 11:53:21 +000
- Development and Validation of Stability-Indicating RP-UPLC Method for
Simultaneous Determination of Related Substances of S(−)Amlodipine and
S(−)Metoprolol Succinate in Fixed Dose Combination Tablet Dosage Form
Abstract: A novel, rapid, accurate, sensitive, precise, and stability-indicating reverse-phase ultra-performance liquid chromatographic (RP-UPLC) method was developed and validated for determination of related substances of S(−)Amlodipine and S(−)Metoprolol Succinate in fixed dose combination tablet dosage form. The chromatographic separation was achieved with the use of Acquity UPLC HSS T3, 1.8 μm, 2.1 × 100 mm analytical column at 45°C employing a gradient elution. Mobile phase consisting of mobile phase-A (solution containing 5.0 gm of sodium dihydrogen phosphate monohydrate per liter of water and Acetonitrile in the ratio of 95 : 5) and mobile phase-B (Acetonitrile) was used at a flow rate of 0.5 mL min−1 with injection volume of 10 μL and the detection was done at 232 nm using UV detector. The retention times of S(−)Metoprolol Succinate and S(−)Amlodipine were found to be 2.8 minutes and 8.1 minutes, respectively. During method validation all the parameters were evaluated as per ICH guidelines, which remained well within acceptable limits. This method can be used for the estimation of related substances of S(−)Amlodipine and S(−)Metoprolol Succinate in fixed dose combination tablet dosage form.
PubDate: Mon, 20 Oct 2014 07:07:47 +000
- Analysis of Selected Amino Acids in Different Varieties of Wheat Available
in Punjab, Pakistan
Abstract: A validated method is proposed to check amino acids variability among eighty-nine wheat samples collected from Punjab province of Pakistan during 2012-2013. Orthophthalaldehyde along with 2-mercaptoethanol was used as a derivatizing reagent that showed florescence at detection wavelength of Ex of 340 nm and Em of 450 nm under suitable pH range of 9-10. RP-HPLC-FLD system employed was Agilent 1100 series equipped with Eclipse XDB C-18 column (2.1 × 150 mm, 5 µ) and column temperature was maintained at 40°C. The maximum concentration of aspartic acid, glutamic acid, leucine, arginine, and histidine was found in Vehari (0.496 g/100 g), Rajanpur (1.292 g/100 g), Rahim Yar khan (0.60 g/100 g), Bahawalpur (0.662 g/100 g), and Narowal (0.377 g/100 g), respectively, while the minimum in Narowal (0.13 g/100 g), Vehari (0.706 g/100 g), Narowal (0.339 g/100 g), Muzaffargarh (0.14 g/100 g), and Rahim Yar Khan (0.088 g/100 g) among the samples obtained from districts. Wheat variety Pb-11 contained relatively high aspartic acid (0.297 g/100 g), glutamic acid (0.897 g/100 g), and leucine (0.484 g/100 g) whereas variety Ass-11 had arginine (0.895 g/100 g) and histidine (0.266 g/100 g). The amino acids were found to vary as follows: aspartic acid 0.130–0.496, glutamic acid 0.706–1.292, leucine 0.321–0.6, arginine 0.118–0.895, and histidine 0.088–0.377 g/100 g flour. The accuracy was in the range of 95.88–100.67%, whereas the RSD for precision was not more than 1.40 for all amino acids.
