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A UPLC (ultra-high performance liquid chromatography) is a variant of HPLC using columns with particle size <2 um (typically, 1.8 um), which provides significantly better separation than the traditional (5 um) columns and enables much faster analysis. Strictly speaking, "UPLC" is Waters Corporation trademark, but is often used as a name for the technique in general.
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What are the advantages of UPLC over HPLC?
UPLC (Ultra Performance Liquid Chromatography) typically provides faster analysis times, higher resolution, and improved sensitivity compared to traditional HPLC (High Performance Liquid Chromatography). UPLC systems use smaller particle sizes in stationary phases and higher pressures, leading to better separation efficiency and lower solvent consumption.
What is Difference Between HPLC UV detector Spectrophotometer uv detector?
HPLC UV detector is a component used in high-performance liquid chromatography (HPLC) to monitor eluent absorbance, while a spectrophotometer UV detector is a standalone instrument used to measure the absorption of light at different wavelengths. HPLC UV detectors are specifically tailored for chromatography applications, whereas spectrophotometer UV detectors are more versatile and used for various analytical purposes.
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What are the key differences between reverse phase and normal phase HPLC techniques?
Reverse phase and normal phase HPLC techniques differ primarily in the polarity of the stationary phase and mobile phase. In reverse phase HPLC, the stationary phase is nonpolar and the mobile phase is polar, while in normal phase HPLC, the stationary phase is polar and the mobile phase is nonpolar. This polarity difference affects the retention and separation of compounds in the sample.
What is difference between high pressure liquid chromatography and high performance lequid chromatography?
High pressure liquid chromatography (HPLC) and high performance liquid chromatography (HPLC) are often used interchangeably. HPLC refers to modern liquid chromatography systems with high resolution and efficiency, while high pressure liquid chromatography specifically highlights the use of higher pressures in the system to improve separation and speed. Both terms generally refer to the same chromatographic technique.
What are the key differences between HPLC reverse phase and normal phase chromatography techniques?
In reverse phase HPLC, the stationary phase is nonpolar and the mobile phase is polar, while in normal phase HPLC, the stationary phase is polar and the mobile phase is nonpolar. This difference in polarity affects how compounds interact with the stationary phase, leading to variations in separation and elution times.
How do you use resolution factor in HPLC?
The resolution factor in HPLC is used to quantify the degree of separation between two adjacent peaks on a chromatogram. It is calculated by dividing the difference in retention times of the two peaks by the sum of their peak widths. A higher resolution factor indicates better separation between the peaks.
What are the key differences between HPLC normal phase and reverse phase chromatography techniques?
In normal phase HPLC, the stationary phase is polar and the mobile phase is nonpolar, while in reverse phase HPLC, the stationary phase is nonpolar and the mobile phase is polar. This difference in polarity affects how compounds interact with the stationary phase, leading to different separation mechanisms and selectivity in each technique.
What are the key differences between normal phase HPLC and reverse phase HPLC in terms of their separation mechanisms and applications?
Normal phase HPLC separates compounds based on their polarity, with the stationary phase being polar and the mobile phase being nonpolar. Reverse phase HPLC separates compounds based on their hydrophobicity, with the stationary phase being nonpolar and the mobile phase being polar. Normal phase HPLC is typically used for separating polar compounds, while reverse phase HPLC is used for separating nonpolar compounds.
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How do you distinguised np-hplc and rp-hplc?
NP-HPLC is "Normal Phase" HPLC, wherein the solvents used are less polar than the substrate in the HPLC column (e.g. using hexane or dichloromethane with a silica HPLC column). RP-HPLC is "Reverse-Phase" HPLC, wherein the solvents used are more polar than the substrate in the HPLC column (e.g. using Water and Methanol with a octadecylsilane (ODS or C18) column).
What is difference between isocratic and gradient hplc?
In isocratic HPLC, the mobile phase composition remains constant throughout the entire run, leading to constant elution times for all analytes. In gradient HPLC, the mobile phase composition is changed during the run, allowing for better separation of complex mixtures by adjusting the solvent strength over time.
