Elution strength and selectivity of the mobile phase in reverse phase high performance liquid chromatography (HPLC)

1976 ◽  
Vol 279 (2) ◽  
pp. 154-155 ◽  
Author(s):  
M. Riedmann
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Mobile Phase ◽  
High Performance ◽  
Reverse Phase

scholarly journals Reverse-phase High-Performance Liquid Chromatography Method Development and Validation for Estimation of Glimepiride in Bulk and Tablet Dosage Form

2020 ◽  
Vol 11 (02) ◽  
pp. 296-302
Author(s):  
Aseem Kumar ◽  
Anil Kumar Sharma ◽  
Rohit Dutt
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Flow Rate ◽  
Retention Time ◽  
Mobile Phase ◽  
High Performance ◽  
Hplc Method ◽  
Reverse Phase ◽  
Rp Hplc ◽  
Development And Validation

The present work demonstrates a simple, rapid, precise, specific, and sensitive reverse-phase high-performance liquid chromatography (RP-HPLC) method for analyzing glimepiride in pure and tablet forms. The present method was developed using a C18 column 150 × 4.6 mm, with 5 μm, and packing L1 maintained at a temperature of 30°C. The mobile phase was prepared by dissolving 0.5 gram of monobasic sodium phosphate in 500 mL of distilled water, pH of the solution adjusted to 2.1 to 2.7 with 10% phosphoric acid, and added 500 mL of acetonitrile. The mobile phase was pumped in the highperformance liquid chromatography (HPLC) system at a flow rate of 1 mL/min, and separation was carried out at 228 nm, using an ultraviolet (UV) detector. The chromatographic separation was achieved with peak retention time (RT) at about 9.30 minutes, and the method was found to be linear over a concentration range of 40 to 140 μg/mL. The specificity of the method represented no interference of the excipients during the analysis, and stability testing after 24 hours also showed that the method is suitable and specific. The accuracy was between 99.93 to 99.96%, with limit of detection (LOD) and limit of quantitation (LOQ) being 0.354 μg/mL, 1.18 μg/mL, respectively. Satisfactory results were found for precision and robustness parameters during the development and validation stage for the analytical method. The proposed method was also adopted for the analysis of glimepiride tablets to improve the overall quality control. Using this method, symmetric peak shape was obtained with reasonable retention time. The retention time of glimepiride for six repetitions is 9.3 ± 0.1 minutes; the run time is 21 minutes. The proposed RP-HPLC method is a modification of the United States Pharmacopeia (USP) method, and it was found to be valid for glimepiride within concentration ranges 40 to 140 μg/mL, using C18 analytical columns, and isocratic elution with UV detection, and at 1 mL/min of flow rate.

scholarly journals Direct Enantiomeric Separation and Determination of Hexythiazox Enantiomers in Environment and Vegetable by Reverse-Phase High-Performance Liquid Chromatography

2020 ◽  
Vol 17 (10) ◽  
pp. 3453
Author(s):  
Ping Zhang ◽  
Sheng Wang ◽  
Dongmei Shi ◽  
Yangyang Xu ◽  
Furong Yang ◽  
...  
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Mobile Phase ◽  
High Performance ◽  
Enantiomeric Separation ◽  
Reverse Phase ◽  
Column Temperature ◽  
Resolution Factor ◽  
Phase Temperature ◽  
Lower Temperature

In the present study, the direct enantiomeric separation of hexythiazox enantiomers on Lux cellulose-1, Lux cellulose-2, Lux cellulose-3, Lux cellulose-4, Lux amylose-1 and Chirapak IC chiral columns were carefully investigated by reverse-phase high-performance liquid chromatography (RP-HPLC). Acetonitrile/water and methanol/water were used as mobile phase at a flow rate of 0.8 mL·min−1. The effects of chiral stationary phase, temperature, thermodynamic parameters, mobile phase component and mobile phase ratio on hexythiazox enantiomers separation were fully evaluated. Hexythiazox enantiomers received a baseline separation on the Lux cellulose-3 column with a maximum resolution of Rs = 2.09 (methanol/water) and Rs = 2.74 (acetonitrile/water), respectively. Partial separations were achieved on other five chiral columns. Furthermore, Lux amylose-1 and Chirapak IC had no separation ability for hexythiazox enantiomers when methanol/water was used as mobile phase. Temperature study indicated that the capacity factor (k) and resolution factor (Rs) decreased with column temperature increasing from 10 °C to 40 °C. The enthalpy (ΔH) and entropy (ΔS) involved in hexythiazox separation were also calculated and demonstrated the lower temperature contributed to better separation resolution. Moreover, the residue analytical method for hexythiazox enantiomers in the environment (soil and water) and vegetable (cucumber, cabbage and tomato) were also established with reliable accuracy and precision under reverse-phase HPLC condition. Such results provided a baseline separation method for hexythiazox enantiomers under reverse-phase conditions and contributed to an environmental and health risk assessment of hexythiazox at enantiomer level.

