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USER MANUAL

English

TEUS00015-03-ING
June-2007
User Manual

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Dear Customer,

Thank you for purchasing our A25 automatic analyser. We are sure that its specifications will make it a valuable instrument
in your laboratory. Although its operation and use is logical and straightforward, using a programme that runs under
Windows, please read this manual carefully. It will help you with both installation and correct programming, use and
maintenance of the instrument, enabling you to get maximum performance from its multiple possibilities.

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User Manual

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TABLE OF CONTENTS
Introduction .............................................................................7
1. Description of the instrument ............................................8
1.1. Components ............................................................................................................... 8
1.1.1. Operating arm ........................................................................................................................ 8
1.1.2. Dispensing system .................................................................................................................. 8
1.1.3. Reactions rotor and reading .................................................................................................. 9
1.2. Working theory ......................................................................................................... 10
1.3. Start-up ..................................................................................................................... 10
1.4. Functional specifications ........................................................................................ 10
2. General operation method................................................ 13
2.1 Program Installation .................................................................................................. 13
2.2. Operation procedure ................................................................................................ 13
2.2.1. Introduction .......................................................................................................................... 13
2.2.2. Start-up procedure, operation and shutdown. .................................................................... 14
2.2.3. Monitor ................................................................................................................................. 15
2.2.3.1. Status of the analyser and the work session. ................................................................... 15
2.2.3.2. Blocking pending samples ............................................................................................... 15
2.2.3.3. Vertical button bar for the control of the analyser ........................................................... 15
2.2.3.4. Horizontal button bar ........................................................................................................ 16
2.2.3.5. Special buttons in monitor screen ................................................................................... 16
2.2.4. Programming ....................................................................................................................... 17
2.2.4.1. Test programming ............................................................................................................. 17
2.2.4.2. Profile programming: ........................................................................................................ 19
2.2.4.3. Programming of Multiple Calibrators ............................................................................... 20
2.2.4.4. Programming of Multiple Controls ................................................................................... 21
2.2.4.5 Calculated tests .................................................................................................................. 21
2.2.4.6. Reagent Rack Programming ............................................................................................ 22
2.2.5. Save/load test file ................................................................................................................ 22
2.2.6. Preparation of the work session .......................................................................................... 22
2.2.6.1. Introduction of New Samples ........................................................................................... 22
2.2.6.2. Positioning of samples and reagents ............................................................................... 24
2.2.7 Memorising sessions .......................................................................................................... 26
2.2.8. Current results ...................................................................................................................... 26
2.2.8.1. Calculated factor modification ......................................................................................... 27
2.2.8.2. Repetitions ........................................................................................................................ 27
2.2.9. Current and past alarms ...................................................................................................... 27
2.2.10. Past results ......................................................................................................................... 28
2.2.11. Patient data ........................................................................................................................ 29
2.2.12. Configuration ...................................................................................................................... 29
2.2.12.1. A25 analyser .................................................................................................................... 29
2.2.12.2. Languages ....................................................................................................................... 31
2.2.12.3. Serial port ........................................................................................................................ 31
2.2.12.4. Header of results reports ................................................................................................. 31
2.2.13. About .................................................................................................................................. 31
2.2.14. Utilities ................................................................................................................................ 32
2.2.14.1. PC-Analyser communications channel test .................................................................... 32
2.2.14.2. Reactions rotor check ..................................................................................................... 32
2.2.14.3. Disassembly of the dispensing needle ........................................................................... 32
2.2.14.4 Reset of the base line historic ......................................................................................... 33
2.2.14.5. Fluid system supply ......................................................................................................... 33
2.2.14.6. Cleaning of the dispensing system ................................................................................. 33

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User Manual

2.2.14.7. Changing the lamp .......................................................................................................... 33


2.2.14.8 Change the rotor type ...................................................................................................... 33
2.2.15. Internal quality control....................................................................................................... 33
2.2. Alarms and alerts ...................................................................................................... 35
3. Calculation and measurement procedures ..................... 37
3.1. End point ................................................................................................................... 38
3.1.1. Absorbance .......................................................................................................................... 38
3.1.1.1. Monoreagent / Bireagent .................................................................................................. 38
3.1.1.2. Monochromatic / Bichromatic........................................................................................... 38
3.1.2. Concentration ....................................................................................................................... 38
3.1.2.1. Factor................................................................................................................................. 38
3.1.2.2. Single calibrator ............................................................................................................... 38
3.1.2.3. Several calibrators ............................................................................................................ 38
3.1.3. Replicates ............................................................................................................................. 38
3.1.3.1. Blank.................................................................................................................................. 38
3.1.3.2. Calibrator........................................................................................................................... 38
3.1.3.3. Sample .............................................................................................................................. 38
3.2. Bireagent differential ................................................................................................ 39
3.2.1. Absorbance .......................................................................................................................... 39
3.2.2. Concentration ....................................................................................................................... 39
3.2.2.1. Factor................................................................................................................................. 39
3.2.2.2. Single calibrator ............................................................................................................... 39
3.2.2.3. Several calibrators ............................................................................................................ 39
3.2.3. Replicates ............................................................................................................................. 39
3.2.3.1. Blank.................................................................................................................................. 39
3.2.3.2. Calibrator........................................................................................................................... 39
3.2.3.3. Sample .............................................................................................................................. 39
3.3. Fixed time .................................................................................................................. 39
3.3.1. Absorbance .......................................................................................................................... 39
3.3.1.1. Monoreagent / Bireagent .................................................................................................. 40
3.3.2. Concentration ....................................................................................................................... 40
3.3.2.1. Factor................................................................................................................................. 40
3.3.2.2. Single calibrator ............................................................................................................... 40
3.3.2.3. Several calibrators ............................................................................................................ 40
3.3.3. Replicates ............................................................................................................................. 40
3.3.3.1. Blank.................................................................................................................................. 40
3.3.3.2. Calibrator........................................................................................................................... 40
3.3.3.3. Sample .............................................................................................................................. 40
3.4. Kinetics ..................................................................................................................... 40
3.4.1. Variation of the absorbance per unit of time ...................................................................... 40
3.4.1.1. Monoreagent / Bireagent .................................................................................................. 41
3.4.1.2. Checking linearity ............................................................................................................. 41
3.4.2. Concentration ....................................................................................................................... 41
3.4.2.1. Factor................................................................................................................................. 41
3.4.2.2. Single calibrator ............................................................................................................... 41

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Introduction
The A25 analyser is an automatic analyser for random diluted. The analyser automatically orders the tests of each
access In Vitro Diagnosis specially designed for performing patient to avoid incompatibilities between reagents and to
biochemical and turbidimetric clinical analyses.The optimise performance. From the Monitor screen, which is
instrument is controlled on-line in real time from an external particularly graphic, the analyser keeps the user informed
dedicated PC. The analyser performs patient-by-patient about its functioning status and the result-obtaining process.
analyses and enables the continual introduction of samples. All these characteristics, together with the variety of possible
The results are shown immediately after each measurement. analysis modes (final monoreagent or bireagent point
The high-speed preparation of the reactions makes it ideal (monochromatic or bichromatic), bireagent differential, fixed
for medium-capacity automated laboratories. The flexibility monoreagent or bireagent time and monoreagent or
of the samples and reagents racks system enables the bireagent kinetic), make the A25 analyser an instrument
perfect adjustment of the capacity of the analyser to the that is highly versatile and effective for clinical analysis
specific needs of each laboratory. laboratories.

In each of the elements of the A25 analyser, BioSystems


has used leading edge technology to obtain optimum
analytical performance, as well as taking into account
economy, robustness, easy use and maintenance. A three-
axis Cartesian operating arm prepares the reactions.
Dispensing is performed by means of a pump with a ceramic
piston via a detachable thermostatised needle with Fuzzy
Logic control. A washing station guarantees that the needle
is kept perfectly clean throughout the process. The
reactions take place in a thermostatised rotor in which
absorbance readings are taken directly by means of an
integrated optical system.

The elaborate computer programme enables the easy con-


trol of the extensive functionality of the analyser. The user
interface is flexible, simple and very graphic, clearly
separating the daily routine tasks from the less frequent
tasks. All the data and results are stored securely, with the
possibility for exportation. Various initialisation, functioning
and shutdown options can be configured for the analyser.
The user can programme an indefinite number of
measurement procedures (techniques), technique profiles
and reagent racks, depending merely on the computer’s
hard disk capacity. The analyser works with 5 sample types:
serum, urine, plasma, whole blood and cerebrospinal. Each
measurement procedure can be adapted to each one. During
a work session, the user can continually introduce normal
or urgent samples in the analyser without interrupting the
measurements in progress. The samples may be pre-
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User manual

1. Description of the instrument


The A25 analyser prepares the reactions by means of a of the instrument. If the user wishes to manually move the
three-axis Cartesian operating arm. This arm supports and arm to its resting position, he or she must ensure that the
displaces a dispensing needle which pre-thermostatises needle is at its highest position. For this, it must be raised
the preparations at 37º. The dispensing is carried out by completely by pressing the vertical chain of the operating
means of a low-maintenance ceramic piston pump. The arm upwards as shown in the photograph. The return system
analyser can carry out one preparation every 15 seconds. prevents the needle from descending and the user can then
The preparations are dispensed in a reactions rotor move the arm with no danger of damaging the needle or
thermostatised at 37ºC. The optical absorbance readings injuring him or herself on it. The operating arm only makes
are taken directly on this rotor. the preparations if the general cover of the analyser is closed.
If the cover is raised while it is functioning, the arm
automatically aborts the task in progress and returns to its
1.1. Components parked position to avoid injury to the user.
The A25 analyser is made up of three basic elements: the
operating arm, the dispensing system and the reading and
reactions rotor.

