Matec CHDF 2000 User manual

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Matec Applied Sciences
56 Hudson St., Northborough, MA 01532 Ph: (508) 393-0155 Fax: (508) 393-5476
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Table of Contents
Frequently Asked Questions I-2
Chapter 1: Introduction to the CHDF 2000
I-6
1.1 CHDF 2000 Components I-7
1.2 CHDF 2000 Operation I-10
1.3 Using the Control Panel I-11
Chapter 2: Theory and Sample Preparation
2.1 Scope II-1
2.2 Safety Precautions II-1
2.3 Intended Use II-2
2.4 Description and Specifications II-3
2.5 Theory of Operation II-7
2.6 Data Analysis II-8
2.7 Sample Preparation II-8
2.8 Eluant II-9
Chapter 3 Setting Up the CHDF 2000
3.1 Performing Power-On Procedures III-2
3.2 Reviewing and Editing Operating Parameters III-7
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Chapter 4 Operating the CHDF 2000
4.1 Priming the Pump IV-1
4.2 Analyzing Samples IV-4
Chapter 5 Troubleshooting the CHDF 2000
5.1 Responding to Error Messages V-1
5.2 Quick Troubleshooting Checklists V-4
5.3 Running CHDF 2000 Diagnostics V-8
Chapter 6 Maintaining the CHDF 2000
6.1 Maintaining the Pump VI-2
6.2 Maintaining the Manual Injector Valve VI-12
6.3 Maintaining the Priming Valves VI-16
6.4 Maintaining the Detector VI-17
6.5 Adjusting the Control Panel Display VI-24
6.6 Dealing with Plugged Cartridges VI-25
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Matec Applied Sciences
CHDF 2000 Hardware Users
Guide
Version 3.8
April 26, 2006
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FREQUENTLY-ASKED QUESTIONS ABOUT
CHDF OPERATION
1. How frequently should the CHDF GR500 eluant be replaced?
The CHDF GR500 eluant degrades with time. It should be replaced every two to three days. Check the
conductivity of the eluant in order to determine if it is suitable. The conductivity ranges for the
different GR500 types are as follows (in microSiemens per centimeter): 1X-GR500: 7-10; 2X-GR500:
22-27; 2X-GR500-LA: 14-18; 1X-GR500 pH9: 13-18. The GR500 eluant absorbs Carbon Dioxide
from the ambient which causes its pH to drop, and its conductivity to increase; this results in
measured particle sizes smaller than expected. The 1X-GR500 pH9 eluant is especially sensitive to
CO2 absorption. Bacterial growth also takes place as the GR500 ages.
2. I obtain smaller than expected CHDF particle size results when I inject calibration standards.
Most likely, your GR500 eluant is either degraded or contaminated. See Question 1 above.
3. How is the GR500 eluant prepared?
Add 950 ml of deionized water (conductivity should be less than 2 micro Siemens per centimeter) to 50 ml
of GR500 concentrate. Eluant degassing by either sonication or vacuum filtration is recommended.
4. How long does the GR500 concentrate last, and how should it be stored?
The GR500 lasts about one month. It should be kept refrigerated.
5. Does the Marker fluid need to be injected at an exact time for every run?
No. In both manual, and auto-sampler operation, the software detects when the marker is injected. The
injection time is recorded; the marker elution time is calculated as the marker exit time minus the
injection time. A marker delay time of zero seconds is recommended for auto-sampler operation (this
is set under the current method file).
6. How should the marker fluid be prepared?
The marker fluid consists of a solution of sodium benzoate in deionized water. Its concentration varies
depending on the type of fractionation cartridge used. Typical concentrations are in the range 0.1% to
0.5%. This concentration should be adjusted in order to obtain a reasonable marker peak height
(displayed on the CHDF software Raw Data graph) upon marker injection into the CHDF. Marker
peaks heights of 10 to 40 mAU’s (milli Absorbance Units) for the CHDF-2000 instrument, and 5,000
to 15,000 counts for the CHDF-1100 are acceptable.
