Copyright: Copyright (C) by Limnological Institute
Citation: IntelliChrom Help File


Contents

Menu and ToolBar description

Analysis window

Run spectrophotometer… window

Set colors, Set font and Set axis windows

Hardware… and Defaults… windows

Analysis sequence

Manual operations

Dual-wavelength detection and spectral ratio

Detector time constant

Gradient elution

The directories and files

Specifications of chromatograph


Menu and ToolBar description

Icon	Function	Description
	File [Open...]	Viewing of Chromatograms (*.chr), Regenerations (*.reg), Specrtrums (*spm) and Detector files (*.det)
	Control [Analysis...]	Reading, setting, saveing parameters of an analysis in window "Analysis" and Subwindow "Text". Run analysis.
	Control [Spectrum...]	Setting of spectrum parameters, zero line supressing and run spectrum at stop flow in window "Spectrum"
	Control [Manual operation...]	Window "Manual operation" (filling and empting of pumps, rinsing of injector port and needle, leak test, regeneration of column, run of detector).
	Control [Resume]	Resume a current procedure after "Pause"
	Control [Pause]	Pause in a current procedure
	Control [Stop]	Pause and then breake of a current procedure
	Process [Swith to...]	Switch to Data Processing Software MLCW
	Options [Auto Scale]	View a whole chromatograml
	Options [Axis...]	Window "Set axis..." for chromatogram and spectrum
	File [Print...] [Print  Setup...]	Print a chromatogram or spectrum in active window and setting printing parameters and printer in windows "Print Setup" and "Print manager"
	Options [Colours...]	Setting colours in window "Set colours..."
	Options [Fonts...]	Setting font in window "Set font..."
	Window [Show info...]	Reading of an information about file in active window
	Help [Help...]	Getting a help in window "Help"
File [Save as...]		Save file as "new" one or convert chromatogram curve to  file *.txt in window "Text Export".
Edit [Copy to Clipboard...]		Copy of chromatogram, regeneration, spectrum or detector curvies in active window to clipboard (window "Copy to Clipboard...").
Options [Communications...]		Setting communication parameters in window "Set Communication Parameters..."
Options [Hardware...]		Setting of chromatograph configuration mode in window "Hardware Setup..."
Options [Column...]		Setting parameters of chromatographic column for including they in a final report (window "Column...")
Options [Defaults]:     [Prereferences...]     [Default Chromtogram Axis...]     [Default Spectrum axis...]     [Default Colours...]     [Default Font...]		Setting named parameters as custom defaults in windows "Prereference", "Set axis...", "Set default axis...", "Set colours..." and "Set default font..."


"Analysis" window

	Open file <*.cha> from directory \I-CHROM\METHODS\ and load parameters of analysis.
	Save file as <*.cha> in directory \I-CHROM\METHODS\
	Go to window "Add text..." and write information additional data which will included in report. For saving of text press bottom "OK".
	RUN an analysis with parameters from loading file <*.cha>.


"Run spectrophotometer..." window

	RUN scanning of spectrum at setting parameters and saving it in file <*.spm>.
	Suppressing of zero line at setting parameters. This procedure must be done before writing of spectrum.


"Set colors", "Set font..." and "Set axis..." windows

	Enter setting parameters for active window.
	Enter setting parameters for all opened files.


"Hardware..." and "Defaults..." windows

	Enter setting parameters as custom defaults.
	Return to Intelli-Chrom defaults.


Analysis Sequence

1.	Filling
2.	Regeneration
3.	Zero Suppression
4.	Filling
5.	Rinsing of Injector (if set)
6.	Suction of Presample in Injection Needle
7.	Suction of Sample in Injection Needle
8.	Injection of Sample and Presample in Column
9.	Elution
10.	Rinsing of Injection Needle (if set)


Fill syringes of  pumps 'A' and 'B'. After filling volumes will be: 'A' : 2380ul, 'B' : 2500ul.


Regenerate column with defined concentration (%B), flow rate and detector settings.


Auto zerroing of baseline in the wavelength range.


Rinsing of the injection port with 40 ul of regeneration solvent with flow rate 200 ul/min.


Elute with gradient and detector settings.


Rinsing of the injection needle three times in solvent 1 and three times in solvent 2.


All operations with sample and presample ary carried out with flow rate 40 ul/min.


Manual operations

Fill	filling syringes of selected pumps from reservoires.
Empty	emptying syringes of selected pumps to reservoires.
Purge solvent	purging syringes of selected pumps to waste.
Change solvent	changing solvents in syringes of selected pumps (purge, 500 ul fill - purge).
Test leak	leakage checkng at 5 MPa. Needle there is in the stopper.
Set temperature	turn on/off heater. Setting of temperature (35-90 °C).
Regenerate column	column regeneration with solvent (%B), at flow rate (5-1000 ul/min) and volume (1-2000 ul).
Run detector	run detector with settings.
Rinse injector	rinsing of the injection port with settings.
Rinse needle	rinsing of the injection needle three times in solvent 1 and three times in solvent 2.


