Comparison of UPLC and HPLC for Analysis of 12 Phthalates

Recent technological advances have resulted in the availability of reversed-phase chromatographic media of particle size 1.7 μm and a liquid-handling system that can be used to operate columns packed with these materials at much higher pressures. This technology, UPLC, has significant theoretical advantages in speed, resolution, and sensitivity of analysis, especially time saving and solvent consumption. The work discussed in this paper with new analytical method used for separation of 12 phthalates and the results were compared with those obtained by use of HPLC. Differences between the techniques, system suitability test data, and advantages and disadvantages of UPLC are discussed.     

Separation of Dietary Folates by Gradient Reversed-Phase HPLC: Comparison of Alternative and Conventional Silica-Based Stationary Phases

A study of ten silica-based stationary phases and gradient elution conditions to separate dietary folates by reversed-phase HPLC was performed. Alkyl-bonded stationary phases (both conventional and alternative) were found to be the most promising for the separation of different folate monoglutamates in terms of selectivity and peak shape. These phases were better than phenyl-bonded phases which lacked selectivity when separating 10-formyl-folic acid and 5-formyl-tetrahydrofolate. Polar-bonded (cyano) stationary phase showed similar retention characteristics as the conventional alkyl-bonded phases, but ranked below those in terms of peak shape. Overall, alternative stationary phases exhibited slightly higher retention of late-eluted folates and greater retention variability for early-eluting tetrahydrofolate and 5-methyl-tetrahydrofolate. Best selectivity was achieved on alternative polar endcapped Aquasil C₁₈ followed by conventional Synergy MAX C₁₂ and Genesis C₁₈ stationary phases.     

UPLC versus HPLC on Drug Analysis: Advantageous, Applications and Their Validation Parameters

Liquid chromatography (LC) is a separation technique used in many different areas to aid the identification and quantification of substances in various matrices. LC techniques with various detection modes have been widely used for the sensitive and selective determination of trace amounts of pharmaceutical active compounds in biological samples and their dosage forms. A completely new system design with advanced technology has been developed, called ultra high performance liquid chromatography, which has evolved from high performance liquid chromatography. The application of LC methods to drug analysis introduces a powerful tool for therapeutic drug monitoring as well as for clinical research. The advantages of short turnaround time, method reliability, method sensitivity, and drug specificity justify the use of LC techniques for various groups of the drug active compounds. This review describes some of the principles of ultra high performance liquid chromatography and high performance liquid chromatography, validation of these methods, system suitability tests for the methods, and application of methods to pharmaceutical analysis in the last 3 years.     

Development and validation of stability-indicating HPLC and UPLC methods for the determination of bicalutamide

Six new process related impurities (Imp-08, Imp-09, Imp-10, Imp-12, Imp-13 and Imp-14) of bicalutamide (BCT) have been reported in this paper. BCT was subjected to oxidative, acid, alkaline, hydrolytic, thermal and photolytic degradation conditions and found to degrade in alkaline condition, yielding Imp-11. Stability-indicating high-performance liquid chromatography and ultra-performance liquid chromatography methods were developed for the determination of BCT in the presence of its 14 process-related impurities and 1 degradant by using Zorbax SB phenyl column (150 × 4.6 mm × 3.5 μm) and HSS T3 column (100 × 2.1 mm × 1.8 μm), respectively. Both the methods were validated as per International Conference on Harmonization guidelines. Quantitation limits (QL) were found be in the ranges of 0.02-0.03% for both the methods. Precision was evaluated by replicate analysis in which % relative standard deviation (RSD) values for areas were found below 2.0. Linearity for the impurities was established in the range of QL to 200% of the specification level and the correlation coefficients derived from of the respective calibration curves were approximately 0.999. The recoveries obtained for purity (90-100%) and assay (98-102%) ensured the accuracy of the developed methods.     

HPLC与UPLC色谱条件转换方法研究

以分离分析化妆品中对羟基苯甲酸酯类防腐剂为内容,对HPLC和UPLC的色谱条件转换方法进行了研究。比较了采取不同方法转换得到的色谱条件参数的分离状况,选出了UPLC法最佳色谱条件。同时,用HPLC法和UPLC法对4种化妆品中的4种对羟基苯甲酸酯类防腐剂进行了定量测定,两种方法的测定结果吻合较好,相互印证了方法的准确性。     

Comparison of UPLC and HPLC for Determination of trans-10-Hydroxy-2-Decenoic Acid Content in Royal Jelly by Ultrasound-Assisted Extraction with Internal Standard

