The waters 2410 refractive index detector is designed for highperformance liquid chromatography applications as either a standalone unit with an integrator or chartreader or otherwise as part of a waters system controller or data system. For example, hplc can be used to determine the amount of morphine in a compounded solution. Refractive index detctor shodex hplc columns, detectors. The single detector supports all applications from highly sensitive measurements to preparative measurements using the three operation modes shown below. Waters 2414 refractive index detector operators guide. The detection principle involves measuring of the change in refractive index of the column effluent passing through the flowcell.
Iii of iv oq and pq operating instructions contents. Next generation technology contained in waters 2414 differential refractive index detector reduces detector baseline noise and drift compared to traditional products. However, seems it could change the refractive index and thus give a detector response. Sometimes the organic solvent used for gpc analysis absorbs uv, and thus uv detector cannot be.
Uv detector is a very commonly used detector for hplc analysis. The detector is used to sense the presence of a compound passing through and to provide an electronic signal to a dataacquisition device. Operational qualification performance qualification for. Naphthalene, acenaphthene, pyrene and perylene in hplc were used as illustrations. Group 3 shariful alam masud rana piash kar sumaiya binta fashiuddin batch. Such a separation requires a refractive index detector since the mobile phase prevents detection of analytes with short wavelength uv. The modern refractive index detector is the result. It provides sensitivity, stability, and reproducibility for the analysis of components with limited or no uv absorption. Baseline stabilization when using refractive index.
The uv, vis, and pda detectors are categorized as absorbance detectors. High performance liquid chromatography hplc introduction different phase in hplc configuration of hplc detector of hplc high performance liquid chromatography high performance or high pressure. The following chromatographic conditions were adopted. This method of monitoring refractive index is called the angle of deviation method and has been used by a number of manufacturers in their detector design. The hplc detectors are used to detect the solute present in the eluent comes from the hplc column.
Waters 2410 hplc refractive index detector marshall scientific. Analysis of glyoxal and related substances by means of. Highperformance liquid chromatography hplc is a type of liquid chromatography used to separate and quantify compounds that have been dissolved in solution. The ability to obtain lower limits of detection and quantitation, when methods are developed and run using the waters 2414 ri detector, are now possible. High performance liquid chromatography hplc uses different types of hplc detectors types comparison principles pdf ppt of ultravioletvisible spectroscopic detectors pda detectors, refractiveindex detector,mass spectrometer, conductivity detector, fluorescence detector. Refractive index detector measures the molecules ability to deflect light in a flowing mobile phase in a flow cell relative to a static mobile phase contained in a reference cell. A differential refractometer dri, or refractive index detector ri or rid is a detector that measures the refractive index of an analyte relative to the solvent. The relation between the refractive index and the concentration depends on the. Development and validation of a novel reversed phase high performance liquid chromatography with refractive index detector method for assay of polyvinyl alcohol in an ophthalmic solution volume. Analysis of carbohydrates, alcohols, and organic acids. Refractive index detector ri responds to nearly all solutes but has poor sensitivity detects changes in refractive index as sample passes through as long as solute has different ri than solvent analogous to tcd in gc. The destructive detectors perform continuous transformation of the column effluent burning, evaporation or mixing with reagents with. The standard solution and the sample contain carbohydrates.
After 30 minutes power on the rid20a reduces the time to. The greater the ri difference between sample and mobile phase, the larger the imbalance will become. The thermal isolated optic with a countercurrent heat exchanger and with its programmable temperature control, results in an extremly stable baseline and an optimal signal noise ratio. Hplc 1 turn on the uv detector for the hplc you plan to use at least 10 minutes before running samples.
Agilent 1260 infinity refractive index detector user manual. Hplc refractive index detectors hplc ri detectors are used with highpressure liquid chromatographs when detecting substances with limited or no uv absorption. These chemical components included alcohols, sugars, fatty acids, polymers and carbohydrates. Refractive index detector liquid chromatography shimadzu. Rid20a inheriting the stability and extensibility that are the strengths of the prominence series, the new rid20a model of differential refractive index detector is designed with a new referencecell autopurge feature and validation support function. Determination of glycols by hplc with refractive index. Analysis of cyclodextrins using hplc with a differential refractive index detector the analysis of cyclodextrin cd is presented below. Introduction to the refractive index detector 1 detection principle detection principle detector design the agilent 1260 infinity refractive index detector is a differential refractometer that measures the deflection of a light beam due to the difference in refractive index between the liquids in the sample and reference cells of a single flow. I recently started with a company and have been given the task of using the hplc more for the analytical work.