1.1.1. Operating arm

This is a three-axis XYZ Cartesian mechanism. The X and


Y axes move the dispensing needle over the analyser
horizontally and the Z axis moves it vertically. It is operated
by three step-by-step motors. In each 15-second
preparation cycle, the operating arm performs the following
actions: first of all, it sucks in the reagent from the
corresponding bottle. Next, the needle is washed externally
in the washing station and sucks in the sample from the
corresponding tube. It is washed externally again and dis-
penses the sample and the reagent into the reactions rotor.
Finally, it is exhaustively washed internally and externally
before proceeding with the next preparation. The arm has a
system for controlling vertical movement to detect whether 1.1.2. Dispensing system
or not the needle has collided into anything on descending. This system consists of a thermostatised needle, supported
If a collision occurs, as may be the case if, for example, a and displaced by an operating arm and connected to a
lid has been left on a bottle of reagent, the arm automatically dispensing pump. The needle is detachable to enable
restarts, verifies the straightness of the needle and continues cleaning and replacement. The analyser has capacity level
working issuing the corresponding alert to the user. A return detection to control the level of the bottles and tubes and
system on the vertical axis automatically raises the needle prevent the needle from penetrating too far into the
in the case of a power failure, preventing it from descending corresponding liquids, thus minimising contamination. An
and colliding with an object at the bottom or with the housing automatic adjustment system informs the user if the needle
is not mounted or if it is too bent. The needle has a
sophisticated Peltier thermostatisation system, with Fuzzy
Logic control, capable of thermostatising the preparations
at approximately 37º in less than 6 seconds. Dispensing
is carried out by means of a low maintenance ceramic
piston pump driven by a step-by-step motor. It is capable
of dispensing between 3 and 1250 µL. The dispensing
system of the analyser uses system liquid which the user
must prepare by adding 6 ml of concentrated system liquid
supplied with the analyser to the corresponding container
filled to the top with distilled water (approx. 2700 ml).
Analyses must never be made using distilled water only in
the dispensing system, since the analytical performance
and the lifetime of the system may notably be lowered.
The exterior of the needle is kept constantly clean by means
of a washing station, which consists of a font specially

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designed to clean and dry the needle, integrated in the take the optical reading is 200 µl. The wells have a maximum
racks tray. A system of membrane pumps supplies the useful capacity of 800 µl. When the reactions rotor is
font with system liquid and transports the waste to its con- completely full, the user must change it for one that is
tainer. empty, clean and dry. The reactions rotors may be reused
if they are appropriately cleaned immediately after use.
The A25 analyser has a tray with 6 free positions for racks The Cleaning the semi-disposable reactions rotor section
of reagents or samples, plus three fixed positions for bottles in the Installation and maintenance manual describes how
opposite the washing station. Each reagents rack can ca- to clean the rotors. The rotors must be rejected if they are
rry up to 10 reagents in 20 ml or 50 ml bottles. Each samples noticeably deteriorated. The user has a test in the computer
rack can contain up to 24 tubes of samples. The samples programme, which he or she may use to check the
can be patients, calibrators or controls. The analyser can condition of the rotor. The rotor is driven by a step-by-step
be configured to work with 13 mm or 15 mm diameter tubes motor with a transmission. A Peltier system with PID con-
of samples with a length of up to 100 mm or with paediatric trol thermostatises the rotor at 37ºC.
wells. Any possible configuration of racks can be mounted
An optical system integrated in the rotor takes the readings
directly on the reaction wells. The light source is a 20 W
halogen lamp. The detector is a silicon photodiode. The
wavelength is selected by a drum with 9 positions available
for optic filters. The filters are easily changed by the user
from the exterior of the analyser, without the need for
disassembling the filter drum. A step-by-step motor
positions the drum. The optical system is capable of taking
5 readings per second, with or without a filter change in
between. The light beam of the lamp passes through a
compensated interferential filter to select the desired
wavelength and through focalisation lenses. It then passes
through the rotor well and finally reaches the photodiode,
where the light signal is turned into an electric signal. A
sophisticated analogical digital integrator-converter system
converts the electric signal into a digital value with which
the analyser obtains the absorbance values. The optical
from 1 rack of reagents (10 reagents) and 5 racks of samples system continues to work when the general cover of the
(120 samples) to 5 racks of reagents (50 reagents) and 1 analyser is open, whereby the analyser can continue to
rack of samples (24 samples). Any reagent may be placed take readings while the user handles, for example, the
in the fixed positions, but it is recommendable to use them sample tubes or the reagent bottles. The rotor cover must
for the bottles of distilled water, saline solution for the be in place for the optical system to work correctly. A de-
automatic pre-dilutions and washing solution. The rack tray tector tells the analyser of the presence of said cover. The
detects and identifies the type of racks. In this way, if the analyser aborts the readings if the user removes the rotor
physical disposition of the racks does not coincide with cover while the optical system is taking photometric
that programmed on the computer, the analyser alerts the measurements. If the rotor is not covered, the analyser
user. informs the user so that he or she places the rotor cover
when it sends samples to be analysed. The analyser also
On the left of the analyser are system liquid containers issues an alert if there is no reactions rotor in it.
(marked in blue) and waste containers (marked in red).
The analyser constantly controls the level of these
containers and issues the appropriate alerts if the system
liquid is nearly empty or if the waste container is full.

1.1.3. Reactions rotor and reading

The preparations are dispensed in an optical quality


methacrylate reactions rotor thermostatised at 37ºC. The
optical absorbance readings are taken directly on this rotor.
Each reaction can be read for 15 minutes. The readings
are taken as they are programmed in each measurement
procedure. The reaction wells have been designed to enable
the mixture of the sample and the reagent during the
dispensing. Each rotor has 120 reaction wells. The length
of the light path is 6 mm. The minimum volume required to

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User manual

1.2. Working theory section for routine start-up. The analyser must be in standby
mode, i.e. plugged into the mains and with the switch in
The A25 analyser is an automatic random access analyser the (I) position. The computer must be started up and the
specially designed for performing biochemical clinical programme launched. From the Monitor screen of the
analyses. The analyser performs patient-by-patient programme, click on the Warm up button to start
analyses and enables the continual introduction of samples. initialisation. The analyser automatically carries out all
The analyser is controlled from a dedicated PC that is the checks required for correct functioning and continually
permanently communicated to the instrument. The informs of its status through this screen. No manual
programme, installed on the computer, keeps the user adjustment is necessary. If, for any reason, user intervention
constantly informed of the status of the analyser and the is necessary, the analyser issues the corresponding alert
progress of the analyses. As results are obtained, the through the computer. Once warm-up has finished, the
computer shows them to the user immediately. analyser is in standby mode, ready for work.
When a Work Session is begun, the analyser proposes
performing the blanks, calibrators and controls programmed 1.4. Functional specifications
for the measurement procedures it is to carry out. The user
may choose between performing the blanks and the All the functions of the analyser are controlled from the
calibrators or not. If they are not performed, the analyser programme installed on the computer. This programme has
uses the last available memorised data. The controls can a flexible, simple and highly graphic interface, with a clear
also be activated or not. During a session, while the analyser separation between routine and less frequent tasks. Its
is working, the user can introduce new normal or urgent flexibility makes routine use of the analyser extremely sim-
samples to be analysed. Each time a new sample is added, ple. It is also possible to access numerous functions to
the analyser automatically proposes the possible new fully customise the functioning of the analyser. All the data
blanks, calibrators or controls to be performed. A work and results are stored securely, with the possibility for
session can remain open for one or more days. When a exportation. The configuration and programming data can
session is closed and another new session is opened be easily updated or stored.
(Reset Session), the analyser again proposes performing
the blanks, calibrators and controls. It is recommended
that the session is reset each working day.

The analyser determines the concentrations of the analytes


based on optical absorbance measurements. To measure
the concentration of a certain analyte in a sample, the
analyser uses a pipette to take a specific volume of the
sample and the corresponding reagent, quickly
thermostatises them in the needle itself and dispenses them
into the reactions rotor. The very dispensing speed together Power off
with the geometry of the reaction well causes the mixture
to be shaken and the chemical reaction begins. In the
bireagent modes, the reaction begins when the analyser
later dispenses a second reagent in the same reaction well. Sleeping
The reactions can be biochemical or turbidimetric. In both
cases, the reaction or the chain of reactions produced
generate substances that attenuate certain wavelengths, Shutdown

either by absorption or by dispersion. Comparing the light


intensity of a certain wavelength that crosses a well when Stand by Warming up
there is a reaction and when there is not a reaction can
determine the concentration of the corresponding analyte.
This comparison is quantified with the physical magnitude
called absorbance. In some cases, the concentration is a Running
direct function of the absorbance, and in other cases, it is
a function of the variation of the absorbance over time,
depending on the analysis mode.

Sampling Stop
1.3. Start-up
Once the analyser has been installed, it can be started up.
The steps to be followed for the first start-up are the same
as those indicated in the Start-up and shutdown procedure Alarm
Mode

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STATUSES OF THE ANALYSER - Several calibrators (Multipoint calibrator). This can be
specific (for a certain test) or multiple (common for several
- Sleeping Mode: The analyser is connected to the mains tests). Up to 8 points and up to 3 replicated points for
with the switch in (l) position, but it is not working. The each one. Calculation of the Calibration Curve (spline,
analyser can be left in this status indefinitely. Power linear regression, quadratic or polygonal regression, with
consumption is minimal linear or logarithmic axes).
- Warm-up: Transitory status during which the analyser
initialises all its elements and performs the checks
required to be able to begin analyses. Once warm-up PROGRAMMING
has finished, the analyser is in standby mode, ready for
performing analyses. - Tests: unlimited number, depending on the hard disk
capacity of the computer. Programmable for 5 sample
- Shutdown: Transitory status during which the analyser types: serum, urine, plasma, cerebrospinal and whole
appropriately switches off all its elements to maintain blood .
them in optimum condition for future analysis work. Once
finalised, the analyser is in standby mode. - Test profiles: unlimited number, with an unlimited number
of tests.
- Standby mode: The analyser is in communication with
the PC. If the initialisation process has taken place, the - Multiple calibrators: up to 10.
analyser is ready to perform analyses.
- Multiple controls: up to 20.
- Running Mode: The analyser is performing analyses
- Contaminations: unlimited number.
(preparations and readings or readings only if there is no
preparation pending). - Reagent racks: unlimited number.
- Sampling stop: The analyser can be taking readings, but OPERATION
the operating arm is parked and the pending reactions
are not prepared. In this status, the user can handle the MONITOR SCREEN
bottles and containers of the analyser with no risk, in
- Warm-up and shutdown process of the analyser .
order to, for example, add a new sample.
- Cancellation of the preparations and readings in process
- Alarm Mode: If during any of the operations, an anomaly
(Abort)
occurs requiring the intervention of the user and whose
correction is essential for the process to continue, the - Real-time monitoring of the status of the analyser and
analyser enters alarm mode and issues the corresponding development of the work session. Display of the tasks
alert through the PC. sent to the analyser and the advance of their execution.
- Mains disconnection: Switch (0) (Power Off): The switch - Access to the results of the measurements, with the
is in the (0) position. The analyser is completely off. It possibility of automatic printing.
can then be disconnected from the mains.
- Alerts and alarms. Access to detailed explanations.
ANALYSIS MODES
- Access to introduction of new samples at all times.
- Final point. Monoreagent or bireagent. Monochromatic
or bichromatic. - Interruption and renewal of the preparation of reactions
(Sampling stop / Continue).
- Bireagent differential.

