Firma:              Pharmacia

Modell:            GPC

Kommentar:   Dokumente engl.

Uvicord SII

Introduction

Pharmacia LKB Uvicord SII Code No. 18-1004-50 is a compact single beam UV-monitor with flow-through cell for monitoring the eluant from various column separations. Available wavelengths, selected by a combination of lamp and interference filter are 206, 226, 254, 280, 313, 365 and 405 nm.

Pharmacia LKB Uvicord SII is housed up in one combined optical and electronic unit. This detector simultaneously produces two absorbance outputs - one fixed at 2 AUFS and one selectable from 0.005 - 2 AUFS in 9 steps. Thus, with the selection of the appropriate absorbance range, Uvicord SII enables monitoring of both high and low concentration substances simultaneously with a dual-channel recorder.

Substances such as non-aromatic peptides, amino acids, lipids, nucleotides and certain carbohydrates have strong absorption at 206 nm whereas at higher wavelengths they show little or no absorption. Furthermore, most proteins also exhibit greater absorption at 206 nm than they do at 280 nm. By employing all fully integrated electronics and the very latest detection principles Uvicord SII performs to specification at ambient temperature and under cold room conditions.

Basic principle

Light from the high frequency excited gas discharge lamp is filtered by the interference filter before passing through the measuring cell. The light transmitted through the measuring cell is focused onto the measuring detector and converted to an electrical signal. This signal is directly proportional to the light intensity and is amplified by-the following linear amplifier, the gain of which is controlled by the "zero" adjust potentiometer.

Filtered light also falls on the Reference Detector. The reference signal is then amplified in the reference amplifier. This signal is directly proportional to the light intensity. It is used to control the lamp driver for obtaining constant and stable light from the lamp. The reference signal is also connected to the Logarithmic Converter. The Log-Converter receives both the reference signal and the measured signal. The ratio between these two signals is transformed by the converter and conveyed to the two output amplifiers. One of the Output Amplifiers has preset fixed gain which produces 1 volt and 100 mV outputs corresponding to 2 Absorbance Units. The other output amplifier has its gain controlled by the range switch. The full scale output signal from this amplifier is 100 mV which corresponds to a full scale absorbance of 0.005 to 2 AU, depending upon the range switch setting. This output signal is also fed through the output filter which time constant is selectable by a front panel switch.

A zero indicator connected to the output signal is used to adjust the baseline to a reading dose to the zero volt. The final zero adjustment is then carried out on the recorder.

In order to ensure stable emission and Jong lamp life the lamp holder is thermostated. The thermostat is set to switch on heating when the lamp temperature goes below ambient temperature (>22°C).

Technical Specifications

Operating wavelengths           206, 226, 254, 280, 313, 365 and 405 nm.

Absorbance ranges                 0.005, 0.01, 0.02, 0.05, 0.1, 0.2, 0.5, 1 and 2 AUFS.

Noise                                       <1x10 ^-4 AU measured peak to peak in a dry cell.

Drift                                          <5x10 ^-4 AU/h at constant temperature.

Operating temperature            0 to +30°C.

Recorder outputs                    100 mV for selected absorbance range and 100 mV for fixed 2A   range.

Integrator output                      1 V = 2 AU.

Power consumption                20 W.

Mains Voltage                          100/115/120/220/240V ±10%, 50/60 Hz.

Dimensions (LxWxH)              313 x 131 x 147 mm.

Weight                                     4.3 kg.

SuperFrac ^TM

Introduction

Fraction Collector SuperFracTM, Code No. 18-1010-19, is a sophisticated, high-capacity preparative instrument enabling a wide variety of users to collect fractions or peaks according to their exact requirements.

To simplify operation an integrated microprocessor helps the user through the basic programming sequence. Optimal functions and parameters can be programmed independently.

SuperFrac fraction collector offers true peak-slope collection, time window collection, and many other advanced features.

Peak-slope collection and time window collection are especially useful for repetitive preparative runs in lab scale, and pilot scale, where the ability of SuperFrac to accept many very large tubes is indispensable.