Two well-known derivatization procedures, H₂SO₄–butanol and H₂SO₄–methanol esterification, were compared for application to GC–MS identification of organic acids in Bayer process liquors. H₂SO₄–butanol and H₂SO₄–methanol derivatization must be combined for analysis of carboxylic acids. Twenty organic acids were identified by gas chromatography–mass spectrometry. Heptanedioic, 3-methylhexanedioic, octanedioic, 1,3-benzenedicarboxylic, nonanedioic, decanedioic, hexadecanoic, 9,12-octadecadienoic, octadecanoic, and phthalic acids were identified for the first time in Bayer liquors. The retention times (y) and carbon numbers (x) of these seven n-dicarboxylic acids (C₄–C₁₀) were fit to a linear relationship by use of Microsoft Excel. These dicarboxylic acids and two benzenedicarboxylic acids were quantified by use of the internal standard method.     

HPLC and UPLC Methods for the Simultaneous Determination of Enalapril and Hydrochlorothiazide in Pharmaceutical Dosage Forms

High efficiency and less elution are the basic requirements of high-speed chromatographic separation. In this study, a new gradient reverse phase chromatographic methods were developed using HPLC and UPLC systems for simultaneous determination of enalapril maleate (ENL) and hydrochlorothiazide (HCZ) in pharmaceutical dosage forms. The chromatographic separations of ENL and HCZ were achieved on a Waters μ-Bondapak C 18, (300 × 3.9 mm, 10 μm) and Waters Acquity BEH C18 (100 × 2.1 mm, 1.7 μm) columns for HPLC within 5.30 min and UPLC within a short retention time of 1.95 min, respectively. A linear response was observed over the concentration range 0.270–399 μg mL^?1 of ENL, 0.260–399 μg mL^?1 of HCZ for HPLC system and 0.270–399 μg mL^?1 of ENL and 0.065–249 μg mL^?1 of HCZ for UPLC system. Also, limit of detection for ENL was 1.848 ng mL^?1 and 31.477 ng mL^?1 for HCZ, 2.804 ng mL^?1 for ENL and 2.943 ng mL^?1 for HCZ using HPLC and UPLC, respectively. The proposed methods were validated according to ICH guideline with respect to precision, accuracy, and linearity. Forced degradation studies were also performed for both compounds in bulk drug samples to demonstrate the specificity and stability indicating power of the HPLC method. Comparison of system performance with conventional HPLC was made with respect to analysis time, efficiency, and resolution.     

HPLC and UPLC Methods for the Simultaneous Determination of Enalapril and Hydrochlorothiazide in Pharmaceutical Dosage Forms

High efficiency and less elution are the basic requirements of high-speed chromatographic separation. In this study, a new gradient reverse phase chromatographic methods were developed using HPLC and UPLC systems for simultaneous determination of enalapril maleate (ENL) and hydrochlorothiazide (HCZ) in pharmaceutical dosage forms. The chromatographic separations of ENL and HCZ were achieved on a Waters μ-Bondapak C 18, (300 × 3.9 mm, 10 μm) and Waters Acquity BEH C18 (100 × 2.1 mm, 1.7 μm) columns for HPLC within 5.30 min and UPLC within a short retention time of 1.95 min, respectively. A linear response was observed over the concentration range 0.270–399 μg mL⁻¹ of ENL, 0.260–399 μg mL⁻¹ of HCZ for HPLC system and 0.270–399 μg mL⁻¹ of ENL and 0.065–249 μg mL⁻¹ of HCZ for UPLC system. Also, limit of detection for ENL was 1.848 ng mL⁻¹ and 31.477 ng mL⁻¹ for HCZ, 2.804 ng mL⁻¹ for ENL and 2.943 ng mL⁻¹ for HCZ using HPLC and UPLC, respectively. The proposed methods were validated according to ICH guideline with respect to precision, accuracy, and linearity. Forced degradation studies were also performed for both compounds in bulk drug samples to demonstrate the specificity and stability indicating power of the HPLC method. Comparison of system performance with conventional HPLC was made with respect to analysis time, efficiency, and resolution.

     

Erratum to: UPLC versus HPLC on Drug Analysis: Advantageous,Applications and Their Validation Parameters

Liquid chromatography (LC) is a separation technique used in many different areas to aid the identification and quantification of substances in various matrices. LC techniques with various detection modes have been widely used for the sensitive and selective determination of trace amounts of pharmaceutical active compounds in biological samples and their dosage forms. A completely new system design with advanced technology has been developed, called ultra high performance liquid chromatography, which has evolved from high performance liquid chromatography. The application of LC methods to drug analysis introduces a powerful tool for therapeutic drug monitoring as well as for clinical research. The advantages of short turnaround time, method reliability, method sensitivity, and drug specificity justify the use of LC techniques for various groups of the drug active compounds. This review describes some of the principles of ultra high performance liquid chromatography and high performance liquid chromatography, validation of these methods, system suitability tests for the methods, and application of methods to pharmaceutical analysis in the last 3 years.

