Advanced Instruments 3300 User manual

The Advanced® Micro-Osmometer
Model 3300
User’s Guide
3305 Rev9 012303

Copyright
This user’s guide is copyrighted by Advanced Instruments, Inc. with all
rights reserved. Under copyright laws, this guide may not be reproduced
in any form, in whole or part, without the prior written consent of
Advanced Instruments.
© 2002 Advanced Instruments, Inc.
Trademarks claimed in this document are the property of Advanced
Instruments, Inc.
Advanced Instruments has reviewed this guide thoroughly. All material
contained within is believed reliable, but the accuracy and completeness
are not guaranteed or warranted, and are not intended to be representations
or warranties concerning the product described.
Hot-Line®Service
If you have any questions or problems regarding the proper operation of
your instrument, please contact our Hot-Line Service department:
•800-225-4034 (toll-free within the USA and Canada)
•+US 781-320-9000 (elsewhere)
•781-320-0811 (fax)
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Advanced®Osmometer Model 3300 User’s Guide

Table of Contents
Parts & Accessories v
Calibrators & Standards vii
Safe Use ix
Symbol conventions ix
General cautions x
Foreword: Theory and Technique xiii
Chapter 1 — Installation & Setup 1
Step 1 — Find a location for your instrument 1
Step 2 — Unpack your instrument 1
Step 3 — Obtain additional items 2
Step 4 — Power up your instrument 2
Step 5 — Set your date and time 3
Step 6 — Check your initial calibration 3
Step 7 — Proceed to instrument operation 4
Chapter 2 — Instrument Operation 5
Hazardous material cautions 5
Function of major components 5
Sample handling 10
Standards & controls 10
Sample test procedure 11
Sample test errors 12
Recall results 13
Repeatability tips 13
Statistics 14
Changing operating settings 15
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Using a barcode scanner with the Model 3300 17
Using a printer with your the Model 3300 18
Using the Model 3300’s RS-232 port 18
Instrument software update 19
Chapter 3 — Calibration 21
Calibration procedure 21
Calibration notes 22
Chapter 4 — Troubleshooting & Service 25
Service & maintainance cautions 25
Obtaining service 27
Troubleshooting checks 28
Internal diagnostics 28
Fuse replacement 33
Chamber cleaning 34
Sample plunger replacement and calibration 34
Appendices
Appendix A: Troubleshooting Table 37
Appendix B: Product Specifications 43
Appendix C: Regulatory Notices 45
Appendix D: Warranty and warranty duties 47
Appendix E: Supplemental RS-232 information 51
Appendix F: Service log 53
Index 55
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Advanced®Osmometer Model 3300 User’s Guide

Parts & Accessories
v
To order parts and accessories, contact the Advanced Instruments
Customer Service Department:
•800-225-4034 (toll-free within the USA and Canada)
•+US 781-320-9000 (elsewhere)
•781-320-3669 (fax)
PART DESCRIPTION
20-µL Sampler
Replacement Sample Probe
Micro-Sample Test Kit (500 tests)
Sampler Plunger Wires (2)
Thermal Printer with Interface Cable, Operation Manual,
Thermal Paper Roll, and Printer Power Supply*
*Note: Contact customer service to order the appropriate
power supply.
Printer Paper (5 rolls)
User’s Guide
Service Manual
Serial Cable
Software Upgrade**
Barcode Scanner
PART NO.
3M0825
330700A
3MA800
3M0828
330555
3D3835
3305
3305SM
RS232-Cable
330990
330016
** Note: 330990 upgrade kit comes with a 3 meter serial cable #330053.

vi
Notes:

vii
Calibrators & Standards
To order calibrators and standards, contact the Advanced Instruments
Customer Service Department:
•800-225-4034 (toll-free within the contiguous 48 states)
•+US 781-320-9000 (elsewhere)
•781-320-3669 (fax)
DESCRIPTION
ClinitrolTM Reference Solution (10 2-mL ampules)
Protinol®3-Level Protein Control Kit (9 3-mL bottles, 3 of
each level)
5-Value Osmolality Linearity Set (10 2-mL ampules, 2 of
each value)
50 mOsm/kg Calibration Standard (10 2-mL ampules)
850 mOsm/kg Calibration Standard (10 2-mL ampules)
PART NO.
3MA029
3MA028
3LA028
3MA005
3MA085

viii
Notes:

