AU Biomaterials Design Lab:Equipment/UV-2550 Shimadzu

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Shimadzu UV-2550
  • Shimadzu® UV-2550 is a ultra-low stray light double monochromator instrument. It provides UV-vis spectral analysis for samples by the embedded double blazed grating double monochromator. High energy optical system, wide dynamic range, small beam size, law stray light are some of its advantages over other double monochromators.
  • Its window-compatible software: UVProbe contains three modes: Spectrum, Kinetics, and Photometric modes, that allows wide range of applications.

Purpose of use


  • Shimadzu™ UV-2550® Spectrophotometer is used to measure spectrum of light absorption with wavelength ranging from 200nm to 1100nm for liquid, gas, and solid samples. However, since this instrument is only equipped with multicell Sample Compartment, the samples measured can only be liquid or solids.

Sample Range

  • With the current multicell Sample Compartment, liquid and solid samples can be run by inserting into a quartz or plastic cuvette that is able to fit inside multicell Sample Compartment.
  • Liquid samples covers wide range including biological and non-biological liquid solutions. Small solid samples can be cut; size of solid samples can be adjusted to fit inside a quartz or plastic cuvette for analysis.
  • Instrument is able to perform test on larger sample with the usage of sample compartment that accommodate larger sizes.


  • Beeghly 207
  • American University
  • Washington, DC, 20016
    • access to the laboratory is restricted; see Dr. Miller, Dr. Hartings or Dr. Fox for access.



  • Instrument has three different methods for analyzing samples: Spectrum, Photometric, and Kinetic methods. The usage for methods are different.
  • When instrument is turned on, lick connect on UVProve software to connect instrument with program.
  • After instruments calibrated and self checked, proceed to toolbar to select methods.
  • Samples can be placed into multicell sample compartment. Door to sample compartment should be closed before samples are run.

Spectrum Method

  • Spectrum mode is used for scanning a range of wavelength onto a sample and can be selected on toolbar.
  • M buttom in PVProbe sets up the acquired parameters.
  • Place sample into sample compartment. Standard samples in reference sides are preferred.
  • Click start to start the run.
  • Table is shown be clicking on Data Table, peaks in spectrum can be labeled by clicking on peak pick table.

Photometric Method

  • Photometric mode is used for measuring absorbance at a specific wavelength.
  • After parameters are set up in "Method", click on photometric method button in tool bar.
  • Select wavelength specified for sample run. Concentration units, type of standard curve can be set up afterwards.
  • Then enter measurement parameters for standards. The measurements are acquired by instrument.
  • Set sample measurement parameters. Measurement acquired by instrument.
  • Standard table is then shown. Fill in the concentration in standard table.
  • Load sample.
  • Click start to run sample.
  • Once data are collected, additional data points are going to shown on standard table.

Kinetics Method

  • Knetics mode collects data at a specific wavelength over a set amount of time.
  • After instrument warmed up, select Kinetics Method in toolbar.
  • Set up the amount of time for samples to run, the amount of time each data point is taken throughout each run, and the range of wavelength the instrument should run samples at under method section.
  • Load sample in sample compartment.
  • Click start to run.
  • Additional data points are shown after each sample run.
  • Graph of absorbance versus time is shown under graphical section of screen.

A copy of the instrument user manual is within the laboratory draw in room 207. See contact for more information.


Electric Shock

  • Do not open the cover of instrument while running. To avoid electric shock, first switch off power, then open instrument.

High Temperature

  • Do not touch or approach instrument too close. Instrument might overheat leading to possible burns on body at small distance or upon contact.


TCC-240A Thermoelectrically Temperature Controlled Cell Holder

  • Uses Peltier effect for controlling the temperatures of the sample and reference sample. No thermostatic bath or cooling water is required, so the operation is quite simple and easy.
    • Number of cells: One each on the sample and reference sides.
    • Temperature control range: 7 to 60°C
    • Temperature display accuracy (difference from the true value): ±0.5°C
    • Temperature control precision (variation of temperature): ±0.1°C

Multicell Sample Compartment

  • Holds up to six 10 mm square cells. No temperature control capability.
    • Number of cells: 6 on the sample side, and 1 on the reference side

UVProbe Software

  • Allows data processing with customized paramaters
    • Multitasking (possible to execute data processing while measurement is being executed)
    • Customizable measurement screen layout (wavelengths, data display font and font size, colors, displayed number of rows)
    • GLP/GMP compliant (security, history)
    • Real-time concentration display


Setting Wavelength Range

  • 190nm to 1100nm

Measurement Wavelength Range

  • 190nm to 900nm, but can measure up to 1100 with choice with detector

Wavelength accuracy

  • ±0.3nm

Wavelength repeatability

  • ±0.1nm

Wavelength Scanning Speed

  • Wavelength slew rate: about 3200nm/min
  • Wavelength scan rate: about 900nm-160nm/min
  • Monitor scan rate: about 2500nm/min

Wavelength setting

  • At 1nm units for scan start and sea end wavelengths, and 0.1nm units for over wavelengths

Lamp Interchange wavelength

  • Auto switching synchronized with wavelength, switching range selectable between 282nm-393nm (0.1nm units)

Spectral bandwidth

  • 6-step switching along 0.1/0.2/0.5/1/2/5nm


  • Optimum response speed automatically set depending on bandwidth, minimum 0.1sec


  • 0.1nm

Stray light

  • Less than 0.0003% (220nm, Nal 10g/L solution)
  • Less than 0.0003% (340nm, UV-39 filter)

Photometric system

  • Double-beam, direct-ratio system with dynode feedback

Photometric modes

  • Absorbance(Abs.), transmittance(%), reflectance(%), energy(E)

Photometric range

  • Absorbance: -4 to 5 Abs
  • Transmittance, reflectance: 0.0 to 999.9%

Photometric accuracy

  • ±0.002 Abs (0-0.5 Abs)
  • ±0.004 Abs (0.5-1.0 Abs)
  • ±0.3%T (0-100% T) filter
  • Tested with NIST 903D standard

Photometric repeatability

  • ±0.001 Abs (0-0.5 Abs)
  • ±0.002 Abs (0.5-1.0 Abs)
  • ±0.1%T

Baseline flatness

  • ±0.001 Abs (excluding noise, using 2nm slit, and slow wavelength scanning speed)

Baseline correction

  • Auto correction using PC (stored baseline is automatically loaded when power is switched on, re-correction is possible)


  • 0.0004Abs/h (after power is on for 2 hours)

Temperature and humidity requirements

  • 15-35°C, 45-80%(no condensation, less than 70% above 30°C)

Light source

  • 50W halogen lamp (2,000 hours life), deuterium lamp (socket type), light source auto position adjustment built in.


  • Grating/Grating type double monochromator
  • Pre-monochromator: double-blazed holographic grating
  • Main monochromator: high-performance blazed holographic grating in aberration-corrected Czerny-Turner mounting


  • R928 Red-Sensitive Photomultiplier

Sample compartment

  • Internal dimensions: 150W x 260D x 120H (mm)
  • Distance between light beams: 100nm
  • Maximum light path length of cell: 100nm

"Power requirements"

  • AC100, 120, 220, 240V, switch selectable
  • 50/60Hz; 250 VA


  • 570W x 660D x 275H (mm)


  • About 36kg


Useful links