PubDate: Thu, 16 Oct 2014 10:47:13 +000
- Extraction of Artemisinin, an Active Antimalarial Phytopharmaceutical from
Dried Leaves of Artemisia annua L., Using Microwaves and a Validated
HPTLC-Visible Method for Its Quantitative Determination
Abstract: A simple, rapid, precise, and accurate high-performance thin-layer chromatographic method coupled with visible densitometric detection of artemisinin is developed and validated. Samples of the dried Artemisia annua leaves were extracted via microwaves using different solvents. This method shows the advantage of shorter extraction time of artemisinin from leaves under the influence of electromagnetic radiations. Results obtained from microwave-assisted extraction (MAE) were compared with hot soxhlet extraction. Chromatographic separation of artemisinin from plant extract was performed over silica gel 60 F254 HPTLC plate using n-hexane : ethyl acetate as mobile phase in the ratio of 75 : 25, v/v. The plate was developed at room temperature 25 ± 2.0°C. Artemisinin separation over thin-layer plate was visualized after postchromatographic derivatization with anisaldehyde-sulphuric acid reagent. HPTLC plate was scanned in a CAMAG’s TLC scanner 3 at 540 nm. Artemisinin responses were found to be linear over a range of 400–2800 ng spot−1 with a correlation coefficient 0.99754. Limits of detection and quantification were 40 and 80 ng spot−1, respectively. The HPTLC method was validated in terms of system suitability, precision, accuracy, sensitivity (LOD and LOQ), and robustness. Additionally, calculation of plate efficiency and flow constant were included as components of validation. Extracts prepared from different parts of the plant (leaves, branches, main stem, and roots) were analyzed for artemisinin content, in which, artemisinin content was found higher in the leaf extract with respect to branches and main stem extracts; however, no artemisinin was detected in root extract. The developed HPTLC-visible method of artemisinin determination will be very useful for pharmaceutical industries, which are involved in monitoring of artemisinin content during different growth stages (in vitro and in vivo) of A. annua for qualitative and quantitative assessment of final produce prior to commercial-scale processing for assessment of cost-benefit ratio.
PubDate: Wed, 01 Oct 2014 12:50:05 +000
- Determination of Strong Acidic Drugs in Biological Matrices: A Review of
Abstract: Strong acidic drugs are a class of chemical compounds that normally have high hydrophilicity and large negative charges, such as organophosphatic compounds and organosulphonic compounds. This review focuses on sample preparation and separation methods for this group of compounds in biological matrices in recent years. A wide range of separation techniques, especially chromatographic method, are presented and critically discussed, which include liquid chromatography (e.g., ion-pair and ion-exchange chromatography), capillary electrophoresis (CE), and other types. Due to the extremely low concentration level of target analytes as well as the complexity of biological matrices, sample pretreatment methods, such as dilute and shoot methods, protein precipitation (PP), liquid-liquid extraction (LLE), solid-phase extraction (SPE), degradation, and derivatization strategy, also play important roles for the development of successful analytical methods and thus are also discussed.
PubDate: Mon, 29 Sep 2014 12:11:40 +000
- HPLC-UV Method Development and Validation of Potato Sprout Inhibitor
1,4-Dimethylnaphthalene Using Different Systems
Abstract: 1,4-Dimethylnaphthalene (1,4-DMN) is effective sprout suppressant used in potato stores in many countries in the world. High residue levels of this compound on the potatoes and in other environmental samples are considered for human health and environmental risks. Determination of the residue requires specific analytical methods to be developed and validated. In this study, HPLC-UV was selected for validating a separation method based on reversed phase for the analysis of 1,4-DMN using 2-methylnaphthalene (2-MeN) as internal standard testing three HPLC systems. Under the same chromatographic conditions, all three systems achieved good separation on a Jones column (Hypersil ODS 5 μm, 250 mm × 4.6 mm) at ambient temperature isocratically using 70% acetonitrile as mobile phase at a flow rate of 1.5 mL min−1, 20 μL injection volume, a run time of 10 min, and a detection wavelength of 228 nm. All three systems showed high precision, good linearity, and low limit of detection (LOD) and quantification (LOQ); particularly, the SpectraSERIES UV100-autosampler system offered lower values of LOD (0.001–0.004 μg mL−1) and LOQ (0.002–0.013 μg mL−1) for both compounds. This system can be used for the quantitative determination of 1,4-DMN residue in potato and environmental samples.
PubDate: Tue, 16 Sep 2014 09:59:51 +000
- Photocatalytic Degradation of Trifluralin, Clodinafop-Propargyl, and
1,2-Dichloro-4-Nitrobenzene As Determined by Gas Chromatography Coupled
with Mass Spectrometry
Abstract: Phototransformation is considered one of the most key factors affecting the fate of pesticides. Therefore, our study focused on photocatalytic degradation of three selected pesticide derivatives: trifluralin (1), clodinafop-propargyl (2), and 1,2-dichloro-4-nitrobenzene (3). The degradation was carried out in acetonitrile/water medium in the presence of titanium dioxide (TiO2) under continuous purging of atmospheric air. The course of degradation was followed by thin-layer chromatography and gas chromatography-mass spectrometry techniques. Electron ionization mass spectrometry was used to identify the degradation species. GC-MS analysis indicates the formation of several intermediate products which have been characterized on the basis of molecular ion, mass fragmentation pattern, and comparison with NIST library. The photocatalytic degradation of pesticides of different chemical structures manifested distinctly different degradation mechanism. The major routes for the degradation of pesticides were found to be (a) dealkylation, dehalogenation, and decarboxylation, (b) hydroxylation, (c) oxidation of side chain, if present, (d) isomerization and cyclization, (e) cleavage of alkoxy bond, and (f) reduction of triple bond to double bond and nitro group to amino.