Numerous combinations of collection modes make SuperFrac the most flexible fraction collector available. When remotely controlled from the LCC-501 Plus, GP-250 Plus or LC Controller 2252, SuperFrac will perform repetitive collection in loops. This feature is useful for large-scale, FPLC® and HPLC preparative runs. SuperFrac will store all parameters for nine complete programs.

The SuperFrac is also designed to work with other equipment from Pharmacia Biotech such as peristaltic pumps, UV-monitors and recorders to form integrated chromatographic systems adapted to your requirements.

Technical Specifications

Tube capacity                     Rack A: 312 tubes, 12-13 mm

                                           Rack B: 180 tubes or scintillation vials, 16-18 mm

                                           Rack C: 84 scintillation vials, 28 mm Rack D: 120 scintillation vials,

                                           16-18 mm

                                           Rack E: 60 scintillation vials, 28 mm

Collection modes

Time collection                   1 s to 99 min 59 s in 1 s intervals or

                                           1 min to 99 h 59 min in 1 min intervals

Volume collection               0.01 ml to 10 l per fraction

Drop collection                   1 to 9 999 drops in 1 drop intervals

Max drop rate                      12 drops/second

Step delay                           1 s to 99 min 59 s (time mode)

                                           1-9 999 drops (drop mode)

                                           0.01 ml to 10 l (volume mode)

Tube change time             Rack A: 250 ms

                                           Rack B: 400 ms

                                           Rack C: 600 ms

                                           Rack D: 440 ms

                                           Rack E: 660 ms

                                           Rack F: 20 ms/mm. Movement in x:y

                                           direction

Wait time                            1 s to 99 h 59 s

Automatic shut-down        Auxiliary equipment can be shut-down after preset number of tubes, after end time, or after end of equilibration time

Event mark                         Open collector output for 1 s ± 10 %

Time windows                   Nine, each with own programmable Start and end time: range 1 to 99:59 hour:min or min:s.

Program memory               Stores nine complete programs

Memory back-up                 Li-battery

End time                             1 min to 99 h 59 min

Equilibration time                1 min to 99 h 59 min, activated by end time

Slope detection                   Defined by threshold (1 to 999 %o FSD) and peak width at half height (1 s to 99 min 59 s)

Asymmetry correction        Adjustable between 0.1 and 99.9, defined as one tenth of the peak height

Isocratic peak‑

broadening correction        Adjustable between 0.00 and 99.59 h/ min.

External control                 Communication with LCC-501 Plus, GP-250 Plus and LC Controller 2252: Run, Return, Pause, Feed, Start loop, New loop and Return loop.

Flow Wing Control              High volume flow meter input

Environment                       +4 to +40 °C, 20–95% relative humidity, 84-106 kPa (840-1060 mbar) atmospheric pressure

Main voltage                        100/115/1201220–230/240 V~

requirements                      50–60 Hz

Power consumption           Max 45 W when all auxiliary equipment is connected

Dimensions (WxDxH)        550x330x430 mm

Net weight                           15.5 kg

Gross weight                      23 kg

LC Controller 2252

Introduction

Using LC Controller 2252, a wide range of individual instruments can be coordinated within a system to completely automate complex chromatographic processes.

Up to ten gradient files or programs can be created and stored in the memory. Individual gradient files can be recalled at any time, and used to reproduce chromatographic runs precisely. Before repeating a run, any of the gradient files can be edited to optimise the experimental conditions. All stored files are retained in the memory which is protected by a lithium battery during mains interruption.

Sophisticated convex or concave elution gradients can easily be created by using one of the four pre-programmed files which are permanently held in the memory. These standard profiles may be used in total or in part, by choosing the complete gradient or a part of it and entering the flow rate. The completed program can then be stored in one of the ten programmable files.

Any program can be looped, enabling an identical chromatographic run to be automatically repeated after completion. The link file facility provides for up to twenty different files to be joined together to form a single complex gradient profile, or to be run consecutively as individual gradients.

