4006073D.
4006073D.
4006073D.qxp 8/17/2009 7:45 AM Page i Table of Contents Section 1 Introduction............................................................1 Section 2 Starter Kit Components ........................................1 Section 3 Buffer and Sample Preparation ............................2 Section 4 System Setup ........................................................3 Section 5 Connecting a Chart Recorder ..............................
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4006073D.qxp 8/17/2009 7:45 AM Page 1 Section 1 Introduction This manual guides you through the installation of your new BioLogic LP chromatography system. You will perform an actual chromatographic separation, demonstrating the simplicity and excellent performance of the BioLogic LP. Section 2 Starter Kit Components This starter kit contains: • One Bio-Scale™ Mini Macro-Prep® High Q anion exchange cartridge, 1 ml • Buffer A, 10x concentrate, 50 ml (when diluted: 25 mM Tris-HCl, pH 8.
4006073D.qxp 8/17/2009 7:45 AM Page 2 Section 3 Buffer and Sample Preparation Buffer “A” 1. Empty the contents of the bottle labeled “Buffer A” into a 500 ml graduated cylinder. 2. Add degassed, deionized water to 500 ml total volume. 3. Place contents in bottle. 4. Stir briefly and gently on stir plate. 5. Label container as buffer “A”. (Pull label off buffer “A” bottle and place on container.) Buffer “B” 1. Empty the contents of the bottle labeled buffer “B” into a 500 ml graduated cylinder. 2.
4006073D.qxp 8/17/2009 7:45 AM Page 3 Section 4 System Setup 1. Assemble the rack as shown in Figure 1. Be sure to install the rack tray drain fitting, and provide a drain tube in case of spills. Column Clamping Set Drain Fitting Horizontal Bar Tapered Collar Side View See Detail Drain Tubing Tray Fig. 1. 2. Position the UV monitor optics module, proportioning valve/mixer, conductivity flow cell/column holder, etc. on the rack.
4006073D.qxp 8/17/2009 7:45 AM Page 4 Model 1327 Chart Recorder System Cable 2 UV Optics Module SV-5 Buffer Select Valve System Cable 4 Controller System Cable 1 SV-3 Column Bypass Conductivity Flow Cell MIXER B A SV-3 Diverter Valver (2110 Fraction Collector) BioFrac Fraction Collector OR Model 2110 Fraction Collector Proportioning Valve and Mixer Module System Cable 15 System Cable 3 Fig. 2. Section 5 Connecting a Chart Recorder 1.
4006073D.qxp b. 8/17/2009 7:45 AM Page 5 System Cable 4 (Mini-DIN/Banana Plugs) connects between COND CHART and the channel 2 banana plug receptacles on the 1327. The red banana plug connects to the “mV, V” terminal, the black to the ground terminal. This cable carries the conductivity signal to the chart recorder channel 2. Note: Be sure to plug the banana plugs into the channel 2 receptacles Section 6 Connecting a Computer Running LP Data View Software 1.
4006073D.qxp 8/17/2009 7:45 AM Page 6 2. Connect the fraction collector to the BioLogic LP system using system cable 3 as shown in Figure 3. Note: Both ends of cable 3 are identical. a. Connect one of the cable’s mini-DIN connector to the REC port on the fraction collector. b. Connect the other mini-DIN connector to the port labeled Diverter Valve on the BioLogic LP system. BioFrac fraction collector Biologic LP system System cable 3 System cable 15 Fig. 3.
4006073D.qxp 8/17/2009 7:45 AM Page 7 a. An SV-3 solenoid valve is used with the Model 2110 fraction collector. b. Plug the SV-3 valve directly into the diverter valve receptacle on the back of the BioLogic LP instrument. Section 9 Power Connections 1. Plug the line power cords into the rear panel of the BioLogic LP and the fraction collector. 2.
4006073D.qxp 8/17/2009 7:45 AM Page 8 3. Install tubing in pump as shown above. 4. PharMed tubing, 1.6 mm ID (furnished with the BioLogic LP system), is recommended for use with the starter kit. 5. Assemble with fittings and lock rings as shown in Figure 4. (Use orange lock rings and “medium” size barb fittings with 1.6 mm tubing.) Section 11 System Tubing Cut tubing and install fittings to make plumbing connections as shown in Figure 5. Tygon tubing, 1.
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4006073D.qxp 8/17/2009 7:45 AM Page 10 2. Connect the cartridge between the injector valve outlet and the UV monitor flow cell, as shown in the system plumbing diagram (Fig. 5). 3. Install 50 cm of 1.6 mm ID tubing for a 1.0 ml sample loop. The male luer fittings on each end of the tubing are used to connect to the injector valve. Section 13 Purging the System 1. Place the inlet tubes for “A” and “B” buffers in their respective buffer containers. 2.