- Fixed time. Monoreagent or bireagent.


PREPARATION OF THE WORK SESSION
- Kinetic. Monoreagent or bireagent. Readings every 15 s,
up to 15 min Linear regression analysis, with linearity - Sample types: Normal patient, urgent patient, blank,
evaluation. Automatically extracted kinetic blank. calibrator and control.

- One patient can have up to 5 different types of samples.

CALIBRATION TYPES - Easy allocation of the tests and profiles to be performed


on one or several samples.
- Factor
- Up to 50 replicates for patient samples. Up to 3 replicates
- Single calibrator (Single-point calibrator). This can be for blanks, calibrators and controls.
specific (for a certain test) or multiple (common for several
tests).

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User manual

- Automatic allocation of blanks and calibrators. Possibility


of storing the results of blanks and calibrators.
CONFIGURATION
- Partial or total sending of work to the analyser.
- Options of the user programme (languages, printing,
- Possibility of introducing patient data while the analyser reports).
is performing the analyses.
- Options for the functioning of the analyser (washing,
- Automatic selection of the reagents required and sample tubes, filter drum, cover detection, rack detection).
indication of the minimum required volumes.
UTILITIES
- Graphic, automatic or manual allocation of sample and
reagent positions in the racks. - Analyser test, preparation and maintenance utilities
(needle disassembly, supply, washing, rotor verification,
- Printable summary of positions to enable the positioning lamp change, PC-analyser communications,
of the samples and the reagents in the racks. demonstration).

- Verification of the correspondence of the programmed


racks with the racks of the analyser.
QUALITY CONTROL
- Automatic reordering of the tests of each sample to
optimise work and avoid contamination between - Analytical limitations control for each test: blank, linearity,
programmed tests. detection, factor.

- Automatic verification of the resources available on the - Internal quality control with up to 2 control materials per
analyser to be able to proceed with the required analyses. test. Manual or statistical calculation. Levey-Jennings
Issuing of alerts with instructions for the user if his or her graphics and Westgard algorithm.
intervention is required.

PROCESSING OF RESULTS

- Current or historical reports, by patients or test, with


possibility of repetition to confirm results.

- Current or past reports, by patients or by tests.

- Patient database.

- Internal quality control reports.

- Alarm and alert records.

- Exportation of results.

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2. General operation method
This chapter gives the basic instructions for using the corresponding blanks, calibrators and controls. The user
instrument. Although the operation of the analyser is very can choose to perform them or reuse the data from the
simple and intuitive, it is recommended you read this chapter previous session. Until the work session is restarted, the
carefully to make the most of its features effortlessly and analyser will not propose the repetition of the blanks,
as quickly as possible. calibrators and controls that have already been carried out,
even if the analyser is switched off, maintaining all the
information of the session in progress. To obtain maximum
2.1 Program Installation quality of the analytical results, it is recommended that
For the program installation, you must carry out the following you perform all the corresponding blanks, calibrators and
steps: controls in each working day. This implies restarting the
work session daily and always choosing to perform new
a) Initiate the computer blanks, calibrators and controls.
b) Uninstall the old version with the Windows option Add
and Remove programs Once the samples have been introduced, the user must
c) Insert the D-ROM with the new version. allocate positions to the samples and to the required
d) Press Start, select Execute and write: name of the CD- reagents and physically place them in the analyser. The
ROM unit:\setup (for example: D:\Setup). Positions screen distributes the samples and the reagents
in the different racks of the analyser tray. This distribution
From version 2.5.0, the program keeps a copy from all the can be made automatically or manually.
data files of the previously installed version, in the
As soon as the required elements have been distributed,
subdirectory \previous of the application directory (generally
the work session can be started. The Monitor screen is
c:\program files\A25).
automatically displayed, containing a summary of the main
Should there be an error during installation, the user can
information received from the analyser and showing the
install the previous version again and recover the old data.
progress of the work session. From this screen, it is possible
Follow the steps to recover a previous version:
to access other screens that offer more detailed information
about results and alarms. The user can cancel the work
a) Uninstall the version of the problem. Do not erase the
session if he or she detects that the results are incorrect.
created subdirectories or the files generated by the
application. At any moment, the user can introduce urgent samples or
b) Install the previous version add new normal samples to the work session in progress
c) Execute it. When initiating the application, it will restore from the Introduction of new samples screen without losing
the old data files. This process will solely be made from the information of the samples that have been sent
version 2.5.0 on. previously. If the sample is normal, it is added at the end of
the list. If it is urgent, and there are no other urgent samples
in progress, the current sample is interrupted and the urgent
2.2. Operation procedure one is introduced in first place. If there are already other
urgent samples, it is added to the end of the group of urgent
2.2.1. Introduction samples. If a new sample is added, with a test that has not
The operation of the analyser is flexible and simple using yet been performed in the current session, the analyser
the user programme from the dedicated PC. This programme will propose new corresponding blanks, calibrators and
is subdivided into different screens that provide specific controls. The user again has the possibility of choosing
functionality. They are described in more detail in the whether to perform them or not.
following sections. The system can automatically block samples or tests in
Firstly, the user must switch on the analyser and launch accord with the information received from the analyser. This
the user programme, and warm up the analyser from the can happen, for example, when a sample or a reagent has
Monitor screen. To start a work session, the samples that run out. When the user corrects the problem and notifies
are to be analysed must be introduced through the the analyser from the Positions screen, the blocked
Introduction of New Samples screen, indicating the samples or tests are automatically unblocked and the
measurement procedures desired. Each measurement affected samples are turned into pending samples, and are
procedure or test is defined by a set of parameters that analysed later.
can be programmed by the user from the Tests programming The user can access the results as they are obtained and
screen. print the results of the patients whose tests have already
In accord with the tests to be carried out, the analyser finalised.
automatically proposes the performance of the The basic screens of the user programme are as follows:

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User manual

- Monitor screen: Main screen that appears when the user issues the corresponding alert through the computer. Once
programme is launched. It displays the status of the warm-up has finished, the analyser is in standby mode,
analyser and the work session in progress. It lets the ready for work.
user intervene in the progress of the work session. It
contains the main buttons for controlling the analyser. Once the work session has been prepared from the
corresponding screens and warm-up has finalised, click
- Programming screens: These enable the programming the Start button on the vertical bar for the analyser to start
of tests, profiles, multiple calibrators, multiple controls, performing the analyses. On beginning the analyses, the
contaminations and reagent racks. analyser enters running mode. If during the running mode
- Work session preparation screens: These enable the you wish to physically introduce a sample or a reagent in
preparation of the work session and the storage of data the analyser, click on the Sampling Stop button on the
that may be used in later sessions. Basically, the user vertical bar menu and the analyser enters running mode
can introduce new samples to be analysed and position with readings only, without the preparation of samples
the corresponding samples and reagents in the analyser. (Sampling stop). The user can then raise the cover and
place the sample or reagent in safety. Closing the cover
- Results screens: These make it possible to view the results and clicking on the Continue button on the same bar, the
and see and print the reports with the results obtained in analyser returns to running mode. When the analyser ends
the current or previous sessions. The reports can be the analyses, it automatically returns to standby mode. If
organised by patients or by tests. It also allows managing you wish to abort the preparation of new reactions as well
sample repetitions as the readings in progress, click on the Abort button
- Alarms screens: These give detailed information about located on the same bar. In this case, the analyser requests
the alarms and alerts occurring in the current session or confirmation and, if the response is affirmative, it aborts
in previous sessions and enable the printing out of reports. the preparations and readings in progress, and returns to
standby mode.
- Patient data screen: This enables the introduction and
modification of the patient data sent to the analyser. To switch off the analyser, click on the Shutdown button on
- Configuration screens: These make it possible to confi- the vertical menu bar from standby mode. The programme
gure the working mode of the user programme and the asks for confirmation and if the response is affirmative, it
analyser. performs the shutdown sequence and returns to sleeping
mode. The analyser can be left in this status indefinitely.
- Utility screens: These let the user access the different
Power consumption is minimal. The user programme can
test, preparation and maintenance utilities of the analyser.
be closed and the computer switched off. If you wish to
- Quality control screens: These enable access to the switch off the analyser completely, for example to carry
internal quality control results of the analyser. out a maintenance operation or repair a breakdown, the
switch must be put in the (0) position (Power off) and the
The following sections describe these screens and their
mains cable unplugged.
functionality in more detail. You can access any of them
easily from the main menu and specific buttons. If during any of the operations, an anomaly occurs requiring
the intervention of the user and whose correction is essential
for the process to continue, the analyser enters alarm mode
2.2.2. Start-up procedure, operation and shutdown.
and issues the corresponding alert through the PC. Once
Before starting up the analyser, the user must check that the user has solved the problem and has indicated this to
the waste container is empty, that the system liquid con-
tainer is full and that a reactions rotor is correctly installed. Statuses of the analyser LED
To start up the analyser, the analyser must be in sleeping
mode, i.e. plugged into the mains and with the switch in Sleeping Mode Orange
the (I) position. Reboot the computer and launch the user
programme. The computer automatically communicates Warm-up Green
with the analyser, which changes to standby mode. The Shutdown Green
screen that appears when the programme is launched is
the Monitor screen. The programme shows horizontal and Standby mode Green
vertical button bars. The horizontal bar enables access to
the main functions of the application, whereas the vertical Running Mode Green
bar enables the control and functioning of the analyser.
Running mode with readings only
Click on the Warm up button on the vertical bar menu to
(Sampling Stop) Red
begin Warming up. The analyser automatically performs
all the checks required for its correct functioning and the Alarm Mode Flashing red
programme continually informs the user of the status of
the analyser. No manual adjustment is necessary. If, for Mains disconnection:
any reason, user intervention is necessary, the analyser Switch (0) (Power Off): Off

14
the analyser, the analyser checks that it has indeed been new rotor is placed in position, which enables the
solved and returns to the previous mode. cancellation of the standby.