Through the integral intelligence of LC Controller 2252, the ancillary equipment connected is monitored when the system is switched on. From the data gathered in this way, the LC Controller automatically configures itself to take full control of the complete system. As soon as a gradient file has been started, the LC Controller continuously monitors and controls each of the individual instruments connected. During the course of the chromatographic run, an audible warning signal will be given if any part of the system causes the pressure to fall outside the limits predetermined in the program.

Operating principles

LC Controller 2252 automates chromatographic processes by controlling individual instruments in a system. Both HPLC and Standard Chromatography systems can be controlled. Systems comprise instruments such as one or two pumps, autosampler, detector, fraction collector and recorder. Solenoid valves and motor valves can be controlled for procedures such as sample injection, column switching and solvent selection.

Technical Specifications

Methods of gradient formation:    A. Two-pump system using high­pressure mixing.

B. One-pump system using programmable mixing valve for low-pressure mixing.

Display:                                        Alphanumeric 16 digits for continuous display of elapsed time, flow rate, composition, auxiliary and valve status.

Available gradient programs:      10 complete, individual gradient programs can be entered in linear segments, by time, flow and % B at breakpoints. Four permanent, pre-programmed, curve-function gradient programs are available. A link file enables profiles to be run in a specified sequence.

Memory capacity:                         2 400 instructions for methods, of which 800 can be programmed by the user.

Scaling functions:                        For modification of time and/or solvent composition axes.

Memory protection:                      Lithium battery.

Maximum run time:                      99.59 hours/minutes.

Plot function:                                For recorder preview of gradient (1 volt = 100 % B)

                                                    Plot time: 30 s/file.

                                                    Gradient also plotted during run.

Printout                                         Status, file and end reports printed an AlphaPlot printer/plotter

Purge function:                             For fast solvent change in pumps.

Auxiliary valves:                            Two available, for switching of 24 V latched or unlatched valves. Further four available for 3 or 8 Position motor valves for automatic injection and column switching.

Auxiliary event functions:             Three available contact closures (ON/OFF).

Programmable parameters and 27 programmable parameters on

parameter display (F-functions):        command:

                                                    - Active file

                                                    - Min. pressure limit: 0-349 bar

-                                                    - Max. pressure Limit: 0-349 bar

                                                    - Compressibility compensation ON/OFF

-                                                    - Valve type: latched/unlatched

-                                                    - Programming modes: advanced/basic

                                                    - Time base: 0.1 min, min:sec, hr:min

                                                    - Operating mode: HPLC/LC

                                                    - Gradient valve 1

-                                                    - Autoinject

                                                    - Delayed initiation

                                                    - Autozero

-                                                    - Stop chart recorder at end of run

-                                                    - Plot delay time: 0-50 000 s

                                                    - Valve cycle volume: 1-1 000 µl

                                                    - Minimum valve time: 0.1-99.9 s

-                                                    - Maximum flow

                                                    - Auto-restart: NO/YES

                                                    - File report

-                                                    - Status report

-                                                    - Date and time

                                                    - Auto reporting

                                                    -Step to next breakpoint

                                                    - Store active file

- Control Net instruments from individual instrument key

   panel

-                                                    - Split function

                                                    - Toggle Pump A & B

External control:                           Autosampler, Pumps: HPLC Pump 2150,

                                                    HPLC Pump 2248,

                                                    HPLC Gradient Pump 2249

                                                    MicroPerpex peristaltic pump,

                                                    Detectors:

                                                    Uvicord SD,

                                                    Variable Wavelength Monitor,

                                                    Fluorescence Detector

                                                    Rapid Spectral Detector,

                                                    Fraction collectors:

                                                    HeliFrac,

                                                    SuperFrac,

                                                    Recording devices:

                                                    Recorder REC-101, REC-102,

                                                    AlphaPlot printer/plotter,

                                                    Integrator

Event marks:                                Indicate activation of auxiliary functions and programmed changes in flow.

System warnings:                       Triggering of pressure units. Error messages for incorrect entries.

