scholarly journals Determination of Cholesterol Content in Butter by HPLC: Up-to-Date Optimization, and In-House Validation Using Reference Materials

Foods ◽  
2020 ◽  
Vol 9 (10) ◽  
pp. 1378
Author(s):  
Lukáš Kolarič ◽  
Peter Šimko
Keyword(s):  
Reference Materials ◽  
Dairy Products ◽  
Mobile Phase ◽  
Cholesterol Content ◽  
Optimal Conditions ◽  
Hplc Separation ◽  
Isocratic Elution ◽  
Validation Criteria ◽  
Double Extraction

This work deals with up-to-date optimization of cholesterol content determination when saponification and extraction procedures as well as HPLC conditions were studied. As found, optimal conditions for saponification process were identified by 15 min heating in the presence of 0.015 L of methanolic KOH solution with a concentration 1 mol/L with subsequent 0.015 L n-hexane–chloroform binary mixture (1:1, v/v) double extraction. HPLC separation consisted of isocratic elution with flow rate of 0.5 mL/min mobile phase composed of acetonitrile/methanol 60:40 (v/v) and stationary phase Zorbax Eclipse Plus C18 column 2.1 × 100 mm, 3.5 μm particle size diameters with detector wavelength 205 nm. The method passed through in-house validation criteria and its suitability was verified by analysis of butter reference materials. In final, the average content of cholesterol content in butter was determined at 2271.0 mg/kg. Thus, the method is suitable for the determination of cholesterol content in butter and probably also in other dairy products.

scholarly journals HPLC determination of hydrochlorothiazide in urine after solid-phase extraction

2005 ◽  
Vol 51 ◽  
pp. 23-28 ◽  
Author(s):  
Violeta Ivanova ◽  
Dragica Zendelovska ◽  
Marina Stefova
Keyword(s):  
Particle Size ◽  
Solid Phase Extraction ◽  
Mobile Phase ◽  
Solid Phase ◽  
Hplc Method ◽  
Phase Extraction ◽  
Hplc Separation ◽  
Isocratic Elution ◽  
Extraction Recovery

A simple, rapid and precise HPLC method has been developed for the assay of hydrochlorothiazide in urine. The clean-up of the urine samples was carried out by solid-phase extraction using HLB cartridges. Extraction recovery was 94.00-100.28 %. HPLC separation was performed with isocratic elution on Hypersil BDS C18 column (100 x 4.0 mm I.D., 3 µm particle size) protected with appropriate guard column. The mobile phase was 18 % acetonitrile and 0.025 mol/L solution of KH2PO4, pH 4 at flow rate of 0.3 mL/min. Detection of the substances was performed at 220 nm. The calibration curves were linear in the range of 2-50 µg/mL. The developed method is validated by checking its accuracy, precision and stability. The detection limit is 2 µg/mL hydrochlorothiazide. The method is proved to be convenient for routine analysis of hydrochlorothiazide in urine.

scholarly journals Validation of developed method by RP-HPLC for simultaneous estimation of famotidine and ibuprofen in human plasma and studying the stability of the drugs in plasma

2018 ◽  
Vol 12 (1) ◽  
pp. 34-48
Author(s):  
K. S Ashutosh ◽  
D. Manidipa ◽  
R. J. V. L. N. Seshagiri ◽  
S. D. Gowri
Keyword(s):  
Human Plasma ◽  
Mobile Phase ◽  
Simultaneous Estimation ◽  
Hplc Separation ◽  
Reverse Phase ◽  
Rp Hplc ◽  
The Stability ◽  
Sodium Dihydrogen ◽  
Isocratic Mode