The three-colour LED indicator (green, orange and red) - The status of the cover of the analyser (open or closed)
located on the front right of the analyser indicates the current and the rotor cover (in position or not in position). The
status of the analyser. analyser also issues an alert if there is no rotor placed in
the analyser.
- Control of the useful lifetime of the lamp.
2.2.3. Monitor
- The status of the thermostatisation systems of the needle
This is the main screen that appears when the user and the rotor.
programme is launched. This lets the user supervise and - The levels of the waste and system liquid containers
intervene in the progress of the work session or any other
task sent to the analyser. It displays the status of the - The status of the racks tray. The analyser alerts the user
analyser and the work session in progress, providing if the configuration of the racks in the analyser does not
graphic information about any alarm produced. The content coincide with the programmed configuration or if some of
of this screen is as follows: the reagents or samples have run out.

- Status of the analyser and the work session. - Other alarms.

- Vertical button bar which enables the control of the - The list of samples in the current work session. This
functioning of the analyser. list presents all the samples to be performed in the
current work session, in accord with the order of
- Horizontal button bar which provides access to the main execution. To indicate the status of each sample, a
function of the application. colour code and icons are used:
- Pending (yellow): Samples that have not yet been
analysed.
- In progress (red): Samples that are being analysed.
- Finished (green): Samples that have been completely
analysed without incidences in any of the tests
performed. The results of these samples are accepted
automatically.
- Finished with incidences (green + attention icon):
Samples with which not all the analyses programmed
have been performed due to an incidence of some kind.
- Blocked (Blocking icon): Samples to which no analysis
has been possible due to an incidence of some kind
(lack of sample, lack of reagent, error in the performan-
ce of the blank or calibrator…).

2.2.3.2. Blocking pending samples


2.2.3.1. Status of the analyser and the work session.
From the monitor screen, you can block samples pending
Real-time monitoring of the status of the analyser and to be analyzed. When pressing the button details, the
development of the work session. monitor screen opens and each patient with all his
programmed test visualizes. If you double-click on any test,
- The total time of the programmed work session and the an icon of a hand will appear showing that this test is
number of rotors necessary for it to be carried out. blocked. The whole patient can also be blocked by double
- The status of the analyser at any given moment: Sleeping, clicking on the patient code. Blanks, calibrators and controls
Warm-up, Shutdown, Standby, Running, Sampling Stop can not be blocked. By double clicking on the same sample
or Alarm. The transitions between the statuses in some a second time, you can unblock it.
cases are automatic and in others they are requested by
the user.
2.2.3.3. Vertical button bar for the control of the
- The warm-up time control, which enables the cancellation analyser
and control of the performance of the photometric base
lines. This bar contains the main buttons for the control of the
- The status of the reactions rotor: number of wells used, functioning of the analyser, which enable the user to change
number of free wells and graphic bar of the percentage of its status and perform different actions. This bar can also
wells used. The thermostatisation time control when a be accessed directly from the positioning screen of the

15
User manual

reagents and samples in the racks. The buttons are as Configuration A25: This enables access to the analyser
follows: configuration screen.
Connection: This re-establishes the connection between Test programming: This enables access to the test
the computer and the analyser if the analyser has been programming screen.
accidentally switched off or the computer has been Profile programming: This enables access to the profile
disconnected (it changes from Sleeping to Standby). If (group of tests) programming screen.
the analyser is in sleeping mode and the computer
programme is launched, the connection is made New sample: This enables access to the introduction of
automatically. new samples screen.
Warm Up: This initialises the analyser (performs Warming Position: This enables access to the screen for the
up and leaves the analyser on standby ready for work). positioning of reagents and samples in the rack tray.
Start: The analyser begins to perform the programmed Monitor: This enables a return to the monitor screen from
analyses (changes from standby to running). any other screen.
Continue: This continues the preparations if they have been Current results: This enables access to the current results
stopped by the analyser or by the user. (It changes screen. These can be displayed in order of patients or
from Sampling Stop to Running). It also continues when tests in real time during the execution of the analyses.
new samples are added to a finished session. (It changes It also allows managing sample repetitions
from Standby to Running). Current alarms: This enables access to the current alarms
Sampling Stop: This enables the interruption of the screen which offers more detailed information about the
preparations without interrupting the readings of the alerts and alarms that have appeared on the Monitor
samples in progress and the reception of results from screen during the current session.
the analyser (it changes from Running to Sampling Utilities: This enables access to the utilities screen.
Stop). Reset Session: This restarts the work session.
Abort: This aborts the work session, both readings and
preparations, i.e. it cancels the execution of the analyses
in progresses (it changes from Running, Sampling Stop 2.2.3.5. Special buttons in monitor screen
or Standby by Rotor End to Standby and considers the Preparation table per well: Button to open a window that
session as finished). shows in each rotor well which sample, test and replicate
New Rotor: This button indicates to the analyser that a have been prepared.
new empty rotor has been installed. This is active only Printing final reports: This button allows printing each
when the analyser is on standby. The analyser controls patient’s results which are finished before ending the
the time required for the thermostatisation of the new whole list. It will only print those finished patients with a
rotor, which is approximately 5 minutes. The user can printer symbol in the printing column.
cancel this waiting time if a previously thermostatised
Legend of icons and monitor colours: This button opens an
to 37ºC rotor is introduced.
extra window that explains what each colour and symbol
New System Liquid Container: This button tells the analyser means.
that the system liquid container has been refilled. The
Details: This button allows opening the monitor screen and
analyser then automatically supply the dispensing
visualizing the programmed tests for each patient. By
system to prevent the accumulation of air, which is
pressing again the same button, you can recover the
detrimental to the execution of the analyses. It is active
previous screen.
when the analyser is on standby, running or sampling
stop mode, but the supply is carried out on standby or
running only.
Shutdown: It performs the shutdown process of the analyser
(it changes from standby to sleeping and performs
shutdown).

2.2.3.4. Horizontal button bar

This button bar provides access to the main functions of


the application and is accessible from all the programme
screens, not only from the monitor screen. These functions
and others whose use is not as frequent can also be
accessed through the dropdown text menu at the top of
the screens. The buttons are as follows:

16
2.2.4. Programming Save: Clicking this button after modifications have been
made provisionally stores the new parameters.
Various screens enable the programming of the parameters
of tests, profiles, multiple calibrators, multiple controls, OK: Clicking this button definitively stores all the changes
contaminations and reagent racks. These parameters are made in the tests file.
usually constant over long periods of time once the analyser Cancel: This enables the recovery of the initial tests file,
has been adapted to the needs of the laboratory where it is without taking into consideration the changes made
being used. Programming requires knowledge of the since the tests programming screen was opened.
analyser and the analysis process. It is not possible to
modify any of the programmed parameters while they are Programmable parameters
being used in a work session.
The parameters programmable for each test are as follows:
2.2.4.1. Test programming General
This screen enables the programming of new tests, the - Test name: Test name with up to 16 characters.
querying or modification of parameters of tests that have - Analysis Mode Final monoreagent point, Final bireagent
already been programmed and the elimination of tests that point, Bireagent differential, Monoreagent fixed time,
are no longer used. It also enables the ordering of the list of Bireagent fixed time, Monoreagent kinetics, Bireagent
tests and the printing of their parameters. The default tests kinetics.
are those pre-programmed by BioSystems to use the line - Sample type: Each test may contain specific parameters
of reagents adapted to the analyser. The total number of for five different sample types: Serum (SER), Urine (URI),
programmable tests is unlimited and depends only on the Cerebrospinal (CSF), Whole blood (WBL) or Plasma
capacity of the computer. The screen is divided into two (PLM). The Delete Type button can delete all the data
parts: related to a certain sample type of a test.
- List of tests.
- Units: Units that are used to express all the concentrations
- Information about parameters, grouped together in five associated with the test. One of the memorised units
folders: General, Procedure, Calibration, Controls, can be selected or a new one introduced.
Options.
- Turbidimetry test: It shows the program that the
The parameters of a test can be queried by selecting it on programmed test is a special one. The analyzer makes
the list with a click. If you wish to modify the parameters of a different cycle with turbidimetry tests.
a test, select it with a double-click or pressing Enter. None
- Reaction type: Increasing or decreasing.
of the programmed parameters of a test can be modified if
it has been programmed in a work session. The list of tests - Decimals: Number of decimals to be used to express
can be ordered by dragging a test to the new position in the concentration in the reports.
the list or alphabetically by clicking on the list header. - Replicates: Number of times each preparation is carried
out for the patient samples (from 1 to 50). The final result
If a test parameter is modified, the results stored for the
obtained is the average of the replicates performed and
blank and calibrator are automatically reset and must be
accepted.
redone.
- Name of the Associated Constituent. Optionally, it is
Buttons possible to introduce the name of the constituent
measured with the test. If introduced, it will appear in the
Various buttons let the user perform the following actions: reports.
New: This enables the creation of a new test and the
introduction of its parameters in the corresponding
folders. The programme introduces some parameters
by default to speed up this process and checks that
the data introduced is within the expected margins. If
this is not so, it sends the user alerts (the alerts appear
on the status bar and do not allow further introduction of
data until the incorrect data has been put right).In order
to identify the created test as a user test different from
the original tests, the colour of the test icon changes to
orange. The original tests are blue.

Print: This prints out the parameters of the selected tests.


Delete: This deletes the selected tests.

17
User manual

Procedure - Calibration curve: For Multipoint Calibrator (Multiple or


- Reading type: Monochromatic or Bichromatic (for End Specific). It can be polygonal, linear regression, spline or
Point only) regression parabola. The x and y axes can be normal or
logarithmic.
- Volumes of Sample and Reagents: Sample volume
(between 3 µl and 40 µl), Reagent 1 volume (between 10 - The calibration curve may be programmed to increase or
µl and 440 µl), Reagent 2 volume (between 10 µl and 200 decrease.
µl). The Reagent 2 volume will be active or not depending - Calibrator replicates: number of times the calibrator is
on the analysis mode used. The programme controls that measured (1, 2 or 3). The final result obtained is the ave-
the total volume of the reaction is between 200 µl and rage of the replicates performed and accepted.
800 µl. - Blank replicates: number of times the blank is measured
- Washing Volume: Internal washing volume of the needle (1, 2 or 3). The final result obtained is the average of the
which the analyser performs automatically after the replicates performed and accepted.
preparation of the test (1200 µl, 1000 µl or 800 µl). - Alternative calibrator: Option that enables the analysis
- Pre-dilution Factor: Pre-dilution factor of the sample. Pre- of a certain type of sample using the results of the
dilution can be carried out automatically or by the analyser calibration of another different sample type. For example,
or manually by the user. it enables the use of calibration for serum to analyse a
- Automatic repetition: It shows the program to repeat that urine sample.
test automatically if the result is out of the limits. Controls
- Postdilution Factor: It is the factor of sample postdilution - Number of controls: Number of controls the analyser
when it makes an automatic repetition. proposes for each work session (0, 1 or 2).
- Filters: One or two filters to be chosen in accord with the - Rejection criteria: Criteria for rejecting an analytical se-
reading type. Only filters that are physically in the filter ries (between 0.1 and 4 standard deviations).
wheel can be chosen (they can be revised from the
Configuration screen of the A25). - Replicates: number of times the control is measured (1,
2 or 3). The final result obtained is the average of the
- Times: Depending on the analysis type, the
corresponding boxes are activated to programme the
different reading or dispensing times of the second
reagent. The times can be introduced in seconds or in
cycles. Each cycle is equal to 15 s.