Safe Use
To reduce the risk of bodily injury, electric shock, fire, and
damage to your instrument, please read and observe the pre-
cautions in this User’s Guide.
• If the product is used in a manner not in accordance with
the equipment design, operating instructions or manufac-
turer's recommendations, the operation of the product may
be impaired to the extent that a safety hazard is created.
• Do not attempt to perform electrical work if you are not
fully qualified. This manual is not a substitute for electrical
training.
Symbol conventions
The exclamation point within an equilateral triangle is
intended to alert the user to the presence of important oper-
ating and maintenance (servicing) instructions in the litera-
ture accompanying this product.
The lightning flash with arrowhead symbol within an equi-
lateral triangle is intended to alert the user to the presence
of uninsulated dangerous voltage within the product's
enclosure that may be of sufficient magnitude to constitute
risk of electric shock to persons.
The static symbol within an equilateral triangle is intended
to alert the user to the presence of internal components that
could be damaged by static electricity.
This static symbol is intended to alert the user to the pres-
ence of a specific component that could be damaged by
static electricity.
ix

This symbol indicates the presence of alternating current (AC).
This symbol indicates the presence of a fuse.
This symbol indicates the presence of protective earth ground.
General cautions
• This product should be operated only with the type of power source
indicated on the product’s electrical ratings label. Refer to the instal-
lation instructions included with the product.
• If the power cord provided is replaced for any reason or if an alter-
nate cord is used, the cord must be approved for use in the local
country. The power cord must be approved for the product’s listed
operating voltage and be rated at least 20% greater than the ampere
ratings marked on the product’s electrical ratings label. The cord
end that connects to the product must have an IEC 60320 connector.
• Plug the product into an approved grounded electrical outlet.
• Do not disable the power cord’s grounding plug.
• If an extension cord or power strip is used, make sure that the cord
or strip is rated for the product, and that the total ampere ratings of
all products plugged into the extension cord or strip do not exceed
80% of the cord’s or strip’s rating limit.
• Route power cords so that they will not be walked on, tripped on, or
pinched by items placed upon or against them. Pay particular atten-
tion to the plug, electrical outlet, and the point where the cord exits
the product.
• Do not pull on cords and cables. When unplugging cords or cables,
grasp the corresponding connector.
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Advanced®Osmometer Model 3300 User’s Guide

• Do not install or use this product in any area subject to extreme
short-term temperature variations, or locations that exceed the speci-
fied operating environment temperatures.
• Never use this product in a wet area.
• To avoid injury or fire hazard, do not operate this product in an
explosive atmosphere.
• Do not install or use the product on an unstable, non-level work sur-
face.
• Do not operate this product with the covers removed or unsecured.
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Safe Use

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Advanced®Osmometer Model 3300 User’s Guide
Notes:

Foreword
Principles of Freezing-Point Osmometry
When a solute is dissolved in a pure solvent, the following
changes in the solution's properties occur:
•the freezing point is depressed,
•boiling point is raised,
•osmotic pressure is increased, and
•vapor pressure is lowered.
These are the so-called "colligative" or concentrative proper-
ties of the solution which, within reasonable limits, change in
direct proportion to the solute concentration; in other words,
the number of particles in solution.
Of the colligative properties, measurement of the freezing
point allows the concentration of an aqueous solution to be
easily determined with great precision.
The freezing point of pure H2O is precisely +0.010°C. One
mole of a non-dissociating solute such as glucose (where the
solute does not dissociate into ionic species, but remains
intact), when dissolved in 1 kilogram (kg) of water will
depress the freezing point by 1.858°C. This change is known
as the freezing point depression constant for water. The
freezing point depression also depends upon the degree of dis-
xiii

