C125-E008H

RF-5301PC

Shimadzu
Spectrofluorophotometer
 World’s Highest Level Performance and Windows Software TM

F ro m R o utin e A n a ly s is to Research Employing t he Highest Level o f

Se n s i t i v i ty in th e Wo rld !

Compared to adsorbance methods, fluorescence sensitivity is tens to thousands times better — this means
that you can analyze nanogram to picogram samples with great results.
Fluorescence can also be used to identify a specific molecule in a complex background.
When the compound of interest does not exhibit natural fluorescence, functional group-specific probes may
be used to label the compound and assist your research. The synchronous scanning mode allows mixtures of
fluorochromes to be analyzed.

A PC directly controls the instrument for data acquisition and processing.

The Windows friendly operating environment allows you to perform measurement, data processing,
TM

editing and recording in one continuous operation with a click of the mouse. Using the Copy Graph function,
measurement data or spectra may be easily transferred to word processing or spreadsheet software for
preparation of documents or additional calculations.

The essence of fluorescence analysis is sensitivity. The high-throughput optical system in the RF-5301PC
employs a blazed holographic grating, photomultiplier and digital circuit to provide the highest level S/N
ratio attainable.

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Open Up a New Spectrum of Fluorescence Analysis.

High-Speed Scanning Versatile Sample Vertical Optics with

High-speed scanning up to Compartment Size Dynode Feedback
5,500nm/min allows you to measure Vertical optics in the RF-5301PC
a spectrum in seconds. And since minimize light loss in measurement
monochromator slewing is about with an LC flow cell, a micro cell or
20,000nm/min, setting of two or a small test tube. This design
more wavelengths can be assures on exceptionally high
performed quickly and easily. signal-to-noise ratio and provides
the ability to attain excellent
High Resolution and analysis results using very small
Extended Range volumes of precious samples.
The bandpass on the RF-5301PC The dynode feedback enhances the
may be set as narrow as 1.5nm, RF-5301PC performance by raising
which makes is possible to The sample compartment or lowering negative high voltage to
distinguish fluorescent peaks from measuring 140mm wide, 170mm the detector in response to different
the background emission. deep and 140mm high, enables use wavelengths. The dynode feedback,
The wavelength range of 220 to of micro cells, high-sensitivity cells, which expands the dynamic range
750nm can be extended to 900nm LC flow cells, etc. for a wide range of the signal detecting system, is
with an optional R-928 of applications. significantly superior to ratio
photomultiplier. methods.
U n iq u e High
Built-In Performance Checks P e rfo rmance Feat ures in Automatic Shutter
th e R F- 5301PC Protects Sample
The wavelength search function An automatic shutter in the
allows the optimum excitation and RF-5301PC excitation path closes
emission wavelengths to be found immediately when measurement
in about two minutes. ceases, thereby protecting the
sample from photo-decomposition.

When the instrument is switched

ON, operating conditions of the
spectrophotometer are
automatically verified. Separately,
noise level (S/N ratio) and light
source (Xenon lamp) usage are
built-in features to help maintain the
instrument in its optimum condition,
providing absolute confidence in the
quality of the data.

3
 Now You Can Really Put Your Data
Se e t h e D iffe re n ce with Windows T M Performance

Working in the Windows TM

Many Convenient and Spectrum Measurement
environment makes operation Time-Saving Features Mode
intuitive. The software is driven by The RF-5301PC software is the Perform emission and excitation
pull-down menus displayed at the ideal software to meet all your scans with ease, or overlay the two
top of your screen that can be fluorescence research needs, from for interpretation. Obtain and
selected with just a click of the teaching to method development differentiate excitation and emission
mouse. For the beginner, on-line and quality assurance. Follow spectra using color and line pattern
HELP is available to guide you changes in intensity over time for assignments. Zoom in using the
through each function. You can kinetic assays or perform mouse or zoom out with the radar
customize the software with quantitative analysis on several function to autoscale all data on-
SpeedBox , an innovative tool that
TM
samples. You’ll save time and effort screen. The software picks and
eliminates keystrokes and enables with features that allow for on- tabulates peaks and valleys
you to assign icons for commonly screen data manipulation. automatically. And if you need a fast
used menu commands. With the spectal scan in any measurement
RF-5301PC software, storage, Quantitative Measurement mode, the PopUp Scan function
TM

retrieval and other file management Mode displays it on-screen in seconds.

is quick and simple. Our quantitative analysis uses 1 to
20 known fluorescence standards to
fit the best calibration curve to your
data. Methods such as DNA
quantification with Ethidium
Bromide or Hoechst dye, OPA-
derivatives of amino acids, cell
viability assays and FITC or
Rhodamine-labeled ligands are
handled with ease. The results are
instantly displayed on-screen,
allowing you to read and average Up to 10 spectra may be displayed
the relative fluorescence intensities together on the screen—each
of unknown samples. Optimize your discriminated by an individual color
analyses with first through third and line style. Then, arithmetic
order curve fits. If desired, the curve operations may be conducted
may be forced through zero. To among the spectra to uncover
facilitate your calculations, the hidden information or smooth a
calibration curve is automatically curve.
generated and displayed graphically
as well as in tabular form.