7. Why is the pump pressure fluctuating more than 200 psi during a run?
Air bubbles are lodged in the pump pistons. Stop the pump; open the pump purge valve (located on the
lower right- hand corner of the CHDF-2000; or on the CHDF-1100 pump front panel); insert a 5-ml
plastic syringe into the purge valve, and slowly draw fluid. Air bubbles in the pump and pump-inlet
tubing should be removed. Tighten the purge valve and start the pump. If the air bubbles re-appear,
the Whatman de-gassing filter on the pump-inlet tubing should be replaced. This filter lasts about one
month. Cavitation, which results in air bubble generation, occurs if the Whatman filter needs
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replacement. For the CHDF-2000, this filter is optional. You may operate your CHDF-2000 without
this filter if you so prefer.
8. Noise and drift are present in the Raw Data baseline.
This is probably due to a defective Whatman filter. Replace this filter and observe the baseline. This filter
must be filled with eluant when in operation. Use the purge knob on the filter is order to draw any air
from the filter. Another possibility is that the UV lamp may need replacement if it has been used for
more than 1000 hours (see the troubleshooting section of the hardware manual).
9. How should samples be prepared for CHDF analysis?
Only aqueous samples can be analyzed on the current CHDF setup. Dry powders must be dispersed in
GR500. Sonication should be applied in order to help suspend the particles in the liquid. Concentrated
samples should be diluted in GR500 eluant. The solids contents are typically in the range 0.1% to 5%
by volume. UV-absorbing samples can be analyzed at lower concentrations while non-UV absorbing
samples (such as colloidal silica) must be run at higher concentrations. Sonication may be used in
order to break-up particle aggregates. Reasonable sample concentrations should produce peak heights
of 20 to 50 mAU’s for the CHDF-2000, and 5,000 to 15,000 counts for the CHDF-1100. A syringe
filter may be used on the sample syringe in order to remove aggregates from samples prior to
injection. Syringe filters may only be used once per sample. Marker, as well as, Rinse filters may be
reused until leaks are observed on the filters.
10. No peaks are detected in the Raw Data graph upon sample, and marker injection.
The fractionation cartridge is likely plugged. Refer to the troubleshooting section of the hardware manual
for cartridge cleaning instructions.
11. The pump pressure is higher than normal.
The filter element in the cartridge in-line filter must be either cleaned or replaced. Refer to the
troubleshooting section of the hardware manual. The CHDF-1100 is affected by ambient temperature.
Low ambient temperatures result in high CHDF-1100 pressures.
12. What Noise Level should be used under Configure System in the CHDF software?
30-40 for the CHDF-2000; 400 for the CHDF-1100.
13. Can any wavelength be used on the CHDF UV-detector?
The CHDF UV-detector wavelength can be varied from 190 to 380 nm. However, the CHDF PC software
only contains particle extinction cross section curves for 200 nm, and 220 nm. One of these values
must be selected in the active Method file. Inaccurate results are obtained if the UV-detector, and the
software wavelengths differ.
14. What is the sharp peak in the Raw Data between the sample peak and the marker peak?
This peak corresponds to UV-absorbing molecular species present in the sample. Molecular species
include surfactants, electrolytes, acids, or bases. These molecules have a smaller size than the particles
themselves, and therefore exit the capillary after the particles.
15. A broad peak after the marker peak is displayed in the Raw Data.
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This peak is likely due to non-ionic surfactant. Non-ionic surfactants can continuously adsorb and desorb
from the capillary wall as they travel down the fractionation capillary. This adsorption/desorption
retards the non-ionic surfactant from exiting the capillary.
16. Can the marker be blended with the sample so that only one injection is made per sample?
Yes, in manual operation only. Simply add maker fluid to your samples, and make only the sample
injection (do not inject the marker).
17. How long should the instrument be allowed to equilibrate (or warm up) at the beginning of the day
before starting sample analysis?
Approximately 30 to 45 minutes.
18. Can the CHDF be turned off at the end of the day, or should it be left running overnight?
The CHDF can be operated in either mode. We prefer to turn it off at the end of the day.
19. How much sample, and marker, should be Loaded into the injection valve?
100 microliters for manual operation. It is recommended that at least 400 microliters be drawn into the
syringe in order to minimize the chances of loading air bubbles into the injection valve. If using a
syringe filter make sure the filter is filled with sample before loading sample into the injection valve.
80-90 microliters is recommended for auto-sampler operation. This is set under System/Configure
Auto-Sampler.
