Determination of gradient elution conditions for the separation of peptide mixtures by reversed-phase high-performance liquid chromatography: bovine β-casein tryptic digest

1986 ◽  
Vol 53 (4) ◽  
pp. 595-600 ◽  
Author(s):  
Christophe Carles ◽  
Bruno Ribadeau-Dumas
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Flow Rate ◽  
High Performance ◽  
Gradient Elution ◽  
Reversed Phase ◽  
Tryptic Digest ◽  
Peptide Mixtures ◽  
Tryptic Hydrolysate

SummaryTaking as an example a tryptic hydrolysate of bovine β-casein, it was shown that the method used for the determination of optimal elution conditions for isocratic systems also applied to the separation of complex peptide mixtures by reversed-phase high-performance liquid chromatography with linear gradients. Using a C18 column and the same solvents, successive studies of the influence of flow rate, pH and temperature allowed a satisfactory separation of the sample in less than 30 min. Valuable information on the specificity of the action of trypsin on β-casein was deduced from the yield of the eluted peptides.

scholarly journals Optimization of Determination of Sucralose in Drink by HPLC

2019 ◽  
Vol 2 (3) ◽  
Author(s):  
Sang Li
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Flow Rate ◽  
High Performance ◽  
Detection Efficiency ◽  
Limit Of Detection ◽  
Reversed Phase ◽  
Operating Conditions ◽  
National Standards

To optimize the method for determination of Sucralose in drink by high performance liquid chromatography (HPLC). Using HPLC with RID, operating conditions were C18 reversed phase chromatograph column, 40:60 = methanol: 0.125% potassium hydrophosphate as mobil phase, measured at a flow rate of 0.8 mL/min. In the range of 20 ~ 400 mg L, with the concentration of Sucralose and corresponding peak area as standard, r = 0.9999, it has good correlation, the recovery of sucralose is 94~108%.The lower limit of detection of Sucralose was 0.0024 g/kg. This method not only meets the requirements of national standards, but also fast, sensitive, and environmentally friendly, improves the detection efficiency and safety of the detection of sucralose in drink by high performance liquid chromatography.

scholarly journals Combined Determination of Succinic Acid and Cetylpyridinium Chloride in Medicinal Films by Gradient High Performance Liquid Chromatography

Medicina ◽  
2021 ◽  
Vol 9 (2) ◽  
pp. 75-88
Author(s):  
O. N. Dvorskaya ◽  
◽  
N. N. Nozhkina ◽  
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Succinic Acid ◽  
High Performance ◽  
Cetylpyridinium Chloride ◽  
Gradient Elution ◽  
Reversed Phase ◽  
Gradient Elution Mode

A technique has been developed based on reversed-phase high-performance liquid chromatography with diode-matrix detection for the joint determination of succinic acid and cetylpyridinium chloride in complex action medicinal films. Efficient chromatographic separation of active drug components was achieved in a gradient elution mode on a Luna C18 (2) 100A column (4.6 × 250 mm, 5 µm) using a mobile phase consisting of a 0.1% solution of phosphoric acid and acetonitrile. The detection wavelength was 210 nm for both compounds. The developed method is validated in terms of specificity, linearity, precision, accuracy and can be used to determine the authenticity and quantitative content of succinic acid and cetylpyridinium chloride in the joint presence in assessing the quality of medicinal films.

scholarly journals SIMULTANEOUS IDENTIFICATION AND QUANTIFICATION OF HYDROQUINONE, TRETINOIN AND BETAMETHASONE IN COSMETIC PRODUCTS BY ISOCRATIC REVERSED PHASE HIGH PERFORMANCE LIQUID CHROMATOGRAPHY

2019 ◽  
pp. 181-185 ◽  
Author(s):  
Baitha Maggadani ◽  
Harmita Harmita ◽  
Yahdiana Harahap ◽  
Hanna Hutabalian
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Flow Rate ◽  
Mobile Phase ◽  
High Performance ◽  
Method Development ◽  
Reversed Phase ◽  
Two Samples ◽  
Simultaneous Identification

Objective: The objective of this study was to obtain a simple and selective analysis method for determination of hydroquinone, tretinoin and betamethasone in whitening creams using reversed phase high-performance liquid chromatography (HPLC). Methods: Reverse Phase HPLC was used for method development, validation studies, and sample analysis. The method was optimized by evaluating several parameters that affects extraction of the sample, composition and types of mobile phase and also flow rate. Chromatographic separation was optimized on a C18 column [Sunfire, 250 x 4.6 mm, 5µm] utilizing a mobile phase consisting acetonitrile, methanol (90:10 v/v) and slightly addition of glacial acetic acid to reach pH 5in the ratio of 30: 50:20 v/v at a flow rate of 0.8 ml/min with UV detection at 270 nm and 350 nm. Results: The analytical methods fulfilled the validation requirements including accuracy, precision, linearity, selectivity, detection limits, and quantitation limits. The results showed the mean levels of hydroquinone, tretinoin and betamethasone in samples A and B were 1.78%; 0.07%; 0.12% and 2.00%; 0.07%; 0.13% respectively. Conclusion: The method was successfully applied for the determination of cosmetic formulation containing hydroquinone, tretinoin and betamethasone simultaneously. There were seven samples analyzed and two samples were positive containing hydroquinone, tretinoin, and betamethasone.

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