EQUIPOS AGILENT

Introduction to Your System

Agilent 8453 Spectrophotometer — Overview 10
General Purpose Agilent ChemStation Software for UV-visible
Spectroscopy — Overview 18

Operation of the system is much easier if you understand the implementation

models. The mind-models of data acquisition, data evaluation and data
handling will help you to run the system successfully.
Your spectroscopy system is based on an Agilent 8453 spectrophotometer and
general purpose Agilent ChemStation software for UV-visible spectroscopy
running on a PC with the supported Microsoft operating system(s). These two
components are linked together by a network connection. This type of link is
very flexible: it can be used for a direct connection between the
spectrophotometer and the PC as well as the integration in a enterprise
network with network access from the PC to the spectrophotometer.
The tasks are split between these devices such that the spectrophotometer
acquires and provides absorbance data which are handled by the PC’s
application software. All of the data display, evaluation and longterm storage
is done under software control on the PC.

This section gives an overview of the optical system and explains the
spectrophotometer front and back panels. It also explains the layout and
construction of the spectrophotometer including the electronic and
mechanical assemblies inside the spectrophotometer.
Optical System Overview
Optical System
The optical system of the spectrophotometer is shown in Figure 1. Its
radiation source is a combination of a deuterium-discharge lamp for the
ultraviolet (UV) wavelength range and a tungsten lamp for the visible and
short wave near-infrared (SWNIR) wavelength range. The image of the
filament of the tungsten lamp is focused on the discharge aperture of the
deuterium lamp by means of a special rear-access lamp design which allows
both light sources to be optically combined and share a common axis to the
source lens. The source lens forms a single, collimated beam of light. The beam
passes through the shutter/stray-light correction filter area then through the
sample to the spectrograph lens and slit. In the spectrograph light is dispersed
onto the diode array by a holographic grating. This allows simultaneous access
to all wavelength information. The result is a fundamental increase in the rate
at which spectra can be acquired.

Lamps
The light source for the UV wavelength range is a deuterium lamp with a
shine-through aperture. As a result of plasma discharge in a low pressure
deuterium gas, the lamp emits light over the 190 nm to approximately
800 nm wavelength range. The light source for the visible and SWNIR
wavelength range is a low-noise tungsten lamp. This lamp emits light over
the 370 nm to 1100 nm wavelength range.
• Source Lens
The source lens receives the light from both lamps and collimates it. The
collimated beam passes through the sample (if one is present) in the sample
compartment.
• Shutter
The shutter is electromechanically actuated. It opens and allows light to
pass through the sample for measurements. Between sample measurements
it closes to limit exposure of sample to light. If the measurement rate is very
fast, you can command the shutter to remain open (Agilent ChemStation
software) or it stays open automatically (handheld controller software).

• Stray-Light Correction Filter

In a standard measurement sequence, reference or sample intensity spectra
are measured without and then with the stray-light filter in the light beam.
Without the filter the intensity spectrum over the whole wavelength range
from 190–1100 nm is measured. The stray-light filter is a blocking filter
with 50 % blocking at 420 nm.
With this filter in place any light measured below 400 nm is stray light only.
This stray-light intensity is then subtracted from the first spectrum to give a
stray-light corrected spectrum. Depending on the software, you can switch
off the stray light correction (Agilent ChemStation software) in case you
want to do very fast repetitive scans or it is switched off automatically.
• Sample Compartment
The spectrophotometer has an open sample compartment for easier access
to sample cells. Because of the optical design a cover for the sample area is
not required. The spectrophotometer is supplied with a single-cell holder
already installed in the sample compartment. This can be replaced with the
Peltier temperature control accessory, the thermostattable cell holder, the
long path cell holder or the multicell transport. All of these optional cell
holders mount in the sample compartment using the same quick, simple
mounting system. An optical filter wheel is also available for use with the
spectrophotometer and most of the accessories.
• Spectrograph
The spectrograph housing material is ceramic to reduce thermal effects to a
minimum. It main components of the spectrograph are the lens, the slit, the
grating and the photo diode array with front-end electronics. The mean
sampling interval of the diode array is about 0.9 nm over the wavelength
range 190 nm to 1100 nm. The nominal spectral slit width is 1 nm.
• Spectrograph Lens
The spectrograph lens is the first of the parts which are collectively known
as the spectrograph. It is mounted on the housing of the spectrograph. The
spectrograph lens refocuses the collimated light beam after it has passed
through the sample.

• Slit
The slit is a narrow aperture in a plate located at the focus of the
spectrograph lens. It is exactly the size of one of the photo diodes in the
photo diode array. By limiting the size of the incoming light beam, the slit
makes sure that each band of wavelengths is projected onto only the
appropriate photodiode.
• Grating
The combination of dispersion and spectral imaging is accomplished by
using a concave holographic grating. The grating disperses the light onto
the diode array at an angle linear proportional to the wavelength.
• Diode Array
The photodiode array is the heart of the spectrograph. It is a series of 1024
individual photodiodes and control circuits etched onto a semiconductor
chip. With a wavelength range from 190 nm to 1100 nm the sampling
interval is about 0.9 nm

EQUIPOS PERKIN

Introduction
The Lambda 25, 35, 45 are versatile spectrometers operating in the ultraviolet
(UV) and visible (Vis) spectral ranges. The spectrometers feature a double-
beam, allreflecting system.
Lambda 25, 35, 45 spectrometers are usable in a wide range of applications as
indicated by their performance specification.
Features

Optical System of the Lambda 25, 35, 45

The Lambda 25, 35, 45 UV/Vis Spectrometers features an all-reflecting optical
system. The optical components are coated with silica for durability. A
holographic
grating is used in the monochromator.
The optical system is depicted schematically in Figure 6.
The monochromator is a holographic concave grating with 1053 lines/mm in
the center.
Two radiation sources, a deuterium lamp and a halogen lamp, cover the working
wavelength range of the spectrometer.
For operation in the visible (Vis) range, mirror M1 reflects the radiation from
the
halogen lamp onto source mirror M2. At the same time M1 blocks the radiation
from the deuterium lamp.
NOTE: In the Lambda 45, M2 is substituted by a pre-monochromator grating,
increasing
the straylight performance of the instrument.
For operation in the ultraviolet (UV) range, mirror M1 is raised to permit
radiation
from the deuterium lamp to strike source mirror M2.
Source change is automatic during monochromator slewing.
Radiation from the source lamp is reflected from source mirror M2 through an
optical filter on the filter wheel assembly.
A stepping motor drives the filter wheel to be in sychronization with the
monochromator.
Depending on the wavelength being produced, the appropriate optical filter is
located in the beam path to prefilter the radiation before it enters the
monochromator.
Filter change is automatic during monochromator slewing

From the optical filter the radiation passes through the entrance slit (Slit 1) of
the monochromator.

The radiation is dispersed at the grating to produce a spectrum. The rotational

position of the grating effectively selects a segment of the spectrum,
reflecting this segment through the exit slit (Slit 2) to mirror M3.
The exit slit restricts the spectrum segment to a near-monochromatic radiation
beam. The slits provide a spectral bandpass of 1 nm on the Lambda 25, and are
selectable to be 0.5, 1, 2, or 4 nm on the Lambda 35 or 45. From mirror M3 the
radiation is reflected onto a beam splitter which allows 50% of the radiation to
pass onto plane mirror M4, and reflects 50% of the radiation onto plane mirror
M5. Mirror M4 focuses the radiation beam in the sample cell. The beam then
passes through a convex lens onto the photodiode detector. Mirror M5 focuses
the radiation beam in the reference cell. The beam then passes through a
convex lens onto the photodiode detector.