Department of Chemistry, Tufts University
If the instrument configuration has changed since the last use (i.e. add or remove autosampler or change detector) run HP Configuration Editor before doing anything else.
1.
Turn on power to modules with ON/OFF switch. Wait for “ready for use” to appear on module screen.
2.
Turn on N
2 tank at 65psi for autosampler. N
2
is not needed if the autosampler is not used. Turn on
He tank @10psi for degassing. Adjust valves A,B on solvent tray for slow bubbling. CW is ON.
Unused valves fully CCW.
3.
Select HP1050 Online from Windows desktop.
1.
Turn on Detector lamp. Use LAMP ON/OFF key on module, or Instrument/More
DAD/Control from Chemstation menu. Allow 20 min warm up before injection.
2.
Turn on pump. Use PUMP ON/OFF key on module, or Instrument/More Pump/Control from Chemstation menu.
3.
If pumps have been idle for several days or more, or if solvent reservoirs have been changed, the pumps should be purged. See below for instructions
4.
Method/Load Method from menu to load an existing method. To edit the method, select
Method/Edit Entire Method from the menu.
5.
Select RunControl/Sample Info from menu. Enter Sample and operator information. Also enter desired file name and data subfolder.
6.
Select View/Online Signals/Sig. Window 1 from menu if no signal window is visible. Select
Change button, and add or remove signals. Select OK .
7.
Select View/Instrument Actuals from menu to display status of each module.
8.
Manually inject the sample, switch the valve to the INJECT, then withdraw the syringe.
Chemstation will start data acquisition automatically.
9.
If using Autosampler, See directions below.
10.
Click on the ▲▼ buttons to adjust vertical scale. Use ◄► buttons to adjust horizontal scale.
11.
To end a run before the time specified in the method, select Run Control/Stop Run from the menu. The data will be saved. Abort will top the run without saving data.
12.
A report will appear on the screen or printer at the end of the run, as specified in the
Report/Specify Report options in the Data Analysis view.
To print click on the printer icon on the tool bar. To print details of the method, choose File/Print/Method from the menu.
13.
To fix vertical scale in Data Analysis mode (e.g. large solvent peaks) use Report/Specify
Report from menu, then Signal Options button.
1.
To enter the Data analysis view, choose View/Data Analysis from the menu, or use the pull down menu on the tool bar.
2.
To change integration parameters, choose Integration/Integration Events from the menu.
Increase Area reject to eliminate small extraneous peaks. See online help files for more options.
3.
To adjust vertical scale in Data Analysis mode (e.g. large solvent peaks) use Report/Specify
Report from menu, then Signal Options button.
4.
To display the spectrum of a peak (DAD detector only) select Spectra/Select Spectrum from the menu. Then use the mouse to select a peak. To print a spectrum, use Graphics/Copy to
Clipboard , then paste to a Word document.
Revised 6/23/03 D. Wilbur

Department of Chemistry, Tufts University
Best results use 10
 l injections from a 50
 l syringe. Use only square cut needles, not beveled ones . With the valve in the LOAD position, insert the syringe until the needle seats in the holder. There will be a slight resistance in the last few mm. Inject the sample, switch the valve to the INJECT position, then withdraw the syringe to put sample on column. Chemstation will start data acquisition automatically.
1.
Select View/Sampling Diagram from menu.
2.
Select single vial or multi-vial icon in tool bar.
3.
To run a single sample: a.
Select the vial in the sampling diagram. b.
Enter sample information, including vial number. You need not start with vial 1. c.
Select Start . Autosampler will load from selected vial.
4.
To run multiple samples: a.
Select Sequence/Sequence Parameters . Enter operator and file information. Select
OK. b.
Select Sequence/Sequence Table . Enter Sample information. You need not start with vial 1. Select OK. c.
Select Sequence/Sequence Output . Select OK. d.
Select Start . Autosampler will load from vials, according to sequence table.
If pumps have been idle for several days or more, if solvent reservoirs have been changed, or if air bubbles are visible in the lines from the solvent reservoirs to the pump, the pump should be purged. Purging is done from the keys on the front of the pump.
1.
Press Purge (do not press Enter). Then press Next to display a channel, press

until the blinking cursor is over the channel (A,B,C,D). Press Next until display reads Channel A Only. Then
Enter .
2.
Open the purge valve. Route the purge line to an appropriate container.
3.
Press Purge , then Enter . Wait until air bubbles are cleared from the line.
4.
Repeat for all other channels to be used.
5.
Close the purge valve
1.
Turn off detector lamps. Use LAMP ON/OFF key on module, or Instrument/More
DAD/Control from Chemstation menu.
2.
Turn off pumps. Use PUMP ON/OFF key on module, or Instrument/More Pump/Control from Chemstation menu.
3.
Exit Chemstation software. Caution: turning off power to modules while Chemstation is running will cause software to lock up.
4.
Turn off degassing master valve on solvent tray..
5.
Turn off power to modules.
6.
Turn off N
2
and He tank valves
If the instrument is to be left unused for more than a few days, the following additional steps should be taken.
1.
The entire system should be flushed with about 100ml acetonitrile/H2O with no buffers. The column should then be tightly capped and put away.
2.
The pump, valve, and detector should then be flushed with deionized water.
Revised 6/23/03 D. Wilbur

Department of Chemistry, Tufts University
File structure
Methods stored in C:\HPChem\1\Methods\<name>.m
Sequences (sample list for autosampler) stored in C:\HPChem\1\Sequences\<name>.s
Data stored in C:\HPChem\1\Data\…, as defined in Run Control/Sample Info window. Each run is a directory consisting of several files, depending on detector.
Vwd1a.ch binary chromatogram data file, UV detector
Dad1.uv
Dad1a.ch
Fld1a.ch
Lcdiag.reg
Acqres.reg binary spectra file, DAD detector binary chromatogram data file, single wavelength. Up to five files, a-e. binary chromatogram data file, fluorescence detector
Instrument curves (gradients, temp, press) and injection vol, solvent.
Column information
Run.log
Sample.mac
Text file run log
Text file with sample information
Report.txt Text portion of report, if print to file is specified in Report/Specify Report
Report00.wmf Chromatogram portion of report, if print to file is specified.
Instrument Configuration
HPLC 1050-1
HP 1050 Solvent tray
HP 1050 Quaternary Pump
Model 79856A SN 3114A03102
Model 79852A SN 3317A02335
Rheodyne front loading injector with 20ul loop
HP 1050 Variable WL detector Model 79856A SN 3114A02496
Generic Celeron 800 computer, Win95.
HP Chemstation 6.03 software
HP 1046A Fluorescence Detector Model 1046AX SN 3137G02410
HPLC 1050-2
HP 1050 Solvent tray
HP 1050 Quaternary Pump
Model 79856A SN 3114A01141
Model 79856A SN 3317A02334
Rheodyne front loading injector with 20ul loop
HP 1050 Autosampler
HP 1050 Diode array detector
Model SN 3404G04003
Model G1306A SN 3319A00205
Generic Celeron 800 computer, Win95.
HP Chemstation 6.03 software
HPIB Addresses
Pumps 16
Diode Array Det 17
Var WL UV Det 10
Fluor Det
Autosampler
12
18
Revised 6/23/03 D. Wilbur