1. Manuals
  2. Brands
  3. biochrom Manuals
  4. Measuring Instruments
  5. Ultrospec 3300 pro
  6. Manual

biochrom Ultrospec 3300 pro Manual

Uv/visible spectrophotometer
Hide thumbs
Table of Contents

Advertisement

Quick Links

This is to certify that the
manufactured by Biochrom Ltd. conforms to the requirements of the following
Directives-:
Standards to which conformity is declared
EN 61 010-1: 1993 Safety requirements for electrical equipment for
measurement, control and laboratory use.
EN 61326-2.3: 1998 Electromagnetic compatibility - Generic emission standard
part 1. Electrical equipment for measurement, control and laboratory use.
EN 61000-4-6: 1992 Electromagnetic compatibility - Generic immunity standard
part 1. Residential, commercial and light industry.
Signed:
David Parr
Managing Director
Biochrom Ltd
Postal address
Biochrom Ltd
22 Cambridge Science Park
Milton Road
Cambridge CB4 0FJ
England
Registered in England No: 974213
Registered Office: 22 Cambridge Science Park, Milton Road, Cambridge CB4 4FJ, England
Declaration of Conformity
Ultrospec 3300 pro UV/Visible Spectrophotometer
Part number 80-2112-33 / 34 / 35 / 36
Serial number 81000 onwards
73/23/EEC & 89/336/EEC
Telephone
+44 1223 423723
e mail: enquiries@biochrom.co.uk
nd
Dated: 2
July 2001
Telefax
+44 1223 420164
website: http://www.biochrom.co.uk
.
Biochrom Ltd
Certificate No. 890333

Advertisement

Table of Contents
loading
Need help?

Need help?

Do you have a question about the Ultrospec 3300 pro and is the answer not in the manual?

Questions and answers

Related Manuals for biochrom Ultrospec 3300 pro

Summary of Contents for biochrom Ultrospec 3300 pro

  • Page 1 This is to certify that the Ultrospec 3300 pro UV/Visible Spectrophotometer Part number 80-2112-33 / 34 / 35 / 36 Serial number 81000 onwards manufactured by Biochrom Ltd. conforms to the requirements of the following Directives-: 73/23/EEC & 89/336/EEC Standards to which conformity is declared EN 61 010-1: 1993 Safety requirements for electrical equipment for measurement, control and laboratory use.

  • Page 2: Table Of Contents

    CONTENTS UNPACKING, POSITIONING AND INSTALLATION Essential Safety Notes OPERATION Introduction Keypad and display Basic Modes Methods Nucleic Protein Wavescan Multiwave Kinetics Standard Curve Substrate Instrument Utilities Output to Printer Download to Spreadsheet Messages ACCESSORIES Multiple Cell Holder Accessories Single Cell Holder Accessories Other Accessories, consumables etc SWIFT II Applications Software MAINTENANCE...

  • Page 3: Unpacking, Positioning And Installation

    Unpacking, Positioning and Installation • Inspect the instrument for any signs of damage caused in transit. If any damage is discovered, inform your supplier immediately. • Ensure your proposed installation site conforms to the environmental conditions for safe operation: Indoor use only Temperature 10°C to 40°C Maximum relative humidity of 80 % up to 31°C decreasing linearly to 50 % at 40°C...

  • Page 4: Essential Safety Notes

    Essential Safety Notes There are a number of warning labels and symbols on your instrument. These are there to inform you where potential danger exists or particular caution is required. Before commencing installation, please take time to familiarise yourself with these symbols and their meaning.

  • Page 5: Operation

    OPERATION Introduction Your UV/Visible spectrophotometer is a stand alone, simple-to-use instrument with a high-resolution liquid crystal display (LCD), and a comprehensive range of spectrophotometry measurements can be undertaken. It fulfils the requirements of the Pharmacopoeia (Appendix). It works on the basis of light from the deuterium and tungsten lamps being directed by a fixed mirror through the monochromator inlet slit.

