HPLC Detectors

High performance liquid chromatograph (HPLC) detectors pass a beam of light through a column effluent as the fluid passes through a low-volume flow cell. Variations in light intensity are recorded and a chromatograph is generated. HPLC detectors use several detection methods. Ultraviolet (UV) detectors measure the ability of a sample to absorb light at one or more wavelengths. Light scattering detectors nebulize the effluent, vaporize the solvent, and then detect droplets in a light scattering cell. Electrochemical detectors measure the current from the oxidation/reduction reaction of an analyte at a suitable electrode. Radiochemical detectors use tritium or carbon-14 to detect the fluorescence associated with beta-particle ionization. Mass spectroscopy detectors ionize a sample and use a mass analyzer to detect the ion current. Nuclear magnetic resonance detectors irradiate nuclei that are placed between the poles of a strong magnet. The radiation is absorbed, the parallel nuclei enter a higher energy state, and each atom produces a spectra specific to its location and chemical composition. Some HPLC detectors measure the change in the refractive index of the column effluent passing through the flow cell. Others detect the fluorescence that occurs when compounds are excited by shorter wavelength energy and emit higher wavelength radiation. 

Important specifications for HPLC detectors include dynamic range, detector wavelength, response time, flow cell volume and flow cell rate. Dynamic range allows for the quantification of unknown samples using a multi-point calibration curve. It indicates when the peak area is directly proportional to the solute amount so that, for example, doubling the solute amount results in a doubling of peak area. Detector wavelength indicates the emission wavelength. Response time is the amount of time between a solute’s entry into an HPLC detector and the generation of a chromatogram. Flow cell volume and flow cell rate are measurements of capacity and efficiency, respectively. Maximum pressure rating is another important specification to consider when searching for HPLC detectors.

HPLC detectors vary in terms of general features, display options, and interface options. General features include self-calibration, self-test diagnostics, data storage, and programmability. Devices with a built-in chromatograph and application software are commonly available. Some HPLC detectors are suitable for extreme temperatures. Others output an analog voltage, analog current, frequency, or pulse. Examples include amplitude modulation (AM), frequency modulation (FM), pulse width modulation (PWN), sine waves and pulse trains. Analog devices usually include an analog meter and a front panel with switches and knobs. Digital devices often provide a serial or parallel computer interface and a video display. Serial protocols include universal serial bus (UBS), RS232, RS422, and RS485.

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