PubDate: Sun, 31 Aug 2014 10:54:31 +000
- Validated HPLC Method for Quantification of Pregabalin in Human Plasma
Using 1-Fluoro-2,4-dinitrobenzene as Derivatization Agent
Abstract: In this study, a sensitive, simple, and reliable HPLC method for quantification of pregabalin in human plasma was developed and validated. 1-Fluoro-2,4-dinitrobenzene was used as precolumn derivatization agent. For chromatography, an analytical reversed phase (C18) column and a mixture of Na2HPO4 10 mM (pH 8.0)—methanol (35 : 65 v/v) were used as stationary and mobile phase, respectively. Detection was performed using a UV detector tuned at 360 nm. The linearity of the method was tested over the concentration range 1–4500 ng/mL in 500 μL of human plasma and satisfactory results were obtained (r2 > 0.999). The method showed good precision and accuracy in terms of within—between days relative standard deviations and percent deviation from nominated values (in the range of 4.3–12.7% and 2.6–8.0%, resp.). The limit of quantification of the method was found to be 1 ng/mL which is better than previously reported method and indicates its potential application for sensitive bioanalysis.
PubDate: Sun, 17 Aug 2014 12:21:00 +000
- Development and Validation of a Rapid Chemometrics Assisted RP-HPLC with
PDA Detection Method for the Simultaneous Estimation of Pyridoxine HCl and
Doxylamine Succinate in Bulk and Pharmaceutical Dosage Form
Abstract: Simple, rapid, precise, and accurate RP-HPLC method was developed and optimized with the help of chemometric tool for the simultaneous estimation of pyridoxine HCl and doxylamine succinate in bulk and pharmaceutical dosage form. Optimization was done by central composite design in response surface methodology. Based on the trial and error, percentage of organic phase (methanol) in mobile phase, flow rate, and molarity of the buffer were selected as factors. Resolution and retention time were used for the estimation of system response during the optimization procedure. The optimized condition was used and the separation was carried out on phenomenex C18 column (150 × 4.6 mm; i.d, 5 μ particle size) using the mobile phase containing 49.37% of methanol and 50.63% of phosphate buffer (45.14 mM) at a flow rate of 1 mL/min. Retention time was found to be 1.884 minutes for pyridoxine HCl and 3.959 minutes for doxylamine succinate. The calibration curves were found to be linear from 10 to 70 μg/mL and 10 to 90 μg/mL for pyridoxine HCl and doxylamine succinate with their correlation coefficient values 0.9995 and 0.9997. LOD and LOQ were found to be 23.5 ng/mL and 71.1 ng/mL for pyridoxine HCl and 99.9 ng/mL and 302.6 ng/mL for doxylamine succinate.
PubDate: Wed, 23 Apr 2014 14:13:40 +000
- A Stability-Indicating High Performance Liquid Chromatographic Assay for
the Simultaneous Determination of Pyridoxine, Ethionamide, and
Moxifloxacin in Fixed Dose Combination Tablets
Abstract: Stability indicating reversed phase HPLC method was developed and validated for the simultaneous quantitation of antitubercular drugs, ethionamide (ETH), and moxifloxacin (MOX) with commonly coprescribed vitamin, pyridoxine (PYR) in tablet dosage form. The method was found rapid, precise and accurate. The separation was performed in Hibar 150-4.6, Purospher STAR, RP-18e (5 μm) column, using mobile phase A (0.03 M sodium citrate adjusted to pH 5 with glacial acetic acid) and mobile phase B (100% methanol), ran at variable proportions at flow rate of 1.0 mL/min. The detection was carried out at 320 nm. The method was observed linearly in the range of 2.5–17.5 μg/mL for PYR, 25–175 μg/mL for ETH, and 40–280 μg/mL for MOX with respective limits of detection/quantitation of 0.125 μg/mL/1.28 μg/mL, 0.25 μg/mL/2.56 μg/mL, and 0.35 μg/mL/3.65 μg/mL. The drugs were also subjected to oxidative, hydrolytic, photolytic, and thermal degradation; the degradation products showed interference with the detection of PYR, ETH, and MOX. The proposed method was observed to be effective to quantitate MOX (400 mg), ETH (250 mg), and PYR (25 mg) in fixed dose combination tablet formulation.