  • Page 6: Keypad And Display

    Keypad and display Navigation around the displays, presented in an index card format, and the options therein is by using the 3456 keys; press enter or 6 to select an option. Number, letter and base entry will become active when appropriate. •...
  • Page 7 There are two standard formats of display, depending on the measurement mode: a) Basic (Absorbance, %Transmission, Concentration), Nucleic (DNA, RNA, Oligo) and Protein (UV methods) have a simple layout, with information presented in a box on the LCD. b) Other measurement modes have a graphical layout with information about the instrument also being provided along a status bar which shows messages, whether a printer is connected or not, time, accessory fitted (cell number for multi-cell holder) and wavelength.

  • Page 8: Basic Modes

    Basic Modes In the basic modes it is possible to change cell position by pressing the required number on the keypad. Absorbance Absorbance mode measures the amount of light that has passed through a sample relative to a blank (this can be air). The procedure is as follows: •...
  • Page 9 Concentration There are two concentration modes, Factor and Standard. Factor Concentration mode is used when a conversion factor is known; this is required to convert the absorbance measurement for a sample at a specific wavelength to a concentration, by a simple multiplication of absorbance * factor. Standard Concentration mode is used when a sample of known concentration is available;...

  • Page 10: Methods

    Methods Recall To recall a stored method from memory Select whether it is between 1-10, 11-20, etc by using 4 Select the method number The appropriate mode is obtained; load reference and samples, and press run Clear To clear a stored method from memory Press C Select whether it is between 1-10, 11-20, etc by using 4 Select the method number...

  • Page 11: Nucleic

    Nucleic Mode Factor A260/A280 DNA quantification and purity checking 50 ng/µl RNA quantification and purity checking 40 ng/µl Oligo Sequence Oligonucleotide quantification and purity checking 33 ng/µl dependent cDNA Fluorescent cDNA and PCR probe quantification 50 ng/µl label for micro-arrays and in-situ hybridisation studies, respectively Scan Spectrum of samples, as well as quantification and...
  • Page 12 Background correction at a wavelength totally separate from the nucleic acid and protein peaks at 260 and 280 nm, respectively, is sometimes used to compensate for the effects of background absorbance. The wavelength used is 320 nm and it can allow for the effects of turbidity, high absorbance buffer solution and the use of reduced aperture cells.
  • Page 13 cDNA Measurement of the labelling efficiency of fluorescently labelled cDNA probes before 2-colour microarray hybridisation ensures that there is sufficient amount of each probe to give satisfactory hybridisation signals. This data also provides an opportunity to balance the relative intensities of each fluorescent dye by adjusting the concentration of each probe before hybridisation.
  • Page 14 After measurement, the scan will autoscale. Typically, the scan will show 2 peaks, at 260nm for cDNA and around that of the fluorophore selected (488, 550 and 650 nm for fluorescein, Cy3 and Cy5, respectively). Buffer carry over may obscure the peak at 260 nm, and Abs260/Abs230 ratio will be affected.
  • Page 15 Scan The integrity of a nucleic acid sample can be established by inspection of its spectrum over the range 200 – 350 nm. This mode is particularly useful for RNA and oligonucleotide samples. The procedure is as follows: • Start wavelength is fixed at 200nm. If using a UV transmitting disposable plastic cell such as UViMicro, strange optical effects between 200 and 220 nm may be observed.

  • Page 16: Protein

    Protein Determination of the amount of protein in solution can be conveniently determined using colorimetric methods in conjunction with a UV/Visible spectrophotometer. A number of methods are available, four of which are included as built in options on the instrument; note that the wavelengths used can vary with manufacturers’ kits. Generic UV methods and information relating to proteins and amino acids are also included.
  • Page 17 Information Information relating to proteins and amino acids. ________________________________________________________________ Issue 02 - 07/2001 English...
  • Page 18 Bradford, Lowry, Biuret, BCA These all follow the same format; wavelengths, numbers of standards and other parameters can be edited, including concentrations, which can vary from one manufacturer’s kit to another. The protocols can therefore be modified to suit your needs;...
  • Page 19 Running Standards • Insert reference and standards, and press run • A reference is always required in position 1, and is assumed to be zero absorbance and zero concentration • To include a zero concentration standard, include this in the number of standards to be entered and enter 0.00 for concentration;...
  • Page 20 UV methods Protein Impurity The presence of nucleic acid in a protein solution can have a significant effect due to strong nucleotide absorbance at 280 nm. To compensate for this by measuring Abs 260, the equation of Christian and Warburg for the protein crystalline yeast enolase (Biochemische Zeitung 310, 384 (1941)) can be applied: Protein (mg/ml) = 1.55 * Abs 280 –...
  • Page 21 280, 205 nm method Protein concentration (mg/ml) can be determined using the empirical formula below (Scopes, R.K. (1974) Measurement of protein by spectrometry at 205 nm, Anal. Biochem. 59, 277 – 282): Protein (mg/ml) = 27 + (120 * Abs280 / Abs205) The procedure is as follows: •...