20. Are samples loaded directly into the fractionation capillary?
No. They are loaded into a sample loop (a piece of stainless steel tubing) which is connected to a vent. As
sample is pushed into the injection valve, the sample loop becomes filled with sample. Sample exits
from the sample loop vent as more sample is loaded. The sample loop becomes connected to the
pump, and the fractionation capillary upon switching to Inject.
21. Can the CHDF software be run under Windows9X, NT, and 2000?
Yes, the CHDF2000 software can be run on any Windows operating system software.
22. What solids concentration should be used for calibration standards?
Calibration standards should not be overly concentrated. Solids concentrations should be such that Raw
Data peak heights are 10 to 30 mAU’s for calibration standards and 20-60 for regular samples.
Calibration standards larger than 300 nm should be sonicated prior to use in order to break up
aggregates.
23. How frequently should the CHDF capillary be calibrated?
Calibration shifts are related mainly to two factors: (i) changes in GR500 eluant composition; and (ii)
changes in ambient temperature (not important for the CHDF-2000). It is recommended that the
conductivity of the GR500 eluant be monitored; replace the eluant if the conductivity is outside its
normal range. Calibration standards can be run prior to regular sample analysis in order to confirm
calibration accuracy. These “control” standards may be added to the calibration curve if re-calibration
is deemed necessary.
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24. How long does a CHDF cartridge (capillary) last?
Fractionation capillaries do not have an exact life span. Cartridges may last several years since no particle
adsorption occurs inside the capillary. Capillary failures are frequently due to plugging right at the
capillary inlet. Cutting-off (“snipping”)a short piece at the capillary inlet frequently removes the plug.
The capillary can also be back-flushed as described in the hardware manual, troubleshooting section.
25. How is the Weight Particle Size Distribution (PSD) calculated?
The particle weight percent is calculated as the Number percent multiplied by the particle diameter raised
to the third power: Wt
1
%= N
i
%(D
p
)
3
. This assumes that the particle density is the same for all
particles in the sample. The area percent is calculated as follows: A
i
% = N
i
%(D
p
)
2
26. What particle Size Reproducibility should be expected from CHDF
About ±5%.
27. What is the mean, 25%, median, 75%, and FWHH in the text PSD report?
The 25%, median, and 75% are the 25th, 50th, and 75th percentiles of the particle size distribution (PSD).
They are calculated as the particle size evaluated at those values of the cumulative PSD. Physically,
for the 25% for example, they indicate that 25% of the particles are below the reported particle size.
The FWHH is the PSD Full Width at Half Height. It is the width of the differential PSD at 50%
height.
28. What is the list of particle sizes at the bottom of the Text Report?
This list contains a description of all the particle size populations (also referred to as modes) automatically
detected by the software in the sample. Each mode is listed by the start and end particle size. The
percentages by Number, Area, and Weight are printed. The Area% is the average particle size of each
mode (Weight-Weighted). The mode detection sensitivity can be changed in the current method under
Optical Mode Detector. Lowering the value of Slope Sensitivity increases mode sensitivity.
29. How can the Particle Size Distribution Report be Customized or Changed?
Open your current CHDF method, and select “Report Setup”. The text report, if used, should be
placed at the bottom of the report.
30. What causes noticeable “tailing” in a fractogram?
The inlet to the fractionation capillary is likely dirty. Snip the fractionation (top) capillary (see
hardware manual section 6.6).
31. How are odd-shape particles, e.g., discs, rods analyzed?
Particles spin in the capillary tube during CHDF fractionation. As a result, particles approach the
capillary wall at different orientations. Some positions result in an exclusion layer equal to the longest
particle dimension while other positions do the shortest or intermediate dimensions. The resulting
elution time is equivalent to an average of all the particle dimensions. Research done at Lehigh
University showed that particle doublets generated from partial latex coagulation produce a CHDF
particle size equal to one particle diameter multiplied by the square root of two, or 1.41*Dp (instead
of 1.5*Dp). Triplets appear to produce one diameter times the square root of 3.
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32. Does the CHDF software extrapolate beyond the two calibration standards at the limits of the
calibration curve?