4006073D.qxp 8/17/2009 7:45 AM Page 11 b. Press the soft key for CALIBRATE. c. Enter the Conductivity Cell Constant displayed on the conductivity flow cell cable. 2. In Manual mode: b. Press the COND soft key. c. Select Min/Max. d. In the Min field, enter the conductivity of “A” buffer (determined above in Step 6 of Section 13). e. Press SETMAX. f. Enter the conductivity of “B” buffer (determined above in Step 5 of Section 13). g. Press OK.
4006073D.qxp 8/17/2009 7:45 AM Page 12 c. Select “BioFrac” as your fraction collector. If your BioLogic LP system does not list the BioFrac, select 2128. The BioFrac communicates with the BioLogic LP using the same communication protocols and cabling as the Model 2128 fraction collector. d. The following question will appear: “Current Model: 2128 with valve Cable, External diverter valve cable connected?” Press the soft key corresponding to YES. e.
4006073D.qxp 8/17/2009 7:45 AM Page 13 In Manual mode: a. Press the PUMP soft key, then press FLOW. b. Set flow rate to 1.5 ml/minute, press OK. Press START. c. Press the UV soft key, press SET RANGE. d. Using the INCREASE and DECREASE soft keys, set UV monitor range to 0.05 AUFS. e. Press OK. f. Press ZERO—the system will set the UV reading to zero. g. Press the PUMP soft key. h. Press the STOP soft key.
4006073D.qxp 8/17/2009 7:45 AM Page 14 Section 19 Setting the Alarm The alarm sounds after 3 minutes to remind you to turn the valve back to the load position. (Proper technique is to switch the sample loop out of the flow path during gradient elution; the starter kit separation will be successful either way.) 1. Press the ALARM soft key. 2. Press ADD. 3. Enter a time of 3 minutes for Alarm 1 and check that Hold Methods is set to NO. 4. Press OK twice.
4006073D.qxp 8/17/2009 7:45 AM Page 15 Sample Loop Inject Port To BioLogic LP or Econo Gradient Pump MV-6 Inject Valve Waste FILL To Column "FILL" POSITION Fig. 7. Section 22 Final Check Before Starting Method 1. Press the MANUAL soft key, then pres the START soft key to start the pump. 2. Make sure that “A” buffer is selected. 3. Check the plumbing connections for leaks, check for air in the lines and purge if necessary. 4. Press the instrument keys to check settings. a. UV monitor: Range 0.
4006073D.qxp 8/17/2009 7:45 AM Page 16 e. Chart Recorder: After zeroing the UV monitor, use the thumbwheels to set both pens near the bottom of the chart recorder paper. Set paper speed (12 cm/hour suggested). Confirm that all switch settings are in the position marked in green. f. Fraction Collector: Confirm that the fraction collector is connected, then place tubes in the fraction collector rack. Check that the drop head is over collection tube #1. On the BioFrac, press Engage.
4006073D.qxp 8/17/2009 7:45 AM Page 17 Example LP Data View Chromatogram Example of chromatogram when running the BioLogic LP starter kit. Section 24 Advanced Users: Automatic Sample Injection If your system is equipped with an SV-5 buffer select valve, you may use it to inject the sample. This feature is important when you have a large sample, or wish to do multiple runs automatically. To demonstrate this feature: 1.
4006073D.qxp 8/17/2009 7:45 AM Page 18 3. Enter a pump program as described above—you may wish to recall your earlier method and edit. Step 1. Buffer “A”, step Length 1 minute, flow rate 1.5 ml/minute Step 2. Buffer “C”, step length 3.5 minutes, flow rate 1.5 ml/minute Step 3. Buffer “A”, step length 1 minute, flow rate 1.5 ml/minute Step 4. Gradient 0% to 50% “B”, step length 10 minutes, flow rate 1.5 ml/minute Step 5. Buffer “B”, step length 6 minutes, flow rate 1.5 ml/minute Step 6.
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4006073D.qxp 8/17/2009 7:45 AM Page 20 Catalog Number Product Information 741-0018 BioFrac Prep-20 Preparative Rack 741-0007 BioFrac Fraction Collector Fittings Kit, includes replacement fittings and tubing for setup of the fraction collector to the BioLogic LP or DuoFlow chromatography systems 741-0088 BioFrac Microplate Drophead Kit, includes preassembled drophead nut with 0.
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4006073D.qxp 8/17/2009 7:45 AM Page 22 Catalog # Description 731-8233 Male Luer Plugs, 25 731-8229 Female Luer T-Connector, 10 732-8302 0.8 mm Barb T-Connector, recommended for minimal dead-volume connection, 25 732-8300 0.
4006073D.qxp 8/17/2009 7:45 AM Page 23 Legal Notices Tygon is a trademark of Norton Co. Windows and Windows 95, 98, ME, NT, 2000, and XP are trademarks of Microsoft Corp.
4006073D.qxp 8/17/2009 7:45 AM Page CVR2 Bio-Rad Laboratories, Inc. Web site www.bio-rad.