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In Its Place.

Time Course Measurement One-touch Operation The obtained spectrum may be

Mode with Speed Box TM
further analyzed using data
Working in Time Course allows you For convenience of operation, processing functions, such as for
to monitor experimental changes frequently used functions can be performing spectral correction and
and measure reaction rates such as assigned to an accelerator button. extraction of hidden information.
dehydrogenase or ß-galactosidase Any of these functions may be
activity while the software provides easily executed graphically using
mathematical support for rate the mouse.
calculations. Data can be collected
every 0.02 seconds for the most Repor t Generation
timely results.

Then these function can be simply

executed by clicking on the as-
signed button. Shape and position
of buttons may be freely customized
by the user.
Before you generate a report, you
Time Course data may be added, Simple, but Powerful for
Extensive Data Processing may choose to preview it on-screen.
subtracted, multiplied or divided by To store files for future use, simply
each other or a factor. High Operations such as first through
save them to a disk or convert them
sensitivity, a stable baseline and a fourth order derivative,
to an ASCII file for accessibility by
20 bit high-resolution A/D converter mathematical smoothing functions,
other software. Finally, when you’re
mean reliable detection of small log conversions and offsets are
ready to print, the software offers
changes in fluorescence, allowing easily performed with the
the option of sending reports to
signal discrimination on small RF-5301PC software. Display up to
either a dot matrix printer for rapid
concentrations. ten spectral curves simultaneously
output of tables or to a graphics
Activity calculations are also or use mathematical
printer/plotter for publishable quality
supported, making this transformations to maximize your
graphics and data.
measurement mode quite suitable results. Select the source and
for enzyme research. destination channels along with the
Using “Copy&Paste”, data may be
desired calculation and the results
pasted into other Windows
are redisplayed on the screen.
applications, such as Word and
Excel, to enable high-level report
generation.

5
 Ve rs a t i l e A p p lica tio n s Mak e t he RF- 5301PC
t h e Pe rfe c t Par tn e r fo r A ny Laborat or y

Pharmaceuticals and Pharmacology

The RF-5301PC is excellent for the analysis of crude drugs; quality
control of drugs; quality control of pharmaceuticals or studies on drug
metabolism. Typically, studies would encompass: barbituric acid,
tetracycline, reserpine, LSD, purine and pyrimidine analogs,
porphyrins, etc.

Biochemistry
The RF-5301PC is the instrument of choice for the detection and
measurement of a wide variety of biological samples, even at trace
quantity levels. Typical applications include: enzymes and coenzymes
such as cytochrome oxidase, protein kinase, derivatized lipids, retionids,
NADH, FAD, vitamins, nucleic acids, amino acids, chlorophyll,
neurotransmitters, cell membranes and enzyme kinetics.

Food Science
The RF-5301PC is especially suitable for the detection and measurement
of trace components in a wide variety of foods. Typical applications
include: amines and amino acids, fats, carbohydrates, coenzymes,
antibiotics, food additives, edible oils and pesticides.

Environmental Monitoring
A growing application of the RF-5301PC includes the monitoring,
analysis and study of water, environmental and food pollutants. Typical
studies include: polynuclear aromatics, phenols, oil contaminants and
ground water effluents.

Chemical Analysis
The RF-5301PC is the perfect alternative for the measurement of trace
components of organic and inorganic compounds, unmeasurable by
absorption spectrophotometry. Typical applications include: polymers,
texiles, plastics, rubber, optical materials research, polynuclear aromatic
compounds, petroleum products and kerosene.

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Spe ci fi cati on s

Light source 150W Xenon lamp. Ozone resolving type lamp housing.

Excitation and emission Concave, blazed holographic grating, F/2.5, 1300 grooves/mm.

monochromators
H a rdw a re Wavelength scale 220-990nm.
S pe c i f i c a t i o n s
Measuring wavelength range 220-750nm and 0 order as standard. 220-900nm with the optional R928 photomultiplier.

Spectral bandwith 6-step selection of 1.5, 3, 5, 10, 15 and 20nm.

(6nm bandwidth is available for half sample height on the excitation side only.)

Wavelength accuracy ±1.5nm.

Light source compensation Dynode feedback system with monochromatic light monitoring.

Sensitivity The S/N ratio is 150 or higher for the Raman line of distilled water

(350nm excitation wavelength, 5nm spectral bandwidth, and 2-second response for 98% of the full scale).