Calibration
- Calibration type: Factor, Multiple Calibrator or Specific
Calibrator. If factor is used, its value must be introduced
in the corresponding field. If the calibrator is specific, the
fields appear to introduce the concentration values,
depending on the number of calibrators. For multiple
calibrators, this data is introduced on the multiple
calibrators programming screen, which is accessed
directly from this screen.

18
Formula applied:

∆T 1 = AbsT 1 − AbsT 1−Cycletime


∆T 2 = AbsT 2 − AbsT 2−Cycletime
∆T 1
∗ 100 < % Pr ozone _ ratio
∆T 2
- Substrate consumed. When this option is activated the
program can detect reactions with the substrate
consumed. These are generally reactions with highly
concentrated samples. To perform this check it is
necessary to enter a value of the absorbance limit with
which the reaction is to be compared. If the reaction
exceeds this limit a warning is given and an automatic
replicates performed and accepted. repetition is carried out after dilution, provided the
automatic repetition option is activated. This option is
- Control type: Multiple or Specific. If the control is specific, only valid for kinetic calculation modes.
the fields appear for introducing Name, Batch, Maximum
Concentration and Minimum Concentration for each of - Slope function (y = ax+b), this option activates the
the controls. For multiple controls, this data is introduced correlation correction. This correction enables the results
on the multiple controls programming screen, which is obtained from the analyser to be correlated with the results
accessed directly from this screen. from other analysers. When this option is activated the
formula Y= aX+b is applied, where X is the concentration
- Calculation Mode: This indicates the method used for obtained by the analyser and Y is the corrected
deciding if a series of results is accepted or rejected. It concentration which will be that appearing in the results.
can be manual or statistical. In statistical mode, the user For each technique it is necessary to enter coefficients a
must introduce the number of series. The Internal Quality and b, as determined experimentally by the user.
Control section gives a detailed explanation of its
functioning. - Factor limits.
- Reference interval.
Options
- Limits of repetition
- Blank absorbance limit. In kinetic or fixed time tests,
this limit refers to the initial reading of the programmed - White with reagent only
absorbance. - White with saline solution
- Kinetic blank limit.
2.2.4.2. Profile programming:
- Linearity limit.
- Detection limit. This screen enables the programming of test profiles. A
test profile is a set of tests identified by a name. Using
- Prozone effect The so-called prozone effect can occur in programmed test profiles simplifies the preparation of the
test based on the principle of the formation of antigen/ work session. On allocating a profile to a sample, all the
antibody complexes (agglutination). This effect is generally tests contained in said profile are automatically allocated
detected in samples with a high antigen concentration. to it. Each profile has one single sample type associated
The excess antigen reverses the direction of the reaction
and can resultin in incorrect measurements of values of
samples. To detect this effect it is necessary to activate
the prozone effect option and enter the 3 parameters:
Time1, Time2 and the ratio in (%). The program calculates
the increases in absorbance in times 1 and 2. It calculates
the ratio of the increases and compares it with the ratio
entered. If the detected ratio is below that defined a war-
ning is given indicating that the sample may be affected
by the prozone effect. The user then has to perform a
manual repetition with a dilution factor to finish determining
the exact value of the sample.

19
User manual

to it. If you wish to create the same profile for different


sample types, a profile must be programmed for each
different type. The total number of programmable tests is
unlimited and depends only on the capacity of the computer.
The screen contains:
- List of profiles
- List of programmed tests
- List of tests in the selected profile

The parameters of a profile can be queried by selecting it


on the list with a click. If you wish to modify the parameters
of a profile, select it with a double-click or pressing Enter. It
is not possible to modify any of the programmed parameters
of a profile while they are being used in a work session.
The list of profiles can be ordered by dragging a profile to
the new position in the list or alphabetically by clicking on the name of the allocated calibrator and its batch. To allocate
the list header. or change the calibrator of a test, click on the name or
batch of the calibrator and choose the desired calibrator
from the list that drops down on the row corresponding to
Buttons the test. If a calibrator that has already been allocated to
one or more tests is modified, the allocations are deleted.
Various buttons let the user perform the following actions:
Once a calibrator has been allocated to a certain test, the
New: Enables the creation of a new profile. user must programme the test calibration parameters:
Add tests (Arrow >) and remove tests (Arrow <): It is possible - Concentration data: Assigned values of calibrator
to add or remove tests to or from a profile that is being (monopoint) or calibrators (multipoint) concentration for
edited by selecting the desired tests from the test or the test, introduced from greater to smaller. The
profile lists and clicking on the corresponding button. corresponding factors are calculated automatically from
Print: Prints the content of one or more profiles. the concentrations introduced.
Delete: This deletes the selected profiles. - Calibration Curve Data: For multipoint calibrators, the
curve type (polygonal, linear regression, spline or
Save: Clicking this button after modifications have been
regression parabola) and the type of x and y axes (nor-
made provisionally stores the new parameters.
mal or logarithmic) must be selected.
OK: Clicking this button definitively stores all the changes
made in the profiles file. By selecting a calibrator with a click or with the cursor
keys, it is possible to query its data and the tests allocated
Cancel: This enables the recovery of the initial profiles file,
to it. The allocated tests appear selected on the tests list.
without taking into consideration the changes made
If you wish to modify the parameters of a calibrator, select
since the profiles programming screen was opened.
it with a double-click or pressing Enter. The data can be
modified on the grid itself. It is not possible to modify any
2.2.4.3. Programming of Multiple Calibrators of the programmed data of a calibrator while it is being
used in a work session. The calibrator grid can be ordered
A multiple calibrator can be used to calibrate more than by dragging a calibrator to its new position in the grid. The
one test. Up to 10 multiple calibrators can be programmed. calibration parameters of a test can be queried by selecting
Each of these calibrators can be monopoint or multipoint, it with a click or using the cursor keys. To edit the calibration
i.e. a single calibrator or several calibrators may be used. parameters of a test, select it with a double click or by
The multiple calibrators programming screen can be pressing Enter.
accessed only from the tests programming screen, from
the Calibration tab. The screen contains: Buttons
- Calibrator grid: Various buttons let the user perform the following actions:
- List of tests with multiple calibrator New: Enables the creation of a new calibrator. You must
- Information about parameters introduce the name, batch and number of calibrators.
Print: Prints out the data of the selected calibrators.
The calibrator grid displays the name of the calibrator, the
batch and the number of calibrators. The number of Delete: Deletes the selected calibrators from the grid.
calibrators ranges from 1 (single calibrator) to a maximum Save: Clicking this button after modifications have been
of 8. The list of tests with multiple calibrator shows the made provisionally stores the new parameters.
tests programmed to be calibrated with a multiple calibrator.
OK: Clicking this button definitively stores all the changes
The display shows the name of the test, the sample type,
made in the calibrators and tests file.
20
Cancel: This enables the recovery of the initial tests and The control grid can be ordered by dragging a control to its
multiple calibrators files, without taking into new position in the grid. The control parameters of a test
consideration the changes made since the multiple can be queried by selecting it with a click or using the
calibrators programming screen was opened. cursor keys. To edit the control parameters of a test, select
it with a double click or by pressing Enter.

2.2.4.4. Programming of Multiple Controls Buttons


A multiple control can be used to control more than one Various buttons let the user perform the following actions:
test. Up to 20 multiple controls can be programmed. The
multiple controls programming screen can be accessed New: Enables the creation of a new control. You must in-
only from the tests programming screen, from the Controls troduce the name and batch.
tab. Print: Prints out the data of the selected controls.

The screen contains: Delete: Deletes the selected controls from the grid.

- Control grid Save: Clicking this button after modifications have been
made provisionally stores the new parameters.
- List of tests with multiple control
OK: Clicking this button definitively stores all the changes
- Information about parameters made in the controls and tests file.
The control grid displays the name of the control and the Cancel: This enables the recovery of the initial tests and
batch. The list of tests with multiple control shows the multiple controls files, without taking into consideration
programmed tests to be controlled using a multiple con- the changes made since the multiple controls
trol. The display shows the name of the test, the sample programming screen was opened.
type, the name of the allocated control and its batch. To
allocate or change the control of a test, click on the name 2.2.4.5 Calculated tests
or batch of the control and choose the desired control from
the list that drops down on the row corresponding to the This screen displays the calculated tests that can be
test. If a test requires 2 controls (generally 2 levels: high performed by the analyser, along with their parameters.
and low), each one must be allocated separately. If a con-
trol that has already been allocated to one or more tests is In order to obtain a calculated test result, the calculations
modified, the allocations are deleted. Once a control has are made using the values for the concentrations of the
been allocated to a certain test, the user must programme associated tests.
the test control parameters: When running a calculated test, the analyser first determi-
- Concentration data: Allocated values of maximum and nes the associated test concentrations, then calculates
minimum control concentration for the test. the result of the operation.
By selecting a control with a click or with the cursor keys, This screen displays the following information for each
it is possible to query its data and the tests allocated to it. calculated test:
The allocated tests appear selected on the tests list. If you
wish to modify the data of a control, select it with a double-
•The tests used in the operation and the formula applied.
click or pressing Enter. The data can be modified on the • The normal reference ranges
grid itself. It is not possible to modify any of the programmed
data of a control while it is being used in a work session. • The units for the calculated test