  • Page 22: Wavescan

    Wavescan A graph of change of absorbance against wavelength is known as an absorption spectrum and is one of the most useful physical characteristics of a compound, both as means of identification (qualitative analysis) and of estimation (quantitative analysis). It arises because of the various electronic transitions that are possible within a molecule, and the peaks are broad (in solution).
  • Page 23 Graph Use this facility to scale data post run for presentation on the display and print out. • Enter the start wavelength • Enter the finish wavelength • Enter if auto-scaling of the absorbance axis is required using 4 • Enter absorbance maximum •...

  • Page 24: Multiwave

    Multiwave The measurement of Absorbance values at specific wavelengths and combining these with appropriate factors is a means of overcoming interference effects in several applications. Abs Ratio This facility enables the determination of Abs λ1 / Abs λ2 and Abs λ1*factor •...
  • Page 25 3 point net This facility enables the determination of true peak height for turbid samples that have a sloping baseline, for example the determination of bilirubin in amniotic fluid. • Enter the first wavelength; this is wavelength on the “UV” side of the peak •...
  • Page 26 MultiWave This facility enables the entry of one or two equations so that post measurement calculations can be done automatically and the end result displayed. This is a very useful for the busy industrial, QC or environmental testing laboratory as the method can be saved.

  • Page 27: Kinetics

    Kinetics A graph of change of absorbance with time is known as a kinetics assay, and gives information about the rate at which a reaction proceeds; further information is given in the Appendix. Note that this mode should be used to check sample stabilisation since this instrument does not have a continuous output source.
  • Page 28 Graph This facility enables scaling of the results, and defines how they are to be presented on the LCD and print out. • Enter the maximum absorbance to be shown on the display during the assay • Enter the minimum absorbance to be shown on the display during the assay •...

  • Page 29: Standard Curve

    Standard Curve The construction of a multi point calibration curve from standards of known concentration in order to quantify unknown samples is a fundamental use of a spectrophotometer. Examples include protein determination using the methods determined previously and the analysis of waste water for metal complexes, salts and disinfectants.
  • Page 30 Running Standards • Insert reference and standards, and press run • A reference is always required in position 1, and is assumed to be zero absorbance and zero concentration • To include a zero concentration standard, include this in the number of standards to be entered and enter 0.00 for concentration;...

  • Page 31: Substrate

    Substrate Reagent test kits are routinely used for the enzymatic determination of compounds in food, beverage and clinical laboratories; an example is measurement of NAD / NADH conversion at 340 nm. The change in absorbance over a specified time period can be used to provide useful information when an appropriate factor is applied;...
  • Page 32 Running Standards • Insert reference and standards, and press run • A reference is always required in position 1, and is assumed to be zero absorbance and zero concentration • To include a zero concentration standard, include this in the number of standards to be entered and enter 0.00 for concentration;...

  • Page 33: Instrument Utilities

    Instrument Utilities Accessory • This identifies the type of cell holder / cell changer that has been fitted • A multi-position cell changer can be made to act as a single cell holder if required • Refer to Accessories section for further information Printer •...
  • Page 34 Set up User This facility enables user parameters for the instrument to be configured. • Enter operator name as described previously • Enter laboratory name as described previously • Enter instrument asset number or preferred description as described previously • Select if sound is required for keypad presses, sipper use and time intervals during kinetics reactions •...

  • Page 35: Output To Printer

    Output to Printer The graphics capability of the instrument means that the following requirements for printer compatibility should be fulfilled: • The printer must not be USB only style; parallel Centronics is required • The printer must not be designed to work with MS Windows only (GDI type); these are less expensive printers and can only function when connected to a PC with the appropriate driver installed If in doubt, check with the printer manufacturer.