sociation of the solute. If the solute is ionic, the freezing point is
depressed by 1.858°C for each ionic species. For example, if one mole
of sodium chloride were to completely dissociate into two ionic species
(Na+ and Cl-) in 1 kg of water, the freezing point would be depressed
by 3.716°C. However, dissociation is never complete. Interference
between solute molecules reduces dissociation by a factor called the
osmotic coefficient.
In a simple solution such as glucose or sodium chloride in water, the
freezing point can be measured and the unit concentration easily deter-
mined from an equation or a reference table. However, the equation is
unique for each solute. In a more complex solution, all ionized and
non-dissociated species contribute to the freezing-point depression and
the concentration of each solute cannot be easily determined.
Each of the colligative properties has a similar problem, and though
each of the colligative properties changes in direct proportion to the
solute concentration, each requires a different mode & unit of measure-
ment. Osmolality is a common unit of concentration measurement that
can be used to relate all the colligative properties to each other, and to
other concentration units. Because of its universality, most osmometry
applications regularly use osmolality, expressed as "mOsm/kg H2O", as
the common unit of concentration rather than applying further conver-
sion factors.
Instrumentation
Advanced Osmometers are devices for the determination of the concen-
tration of solutions by means of freezing-point measurement.
Advanced Osmometers utilize high-precision thermometers to sense the
sample temperature, to control the degree of supercooling and freeze
induction, and to measure the freezing point of the sample. They can
routinely determine differences of ±1 mOsm/kg H2O.
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Advanced®Osmometer Model 3300 User’s Guide

Freezing-Point Thermodynamics
The quickest and most precise way to measure the freezing point of a
solution is to supercool it several degrees below its freezing point. It is
unstable in this state, and a mechanical agitation induces crystallization.
The heat of fusion suddenly liberated causes the sample temperature to
rise toward a plateau temperature, where a liquid/solid equilibrium
occurs. The equilibrium temperature is, by definition, the freezing point
of the solution. Managing the plateau temperature for precise measure-
ment is the basis for several patents issued to Augustus Fiske.
The time over which liquid/solid equilibrium develops and is main-
tained, is a function of the speed with which the heat-of-fusion is liber-
ated vs. the speed it is transferred away, or absorbed, by the surrounding
environment. This ratio can be slowed and the equilibrium time
stretched, to give a distinct plateau height measurable to 0.001°C.
Sensitive thermistor probes monitor the sample temperature and control
the thermoelectric cooling element. Microprocessor control and auto-
mated operation minimize imprecision due to operator technique.
Figure 1 illustrates the temperature of a sample as it progresses through
the freezing cycle and shows the action of the instrument at each stage
of the cycle.
xv
Foreword
Figure 1: Standard Freezing Curve

Definitions
Solution: A homogeneous mixture of solute and solvent in which the
solvent is usually the major component, and the solute is the minor one.
Concentration: The ratio of solute to a given amount of solvent
(molal), or ratio of solute to solution (molar).
The amount of solute is usually expressed in terms of moles, i.e., gram
molecular weights. One mole = 6.028 x 1023 molecules (Avogadro's
number). One mole of glucose (180.2 g) and one mole of sodium chlo-
ride (58.4 g) each contain Avogadro's number of molecules.
Common units of concentration are:
•Molality: Moles of solute per kilogram of pure solvent.
•Osmolality: Osmols of solute particles per kilogram of pure sol-
vent. As noted above, most ionic solutes do not completely dis-
sociate. Osmolality is a unit of concentration that takes into
account the dissociative effect. Osmolality is usually expressed
in mOsm/kg H2O. One milliosmol (mOsm) is 10-3 osmols.
Osmolality is defined as:
where:
ø = osmotic coefficient, which accounts for the degree of mole-
cular dissociation.
n = number of particles into which a molecule can dissociate.
C = molal concentration of the solution.
•Molarity: Moles of solute per liter of solution.
•Osmolarity: Osmols of solute particles per liter of solution.
Although molarity and osmolarity may be common units of
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Advanced®Osmometer Model 3300 User’s Guide
OHkg
osmol
nCOsmolality
2
== φ