The chromatographic system coupled to the refractive index detector hplcri was equipped with a quaternary pump waters 2695 alliance, milford ma, usa, degasser, auto injector, and waters ri 2414 refractive index detector milford ma, usa, and the chromatographic data were acquired using the empower 2 software. A simple and fast hplc method for the determination of glycols is described. In contrast, change in reflective index occurs for all analyte, thus a ri detector can be used to measure all analyte. The refractive index is affected significantly by the solution temperature.
As light passes through the interface between two mediums of different refractive indices, it bends as shown in fig. Harun ergen, muge guleli, cigdem sener, cem caliskan, sercan semiz and mahmut ozbek affiliation. Waters 2410 hplc refractive index detector marshall. After comparison of all detector, find out which is the best detector for our solute analysis. A simple and rapid method is described for the analysis of glyoxal and related substances by highperformance liquid chromatography with a refractive index detector. Development and optimization of a hplcri method for the. Waters 2414 differential refractive index detector. The power switch is located on the front of the detector. During the analysis, sample goes through a clear colorless glass cell, called flow. This procedure is recommended for testing newly installed azura analytical hplc systems as well as for regular monitoring of system performance. Hplc detector refractive index detctor conductivity detector shodex cd200. Refractive index detector measures the overall ability of the mobile phase and its solutes to refract or bend light. Change in refractive index in response to change in cell temperature.
Countercurrent heat exchangers keep the optical unit and the flow cells at constant. Reduced stabilization time and improved baseline stability. The main types of detectors used in hplc are refractive index ri, ultraviolet uvvis and. Vsp0012n performance verification of azura systems with. Refractive index detector ri responds to nearly all solutes but has poor sensitivity detects changes in refractive index as sample passes through as long as solute has. World medicine pharmaceutical industry and trade inc. Page 81 span adjustment set the detector that the plot full scale of the chromatopac becomes 128 1 06 auxrange of the detector flow distilled water from the inlet port using the syringe and the adapter. The fourpartition photodetector in the rid20a allows a wide refractive index range 0.
Hplc and uhplc detectors thermo fisher scientific us. Baseline stabilization when using refractive index detectors. Inheriting the stability and extensibility that are the strengths of the prominence series, the new rid20a model of differential refractive index detector is designed with a new referencecell. Refractive indexbased detection of gradient elution liquid. Refractive index or the index of refraction is the ratio of the speed of light in vacuum or air to the speed of light in a transparent material. Aminex hpx87h column, mobile phase consisting of 0. The detector measures a change in refractive index, so the reference cell is filled with the eluent, and the detector subtracts the reading from the reference cell from that of the sample cell. Refractive index hplc detector the ri403035 refractive index detectors include an optical system mounted in a precise temperatureregulated housing to eliminate the influence of changes in ambient temperature.
Development and validation of a novel reversed phase high. It is applicable for the use with solvent that has uv absorbance. The 1260 infinity ii rid is an hplc refractive index detector rid detector that provides excellent performance for entrylevel uhplc and hplc differential refractometer dri experiments. Both detectors are fully stable just one hour after powering on. A chromatography detector is a device used in gas chromatography gc or liquid chromatography lc to detect components of the mixture being eluted off the chromatography column.
Photodiode array detector refractive index detector. They are easy to operate and provide good stability. Change in refractive index when degassing unit is switched from off to on with no column, 1. What are the applications of refractive index detector in. Concentration in most solutions, the concentration of solute in a solvent can be determined by measuring the refractive index n d. Detectors waters 2414 refractive index ri detector 2. There are various types of detectors available, but detector selection in hplc system is most valuable part in the solute detection. One of the most common bulk property detectors is the refractive index detector. The agilent 1260 infinity refractive index detector rid is the ideal detector for fast and reliable lc results in the routine analysis of nonuv absorbing substances, such as carbohydrates, lipids, and polymers. Refractive index detector questions chromatography forum. The thermal isolated optic with a countercurrent heat exchanger and with its programmable temperature control, results in an extremly stable baseline and an optimal signal. Highperformance liquid chromatography hplc is a type. Jan 29, 2017 high performance liquid chromatography hplc uses different types of hplc detectors types comparison principles pdf ppt of ultravioletvisible spectroscopic detectors pda detectors, refractiveindex detector,mass spectrometer, conductivity detector, fluorescence detector.