The CHDF reports particle size populations in the range specified in the CHDF2000 software under
system/Configure-System. On the small particle size end, the CHDF interpolates below the smallest
standard and the data point Rf=1, Dp=0, where Rf is the separation factor for a near-zero particle size
(Dp) species such as the marker fluid. In fact, if you run the marker as a sample, you obtain Rf=1. Rf
is calculated as the particle elution time divided by the marker elution time. Rf increases with
increasing particle size as shown in the calibration curve. Elution time is peak exit time minus
injection time. For larger particles, the software does extrapolate using a curve-fitting expression of
the calibration-standard data points (up to the upper limit specified under System/Configure-System.
This curve fit can be a sigmoidal or polynomial fit. Both options have sigmoidal shapes.
33. What is Particle Frequency in the PSD graphs?
"Particle Frequency" and "Volume Percentage" are different. The CHDF produces relative PSD's.
An absolute PSD shows the actual particle volume percentage. However, absolute PSD's are
sometimes difficult to compare between different samples (overlay). The reason is that samples with
different percent solids will show different PSD-peak heights.
Relative PSD's as currently shown by the CHDF are normalized so that the tallest peak or peaks have
a height of 100 "frequency units". These are not percentage values because there can be two or more
peaks with the same 100 value. Also, next to the 100 point, there are values of 98, 92, 86, and so on. If
you add all these together, you would obtain much more than 100% for the sample.
The particle frequency can be used to compare two populations. For example, a 37% peak has half the
percentage of a 74% peak.
If you need to know the percentage of a population, you should use the cumulative distribution (CD).
You can subtract the CD values at both ends of each peak. This gives you the percentage of this peak.
See example graph below.
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34. How can the percentage of a particle-size population be calculated?
See “Particle Frequency” question above.
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35.
1 Introduction to the CHDF 2000
The CHDF 2000 (Figure 1-1) uses the principle of Capillary Hydrodynamic Fractionation
(CHDF) to measure particle sizes from 15 nanometers to 1.1 microns in diameter.
Figure 1-1 CHDF 2000
The CHDF 2000 provides the following features
•Simple front panel control of all system operations
•An eluant delivery system
•UV absorbance detection
•Reduced workbench space requirements
Control panel
Front door
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1.1 CHDF 2000 Components
1.1.1 System Components
The CHDF 2000 contains the following components:
•Control panel
•Manual injector valve
•Eluant Pump
•Fractionation Cartridge
•Absorbance detector
•Signal connector module
Control Panel
The CHDF 2000 control panel (Figure 1-2) provides a simple icon-based interface that allows
you to directly configure, operate, and troubleshoot the CHDF 2000 . The control panel is located
on the CHDF 2000 front door. Refer to Section 1.3, Using the Control Panel, for detailed
instructions on using the CHDF 2000 control panel.
Figure 1-2 Control Panel and Manual Injector Valve
Manual injector valve
Control panel
pH Sensor
Autosampler
Connections
In
Out
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Manual Injector Valve
The CHDF 2000 manual injector valve (see Figure 1-2) provides a port for loading a sample and a
manual rotation handle to inject the sample into the eluant stream for analysis. The injector valve
is mounted to the right of the front door.
An autosampler option is also available for automated sample injection.
Eluant Pump
The base system pump supplies the eluant for CHDF 2000 operation. You can access the pump behind the
front door (Figure 1-3). A priming valve is connected to the pump outlet and is mounted directly below
the manual injector valve.
Figure 1-3 Eluant Pump and Fractionation Cartridge
Fractionation Cartridge
In the CHDF2000, a capillary column provides hydrodynamic separation of the sample
Instructions for installing the column are provided in Chapter 6.
Absorbance Detector
The CHDF 2000 includes a multi-wavelength, tunable, ultraviolet (UV) absorbance detector
(Figure 1-4). The UV detector measures the Ultraviolet light extinction of particles in the flow
stream from the column outlet.
Pressure
xducer
Access hole for
offset-fine-
adjust pot
Inlet Check Valves (2)
Outlet
check
valves
(2)
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Rear
Panel
Com puter
Connection
Absorbance
Detector
Figure 1-4 Absorbance Detector
Computer Connection
A 9 pin, RS-232 connection is used to provide data transfer between the CHDF 2000 and a
personal computer. A Pentiumprocessor and Windows ‘95are recommended for the
personal computer.