     

Use of Principal Component and Cluster Analysis for the Comparison of Reversed-Phase HPLC Columns

Abstract

The retention of ethoxylated nonylhylphenyl surfactants was determined in reversed-phase high-performance liquid chromatography (RP-HPLC) using various supports (C1, C2, C6, C8, C18, polyethylene-coated silica, and polyethylene-coated alumina). The retention data matrix was evaluated both by principal component analysis and cluster analysis. The retention characteristics of both polyethylene-coated supports were similar to that of C1, the retention capacity of RP columns for surfactants increased with the increasing length of the covalently bonded hydrocarbon chain. The retention of ethoxylated surfactants depended nonlinearly on the length of ehtyleneoxide chain suggesting that the polar ethyleneoxide chains are in folded state under the chromatographic separation.     

Green Spectrophotometric Method for the Quantitative Analysis of Vancomycin in Pharmaceuticals and Comparison with HPLC

Abstract

A spectrophotometric method for the determination of vancomycin base, (VCM), and vancomycin hydrochloride, (HVCM), based on the reaction with copper (II) ions, is presented. The obtained detection limit is about 4.5×10^?5 mol L^?1. The working analytical range falls between 1.0×10^?3 mol L^?1 and 1.0×10^?2 mol L^?1. Recovery studies in presence of excipients were performed. The recovery results were compared with HPLC. For HVCM the proposed method presented similar recovery to that of HPLC, 100.4% vs. 100.2%, but better precision, 1.9% vs. 6.1%. In the VCM case the recovery is quite better, 100.5% vs. 89.6%, with a little smaller precision, 2.1% vs. 1.3%.     

Comparison of Various Columns for the High-Speed HPLC Analysis of Drugs of Forensic Interest

Abstract

A comparison of the use of various commercially available columns for the high-speed reverse-phase ion-pair high performance liquid chromatographic separation of drugs of forensic interest is discussed. The columns include a Partisil 5 ODS-3 RAC, a Partisil 5 C8 RAC, a Radial Pak microBondapak C18 cartridge, a Perkin-Elmer HS/5 C18 and a Perkin-Elmer HS/3 C18. The mobile phases employed contain water, acetonitrile, phosphoric acid, and sodium hydroxide, with or without hexylamine. When a mobile phase without an amine modifier is employed, retention times were at least halved, except with a HS/3 C18 column, over those obtained with conventional columns. Basic drugs did not elute when the above mobile phase is used with a HS/3 C18 column. In addition, the selectivities of the other high speed columns were similar. Further reductions in retention times and different selectivities were obtained when an amine modifier is utilized. Column performance parameters such as n, V and v are presented for the colupns examined. A new column performance parameter S which is (n/V)^1/2 is introduced and discussed.     

Comparison of GC and HPLC Methods for Quantitative Analysis of Tinuvin 622 After Saponification in Polyethylene

This study describes the sample preparation and two chromatographic techniques for determination of Tinuvin 622 in polyethylene. The first part of the two methods consisting of dissolving the polyethylene in boiling xylene is followed by addition of a methanolic solution of potassium hydroxide. The polymeric light stabilizer, Tinuvin 622, is thereby saponified to 4-hydroxy-2,2,6,6-tetramethyl-1-piperidineethanol (diol). Addition of the methanolic solution of the saponification reagent simultaneously precipitates the polyethylene matrix. Then the diol is quantified using either gas chromatography (GC) or high performance liquid chromatography (HPLC). For GC, a Macherey Nagel Optima-17 capillary column (30m×0.25mm ID, film thickness 0.25µm) is used. Nitrogen is used as carrier gas and make-up gas. The detection system is a flame ionization detector. For HPLC, an octadecyl silane (ODS) column (30cm×4mm, particle size 5µm) and a mobile phase methanol: water mixture (3:97, v/v) are used. Detection of analyte is carried out at 215nm. Both methods can be used to determine Tinuvin 622 in polyethylene in the concentration range of 0.02–1%, which represents the usual application concentration.     