scholarly journals Stability-Indicating Simultaneous Method Development and Validation of Guaifenesin and Dextromethorphan HBr by Reverse-Phase High-Performance Liquid Chromatography

2020 ◽  
Vol 11 (02) ◽  
pp. 262-270
Author(s):  
Nathi Rathnakar ◽  
Dannana Gowri Shanker
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Mobile Phase ◽  
High Performance ◽  
Drug Product ◽  
Simultaneous Estimation ◽  
Reverse Phase ◽  
Stability Indicating ◽  
R Value ◽  
The Stability

The developed method was validated according to ICH guidelines with respect to specificity, linearity, limits of detection, quantification, accuracy, precision, and robustness. The stability-indicating reverse-phase high-performance liquid chromatography (RP-HPLC) method is precise; it has been developed for the simultaneous estimation of assay of guaifenesin (GN) and dextromethorphan hydrobromic (HBr) (DN) in drug substance and drug product. The chromatographic separation was done in an isocratic mode using the Syncronus C8 (250 × 4.6 mm, 5 μ particle size) column with mobile phase containing a 10 mM ammonium acetate in water (modulated pH 4.30 with orthophosphoric acid) and acetonitrile in the ratio of 60:40 (% v/v) used for efficient chromatographic separation. The flow rate of the mobile phase was 1 mL/min with ambient column temperature and detection of wavelength at 279 nm; injection volume 10 μL was fixed for achieving good elution of eluents. The retention time for GN was found to 3.46 minutes and DN was found to 7.58 minutes. GN and DN were linear in the concentration range from 357 to 1,428 and 19 to 75 μg/mL, respectively. Regression analysis showed that the r value (correlation coefficient) greater than 0.999 for GN r value was found to be 0.999, GN r value was found to be 0.999, DN r value was found to be 0.999. Limit of detection (LoD) and limit of quantification (LoQ) of GN was found to be 0.151 and 0.904 μg/mL, DN was found to be 0.241 and 0.726 μg/mL. The developed method was validated and found to be accurate, specific, and robust. Both the drugs were subjected to the stress conditions like acidic, basic, oxidative, photolytic, and thermal conditions. The degradation results were found to be satisfactory. In peroxide stress condition, GN was found stable over DN, and DN was found to degrade significantly. The degradation products were well resolved from GN, DN, and their impurities. The peak purity test results confirmed that the GN and DN peak were homogenous and pure in all stress conditions, thus, proving the stability-indicating nature of the method. This method could be applied for the simultaneous estimation of GN and DN in drug substance and drug product.

scholarly journals Analysis of commercial samples of acridine orange using high-performance liquid chromatography.

1988 ◽  
Vol 36 (12) ◽  
pp. 1489-1493
Author(s):  
R D Paulsen ◽  
G S Nettleton
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Acridine Orange ◽  
Sulfonic Acid ◽  
Mobile Phase ◽  
High Performance ◽  
Principal Component ◽  
Reverse Phase

We analyzed commercial samples of acridine orange using reverse-phase high-performance liquid chromatography. The mobile phase of 90:10:2.5 acetonitrile:de-ionized water:pentane sulfonic acid gave baseline separations of components of acridine orange samples in 15 min. Many of the samples were fairly homogeneous; the absorbance due to the acridine orange component ranged from 83-97%. Among the 11 samples tested, one was not acridine orange, reputedly different samples were identical, and the principal component of one sample was not acridine orange.

scholarly journals STABILITY INDICATING REVERSE-PHASE HIGH- PERFORMANCE LIQUID CHROMATOGRAPHY METHOD DEVELOPMENT AND VALIDATION FOR SIMULTANEOUS ESTIMATION OF TELMISARTAN AND BENIDIPINE HYDROCHLORIDE IN PHARMACEUTICAL DOSAGE FORM

2018 ◽  
Vol 11 (5) ◽  
pp. 342
Author(s):  
Majan Naim ◽  
Aejaz Ahmed ◽  
Khan Gj
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Mobile Phase ◽  
High Performance ◽  
Dosage Form ◽  
Simultaneous Estimation ◽  
Reverse Phase ◽  
Stability Indicating ◽  
Rp Hplc ◽  
Development And Validation

 Objective: Development and validation of stability indicating reverse-phase high- performance liquid chromatography (RP-HPLC) method for simultaneous estimation of telmisartan (TEL) and benidipine hydrochloride (BND) in pharmaceutical dosage form.Methods: Reverse phase chromatography was selected because of its suggested use for ionic and moderate to non-polar compounds. Reverse phase chromatography is simple, suitable, better regarding efficiency, stability, and reproducibility. C18 column, a 250×4.6 mm column of 5.0 μm particle packing, was selected for separation of TEL and BND. Different solvent systems were tried and optimized in combinations as mobile phase. TEL (40 μg/ml) and BND (4 μg/ml) in buffer, pH 4.0: Methanol (50:50) was developed as it was showing good peak shapes and a significant amount of resolution. The mobile phase was flowed at 1.0 ml/min with detection of both the analytes at 210 nm using photodiode array detector.Result: Development of method was done, and validation was accomplished using specificity, linearity, accuracy, precision, robustness, limit of detection, and limit of quantitation. The method was found linear from 20 to 60 μg/ml and 2–6 μg/ml for TEL and BND individually. The percentage recoveries of TEL 100.46% and BND100.08% were, respectively.Conclusion: This stability indicating RP-HPLC methods were developed by degradation of sample and compared with standard. The percentage relative standard deviation was also <2 % showing high degree of precision of the proposed method. The proposed method can be used for routine analysis of benidipine HCl and TEL in combined dosage form and quality control in bulk manufacturing.