Signal Connector Module
1V/AU OUT
PRESS. OUT
% B OUT
INJ. OUT
START IN
+
–
+
–
+
–
+
–
The signal connector module (Figure 1.5) on the CHDF 2000 rear panel provides signal
connections for external devices such as a chart recorder or integrator. You can also use these
signals to synchronize the operation of the CHDF 2000 with other devices.
Figure 1-5 Signal Connector Module
Signal connector
module
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1.2 CHDF 2000 Operation
1.2.1 Operation
Figure 1-6 is a fluidic diagram illustrating CHDF 2000 operation.
Figure 1-6 Fluidic Diagram of Operation
Tubing ID Legend
Waste outlet
Strain gauge
pressure transducer
In-line filter
Pulse
dampener
(Pump A)
0.009 SS
0.020 SS
0.040 SS
0.062 TFE
20-µL loop
Unions
Manual injector
valve (7725i)
Drain tubes
SSI priming
valve
Optics bench
Pump A
solvent reservoir
Whatman
filter
(optional)
Pump A
Eluant
Reservoir
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1.3 Using the Control Panel
The CHDF 2000 control panel provides easy control of system configuration, operation, and
diagnostic functions.
The control panel components (Figure 1-7) include:
•Control panel display
•System controls
Figure 1-7 Control Panel Components.
Selector knob
Keypad
Control panel
display
Function keys
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1.3.1 Control Panel Display
The CHDF 2000 control panel display consists of four sections (Figure 1.8)
Figure 1-8 Control Panel Display Sections
Screen name – A screen name appears at the top of each screen to identify the specific
program screen.
Screen body – The main body of each screen contains one or more of the following types
of information:
−Control options, including system tasks and navigation options
−Status of current operation
−Instructions
Secondary options or instructions – Secondary options or instructions appear at the
bottom of most screens. Secondary options allow you to choose alternate navigation or
control options.
Screen
Name
Screen
Body
Secondary
Options
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1.3.2 System Controls
System controls are just below the control panel display on the CHDF 2000 front door.
Table 1-1 identifies the function of each CHDF 2000 control.
Table 1-1 Control Panel
Control Function Description
0 – 9 Number values Used in combination to insert a value for
a selected parameter.
. Decimal point Inserts a decimal point in a value for a
parameter.
CL Clear Clears an entered value.
OK Execute Selects a highlighted option. Alternative
to pressing the selector knob to select a
highlighted option.
∅AU Autozero Detector Resets the absorbance detector output
signal to the zero offset value.
Home Go Home Returns the control panel display to the
Home screen.
Go Back Returns the control panel display back to
the preceding program screen.
Selector knob Rotate to change highlighted screen
option or to increase or decrease a value.
Press to select a highlighted option or to
accept a displayed value.
1.3.3 Moving Within and Between Screens
The default option for a screen is highlighted when the screen first appears.
Moving Among Screen Options
To highlight an option on the screen, rotate the selector knob to move the cursor from screen
option to screen option. As you move the cursor to a new screen option, the option appears
highlighted.
For example, when you first access the Home screen, the Analyze/Monitor screen option appears
highlighted (see Figure 1.8). To highlight the Configure System option on the Home screen,
rotate the top of the selector knob to the right two times. The Configure System option appears
highlighted.
Moving Between Screens
When you choose a screen option, the control panel display shows the screen for the option you
chose. The default screen option for the new screen appears highlighted.
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1.3.4 Choosing a Screen Option or Parameter
To execute a screen option or edit a parameter, you must first choose it. Choosing a screen option
or parameter requires that you:
1. Highlight the screen option or parameter.
2. Accept the screen option or parameter.
To choose a screen option or parameter:
1. Rotate the selector knob to highlight the screen option or parameter.
2. Press the selector knob to accept the screen option or parameter.
or
Press OK on the keypad.
1.3.5 Entering Parameter Values
You can enter parameter values by using either the selector knob or the keypad.
Using the Selector Knob
To enter a value using the selector knob:
1. Choose the parameter you wish to change.
2. Rotate the selector knob to change the parameter value.
3. Press the selector knob to accept the displayed value.
Using the Keypad
To enter a value using the keypad:
1. Choose the parameter you wish to change.
2. Enter the desired value using the keypad.
3. Press OK to accept the displayed value.
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