PubDate: Wed, 23 Apr 2014 10:02:33 +000
- Development and Validation of a Novel RP-HPLC Method for Estimation of
Losartan Potassium in Dissolution Samples of Immediate and Sustained
Abstract: A simple, rapid, selective, and reproducible reversed-phase high performance liquid chromatographic (RP-HPLC) method has been developed and validated for the estimation of Losartan potassium in dissolution samples of Losartan potassium immediate and sustained release tablets. Analysis was performed on an Agilent, Zorbax Eclipse XDB C18 column (150 mm × 4.6 mm, 5 μm) with the mobile phase consisting of orthophosphoric acid (0.1% v/v)—acetonitrile (55 : 45, v/v) at a flow rate of 1.0 mL/min. UV detection was performed at 225 nm and the retention time for Losartan was about 2.6 minutes. The calibration curve was linear (correlation coefficient = 0.999) in the selected range of analyte. The optimized dissolution conditions include the USP apparatus 2 at a paddle rotation rate of 50 rpm and 900 mL of pH 6.8 phosphate buffer as dissolution medium, at ∘C. The method was validated for precision, linearity, specificity, accuracy, limit of quantitation, and ruggedness. The system suitability parameters, such as theoretical plate, tailing factor and relative standard deviation (RSD) between five standard replicates, were well within the limits. The stability result shows that the drug is stable in the prescribed dissolution medium.
PubDate: Wed, 09 Apr 2014 11:36:01 +000
- Stability Indicating Liquid Chromatographic Method for Estimation of
Trihexyphenidyl Hydrochloride and Risperidone in Tablet Formulation:
Development and Validation Consideration
Abstract: This paper describes validated reverse phase high-performance liquid chromatographic (RP-HPLC) method for simultaneous estimation of trihexyphenidyl hydrochloride (THP) and risperidone (RSP) in the pure powder form and in combined tablet dosage form. The HPLC separation was achieved on a core shell C18 (100 mm length × 4.6 mm, 2.6 μm particle size) using methanol : ammonium acetate buffer 1% (85 : 15 v/v; pH-6.5) as mobile phase and delivered at flow rate of 0.8 mL/min. The calibration plot showed good linear relationship with r2 = 0.997 ± 0.001 for THP and r2 = 0.998 ± 0.001 for RSP in concentration range of 50–175 μg/mL and 50–175 μg/mL, respectively. LOD and LOQ were found to be 0.40 and 1.29 μg/mL for THP and 1.24 and 3.92 μg/mL for RSP. Assay of THP and RSP was found to be 100.16 ± 0.03% and 99.83 ± 0.02%, respectively. THP and RSP were subjected to different stress conditions (acidic, basic, oxidative, thermal, and photolytic degradation). The degraded product peaks were well resolved from the pure drug peak. The method was successfully validated as per the ICH guidelines. The developed RP-HPLC method was successfully applied for the estimation of THP and RSP in tablet dosage form.
PubDate: Wed, 19 Mar 2014 08:40:26 +000
- Estimation of Diafenthiuron Residues in Cardamom (Elettaria cardamomum
(L.) Maton) Using Normal Phase HPLC: Dissipation Pattern and Safe Waiting
Period in Green and Cured Cardamom Capsules
Abstract: Diafenthiuron is an effective insecticide used for pest management in cardamom. Residues of diafenthiuron and its degradation/dissipation pattern in cardamom were determined to work out safe waiting period. Samples were collected after three sprays of diafenthiuron @ 400 and 800 g a.i ha−1 and the residues extracted in acetonitrile and quantified in normal phase HPLC in UV detector. Diafenthiuron was detected in min. The limits of detection (LOD) and limits of quantification (LOQ) were determined to be 0.01 and 0.05 μgmL−1. The initial deposits were found to be 3.82 and 4.10 μg g−1 after sprays of diafenthiuron @ 400 g a.i ha−1 in the first and second experiments, respectively. Nearly cent percent of residues dissipated at 10 days after treatment in the recommended dose of diafenthiuron 400 g a.i ha−1 and the half life varied from 2.0 to 2.8 days with a waiting period of 5.5 to 6.7 days in green capsules of cardamom. The waiting period was 5.4 to 7.0 days in cured capsules of cardamom. With harvest being the focal point for enforcement of residue tolerances, the suggested waiting period of seven days is safe without the problem of pesticide residues in harvestable produce.