Operating temperature:               4 - 40 °C

Power requirements:                   110, 115, 120, 220 or 240 V, 50/60 Hz

Dimensions (WxDxH):                 365 x 265 x 145 mm

Weight:                                         6.2 kg

Superose 12HR 10/30

Start to equilibrate at a low flow rate (0.2-0.5 ml/min) and keep the pressure below 2 MPa (20 bar, 300 psi) undl two column volumes (50 ml) have passed. After equilibration, we recommend a flow rate of 0.5-1 ml/min. However, if buffers and samples are rather viscous, you may need to choose a lower flow rate to keep the pressure below 2.4 MPa.

Note:    A precipitation of the sample may block the filter and cause it to comp­ress the gel. To avoid this, we advise you neuer to set the pressure limit control at more than 0.2 MPa above your actual operating pressure.

Gel properties

Superose is a cross-linked, agarose-based medium optimised for high performance gel filtration of biomolecules. The narrow particle size distribution of Superose enables high flow rates at low back­pressures.

Property                            Superose 12

Matrix

Exclusion limit                    ~2x108 M _r

(globular proteins)              (approx.)

Optimal separation range 1 000-3x10 ⁵ M _r

(globular proteins)

Matrix composition             Composite of cross-linked agarose

Bead diameter (µm)           8-12

Prepacked column

Bed dimensions (mm)       10 x 300-310

Bed volume (ml)                 24 (approx.)

Max bed pressure               3 MPa

                                           (30 bar, 430 psi)

Operation bed pressure#   2.4 MPa

                                           (24 bar, 350 psi)

Max flow rate (ml/min)#      1.5

Recommended flow rate

(ml/min)#                            0.5-1.0

Column efficiency (H ^-1 )      > 40 000

pH stability

long term                            3-12

short term                           1-14

Temperatur                         4-40 °C

# At room temperature in aquos buffer

Hydrophobic interactions: Some hydrophobic interactions have been recognized, i.e. some com­pounds may be eluted somewhat later than predic­ted. These interactions can be of considerable value to the resolution.

Chemical and physical stability

The columns with Superose are resistant to all solutions commonly used in gel filtration, inclu­ding 8 M urea, 6 M guanidine HCl and 30% acetonitrile in the temperature range 4-40 °C. With aqueous Salt solutions, Superose is stable in the pH range 3-12 during operativ and pH range 1-14 for cleaning procedure. All detergents, nonionic or ionic, such as SDS, may be used. Limited degradation of the polysaccharide chains may occur under oxidizing conditions. If the column is used for Jong periods at pH 1, die gel matrix may be subjected to limited hydrolysis.

The back-pressure should not exceed 1.2 MPa for Superose 6 or 2.4 MPa for Superose 12. The column materials are glass and plastics which are all biocompatible. This means that there is no risk for sample contamination by metal ions.

Eluent and sample preparation

Degas and filter all solutions through a 0.22 .tm sterile filter. Water should be of Milli-Q * or corres­ponding quality. Use HPLC grade solvents, salts, and buffers. Centrifuge (10 000x g for 10 min) or filter samples through a 0.22 µm filter. Be sure to select a solvent resistant filter if samples are dissol­ved in organic solvents. If your sample is of high viscosity, dilute it with die eluent.

Column equilibration

Before applying die sample, equilibrate the column with two column volumes (50 ml) of eluent buffer. Equilibration is not necessary between runs with the Same eluent.

Sample application and eluation

Make sure the sample is recently filtered or centri­fuged before applying it to the column. A prefilter between injector and column is not recommended unless automated injections are performed. A pre­filter reduces the resolution by approximately 10%. Flow rates above 0.5 ml/min are not recommended for best resolution.