The RP-HPLC separation was carried out by reverse phase chromatography on a Symmetry C18 (4.6 x 150 mm, 3.5 μm, make: XTerra) with a mobile phase composed of sodium dihydrogen ortho phosphate [pH 2.5] and acetonitrile in the ratio of 30:70 v/v in an isocratic mode at a flow rate of 1.2 mL/min. The run time was maintained for 8.0 min. The detection was monitored at 236 nm. The accuracy was calculated in human plasma and the % recovery was found 99.80 - 99.85 for famotidine and 99.56 -99.85.5 for ibuprofen and reproducibility was found to be satisfactory. The calibration curve for famotidine in human plasma was linear over 3.32 to 6.65 μg/mL and 100- 200 μg/mL for ibuprofen in human plasma respectively. The inter-day and intra-day precision in human plasma was found within limits. The proposed method has adequate sensitivity, reproducibility, and specificity for the determination of famotidine and ibuprofen in plasma. The LLOQ obtained by the proposed method in human plasma were 1.24 and 5.0 μg/mL for famotidine and ibuprofen respectively. The proposed method is simple, fast, accurate, and precise for the quantification of famotidine and ibuprofen in plasma as per the ICH guidelines.Kathmandu University Journal of Science, Engineering and TechnologyVol. 12, No. I, June, 2016, Page: 34-48

Determination of Serum Diclofenac by High-performance Liquid Chromatography by Electromechanical Detection

1985 ◽  
Vol 4 (3) ◽  
pp. 317-322 ◽  
Author(s):  
F. Plavsic ◽  
J. Čulig
Keyword(s):  
Entire Range ◽  
Mobile Phase ◽  
High Performance ◽  
Dose Interval ◽  
Phase Flow ◽  
Electrochemical Detector ◽  
Mobile Phase Flow Rate ◽  
Coefficients Of Variation ◽  
Double Extraction

Diclofenac was isolated from plasma by double extraction with organic solvent. Chromatography was performed on Lichrosorb NH2 column with acetonitrile and perchloric acid (0.0025 mol/l; 35:65, v/v) as the mobile phase (flow rate 0.9 ml/min). The electrochemical detector was operated at +900 mV and sensitivity of 5-10 nA. The sensitivity of the method was 10 μg/1 and the coefficients of variation below 10% in the entire range of the concentration in a dose interval.

Validation of HPTLC and HPLC Methods for the Quantitative Determination of Allyl Disulfide in Some Polyherbal Oils

2012 ◽  
Vol 95 (6) ◽  
pp. 1574-1578
Author(s):  
Nitin Dubey ◽  
Nidhi Dubey ◽  
Rajendra Mehta
Keyword(s):  
Particle Size ◽  
Quantitative Determination ◽  
Silica Gel ◽  
Vegetable Oil ◽  
Flow Rate ◽  
Mobile Phase ◽  
Allium Sativum ◽  
Hplc Separation ◽  
Reliable Marker

Abstract Allium sativum L (garlic) is an essential component of many polyherbal oils used in traditional systems of medicine. Allyl disulfide has been a major component found in vegetable oil macerate of garlic, and can be used as reliable marker for determination of garlic in oil macerates of garlic. The HPLC separation of allyl disulfide was achieved on a Phenomenex Luna C18 (25 cm × 4.6 mm id × 5 μm particle size) column using acetonitrile–water–tetrahydrofuran (70 + 27 + 3, v/v/v) mobile phase at a flow rate of 1.0 mL/min. Quantitation was achieved with UV detection at 298 nm over the concentration range 8–48 μg/mL. HPTLC separation of allyl disulfide was achieved on an aluminum-backed layer of silica gel 60 F254 using n-hexane mobile phase. Quantitation was achieved by densitometric analysis at 298 nm over the 200–1200 ng/band concentration range. The methods were validated according to International Conference on Harmonization guidelines.

scholarly journals Simultaneous HPLC estimation of Amphetamine and Caffeine abuse drugs in Iraqi human addicts

2021 ◽  
Vol 4 (1) ◽  
pp. 25-31
Author(s):  
Rulla Sabah
Keyword(s):  
Particle Size ◽  
High Performance Liquid Chromatography ◽  
Mobile Phase ◽  
High Performance ◽  
Linear Regression Analysis ◽  
Analysis Data ◽  
Hplc Separation ◽  
Isocratic Elution ◽  
Diethyl Amine ◽  
Buffer Value