Wavelength scanning 7-step selection of Survey (about 5500nm/min), Super (about 3000nm/min)

Very Fast, Fast, Medium, Slow and Very Slow.

Wavelength slewing speed About 20,000nm/min.

Response 8-step selection of 0.02, 0.03, 0.1, 0.25, 0.5, 2, 4 and 8 seconds for 98% of the full scale.

Sensitivity selection 2 steps of HIGH and LOW. (The sensitivity at HIGH is about 50 times that of LOW.)

Interface RS-223C interface, interface for autosampler, and interface for sipper unit.

Dimensions and weight 667W x 530D x 270H mm; 43kg.

Power requirements 100, 120, 220, 240V; 50/60Hz; 400VA.

Operational temperature range 15-35°C

Operational humidity range 40-80% (Below 70% with temperature higher than 30°C).

Measurement Excitation, emission and synchronous spectrum measurement, time-course measurement,

quantitation, automatic search of optimal excitation and emission wavelengths, PopUp Scan . TM

Data processing Arithmetic calculation between spectrum and a constant, smoothing, 1st through
S oft w a re 4th derivatives, 1/Y, logarithmic conversion, data printout (with or without activity
S pe c i f i c a t i o n s value computation), peak pick, point pick, area calculation, averaging

(in quantitation), generation of calibration curves of 1st through 3rd order.

Filing Save, recall, and delete of data. Conversion into ASCll and DIF formats.

Data output Automatic scale adjustment, readout of data at user-specified point, data printout

(preview function provided), selection of colors and types of curves.

Maintenance Automatic monitoring of signal-to-noise ratio, monitoring of the run time of light source lamp.

User interface SpeedBox (assign icons for commonly used menu commands).
TM

Other functions Data exchange via clipboard, auto response control, automatic shutter.

PC requirements IBM-PC/AT or 100% compatible; i486 or higher CPU; 8 Mbyte or larger main

memory. Operates on MS-Windows version 3.1 or higher.

Windows is a registered trademark of Microsoft Corp. SpeedBox and PopUp Scan are registered trademarks of Shimadzu Scientific Instruments.

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 A W i d e Va rie ty o f Op tio n a l Accessories for t he RF- 5301PC

C on s t a n t - T e m p era tu re N o n - Fluorescent High Sensit ivity

F our C e l l H o ld e r C e ll Cell Holder
Cat. No. 206-24940-91 Cat. No. 200-34594-03 Cat. No. 204-26841-01

Keeps four sample cells at the Provides a 10mm square non- This cell holder enhances the
desired constant temperature via fluorescent cell, polished on four sensitivity two to three times or
connection to a water bath. sides. This cell is recommended for more. This is recommended for low
Note: Requires the Temperature trace analysis particularly when low concentrations. The excitation and
Controlled Water Bath. UV wavelengths are used for emission beams are repeatedly
excitation. Does not fluoresce when reflected through the sample to
excitation wavelengths from 240 to increase the excitation efficiency.
260nm are used.

Mi c r o C e l l H o ld e r Q u a rt z Cell Polished R- 928- 08

w i th M i c ro C ell o n F our Sides Phot omult iplier
Cat. No. 204-27125 Cat. No. 200-34441 Cat. No. 200-75021

Provides a 10mm square cell, polished

Permits measurement of samples on four sides. Recommended for Extends the measuring wavelength
as small as 400μL in volume. work in the visible range. range from 750 to 900nm.
The cell is installed in an adapter of This cell has a slight absorption at
the same dimensions as an ordinary 260nm and shows small
10mm square cell, so it can be fluorescence intensity at 400nm.
placed in the standard cell holder.

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So l i d ( Po w d e r) F lo w-Thru Cell Unit 8mm dia.
Sa m pl e H o l d e r fo r H PLC ( 12μ L) Test Tube Holder
Cat. No. 204-26836-01 Cat. No. 204-05566 Cat. No. 204-05853

Provides a means to examine the With this attachment, the RF- Holds a test tube, 8mm outer
fluorescence of a solid sample. The 5301PC is used as a highly diameter and 45mm~100mm long.
angle of the holder is designed to sensitive spectral fluorescence The minimum sample volume
limit the reflected excitation beam monitor in HPLC. required is 400μL.
from the emission monochromator. Free selection of the excitation and
Standard accessories include a emission wavelengths permits
sample holder, quartz plate, powder selective detection. The mobile 12mm dia.
sample plate, spacers and a sample phase stream can be stopped for Test Tube Holder
fixing bar. Cutoff filters are included spectral measurement, so that Cat. No. 204-03293
to reduce scattering further (UV-31, chromatographic peaks can be
UV-35, UV-39, UV-Y43, UV-Y45, qualitatively determined with higher
UV-Y47). reliability. The flow-thru cell is a
square type with an inner volume as
small as 12μL and is made of silica,
ensuring minimized light scattering.