  • Page 36: Download To Spreadsheet

    Parameters and graphics are printed automatically if Auto Print is selected (see User). Pressing print from within a mode set up page will print the parameters for that mode, whereas pressing print after experimental results are displayed will print both parameters and graphics.
  • Page 37 Beam blocked Not enough light getting to detector; check light beam is not blocked by a cell > 3.0 Sample too concentrated or something blocking light path Accessory not initialised properly ________________________________________________________________ Issue 02 - 07/2001 English...

  • Page 38: Accessories

    ACCESSORIES If an accessory is changed, press Function > Accessory to initialise the instrument in order that the appropriate accessory can be identified. Depending on the accessory type, a list of options is presented. Multiple Cell Holder Accessories • Install by removing accessory in place, replacing with the new one, turning the central mounting screw until it is finger tight and initialising as above.

  • Page 39: Single Cell Holder Accessories

    Single Cell Holder Accessories • Install by removing accessory in place, replacing, if necessary, the baseplate plug supplied and positioning the single cell holder so that the arrow is on the front face and it locates in place. Then push the finger locks backwards so that they lock into position.

  • Page 40: Other Accessories, Consumables Etc

    Other Accessories, consumables etc Description Part number Comments Sipper 80-2112-15 Use if a large number of samples for single readings are required. Requires single cell holder (80-2106-05 or 80-2106-13). 10mm flowcell and tubing supplied, together with separate user instructions. Temperature Control 80-2105-49 Required to supply the extra power Unit...

  • Page 41: Swift Ii Applications Software

    SWIFT II Applications Software SWIFT II comprises application modules for wavelength scanning, reaction kinetics, quantification, multi wavelength, time drive and fraction analysis, and can be used to enhance the software already included on the spectrophotometer. There are two application packages available; SWIFT II - LAB and SWIFT II - METHOD. 80-2108-26 SWIFT II - LAB - for general analytical purposes, Wavelength Scanning, Reaction Kinetics, Quantification, Time...

  • Page 42: Maintenance

    MAINTENANCE After Sales Support We supply support agreements that help you to fulfil the demands of regulatory guidelines concerning GLP/GMT. • Calibration, certification using filters traceable to international standards • Certificated engineers and calibrated test equipment • Approved to ISO 9001 standard Choice of agreement apart from break down coverage can include •...

  • Page 43: Lamp Replacement

    Lamp Replacement Replacement lamps are available from your supplier using the following part numbers: Deuterium lamp 80-2106-17 (includes tungsten lamp as well) Tungsten lamp 80-2106-16 The deuterium lamp is supplied fitted into a mounting and pre-adjustment plate; a new tungsten lamp is also included. NOTE: •...
  • Page 44 To change a lamp, proceed as follows: Switch off the instrument, remove sample from cell holder and disconnect the power supply cord. Allow time for lamps to cool. Locate the lamp access cover on the left-hand rear side of the instrument, remove the screw and push the cover back using the insert at its front.

  • Page 45: Deuterium Lamp Warranty

    Deuterium Lamp Warranty Criteria for lamp replacement are that it must: be less than 15 months old AND have had less that 750 hours use. Fuse Replacement Switch off the instrument and disconnect the power supply cord. The fuse holder can only be opened if the power supply plug has been removed, and is located in the power input socket on the back panel of the instrument.

  • Page 46: Cleaning And General Care

    Cleaning and General Care External cleaning • Switch off the instrument and disconnect the power cord. • Use a soft damp cloth. • Clean all external surfaces • A mild liquid detergent may be used to remove stubborn marks. • Sample compartment spillages •...

  • Page 47: Appendix

    APPENDIX Pharmacopoeia In general, there has been an increase in laboratory requirements to conform with Good Laboratory Practice techniques; this is particularly the case in Pharmaceutical companies and in Biotechnology facilities, where the interest in finding solutions to gene therapy opportunities is great. Typically, scientists working in pharmaceutical and bio-pharmaceutical research, be it University or Industry, require a high specification instrument with the ability to develop methods.