measurement in other branches of chemistry, they are not used in
osmometry because the ratio of solute to solution is not linear.
Molality and osmolality are linear, independent of the effect of
temperature and volume displaced by solute. A calculated con-
version between units of molality and molarity is complex and
generally unnecessary when the terms are properly understood.
Freezing Point/Melting Point: The temperature at which the liquid
and solid phases of a substance will remain together in equilibrium.
Freezing-Point Depression: When a solute is added to a solvent, the
freezing point of the solvent is lowered. In aqueous solutions, one
mOsm of solute per kilogram of water depresses the freezing point by
1.858 millidegrees Celsius (m°C).
Supercooling: The tendency of a substance to remain in the liquid state
when cooled below its freezing point.
Crystallization Temperature: Aqueous solutions can be induced to
freeze (i.e., crystallize) most reliably when supercooled. When super-
cooled, agitating the solution (freeze pulse) induces crystal formation.
The crystallization temperature is the temperature at which crystalliza-
tion is induced. During crystallization, the heat of fusion raises the tem-
perature of the sample to an ice/water freezing-point plateau.
Heat of Fusion: The heat released when the mobile molecules of a liq-
uid are frozen into rigid ice crystals.
Freezing-Point Plateau: The constant temperature maintained during
the time that ice and liquid exist in isothermal equilibrium after crystal-
lization occurs.
xvii
Foreword

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Advanced®Osmometer Model 3300 User’s Guide
Notes:

1Installation & Setup
In order to set up your instrument properly, it is important that
you read and follow the steps in this section. Please follow these
steps carefully and be sure to read Chapter 2 — Operating Your
Osmometer before attempting to run tests on your instrument.
Step 1 — Find a location for your instrument
When choosing a location for your new osmometer, be sure to
meet the following criteria.
• Adequate space. The dimensions of the instrument are
13 × 10.5 × 15.5 inches (26.7 × 33 × 39.4 cm). Be sure to
keep your workplace free of debris, especially near the
front of the instrument where proper ventilation is needed.
•Electric outlet availability. Your instrument will need to
operate within five feet of a properly grounded, three-
prong electrical outlet capable of continuously supplying 1
ampere at 200-250V to 1.25 amperes at 100-130V. If the
instrument is not grounded properly, its operation may be
impaired and a safety hazard may exist. Therefore, be sure
to test the outlet and record the results before operating
your instrument.
Step 2 — Unpack your instrument
To unpack your osmometer, take the following steps.
a. Carefully unpack your osmometer, accessories and supplies
and inspect them for shipping damage. Use the enclosed
packing list to verify that all items have been received.
1

b. Save your osmometer's shipping boxes and packaging material in
case future transport of the instrument becomes necessary.
c. If any item on the packing list appears to be missing from your ship-
ment, please search carefully through and under all packing materi-
als. If the item is not found, notify your receiving department imme-
diately. Advanced Instruments can only be responsible for items
reported missing within 10 days of a shipment’s arrival.
d. If you receive any damaged items, save the cartons and packing
material those items came in for inspection by the insurer. The carri-
er, dealer, and Advanced Instruments must be notified within 24
hours in order for your warranty and insurance to apply. Have the
transportation company inspect items, fill out a “Report of Concealed
Damage,”and file your claim. Then, notify Advanced Instruments
immediately for repair or replacement.
e. Fill out the postage-paid “(U.S.A. Only)”warranty card enclosed.
Mark the appropriate boxes if you wish to receive additional informa-
tion. Customers outside of the United States may fax the warranty
card to 781-320-8181.
Step 3 — Obtain additional items
Soft, no-lint, non-ionic paper tissues are needed for wiping the sample
cells prior to testing. Please be sure that you have an adequate supply on
hand before attempting to run tests on your instrument.
Step 4 — Power up your instrument
Turn your instrument on by pushing the rocker-style power switch on
the instrument’s back panel into the on ( |) position.
Each time your instrument is turned on, it automatically runs a one-
minute setup and self-diagnostic program. During these diagnostics, your
display and printer (if in use) display critical instrument information, such
as software version and block and sample probe bin numbers. The first
time you power up your osmometer, record the software revision and
Advanced®Osmometer Model 3300 User’s Guide
2