High Performance Liquid Chromatography of Furfural and Hydroxymethylfurfural in Spirits and Honey

1980 ◽  
Vol 63 (6) ◽  
pp. 1215-1218 ◽  
Author(s):  
Hans J Jeuring ◽  
Frans J E M Kuppers
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Sample Preparation ◽  
Mobile Phase ◽  
High Performance ◽  
Colorimetric Method ◽  
Reverse Phase ◽  
Effluent Flow Rate ◽  
Different Levels ◽  
Phase Column

Abstract Furfural (2-Jfuraldehyde) and hydroxymethylfurfural (S-hydroxy-2-furaIdehyde, HMF) are determined in brandy and honey by reverse phase high performance liquid chromatography. Brandies and other spirits are injected without sample preparation; honey is diluted with water and the solution is filtered before injection onto a reverse phase column with detection at 285 nm. The mobile phase is methanol–water (10+90) and the effluent flow rate is maintained at 1.0 mL/min. External standardization is used for quantitative determination. Recoveries from cognac and honey spiked at different levels ranged from 95 to 99% (furfural) and 95 to 100% (HMF). The furfural content of the brandies was also determined by the existing colorimetric method of the Bureau National Interprofessionel du Cognac. The HMF content of the honey was correlated to the results of the classic method of Winkler.

scholarly journals Chiral Separation and Determination of Etoxazole Enantiomers in Vegetables by Normal-Phase and Reverse-Phase High Performance Liquid Chromatography

Molecules ◽  
2020 ◽  
Vol 25 (14) ◽  
pp. 3134
Author(s):  
Ping Zhang ◽  
Yuhan He ◽  
Sheng Wang ◽  
Dongmei Shi ◽  
Yangyang Xu ◽  
...  
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Chiral Separation ◽  
Mobile Phase ◽  
High Performance ◽  
Normal Phase ◽  
Reverse Phase Hplc ◽  
Reverse Phase ◽  
Normal Phase Hplc ◽  
Chiralpak Ic

The chiral separation of etoxazole enantiomers on Lux Cellulose-1, Lux Cellulose-3, Chiralpak IC, and Chiralpak AD chiral columns was carefully investigated by normal-phase high performance liquid chromatography and reverse-phase high performance liquid chromatography (HPLC). Hexane/isopropanol, hexane/n-butanol, methanol/water, and acetonitrile/water were used as mobile phase at a flow rate of 0.8 mL/min. The effects of chiral stationary phase, mobile phase component, mobile phase ratio, and temperature on etoxazole separation were also studied. Etoxazole enantiomers were baseline separated on Lux Cellulose-1, Chiralpak IC, and Chiralpak AD chiral columns, and partially separated on Lux Cellulose-3 chiral column under normal-phase HPLC. However, the complete separation on Lux Cellulose-1, Chiralpak IC, and partial separation on Chiralpak AD were obtained under reverse-phase HPLC. Normal-phase HPLC presented better resolution for etoxazole enantiomers than reverse-phase HPLC. Thermodynamic parameters, including ΔH and ΔS, were also calculated based on column temperature changes from 10 °C to 40 °C, and the maximum resolutions (Rs) were not always acquired at the lowest temperature. Furthermore, the optimized method was successfully applied to determine etoxazole enantiomers in cucumber, cabbage, tomato, and soil. The results of chiral separation efficiency of etoxazole enantiomers under normal-phase and reverse-phase HPLC were compared, and contribute to the comprehensive environmental risk assessment of etoxazole at the enantiomer level.

Analysis for indole compounds in urine by high-performance liquid chromatography with fluorometric detection.

1976 ◽  
Vol 22 (2) ◽  
pp. 184-187 ◽  
Author(s):  
A P Graffeo ◽  
B L Karger
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Fluorescence Detection ◽  
Mobile Phase ◽  
High Performance ◽  
Fluorometric Detection ◽  
Chromatographic System ◽  
Reverse Phase ◽  
Indole Compounds ◽  
Phase Column

Abstract We describe a chromatographic system involving a high-performance chemically-bonded reverse-phase column and fluorescence detection for measurement of indoles in urine. We controlled retention and selectivity by optimizing the methanol content and pH of the mobile phase. Six reference indoles were separated in less than 20 min; three 5-hydroxyindoles were eluted in less than 7 min. About 5-15 ng of aqueous solutions of these compounds can be detected. The combination of selectivity (from use of the chromatographic column) and fluorescence detection permitted analysis for five of the six indoles after a single urine-deproteinization step.

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