The main aim of this research is to establish and validate a high performance liquid chromatography (HPLC) process for the separation and estimation of Amphetamine (AM) and Caffeine (CAF) in its illegal formula and in sera of addicts. This method is established on the HPLC separation of the two drugs on the ZORBAX ODS column (250×4.6×5µm particle size). The mobile phase contained 1% ortho-phosphoric acid 85% and 1%of diethyl amine 99%, acetonitrile and methanol ratio was 85:10:5 v/v/v. The flow rate is 1.2 mL.min-1, buffer value pH of 2.5 via isocratic elution also UV detection at 210 nm. The retention times for the two drugs AM and CAF were obtained at 4.425 and 6.456 min, respectively. The calibration curves founded that the linear regression analysis data gave a good linear relationship for the concentration range 1 to 100 µg.mL-1 for AM and CAF. The values achieved for correlation coefficient, slope and intercept were 9999, 8104.2 and 5012 for AM and 0.9999, 9698.5 and 6342.9 for CAF, whereas the LOD and LOQ was 0.51, 1.64 µg.mL-1 for AM and 0.60, 1.32 µg.mL-1 for CAF  

APPLICATION OF HPTLC AND RP-HPLC METHODS FOR SIMULTANEOUS ESTIMATION OF OFLOXACIN AND PREDNISOLONE ACETATE IN OPHTHALMIC DOSAGE FORM

INDIAN DRUGS ◽  
2015 ◽  
Vol 52 (05) ◽  
pp. 17-26
Author(s):  
M. L Chavhan ◽  
◽  
A. S Patil ◽  
S. J Surana ◽  
A. A. Shirkhedkar
Keyword(s):  
Mobile Phase ◽  
Dosage Form ◽  
Internal Standard ◽  
Prednisolone Acetate ◽  
Simultaneous Estimation ◽  
Hplc Separation ◽  
Potassium Dihydrogen ◽  
Rp Hplc ◽  
Ophthalmic Formulation

Two simple, rapid, precise, accurate and robust (HPTLC and RP-HPLC) methods have been established for simultaneous determination of ofloxacin and prednisolone acetate in its combined ophthalmic formulation. HPTLC separation of two drugs was accomplished on a HPTLC aluminum-backed layer of silica gel 60 F254 using n-butanol: methanol: ammonia (60:10:30% V/V/V) as a mobile phase. The densitometric scanning was performed at 275 nm which showed Rf 0.43 for ofloxacin and 0.78 for prednisolone acetate, respectively. RP-HPLC separation of the two drugs was achieved on LC-GC Qualisil BDS C-18 column using mobile phase methanol: acetonitrile: 0.02 M potassium dihydrogen ortho-phosphate anhydrous (50:15:35% V/V/V), pH adjusted to 4.5 with triethylamine. Detection of both drugs was done at 275 nm. Aspirin was used as an internal standard (IS). The retention time for ofloxacin, prednisolone acetate and aspirin (IS), was found to be 4.36 min. and 6.60 min and 3.55 min, respectively. The proposed methods were successfully applied for the determination of both drugs in bulk and in combined ophthalmic formulation. Assay of both these methods were compared using student t-test.

scholarly journals Simultaneous Determination of Phenolic Compounds in Leptadenia pyrotechnica (Forssk.) Decne. by Using High-Performance Liquid Chromatography (HPLC-DAD-UV)

2018 ◽  
Vol 2018 ◽  
pp. 1-4
Author(s):  
Raman Preet ◽  
Raghbir Chand Gupta
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Phenolic Compounds ◽  
Mobile Phase ◽  
High Performance ◽  
Isocratic Elution ◽  
Methanolic Extracts ◽  
Aerial Parts ◽  
Whole Plant

During the present study, an endeavor has been made to produce a simple, rapid, and simultaneous method for determination of phenolic compounds by using high-performance liquid chromatography in aerial parts of Leptadenia pyrotechnica (Forssk.) Decne. collected from the Indian Thar Desert. The optimized process was used for the quantification of ten phenolic compounds. The chromatographic separation was accomplished on an Atlantis T3 column at 25°C with isocratic elution. A mixture of acetonitrile and water was used as the mobile phase at a flow rate of 0.8 mL/min. The linear regression examination data for the calibration plots displayed a good linear relationship with r2 > 0.999 in the concentration range of 2–20 µL. In the methanolic extracts of the whole plant of L. pyrotechnica, the content of caffeic acid (3.3%) was reported to be the highest concentration.

scholarly journals Determination of Magnesium Valproate and Its Process Related Impurities by Ultraperformance Liquid Chromatography