F lo w Cell Unit ( 120μ L)

fo r L C Applicat ion Holds a test tube, 12mm outer
Cat. No. 204-06249 diameter and 60mm~100mm long.

The square micro-flow cell has an

inner volume of 120μL and is made
of silica to ensure minimum light
scattering. The inlet and outlet
tubes are both made of vinyl resin
and are 0.5mm I.D. and 2mm O.D.

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 A W i d e Va rie ty o f Op tio n a l Accessories for t he RF- 5301PC

B ott o m R i s e C o n st ant - Temperat ure Polarizer At t ac h m en t

S a m p l e St a g e S in gle Cell Holder ( f or UV/Vis regio n )
Cat. No. 204-04811 w ith St irrer Cat. No. 204-03290
Cat. No. 206-24930-91 (100V)
Cat. No. 206-24930-92 (120V)
Polarizer At t ac h m en t
Cat. No. 206-24930-93 (220/240V) ( f or Vis region)
Cat. No. 204-03290-01

Raises the sample cell position in

order to reduce the minimum
sample volume to 1.5mL.
Measurement of fluorescence
Keeps the sample at the desired polarization provides information on
constant temperature during the size, fluidity and environments
measurement by circulating of fluorescent molecules. Permits
temperature-controlled water reliable measurement of
(5~70°C) around the sample cell. polarization characteristics over a
The stirring option has variable wide wavelength range from 240nm
speed control and the minimum to 800nm.
sample volume is 2mL. Effective for
fluorometry of suspended cell
samples.
Note: Does not include the
Temperature Controlled Water Bath.
Not applicable to samples having a
specific gravity greatly different from
that of the solvent.
Note: The hose joint is applied to
the 6~9mmID tube.

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NT T-2 2 0 0 P C o nstan t- F ilter Set ASC- 5 Aut o Sam p le
Te m p e ra t u re W ater Cat. No. 204-04691 Changer
Ci r c ul a t o r Cat. No. 206-23810-xx
Includes 8 filters: two broad-band
Cat. No. 208-97263
interference filters U-340 and
B-390 for isolation of excitation
wavelengths of 300-380 and 350-
450nm, respectively. Six cutoff
filters: UV-29, L-41, Y-46, Y-50,
O-56 and R-60 with approximately
50%T at 300, 420, 460, 500, 560
and 600nm, respectively.

S p e c tr a Correct ion Builds an automatic multi sample

A c c e s sory spectrophotometry system in
Cat. No. 204-07550 combination with a sipper unit.
Circulates temperature-controlled Up to 100 test tubes are set
water to a constant-temperature cell Used to correct excitation and together on the rack. The aspirating
holder. emission spectra in the wavelength nozzle is programmable to move X,
-Temperature range: Ambient + 5°C to + 80°C region from 250nm to 600nm, using Y and Z (vertical) direction.
-Temperature control precision: ±0.05°C~ rhodamine solution as the reference
-Max. pumping rate: 27/31 L/min, (quantum counter).
9.5/13m (50/60Hz) Sipper Unit 5300
-External circulation nozzle: 10.5mm Cat. No. 206-24950-91
R e c tangular Flow Cell,
OD (both outlet and return)
12 0 μL
-Tank capacity: About 10L (9L during use)
Cat. No. 204-03285-04
-Safety features: Detection of over-
temperature of Upper or Lower Used for fluorescence monitoring of
limits, Detection of heater wire effluents in column chromatography.
malfunction, Protection of heating Made of fused silica. Requires
too little circulating water, adapter (Cat. No. 206-83934-91).
Detection of sensor malfunction,
Independent over heat protection,
Over current circuit protector
-Standard accessories: Lid with Aspirates solution samples using a
handles, Rubber hose (4m, 9mm peristaltic pump driven by a
ID), Hose clamps (4pc.), Instruction stepping motor, to be measured
manual successively. Minimum sample
-Dimensions: 270W x 560H x 400D (mm) volume is 2mL, linked operation
-Power requirements: 100VAC, with the ASC-5 Auto Sampler
1250VA, with 1.7m power cord and possible.
grounded plug
Note: Piping to Constant-
Temperature Cell Holder (Cat. No.
206-24930-xx) is possible by the
hose (9mm ID).

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 JQA-0376

Founded in 1875, Shimadzu Corporation, a leader in the

development of advanced technologies, has a distinguished
history of innovation built on the foundation of contributing to
society through science and technology. We maintain a
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our Web site at www.shimadzu.com

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Phone: 81(3)3219-5641 Fax. 81(3)3219-5710
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The contents of this brochure are subject to change without notice.

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