21
User manual

• Whether the partial values for the experimental tests are


to be printed in the final report for each patient. Buttons

When a calculated test is allocated to a sample, all the Various buttons let the user perform the following actions:
associated tests are automatically allocated in the New: Enables the creation of a new programmed rack.
calculation, exactly as for a profile.
Print: Prints out the data of the selected programmed racks.
Delete: Deletes the programmed racks selected on the list.
2.2.4.6. Reagent Rack Programming
Save: Clicking this button after modifications have been
Programming a reagent rack consists of saving a certain made provisionally stores the programmed rack.
configuration of reagent bottles of a rack, identified by a
OK: Clicking this button definitively stores all the changes
name. When preparing a work session, to position the
made in the programmed racks file.
reagents, you can load a programmed rack and the saved
reagent distribution is automatically allocated. If the user Cancel: This enables the recovery of the initial programmed
frequently uses the same reagents, it is very practical to racks file, without taking into consideration the changes
use programmed racks to speed up the work. An unlimited made since the racks programming screen was opened.
number of reagent racks can be programmed, depending
on the capacity of the computer only. The rack programming
2.2.5. Save/load test file
screen can be accessed directly from the programming
menu of the main programme menu. From the positioning The user can save the test programming in an external file,
screen, it is also possible to save sample racks with a he has only to execute the option Save Test File of the
specific identifying name. It is not possible to access sample programming menu. This option saves all the test data in a
racks from the reagent racks programming screen. It may compressed file. It is the program itself the one that
be useful, for example, to save a calibrator and control rack compresses and decompresses the file with no need of
that is used frequently. The reagent rack programming the user’s intervention.
screen contains:
- List or programmed reagent racks In order to load a new test file, the user has to execute the
option Load Test File of the programming menu and then
- Information about parameters all the tests of the application are automatically replaced.
The parameters interface contains a list with all the reagents This process is only active with the following options:
associated with the tests programmed in the analyser, the
name of the selected rack and an image of this rack showing - When working without passwords or as Supervisor user.
the real positions of the reagents, their names and volumes.
The bottles on the racks can be added, removed or reordered - When the analyzer is in SLEEPING or STANDBY state
by dragging them with the mouse. The bottle type (50 or 20 - When there is no working list. If you have a programmed
ml) may be changed using the corresponding button. Its list, make a RESET.
parameters may be queried by selecting a programmed
rack with a click or using the cursor keys. If you wish to
modify a programmed rack, select it with a double-click or 2.2.6. Preparation of the work session
pressing Enter. A programmed rack cannot be modified while
The Introduction of New Samples and Reagents and
it is being used in a work session. The list of programmed
Samples Positioning screens help the user to prepare the
racks can be ordered by dragging a rack to the new position
work session conveniently, easily and quickly. Furthermore,
in the list or alphabetically by clicking on the list header.
it enables the storage of data that can be used in later
sessions.

2.2.6.1. Introduction of New Samples

This screen contains the interface for introducing the data


of each sample and the lists of New Samples and Sent
Samples.

Interface for introducing sample data

The user can select the sample class (normal patient, urgent
patient, blank, calibrator or control) and the sample type
(serum, urine, cerebrospinal liquid, whole blood or plas-
ma). A tree diagram shows all the tests, calculated tests
and profiles programmed in the analyser. The user can select

22
or introduce the patient code (alphanumeric). If the patient Add (Arrow >) button.
code introduced does not exist in the current work session,
the new patient is added together with his or her sample. If The code of a patient can be changed on the list itself by
the patient already exists, the sample of the patient in clicking the right button of the mouse on the existing code.
question is selected and, on the tests tree, the tests Once you have made the desired changes, press Enter. It
previously allocated are selected for possible edition. If the is possible to add tests or profiles to a patient sample that
user does not introduce the patient code, the programme has already been introduced. Different sample types can
adds the patient sample with a correlative numeric identifier. be introduced from the same patient and the programme
If several normal or urgent samples are to be introduced groups them together automatically. It is also possible to
with the same tests to be carried out, the desired number add, in one go, tests, calculated tests or profiles to several
must be introduced in the Number of Patient Samples field samples of the same type for different patients.
and the programme automatically adds all the samples,
allocating them a correlative patient code. By default, the
Number of Patient Samples is 1. The maximum value is Lists of samples
120. If the user has got a code bar reader, he can use such The lists of New Samples and Sent Samples show the
reader to insert the patient code. Place the cursor in the information grouped together by sample class. Each one
box of the patient code and read the code to be inserted contains a patient grid, one of blanks and calibrators and
with the reader. one of controls. By default, the patient grid of the list of
The list of New Samples shows all the samples introduced, new samples is displayed.
grouped together by classes, which the user can then send The grids of New Samples enable the modification of the
to the analyser. The list of Sent Samples shows those the information they contain. The grids of blanks and calibrators
user has sent to the analyser, also grouped together by and controls are completed automatically as patients are
classes. To introduce a new sample into the list of new added. If the user wishes, he or she can also introduce
samples, proceed as follows: blanks, calibrators or controls manually. From the
1. Select the sample class corresponding grids, it is possible to activate or deactivate
2. Select the sample type the performance of blanks, calibrators and controls,
changing the value of the fields New or In Use, respectively
3. If desired, introduce the patient code. (as long as there are results stored from a previous session).
4. Select the profiles, tests and calculated tests to be If the blanks and calibrators are deactivated, the analyser
performed. will use the data of the last work session. If the user wishes
to send certain samples for positioning on the racks and
5. Add the sample to the list of samples by clicking on the
analysis, they must be selected and the Position button
Add (Arrow >) button.
pressed. It is also possible to send certain tests of a sample
If you wish to introduce a certain number of samples n with only. The associated and activated blanks, calibrators and
the same set of tests to be performed, proceed as follows: controls are sent automatically. By clicking on the Position
button, with no previous selection, all the introduced and
1. Select the sample class
activated samples of the corresponding grid are sent.
2. Select the sample type Clicking the Delete button deletes the samples or the tests
3. Select the profiles and tests to be performed. of a sample that have been selected.
4. Introduce the number of samples n The Sent Samples grids enable the query of the samples
5. Add the sample to the list of samples by clicking on the sent to the analyser and the tests to be performed on each

23
User manual

Position: This sends a selection or a complete list of


samples to be positioned in the analyser for later
analysis. This is active for all the new samples grids. It
enables:
- Sending of groups of patient samples. In this case,
the associated and activated blanks, calibrators and
controls are also automatically sent.
- Sending of control groups. In this case, the
associated and activated blanks and calibrators are also
automatically sent.
- Sending of groups of blanks and calibrators for
making lists for calibration only. In this case, the
associated and activated blanks are also automatically
sent.
Patient data: This enables access to the screen for
sample, but they do not allow modifications or the resending introducing the data of each patient. This data is not
of samples to the analyser. necessary for the performance of analyses and can be
The Reset Session button on the horizontal bar restarts introduced once the analyser is running.
the work session and deletes all the sample lists.

When you reuse a Calibrator, you can modify the memorized 2.2.6.2. Positioning of samples and reagents
factor from the New Samples screen, within the Blanks This screen contains a tree diagram showing all the
and Calibrators folder in order to recalculate the results elements to be positioned, an image of the analyser tray
with the new inserted value. indicating the current programmed distribution of the racks
- In order to edit the factor: double click its cell, insert a and an enlarged image of the racks showing the details of
new value and accept by pressing Enter. This action all the elements in the selected rack.
recalculates all the affected Concentration results.
- In order to recover the previous factor value: click any Tree diagram of elements to be positioned
crossed cell. This action recalculates all the affected This tree contains all the reagents and samples to be
Concentration results. positioned on the racks to execute the work session. For
Buttons each of the reagents, the tree shows the name, the volume
required and the minimum number of bottles required for
Various buttons, some of which have already been each type (20 or 50 ml). The samples are grouped together
mentioned, let the user perform different actions. The by class: calibrators, controls and patients. For the
following is a brief description of the functionality of each calibrators, the tree indicates the name, the number of
one. calibrators if it is multipoint and, if the test or tests allocated
Add sample (Arrow >): This dumps the information of the have pre-dilutions programmed, the pre-dilution tubes
interface to introduce sample data in the corresponding required, together with the corresponding factor of each
new samples grid. one. For the controls, the tree indicates the name and, if
Edit patient (Arrow <): This enables the edition of a patient the test or tests allocated have pre-dilutions programmed,
sample or of several of the same type, of the new
samples grids. The edition enables the addition of tests
or profiles to the group of samples selected. It is also
possible to change the priority of the edited samples,
i.e. change from urgent to normal or vice versa. The
patient code of a sample is changed during the edition,
the programme adds a new patient, which is a copy of
the previous one. If you wish to change the patient code
only without adding another new patient, make the
change directly on the samples grid with the right button
of the mouse.
Delete: This deletes the patient sample or control data
selected. It is possible to delete tests, test or sample
profiles of a patient or patients that have been completed.
This is active for the new samples grids only.