2014 ◽  
Vol 2014 ◽  
pp. 1-6
Author(s):  
Rakshit Thakkar ◽  
Hitesh Saravaia ◽  
Madhavi Patel ◽  
Anamik Shah
Keyword(s):  
Mobile Phase ◽  
Reversed Phase ◽  
Isocratic Elution ◽  
Related Impurities ◽  
System Suitability ◽  
Liquid Chromatographic ◽  
Ammonium Dihydrogen Orthophosphate ◽  
Dihydrogen Orthophosphate

A selective ultraperformance liquid chromatographic (UPLC) method for the determination of magnesium valproate and its process related impurities has been developed. The method includes reversed-phase Acquity BEH C18 column with 100 mm × 2.1 mm i.d. and 1.7 µ particle size. The mobile phase consists of acetonitrile and 5 mM ammonium dihydrogen orthophosphate with pH = 3.0 at 45 : 55 isocratic elution. The flow rate was set at 0.3 mL/min and UV detection was performed at 215 nm. A system suitability test (SST) was developed to govern the quality of the separation. The developed method has been validated further with respect to linearity, accuracy, precision, selectivity, LOD, LOQ, and robustness. Different batches of samples were examined using this method; the method proved to be successful when applied to analyze a marketed magnesium valproate formulation.

scholarly journals Determination of Retinol and Tocopherols in Human Serum using Ultra-Performance Liquid Chromatography with Photodiode Array

2020 ◽  
Vol 36 (05) ◽  
pp. 819-824
Author(s):  
Augosto Asor Misolas
Keyword(s):  
Liquid Chromatography ◽  
Human Serum ◽  
Mobile Phase ◽  
Ultra Performance Liquid Chromatography ◽  
Tocopheryl Acetate ◽  
Isocratic Elution ◽  
Retinyl Acetate ◽  
Internal Standards ◽  
Photodiode Array

Method that can simultaneously determine retinol, γ-tocopherol and a-tocopherol in human serum was developed utilizing ultra-performance liquid chromatography. Retinyl and tocopheryl acetates were employed as internal standards. The reverse-phased method utilizes isocratic elution with a mobile phase consisting of 20% acetonitrile and 80% methanol at a flow rate of 0.800 mL/minute. Separation was attained using an ethylene bridged hybrid (BEH) C18 column. Retinols and tocopherols were detected by photodiode array at wavelengths 325 nm and 295 nm, respectively. The retention times for retinol and retinyl acetate were 0.42 and 0.49 minutes respectively. γ-Tocopherol, α-tocopherol and tocopheryl acetate eluted at 0.86, 0.94 and 1.1 min, respectively. The limits of quantification were determined and found to be 0.025 μg/mL, 0.50 μg/mL and 1.0 μg/mL for retinol, γ-tocopherol and α-tocopherol, respectively. The method has been found to be suitable for the determination of retinol and tocopherols in human serum.

A SIMPLE AND RAPID HPLC METHOD FOR ESTIMATION OF DIMETHICONE FROM FORMULATIONS

INDIAN DRUGS ◽  
2013 ◽  
Vol 50 (03) ◽  
pp. 26-29
Author(s):  
J. J Jadhav ◽  
◽  
S Mungekar ◽  
J. V. Velada ◽  
H. A. Doshi ◽  
...  
Keyword(s):  
Mobile Phase ◽  
High Performance ◽  
Dosage Forms ◽  
Hplc Method ◽  
Normal Phase ◽  
Hplc Separation ◽  
Tablet Dosage ◽  
Refractive Index Detector ◽  
Isocratic Mode

A simple, sensitive, precise and specific normal phase high performance liquid chromatography (HPLC) method was developed and validated for the determination of dimethicone from tablet dosage forms. It was found that the excipients used in the tablet dosage form did not interfere in the quantification of dimethicone. The HPLC separation was carried out by normal phase chromatography on Princeton Sphere Cyano, 250 x 4.6mm, 5µ with a mobile phase composed of hexane : ethanol : ethyl acetate (80:20:0.2) in isocratic mode at a flow rate of 0.5mL/min. Dimethicone was quantified using a refractive index detector. The calibration curve for dimethicone was linear from 1.75 to 3.25 mg/mL. The inter-day and intra-day precisions were found to be within limits. The proposed method has adequate sensitivity, reproducibility and specificity for the determination of dimethicone from tablet dosage forms.

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