FPLC Pumpe P-500

PRÄZISIONS-KOLBENPUMPE P-500

Einführung

Die Pumpe P-500 von Pharmacia ist eine äußerst präzise Laborpumpe für die Flüssigchromatographie oder andere Anwendungen, wo konstante Flußraten bei Drücken bis zu 40 bar erforderlich sind. Die P-500 arbeitet sehr genau und reproduzierbar, be­sonders bei niedrigen Flußraten im gesamten Druckbereich. Die hervorragende chemische Resistenz der Pumpe macht es möglich, mit korrosiven Flüssigkeiten, organischen Lösungsmitteln sowie wäßrigen Lösungen zu arbeiten. Die P-500 ist für den täglichen Gebrauch im Labor konstruiert. Die Flußrate wird einfach und exakt mit einem dreistelligen Schalter eingestellt. Durch den großen Flußratenbereich ist die Pumpe sowohl für analytische als auch für präparative Chromatographie geeignet. Ein einzig-artiges Waschprogramm ermöglicht schnellen und sicheren Lö­sungsmittelwechsel, und eine Drucklimitkontrolle schützt die Säule vor zu hohen Drücken. In Verbindung mit dem Gradienten­programmer GP-250 oder dem Liquid Chromatography Controller LCC-500 kann die P-500 zur Bildung von reproduzierbaren Gra­dienten eingesetzt werden. Die P-500 ist vollkommen kompatibel mit anderen Geräten von Pharmacia, z. B. den Fraktionssammlern FRAC-100 und FRAC-300 oder dem Autosampler ACT-100. Mit der Pumpe P-500 kann das gesamte Spektrum an Säulen und Gelen, das von Pharmacia Biotechnology angeboten wird, betrieben werden.

Funktionsweise

Die Pumpe wird durch einen Schrittmotor angetrieben. Die Flüs­sigkeit wird mit Hilfe zweier Kolben gefördert, die sich in je einem Glaszylinder hin und her bewegen. Die Bewegung der Kol­ben ist so eingerichtet, daß jeweils einer fördert, während der andere neue Flüssigkeit ansaugt. Die Kolben bestehen aus vier Teilen: dem Kopf (piston head), einem Dichtungsring (sealing ring), einer Lochscheibe (plate) und einer Kolben­stange (piston rod) (s. Abb. 2). Die Dichtung kann sehr ein-fach ausgewechselt oder nachgespannt werden.

Der Weg der Laufmittel durch die Kolben wird durch das 4-Wege-Ventil an der Frontseite der Pumpe gesteuert. Das Ven­til schaltet automatisch, wenn die Kolben ihre Endposition er-reichen und die Bewegungsrichtung ändern.

Bevor die geförderte Flüssigkeit die Pumpe verläßt, fließt sie durch einen Drucksensor. Dieser besteht aus einem Diaphragma, das bei Druckänderungen kontrahiert bzw. expandiert. Dabei wird in einer elektronischen Meßbrücke ein Widerstand ver-

ändert. Dadurch wird eine Spannung erzeugt, die dem Druck pro­portional ist. Diese Spannung wird verstärkt und zum Meßin­strument an der Vorderseite der Pumpe und zum Druckmess-Aus­gang (pressure output) auf der Rückseite geführt.

Der Drucksensor erfüllt zwei Aufgaben: einmal die Anzeige des aktuellen Druckes und zum andern Schutz von druckempfindlichen Trennsäulen.

Technische Daten

Flußraten                                 1-499 ml/h, in Schritten von 1 ml/h veränderbar

Druckbereich                           bis        200 ml/h           4.0 MPa           (40 bar),

                                               darüber 2.5 MPa (25 bar)

Reproduzierbarkeit                  besser als 0.5 % der eingestellten Flußrate

Flußkonstanz                         besser als 1 % über 8 h bei 160 ml/h mit gleichem Lösungsmittel bei kon­stanter Temperatur

Teile in Kontakt                        Kel-F(R), Tefzel(R), Lubriflon(R),

mit Lösungsmitteln                  Titan, Borsilikatglas

Ausgang für externe                0-5 MPa (0-50 bar); 0.5 MPa/mV

Druckanzeige

Drucklimit                               Pumpe schaltet bei vorgewähltem Maximaldruck (0-5 MPa bzw. 0-50 bar) automatisch ab

Netzanschluß                          200-240 V + 10 %

                                               50-60 Hz

Energieverbrauch                    18 W bei Normalbetrieb

Temperaturbereich                 0-40°C

Abmessungen                         410 x 217 x 165 mm

Gewicht                                   9 kg

Motor                                       Schrittmotor

Sicherheit                                entspricht IEC- und UL-Anforderungen