I have known people to put those detectors under a cardboard box bc hoods or the heat coming on can blow. High performance liquid chromatography hplc uses different types of hplc detectors types comparison principles pdf ppt of ultravioletvisible spectroscopic detectors pda detectors, refractive index detector,mass spectrometer, conductivity detector, fluorescence detector. Bulk property detectors are the most universal detectors for hplc as they measure properties common to all analytes by measuring differences in the mobile phase with and without the sample. However the change which is measured is very much smaller than the example given above, typically of the order of 0. The ri4030 can be used for analytical and preparative scales. This property is very helpful in designing optical lenses as it allows to calculate the bend angle of beam of light when passes. They are considered to be universal detectors because they can detect anything with a refractive index different from the. Rapid equilibration times and stable baselines make it easy to analyze carbohydrates, triglycerides, organic acids, and pharmaceutical excipients.
The s 3580 differential refractive index detector series offers the sensitivity, stability and reproducebility required for optimal ri detection. Intended use of the waters 2414 refractive index detector the 2414 refractive index ri detector is designed for high performance liquid chromatography hplc applications. The use of a refractive index detector precludes the use of a gradient program. Therefore, many commercially available rids are designed to control the temperature around the flow cell to ensure its temperature remains as constant as possible. The problem is that a chromatogram shows negative peaks after positive peak exiting, working with hplc ri detector. Continuously measures the refractive index of the effluent. Applications where hplc ri detectors are used include polymer analysis. Inject the standard solution whose refractive index is known, and adjust the detector output so that the signal intensity can be verfied.
Refractive indexbased sensors offer attractive characteristics as nondestructive and universal detectors for liquid chromatographic separations, but a small dynamic range and sensitivity to minor thermal perturbations limit the utility of commercial ri detectors for many potential applications, especially those requiring the use of gradient elutions. Different flavors of optical detectors are available. They are often used as detectors for highperformance liquid chromatography and size exclusion chromatography. Electrochemical detector ecd refractive index detector refractive index detector. Index introduction types characterization absorbance detector fluorescence detector refractive index elsd electrical conductivity detector electrochemical detector 2 3. The rid20a differential refractive index detector offers excellent stability and reproducibility while achieving a wide refractive index range. Offline ankur choudhary is indias first professional pharmaceutical blogger, author and founder of pharmaceutical guidelines, a widelyread.
You have a wide array of lc detectors for the thermo scientific vanquish and ultimate 3000 hplc and uhplc systems to choose from. Other types of hplc detectors are the evaporative light scattering detectors, ir detector, transport detectors, aerosolbased detectors, chiral detectors and the pulsed amperometric detectors. The refractive index ri detector is the only universal detector in hplc. The method was applied to investigate the biodegradation of glycols in a laboratory activated sludge plant and to determine the content of glycols or alcohols in detergents. The modern refractive index detector is the result of considerable research which has been extended by. Is something wrong with my refractive index detector for hplc. It is characterized by a reversedphase separation using water as eluent and a refractive index detection. A uv detector cannot be used with solvent which has uv absorbance. Used hplc equipment guide to refractive index detectors. Refractive indexbased detection of gradient elution. However the change which is measured is very much smaller than the example given. Hplc detectors types comparison principles pdf ppt.
I have been able to hook it in line with the system and configure it. The refractive index detector seton hall university. We have an agilent 1260 binary pump setup with an diode array detector and a refractive index detector. Often used in size exclusion chromatography for polymer analysis. They provide good sensitivity for lightabsorbing compounds at pg level. The ability to obtain lower limits of detection and quantitation, when methods are developed and run using the waters 2414 ri detecto.
375 1227 204 109 395 918 987 919 1430 307 1105 858 646 1540 1408 1276 1113 748 859 1355 11 396 1027 963 1132 142 529 1596 122 705 237 126 1490 941 237 12 446 475 974 599 387 237