  • Page 48: Good Laboratory Practice

    Good Laboratory Practice Good laboratory Practice (GLP) concerns being able to trace experimental results to an instrument, an operator and the time the result was obtained so that a laboratory can prove that the instrument was functioning correctly or not. Laboratory, operator and internal instrument reference names can be entered on the spectrophotometer.
  • Page 49 GLP print out at instrument calibration (GLP enabled) Ultrospec 3300pro UV/Vis Spectrophotometer Lab name ....Instrument ....Serial no : 81012 Software : 4197 V1.4, Slave 4194 V1.0 Last serviced : 14/08/00 13:36 Instrument state at calibration GLP Calibrated 09/10/00 at 12:56 Calibration Full UV/Visible Bandwidth (1.3- 1.8nm): 1.7nm...
  • Page 50 baseline stored: 14/08/00 13:38 GLP enabled causes printed results to have calibration status included. GLP results can also be sent to PC for electronic archiving (Spreadsheet Interface Software required). ________________________________________________________________ English Issue 02 - 07/2001...

  • Page 51: Equation Entry Using Multiwave

    Equation Entry using MultiWave Always write out the equation(s) in front of you before using this mode. Step by step entry of the following equations is shown in the example below: Cobalt (g/l) = ((12.26 * A511) - (0.30 * A720)) * 100 Nickel (g/l) = (( -0.40 * A511) - (27.41 * A720)) * 100 Enter numbers using the keypad;...
  • Page 52 title) < > litre (unit will appear in print out, not in title) ü Enable > Check the equation!! equation ________________________________________________________________ English Issue 02 - 07/2001...
  • Page 53 Equation 2 If there is no equation 2, go directly to save method. Press enter or V after each parameter entry; use C to remove an incorrect parameter entry. Description Nickel > alphanumeric keypad; press stop after entry of name Equation press <...

  • Page 54: Kinetics

    Kinetics The usual way of measuring the rate of an enzyme reaction is to monitor the change in concentration of one of the substrates involved in, or of one of the products produced by, the reaction. Take for example the alanine transaminase (ALT) enzyme reaction: - α–oxoglutamate + alanine pyruvate + glutamate.
  • Page 55 where dC/dt = rate of change in concentration (mol/litre), L = cell path length (normally 1 cm) and E = molar absorptivity (molar extinction coefficient) of the compound being measured (for NADH, E = 6300 litres/mol/cm). ________________________________________________________________ Issue 02 - 07/2001 English...
  • Page 56 The rate of change of concentration can then be used to calculate enzyme activity, which is defined as: Enzyme activity = (dC/dt) x (Vt/Vs) where Vt = total volume of the reaction mixture and Vs = volume of sample. There are two internationally accepted units for enzyme activity: International Unit of Enzyme Activity, U or IU, defined as that amount of enzyme activity which will convert 1 micromole of substrate per minute at 25 o C.
  • Page 57 working in the IU unit, but to work in microkatals the conversion factor must be divided by 60. ________________________________________________________________ Issue 02 - 07/2001 English...

  • Page 58: Least Squares Regression Analysis And Linearity

    To calculate the enzyme activity, multiply the rate of change in absorbance by the conversion factor: Enzyme activity (IU/litre) = dA/dt x Factor. Least squares regression analysis and linearity The slope (or best straight line) and intercept in a kinetics assay or standard curve determination is calculated from a least squares linear regression of the data.

  • Page 59: Specification And Warranty

    SPECIFICATION AND WARRANTY Wavelength range 190 -1100nm in 0.1nm data intervals Monochromator 1200 lines/mm Aberration corrected concave grating Maximum scanning speed 7300 nm/minute at 2 nm intervals Spectral bandwidth < 1.8nm Wavelength accuracy ± 0.7nm Wavelength reproducibility ± 0.2nm Light source Tungsten halogen and deuterium lamps Detectors silicon photodiode...
  • Page 60 They can accept no liability for loss or damage, however caused, arising from the faulty or incorrect use of this product. This product has been designed and manufactured by Biochrom Ltd, 22 Cambridge Science Park, Milton Road, Cambridge CB4 0FJ, UK.

Table of Contents

Save PDF