Specifications of chromatograph

COLUMNS: 2 mm i.d.; 60-80 mm length; [image]; void volume
DETECTOR: double-beam multi-wavelength UV-spectrophotometer
Wavelength range in steps 2 nm (nm):	190 [image] 360
Spectral bandwidth (nm)	10
Wavelength accuracy (nm)	±0.5
Wavelength reproducibility (nm)	±0.005
Time constant and time of measurement at one wavelength tau (sec)	0.06 [image] 10
Number of wavelengths in one cycle	Up to 8
Noise level with cell pathlength 1.6 mm at tau =0.3 sec and lambda=260 nm (AU)	<0.0002
Drift with cell pathlength 1.6 mm at tau=0.3 sec and lambda=260 nm (AU/hour)	<0.0001
Linear absorbance range with cell pathlength 1.6 mm (AU)	-10 [image] 10
Flow cell pathlength (mm)/volume (µl)	1.6/1.3
PUMP: two syringe high pressure gradient metering pump with switching 6-port valve
Volume of one syringe (µl)	2500
Flow rate setting range (µl/min)	5 [image] 999
Flow rate accuracy at 100 µl/min (%)	±1
Flow rate reproducibility at 100 µl/min (%)	±0.3
Maximum pressure (MPa)	7
COLUMN HEATER: solid type
Temperature (ambient temperature +10°) control range in 1° C steps (°C)	35 [image] 90
Temperature control precision (°C)	±0.1
AUTOSAMPLER: stop-flow sample auto injection programmed method
Number of vials/number of injections/volume of vial (µl)	46/200/200
Sample volume setting range in 1 µl steps (µl)	1 [image] 100
Injection reproducibility with 2 µl sample (%)	±0.5
DIMENSIONS & WEIGHT (without computer): W530xD200xH300 mm; 17 kg
POWER REQUIREMENTS (without computer): 100-120/200-240 V; 100 VA; 50/60 Hz

See also: Portable liquid chromatograph


Portable liquid chromatograph


Dual-wavelength detection and spectral ratio

	An integral spectral ratio ( [image]) for a peak of a substance can be calculated as:      A current spectral ratio (R) for a peak of a substance can be calculated as:

R and [image] are unic characteristics of a substance. They are not depend on it concentration.

For a homogeneous peak of a substance R=const and it is criterion of a peak homogenity.

Retention time and [image] are two independence criterions for peak identification.


 Detector time constant

	Remarks: 1. A Peak Square is not depend on . 2. A noise level =
In a routine HPLC we are recommending to set tau=0.34 [image] 1.2 sec


Gradient elution

n - No. of linear gradient step (0...20)

V - current volume (µl)

%B - concentration of eluent B at the end of the step

Reg - isocratic step "Regeneration of Column"

Vreg - volume of step "Regeneration of Column"

Breg - concentration of eluent B at the step "Regeneration of Column"


The directories and files

Tree and Directories:

After software installation on hard disk the following directories were created:

\I-CHROM\ - main directory;

\I-CHROM\METHODS\

\I-CHROM\REPORTS\

After the first analysis will be RUN the following subdirectories will be automatically created:

\I-CHROM\DATA\METHOD\YY-MM-DD\. The last subdirectory is date of RUN of analysis (YY - Year; MM - Month; DD - Day). It will be created every new day when analyses will run.

\I-CHROM\SPECTRA\YY-MM-DD\.

\I-CHROM\TEMP\

Types of Files:

There are 7 types (extensions) of files:

<*.cha> - files "method of chromatographic analysis" are placed in: \I-CHROM\METHODS\

<*.mtw> - files "method of data processing" are placed in: \I-CHROM\METHODS\

<*.chr> - files "chromatogram" are placed in: \I-CHROM\DATA\METHOD\YY-MM-DD\

<*.chw> - files "chromatogram ready for data processing" are placed in: \I-CHROM\DATA\METHOD\YY-MM-DD\

<*.reg> - files "regeneration of column" are placed in \I-CHROM\DATA\METHOD\YY-MM-DD\

if regenerations of column are parts of analyses and files are placed in \I-CHROM\TEMP\

if regenerations were made as manual operations.

<*.spm> - files of "spectra" are placed in \I-CHROM\DATA\METHOD\YY-MM-DD\

if spectra were written during chromatographic analysis and files are placed in \I-CHROM\SPECTRA\YY-MM-DD\

if spectra were written in Manual Mode. The last subdirectory is date of RUN of spectra (YY - Year; MM - Month; DD - Day). It will be created every new day when spectra will run.

<*.det> - files of detector writing in \I-CHROM\TEMP\

Names of Directories and Files:

<xxxxxxxx.cha> - this name of file of METHOD is writing by operator after edition of parameters of analysis. Default name is <new.cha>. The name of file <*.cha> is repeated in automatically created the same name of directory \I-CHROM\DATA\XXXXXXXX\YY-MM-DD\

<hhmmnnn0.reg> - this name of file is created automatically at a moment of run of chromatographic analysis. hh - hour; mm - minute; nnn - number of run of analysis (1...200).

<hhmmnnn0.chr> - this name of file is created automatically at a moment of start of elution. hh - hour; mm - minute; nnn - number of run of analysis (1...200).

<hhmmnnn0.spm> - this name is created automatically at a moment of run of spectra during chromatographic analysis. hh - hour; mm - minute; nnn - number of run analysis (1...200).

<hhmmxxxx>.spm - this name is created automatically at a moment of run of spectra in Manual Mode. hh - hour; mm - minute.

<dddsssss.reg> - this name is created automatically at a moment of run of regeneration of column in Manual Mode. ddd - day of year + 31; sssss - second of the day.

<dddsssss.det> - this name is created automatically at a moment of run of detector in Manual Mode. ddd - day of year + 31; sssss - second of the day.