24
the pre-dilution tubes required, together with the and place the elements by dragging them with the mouse
corresponding factor of each one. For the patients, normal from the tree to a free position in the corresponding enlarged
or urgent, it indicates the patient code, the sample type rack. It is possible to position individual bottles of reagent
and, if the test or tests allocated have pre-dilutions or all the required reagent in one bottle. If a sample is
programmed, the pre-dilution tubes required, together with dragged and it has programmed pre-dilutions, the pre-dilution
the corresponding factor of each one. tubes required are also dragged. If you wish to use different
sample rack types in one session, the user must manually
The colour blue indicates which reagents and samples are position each sample in the corresponding rack in accord
completely positioned in the racks tray. The colour black with the type of tube used.
indicates the reagents and/or samples that are not
positioned completely or whose volume has finished during Once all the elements have been positioned on the screen
the work session. and physically in the analyser, the user can begin the
analyses.
Rack tray and extended rack
Buttons
The image of the rack tray shows the current rack
distribution in the analyser. In each of the 6 rack positions, Various buttons, some of which have already been
there may be a reagent rack or a sample rack or the mentioned, let the user perform different actions. The
position may be empty. In the reagent racks, the image following is a brief description of the functionality of each
shows which positions are occupied by 20 and 50 ml one.
bottles. In the same way, in the sample racks the image Position Reagents Automatically: The programme places
shows the positions occupied by sample tubes. The 3 cen- the reagents from the tree that are not yet positioned
tral positions, preferably used for bottles of distilled water, (black colour) in the free positions on the reagent racks
saline solution and washing solution, are also shown. on the tray. If required, it allocates new reagent racks
The user can choose the rack type at each position on the to the free positions on the tray and fills the positions. A
tray. The limit configurations are 5 racks of reagents and 1 maximum of 5 reagent racks is permitted. If there are
of samples (50 reagents / 24 samples) and 1 rack of reagents elements on the racks that are not required for the current
and 5 of samples (10 reagents / 120 samples). In the work session, the programme alerts the user to remove
analyser options, the user can choose which type of them. If there is no room for all the reagents, the user is
samples rack he or she wishes to use for all the samples also alerted.
in the current session (13 mm diameter sample tube, 15 Position Samples Automatically: The programme places
mm diameter sample tube or 13 mm diameter well). If he or the samples from the tree that are not yet positioned
she wishes to use racks of samples of different types in (black colour) in the free positions on the samples racks
the same session, the user must deactivate the rack on the tray. If required, it allocates new samples racks
detection on the analyser configuration screen. It must also to the free positions on the tray and fills the positions. A
be taken into account that the analyser will not check that maximum of 5 samples racks is permitted. If there are
the physical disposition of the racks on the tray is the same elements on the racks that are not required for the current
as the programmed disposition. Besides empty racks, it is work session, the programme alerts the user to remove
also possible to position reagent racks programmed on them. If there is no room for all the samples, the user is
the rack programming screen or racks previously saved also alerted. If different types of sample racks are used
from the positioning screen itself. in one session, the user must manually reposition each
sample on the corresponding rack in accord with the
The screen displays an enlarged view of the rack selected tube type used, since the analyser does not have
on the tray, enabling easy manipulation of its content. The information about the tube type used for each sample.
selected elements can be unloaded from the rack or the
rack can be completely emptied. It is possible to change Remove: This removes the selected elements from the
the position of any element by dragging it to a free position enlarged rack. If no selection has been made, all the
on the enlarged image of the rack. The volume of a bottle of elements are removed from the rack. It is also possible
reagents can also be changed by double-clicking on it. to remove the selected elements using the delete button.
When any change is made, the programme checks if all Save Rack: This saves the enlarged rack (reagents or
the required volume of said reagent is in position and alerts samples) for later use.
the user if it is not. Summary of positions: This enables easy verification of all
The positioning of the reagents and the samples on the the allocated positions. The user accesses a screen
racks can be carried out automatically or manually. Two showing the current distribution of the racks tray in text
buttons make it possible to automatically position the mode, indicating the content of each position rack by
reagents or the samples, respectively. With automatic rack. This information may be printed out.
positioning, the programme allocates the rack types that OK: By clicking OK, the programme checks that all the
are required. If the user wishes to position the elements required elements are in position and requests
manually, he or she must choose the required rack types
25
User manual

confirmation from the user to save the information as


definitive. When all the elements have not been
positioned, an auxiliary alerts screen opens. If the
analyser is ready for work, the Start button of the verti-
cal buttons bar is enabled and the user can press it for
the analyser to start the analyses. The programme
automatically changes to the Monitor screen, from which
the execution of the work session can be supervised.
When the analyser is carrying out the analyses, the
user can press the Sampling Stop button on the vertical
button bar to introduce new elements into the analyser
physically. Once the positions have been programmed
on the Positions screen, by clicking OK, the programme
checks that all the new required elements are in position
and requests confirmation from the user to save the
information as definitive. The Continue button on the
vertical bar is enabled and when the user presses it,
the programme changes to the Monitor screen and the
analyser continues the pending analyses and the new
ones that have been sent.
Cancel: This exits the screen without updating any of the
changes made and keeps the previous distribution.
Alerts: This button appears when there are elements not
positioned in the prepared work session and it informs
the user of everything that cannot be analysed. When
nothing is in position, the Start button is not enabled.

2.2.7 Memorising sessions

When Memorizing a working session, the assigned


samples and tests are saved, but the positions are not. If
you wish to memorize the current session, you should save of the reaction as a function of time. Time is expressed as
it before starting its execution. When loading a memorized a number of cycles and the absorbance values are measured
session, the tests are loaded such as they are programmed with respect to the baseline. The screen also shows the
at that moment, and the tests that have been deleted from numerical values of the measurements. The graph is shown
the test file are eliminated. for each of the filters used in the reaction. The main filter
and/or reference with which the measurements were made
In order to Load a memorized session, it is necessary to can be selected.
carry out a RESET one previously. Once it is loaded, the
session can be freely modified (modify identifiers, eliminate There are some screens where the observations field is not
tests, add samples or tests...) displayed. This indicates that there are warning messages
about the results in this field. An asterisk is displayed in a
The memorized sessions can be also Eliminated. more visible column.

2.2.8. Current results


print buttons
The results can be consulted as they are obtained on the
Current results screen, grouped together by patients or by there are 4 buttons enabling printable reports to be
tests. The results are updated automatically in real time generated. Each button displays a screen with a preview of
as information is received from the analyser. For each the report before it is printed.
analysis, the results of the replicates performed are
displayed together with the corresponding average. Once Three reports can be accessed from the patients tab:
the analyses have finalised, the user can discard the - Per-patient printout:
replicates of samples (blanks, calibrators, patients or Report showing all the results sorted by patient. This
controls) he considers aberrant and the programme includes the absorbance and concentration values with
automatically recalculates and displays the new results. replications and the results that could not be calculated.
Clicking on the graph icon for one of the results displays a - Final patient printout:
screen with the absorbance/time graph for the selected
result. The graph represents the progress of the absorption
26
Report showing only the results for each patient that are
correct. Only concentration results are shown. The report
header includes the patient data.

- Table summarising the results:


Simplified results report. The report shows all a list’s
concentration values in table format. The rows show the
patients and the columns the techniques. These data can
be transferred to an Excel spreadsheet from this report.

A report can be accessed from the techniques tab:

Per-technique printout:
Report showing all the results sorted by technique. This
is similar to the patient report, but is sorted by technique.

the working list. It will be added (rep) to the end of the


sample name to distinguish it from the initial sample. In
the Current results screen, the result of the repetition
will appear under the initial sample. So the user will be
able to choose, by means of the OK column, which of
both results will comprise the patients’ report and
become historical. The repetition result will be selected
by default. If repeating blanks and calibrators, all the
associated values will be recalculated.
Automatic repetitions: In the test programming, some limits
or intervals of reference can be programmed to carry
out automatic repetitions if the sample result is outside
these intervals. In order to carry out the repetitions
automatically, the automatic repetition tick must also
be selected in the test programming:
- If the patient concentration is higher than the linearity
2.2.8.1. Calculated factor modification limit, the sample is repeated with the decreased
Factor of postdilution.
The calculated Factor can be experimentally modified by
the user in the Current Results screen, once the present - If the patient concentration is lower than the detection
session is finished. limit, the sample is repeated with the increased Fac-
tor of postdilution.
- In order to edit the factor: double click its cell, insert a
new value and accept by pressing Enter. This action - If the patient concentration is within the Rank of
recalculates all the affected Concentration results. It repetition, the sample is repeated with the same
also changes the test programming and becomes relation of sample-reagent.
programmed with factor - If the absorbances of the kinetic in blanks, calibrators,
- In order to recover the factor experimental value: click any controls and samples are not linear, it is repeated
cell crossed at the average or replicate level. This action with the same relation of sample-reagent volumes.
recalculates all the affected Concentration results.
2.2.9. Current and past alarms
2.2.8.2. Repetitions The Monitor also displays information about the alarms or
The User can make repetitions of blanks, calibrators, alerts that may appear throughout the work session in the
controls and samples. Repetitions can be manual or shape of identifying icons, together with a brief informative
automatic. text. By clicking on the corresponding button, the user can
access the Current alarms screen, which offers more
Manual Repetitions: In order to make manual repetitions, detailed information about the problems and their possible
the own user has to select the samples that he wants solution. The alarms and alerts occur when certain detectors
to repeat. Once the sample measurement is finished or sensors are triggered in the analyser or when the analyser
and the result is in the Current results screen, select detects anomalous functioning. When necessary, the
the results to be repeated in the Rep column and then analyser acts to avoid continuing working in unsafe
press the Repeat Selected Samples button. The program conditions and notifies the computer. The alarms screen
will automatically insert the sample to be repeated in

27
User manual

displays a list with all the alarms and alerts generated by List of elements
the analyser in the current session. It is also possible to
access the Past alarms and alerts, which contains a list This list contains all the elements of the selected sessions.
with the alarms and alerts generated during previous work In the past results by patient, the list shows the patient
sessions. For each alarm or alert, the following can be code and the patient names introduced, corresponding to
displayed: the selected session or to all the sessions. In the past
results by test, the list contains the name of the tests
- Type of alarm or alert performed in the specific selected session. This list enables
- Date of alarm the selection of elements for query, deletion or printing out
- Brief definition of the alarm (name) the associated data by means of the corresponding buttons.
The list can be ordered alphabetically by clicking on the
- Detailed description title. In the past results by patient, the Allocate button
- Proposed solution enables the allocation to the selected patient previously
- Observations (lets the user introduce information: applied introduced patient data. The Delete button enables the
solution, date of solution, implicated personnel…) deletion of the elements selected on the list. The Print button
enables the printing or exportation to file of a report with
The lists are updated automatically in real time as information the data and results of the selected elements. The
is received from the analyser. The alarms pending correction programme always displays a preview of the report.
can be edited for text introduction indicating the
implemented solution, the date of the solution and the Data Interface
personnel implicated. If so desired, the corrected alarms
can be eliminated. The lists can be ordered according to In the past results by patient, the data interface contains
the field type, date or name. Problem solution reports can the following patient information: name of the selected
be printed with the selected alarms. The Alarms and alerts patient, sex, date of birth, person who has requested the
section of this manual lists the main alarms and alerts the analysis and a field for comments. A button enables access
analyser can display and which require user intervention. to the Patient Data screen, where the data can be
introduced or modified. In the past records by test, the
2.2.10. Past results data interface contains the absorbance of the blank and

The Past results screen displays the results obtained in all


the work sessions prior to the current session and prints
out reports with said results. The results can be organised
by patients or by tests. It is also possible to make reports
of patients per session or with all sessions. Reports by
tests can be made only by previous selection of a specific
session. The Delete button deletes all the data of the
selected work session.