PubDate: Mon, 24 Feb 2014 08:08:09 +000
- Quantification of Gymnemagenin and -Sitosterol in Marketed Herbal
Formulation by Validated Normal Phase HPTLC Method
Abstract: This research study describes development and validation of new, rapid, accurate, robust, and precise, high performance thin layer chromatographic (HPTLC) method for concurrent quantitative determination of gymnemagenin and β-sitosterol in herbal formulation with densitometric detection. Chromatographic separation was achieved on Merck aluminum HPTLC plates precoated with silica gel 60 F254. The optimized solvent system consisted of toluene : ethyl acetate : methanol (6.5 : 2.5 : 1.4, v/v/v). Developed plates were derivatized with 5% sulphuric acid reagent followed by heating at 110°C for 4 min in a preheated oven and scanned at 423 nm in reflectance-absorbance mode. The retention factor for gymnemagenin and β-sitosterol was found to be and , respectively. The proposed densitometric method was validated according to ICH Q2 (R1) guidelines. Results were found to be linear over a range of 100–1200 ng band−1 and 200–1200 ng band−1 for gymnemagenin and β-sitosterol, respectively. The percent content of gymnemagenin and β-sitosterol in the marketed polyherbal formulation was found to be 0.0405% and 0.1377%, respectively. The proposed HPTLC method can be used for quantification of gymnemagenin and β-sitosterol in marketed polyherbal formulation used in the study in quality-control laboratories.
PubDate: Thu, 06 Feb 2014 06:47:09 +000
- Simultaneous Quantification of Glibenclamide, Simvastatin, and Quercetin
by Using LC-UV Method and Its Application to Pharmacokinetic Study in Rats
Abstract: A sensitive, precise, and simple LC method for the simultaneous quantification of glibenclamide, simvastatin, and quercetin in rat plasma has been developed and validated. The chromatographic separation was achieved on a cyano column (250 mm × 4.6 mm, 5 µm) maintained at room temperature, using isocratic elution with methanol : acetonitrile : 10 mM potassium dihydrogen orthophosphate, pH adjusted to 4.5 with o-phosphoric acid (8 : 32 : 60, v/v) and detected using UV-VIS detector. Plasma samples were deproteinated with 0.1% perchloric acid and acetonitrile for extraction of the glibenclamide, simvastatin, and quercetin which resulted in their high recoveries. LC calibration curves based on the extracts from the rat plasma were linear in the range of 50–1000 ng mL−1 for all the three drugs. The limit of quantification was 50 ng mL−1. The described method was successfully applied to study the pharmacokinetics of glibenclamide, simvastatin, and quercetin following oral administration, in combination to Sprague-Dawley rats.
PubDate: Mon, 16 Dec 2013 08:53:46 +000
- RP-HPLC Method for Determination of Several NSAIDs and Their Combination
Abstract: An RP-HPLC method for simultaneous determination of 9 NSAIDs (paracetamol, salicylic acid, ibuprofen, naproxen, aceclofenac, diclofenac, ketorolac, etoricoxib, and aspirin) and their commonly prescribed combination drugs (thiocolchicoside, moxifloxacin, clopidogrel, chlorpheniramine maleate, dextromethorphan, and domperidone) was established. The separation was performed on Kromasil C18 (250 × 4.6 mm, 5 μm) at 35°C using 15 mM phosphate buffer pH 3.25 and acetonitrile with gradient elution at a flow rate of 1.1 mL/min. The detection was performed by a diode array detector (DAD) at 230 nm with total run time of 30 min. Calibration curves were linear with correlation coefficients of determination . Limit of detection (LOD) and Limit of quantification (LOQ) ranged from 0.04 to 0.97 μg/mL and from 0.64 to 3.24 μg/mL, respectively. As an application tool of quality by design, full factorial experimental design was used for the testing of robustness of the method. The prediction profiler correlating various parameters and responses was established from the results of design of experiments (DOE).