Comparison of Different Cartridges of Solid Phase Extraction for Determination of Polyphenols in Tobacco by UPLC/MS/MS and Multivariate Analysis

The comparison of solid phase extraction(SPE) for the preconcentration and isolation of polyphenols in tobacco samples was carried out by ultra-high performance liquid chromatography/tandem mass spectrometry (UPLC/MS/MS) and multivariate analysis.Several adsorbing materials of SPE(C18,NH2,SAX and OASIS) were investigated.It was found that the C18 and OASIS cartridges can not only speed up the purification process,but also simplify the SPE operation.A UPLC/MS/MS was used for the determination of polyphenols ...     

An Improved UPLC Method for Rapid Analysis of Levofloxacin in Human Plasma

A rapid, specific, and sensitive ultra-performance liquid chromatographic method for analysis of levofloxacin in human plasma has been developed and validated. Plasma samples were spiked with the internal standard (enoxacin) and extracted with 10:1 (v/v) ethyl acetate–isopropanol. UPLC was performed on a 100 × 2.1 mm i.d., 1.7 µm particle, C₁₈ column with 88:12 (v/v) 0.4% triethylamine buffer (pH 3)–acetonitrile as mobile phase, pumped isocratically at a pressure of 11,000 psi (758 bar) and a flow-rate of 0.3 mL min^?1. Ultraviolet detection was performed at 300 nm. The retention times of levofloxacin and enoxacin were 3.4 and 2.8 min, respectively, and the run-time was 5 min. Calibration showed that response was a linear function of concentration over the range 0.05–10 µg mL^?1 (r ² ≥ 0.99) and the method was validated over this range for both precision and accuracy. The relative standard deviation was <15% for both intra-day and inter-day assay (n = 5). Levofloxacin and enoxacin were stable in plasma; there was no evidence of degradation during three freeze–thaw cycles, post-preparative stability at 20 °C was ≥24 h, short-term stability at room temperature was ≥6 h, and long-term stability at ?70 °C was ≥30 days. The method was successfully used in a study of the bioequivalence of two levofloxacin tablet formulations in healthy volunteers.     

Comparison of HPLC and MEEKC for Miconazole Nitrate Determination in Pharmaceutical Formulation

A simple solid phase extraction (SPE) method coupled with high performance liquid chromatography (HPLC) using UV detector and microemulsion electrokinetic chromatography (MEEKC) has been developed and compared for the quantitative determination of miconazole nitrate in pharmaceutical formulation. For HPLC method, two parameters were optimized, namely, the wavelength and the mobile phases. The optimized condition was at the 225 nm wavelength and the mobile phase of ACN:MeOH (90:10 v/v). There are seven MEEKC parameters that were optimized, in this research, which were applied to voltage, temperature, wavelength, sodium dodecyl sulfate (SDS) concentration, buffer pH, buffer concentration and butan-1-ol concentration. The optimum MEEKC condition was obtained using 86.35 % (w/w) 2.5 mM borate buffer pH 9, 0.25 % (w/w) SDS, 0.8 % (w/w) ethyl acetate, 6.6 % w/w butan-1-ol and 6.0 % (w/w) acetonitrile. The combination of SPE using a diol column with HPLC–UV and the MEEKC methods were successfully applied for the determination of miconazole nitrate in a pharmaceutical formulation with the recovery percentage of 98.35 and 92.50 %, respectively.

     

Comparison of HPLC and MEEKC for Miconazole Nitrate Determination in Pharmaceutical Formulation

A simple solid phase extraction (SPE) method coupled with high performance liquid chromatography (HPLC) using UV detector and microemulsion electrokinetic chromatography (MEEKC) has been developed and compared for the quantitative determination of miconazole nitrate in pharmaceutical formulation. For HPLC method, two parameters were optimized, namely, the wavelength and the mobile phases. The optimized condition was at the 225 nm wavelength and the mobile phase of ACN:MeOH (90:10 v/v). There are seven MEEKC parameters that were optimized, in this research, which were applied to voltage, temperature, wavelength, sodium dodecyl sulfate (SDS) concentration, buffer pH, buffer concentration and butan-1-ol concentration. The optimum MEEKC condition was obtained using 86.35 % (w/w) 2.5 mM borate buffer pH 9, 0.25 % (w/w) SDS, 0.8 % (w/w) ethyl acetate, 6.6 % w/w butan-1-ol and 6.0 % (w/w) acetonitrile. The combination of SPE using a diol column with HPLC–UV and the MEEKC methods were successfully applied for the determination of miconazole nitrate in a pharmaceutical formulation with the recovery percentage of 98.35 and 92.50 %, respectively.     

Comparison of HPTLC and HPLC for Determination of Isoflavonoids in Several Kudzu Samples

JPC – Journal of Planar Chromatography – Modern TLC - HPTLC and HPLC methods have been established for separation and quantitative determination of puerarin, 3’-methoxypuerarin,...