PubDate: Wed, 11 Dec 2013 13:16:39 +000
- Stress Degradation Studies on Flupirtine Maleate Using
Stability-Indicating RP-HPLC Method
Abstract: With the objective of developing an advanced method for rapid separation with shorter runtime, a simple, precise, and accurate stability-indicating isocratic RP-LC method coupled with PDA detector was developed for the quantitative determination of flupirtine maleate in bulk and in capsule dosage form. Good resolution between the peaks for degradation products and the analyte was achieved on a Waters Agilent XDB C18 ( mm, 5 μm) column using mobile phase containing a mixture of phosphate buffer pH 3.36 and acetonitrile in the ratio of 65 : 35. The eluted compounds were monitored at 344 nm and the flow rate employed for the present investigation was 1 mL/min. The newly developed method was validated as per ICH guidelines with respect to specificity, linearity, limit of detection, limit of quantification, accuracy, precision, and robustness. The method may be employed for the assay determination of flupirtine maleate in pharmaceutical dosage forms.
PubDate: Wed, 04 Dec 2013 18:00:51 +000
- A Comparative Validation Study of Fluconazole by HPLC and UPLC with Forced
Abstract: The simplest stability indicating reversed phase Isocratic HPLC and UPLC methods has been developed and validated for the determination of fluconazole in bulk and solid pharmaceutical dosage form. A SunFire C18 (250 × 4.5 mm, 5 μm particle size) column has been used for HPLC and BEH C18 (100 × 2.1 mm, 1.7 μm particle size) column used for UPLC. The Mobile phase consisted of Methanol : Water (70 : 30) for HPLC and Methanol : Water (55 : 45 v/v) for UPLC. Isocratic flow was set at 1 mL/min and 0.30 mL/min, respectively, for HPLC and UPLC. For both HPLC and UPLC system detection has been performed at 211 nm with 30°C column oven temperature (good elution was obtained at 30°C) and injection volume, respectively, 2 μL and 20 μL for HPLC and UPLC.
PubDate: Wed, 04 Dec 2013 13:30:47 +000
- Recycle HPLC: A Powerful Tool for the Purification of Natural Products
Abstract: Natural compounds occur as various isomeric or closely related structures in biological matrices. These compounds are difficult to separate from the complex mixtures, and hence, the need for effective and innovative separation techniques arises. Recycle HPLC allows the recycling of sample, in part or full, and increases the separation efficiency of the process while keeping the peak dispersion to a minimum. Recycling in an HPLC system has been used in the isolation and purification of different types of natural products including enantiomers, diastereomers, epimers, positional isomers, and structurally related or unrelated compounds having similar retention characteristics. The present paper overviews the development of instrumentation and setup of recycle HPLC and its applications in the separation of natural products.
PubDate: Wed, 06 Nov 2013 14:48:28 +000
- Trends and Advances in Separation and Detection of SSRIs and SNRIs in
Abstract: Nowadays antidepressant drugs like selective serotonin reuptake inhibitors (SSRIs) and selective norepinephrine reuptake inhibitors (SNRIs) represent the first choice in the treatment of moderate to severe depressive illness, various phobias, and personality disorders. In spite of the therapeutic aspects, they often produce very severe and toxic effects in deliberate and accidental cases of poisoning. These are also considered as date-rape drugs used for drugged victims for raping or robbing. Therefore, in recent years, their analyses in different biological matrices for clinical and toxicological analysis purposes has been a target worthy of interest. Thus, the review focuses on recent advancements of various separation techniques like chromatography and electrophoresis that are concernd with the determination of selective serotonin reuptake inhibitor and selective norepinephrine reuptake inhibitor drugs and their metabolites in various biological matrices. In addition to this, a critical discussion on analytical approaches has also been incorporated, suggesting their applicability and limitations for further implementations. Thus, this paper will definitely help in the selection and development of proper analytical methodologies to achieve satisfactory results, better scientific understanding, and test interpretation.
PubDate: Thu, 31 Oct 2013 18:35:58 +000