The screen contains:


- List of completed session dates
- List of elements: patients or tests
- Interface of patient or test data selected on the list
- Interface of patient or test results selected on the list
28
the calibrator or calibrators used in the selected session, List of patients
the date the blanks and calibrators were performed and the
reference range of the test. This list contains the code and the name and surnames of
all the patients with allocated data. To query patient data,
Results Interface the user must select it on the list. The list can be ordered
alphabetically by title. To edit the data of a patient, double
This interface shows the results of the element selected click on the patient on the list. The New button enables the
on the list, whether it is a patient or a test. Each grid re- introduction of new patient data. The Delete button deletes
cord of results by patient contains: name of the test and the data of the selected patient. The Print button prints out
sample type on which it has been carried out, value of the the data of the selected patient.
result, units, reference range programmed in the test
(optional), date and time of the result and observation about
the result obtained. Each grid record of results by test Patient data interface
contains: patient code, result value, units, date and time of
The data interface shows the code of the selected patient,
the result and observations. By clicking on the
his or her name and surnames, sex, date of birth, the person
corresponding column, the results can be ordered by
who has requested the analysis and a field for introducing
session date, by test or by patient name.
comments. The user can consult current data, modify them
or introduce new data.
General buttons

The past results screen contains two general buttons:


General buttons
Delete: This deletes completed past sessions.
The patient data screen contains two general buttons:
OK: This saves the changes made to patient data and to
the results and exits the screen. Save: Saves all the changes made to the data.
Cancel: This exits the screen without saving the changes Cancel: Recovers all previous data without saving any
made. change.

2.2.12. Configuration
2.2.11. Patient data
These screens enable the user to configure the running
This screen enables the introduction and modification of
mode of the analyser and the user programme. The different
the data of patients who are analysed frequently. This data
parameters can be configured from four dedicated screens:
is not necessary for the performance of analyses and can
be introduced once the analyser is running. This screen - A25 analyser
can be accessed directly from the Introduction of New - Languages
Samples or Past Results screen by patients using the
- Configuration of the serial port
Patient Data button or from the Past results menu. It also
enables the allocation of previously introduced data to a - Header of results reports
patient. This screen contains a list of patients and a patient
data interface.
2.2.12.1. A25 analyser

Clicking on the A25 button on the horizontal button bar


directly accesses the analyser configuration screen. The

29
User manual

different options are separated in three tabs: Washes, When the analyser cannot avoid contamination between
Session and Analyser incompatible tests by reordering the tests of each patient,
it automatically performs and intermediary special washing
Washes cycle. The user can choose to perform these special washes
- Initial, final and special washes between incompatible with system liquid or with washing solution. If the second
tests option is chosen, a 50 ml bottle of washing solution must
be placed in the analyser.
Session
- Carrying out of automatic repetitions Carrying out of repetitions

- Calibrators and blanks in paediatric racks This option allows to activate or to deactivate all the
- Automatic printing of patient reports automatic repetitions.

- Export session when reset is selected Separate Calibrators and Controls in Paediatric Racks
- Used sample tube In the A25 Configuration–Session screen, you can select
the option: “Calibrators and Controls with paediatric racks”.
Analyser
- Container level control scales This option is intended for the Automatic positioning of
samples. When pressing the button Place Samples
- Rack detection
Automatically of the “Positioning of Samples and Reagents”
- Cover detection screen, the samples of Calibrator and Control type are
- Configuration of the filter wheel placed in paediatric racks and the patient samples in the
type of rack selected in the A25- Configuration-Session
screen.
Initial, final and special washes between
Automatic printing of patient information
incompatible tests
This option allows activating and deactivating the automatic
The analyser automatically performs a wash with washing
printing by patient during a current session without waiting
solution during the warm-up and during the shut-down.
for the list to finish.
The user must replace the system liquid container with the
Export session when reset is selected
washing solution container (supplied with the analyser,
bottle with the green dot), when required to do so by the This operation permits automatic activation or deactivation
analyser, during warm-up (initial wash) or during shut-down of results export file generation when reset is selected.
(final wash). Once the wash has been performed, the
analyser asks the user to replace the container with system Used sample tube
liquid (only in the initial wash) and automatically performs
a wash and rinse of the dispensing system with system This enables the selection from a dropdown list of the type
liquid. With the initial wash, the system is ready for working of sample tube to be used in the work session. To access
in optimum conditions during the entire working day, offering this list, the work session must be restarted. The possible
maximum performance. In the final wash, the dispensing types are: 15 mm diameter tube with a maximum height of
circuit is left full of washing solution until the next warm-up 100 mm, 13 mm diameter tube with a maximum height of
is performed, in order to condition the dispensing circuit 100 mm and a 13 mm diameter well. If he or she wishes to
during the rest period of the analyser. use racks of samples of different types in the same session,

30
the user must deactivate rack detection. It must also be installed. If the filter is covered, the value 0 must be
taken into account that the analyser will not check that the introduced. The programme requests confirmation to know
physical disposition of the racks on the tray is the same if the filter in the analyser has been physically changed. If
as the programmed disposition. it has, the programme resets certain reference values used
to issue alerts and alarms from the optical system.
Container level control scales
An alert is issued to the user if any of the tests has
It enables the deactivation of the functioning of the scales programmed filters that do not physically exist in the wheel.
the analyser uses to control the level of the waste and
system liquid containers. In this way, the user can continue
working with the analyser in case of breakdown of any of 2.2.12.2. Languages
the scales, until the technical assistance service arrives to The user can change the language of the user programme on
repair it. In this case, the user must ensure that the system a dropdown list that offers all the languages available. When
liquid does not run out and that the waste container does the language has been selected and OK clicked, the
not fill while the analyser is running. programme updates all the texts to the selected language.

Rack detection 2.2.12.3. Serial port


This enables the deactivation of the rack tray that identifies This screen enables the configuration of the computer se-
the rack in each position. In this way, the user can continue rial port used by the programme. The user can choose for
working with the analyser in case of breakdown of any of the configuration to be performed automatically or manually.
the detectors, until the technical assistance service arrives If manual, he or she can choose the computer serial port
to repair it. If he or she wishes to use racks of samples of and the transmission speed of the communications.
different types in the same session, the user must
deactivate rack detection. It must also be taken into account
that the analyser will not check that the physical disposition 2.2.12.4. Header of results reports
of the racks on the tray is the same as the programmed
disposition. From this screen, the user can customise the header of
the results reports. It is possible to introduce the name of
the laboratory, the address, telephone number and a bitmap
Configuring the acoustic alarm. (.bmp) image. The format of the texts introduced can be
varied.
This option configures the enabling or disabling of the
acoustic alarm. If this option is enabled, an acoustic alarm
sounds when the analyser is running and one of the following 2.2.13. About
alarms appears: lack of reagent, lack of sample, waste
bottles full, system liquid bottles empty, etc., together with This screen displays information about the User Programme
the corresponding icon. Said acoustic signal sounds until version being executed. The Firmware version also appears
the user stops it by pressing the end alarm button or any of (program executed directly by the analyser) and the serial
the star, sampling&stop, continue, new rotor buttons. number of the analyser. In order to see the Firmware and
serial number, the program must be connected to the
analyser.
Configuration of the filter wheel

This screen enables the modification of the analyser filter


wheel. To access this screen, the work session must be
restarted. The wheel has 10 positions. Position 0 must
always contain a covered filter so that the analyser can
perform the darkness adjustment. Positions 1 to 9 can be
used for optical filters. All the positions of the wheel must
be occupied for it to work correctly. The positions that do
not contain an optical filter must be occupied by a covered
filter. The analyser includes as standard 8 optical filters in
positions 1 to 8 and two covered filters in positions 0 and
9. If one of the filters is to be changed, select the desired
position of the wheel and press the Change Filter button.
The analyser automatically positions the wheel filter
appropriately so that the user can change the filter through
the window of the optical system. Next, if it is different,
introduce the wavelength of the new filter that has been

31
User manual

2.2.14. Utilities

Clicking on the utilities button on the horizontal button bar


directly accesses the main utilities screen. This screen
lets the user access the different test, preparation and
maintenance utilities of the analyser, by clicking the
corresponding button. For some of the utilities, the
programme displays a corresponding screen. The available
utilities are:
- PC-Analyser communications channel test
- Reactions rotor check
- Disassembly of the dispensing needle
- Reset of the base line historic
- Fluid system supply
- Cleaning of the dispensing system
- Changing the lamp performs a base line in each well with the selected filter.
The analyser presents a graph of the absorbances relative
- Change rotor type to the average of the light intensity of the wells together
All the utilities require the analyser to change to specific with the standard deviation of said standards. In accord
mode, called Test mode, and for this reason it is not possible with these results, the user can decide whether or not the
to access this screen when a work session is not being rotor can be reused or if it should be replaced by a new
prepared or performed. When running test utilities, the rest one. It is important to point out that the test checks the
of the programme’s functionality is blocked until the test optical quality of the wells but not the status of chemical
ends and the utilities screen is closed. cleansing. Depending on the analysis made with the rotor,
the chemical residues that may be left in the wells can
have a noticeable effect on the following analyses performed
with the rotor. In case of doubt, or when very sensitive tests
are to be performed, it is always recommendable to use a
new rotor. After the test, the user must remove the rotor of
the analyser, empty it and dry it completely before using it
for analyses.

2.2.14.3. Disassembly of the dispensing needle

On clicking on the Disassemble Needle button, the


operating arm positions itself over the rack tray. The
programme alerts the user to remove any object positioned
under the arm. On clicking OK, the needle descends and
the user can remove it to work with it or change it. To remove
the needle, unscrew it by holding the top fitting. If, while

2.2.14.1. PC-Analyser communications channel test

On pressing the test button, the computer attempts to


establish communication with the analyser. The programme
tells the user if communication with the analyser has been
possible or not.

2.2.14.2. Reactions rotor check

The user can use this test to check the optical status of a
reactions rotor. He or she can choose the optical filter with
which the test is to be performed. The rotor must be placed
in the analyser, the rotor cover replaced and the analyser
cover closed, then press the Test button. The analyser fills
the 120 wells of the rotor with system liquid and then
32
handling the needle, the carriage rises due to the pressure
made by the user, press the Lower Needle button for the
needle to descend once again. Once the needle has been
reassembled on the analyser, press the OK button for the
needle to rise. It performs the self-centring test and the
arm finally returns to its parked position. These operations
must be done with utmost care since they are carried out
with the analyser cover open and the needle may be
contaminated. Laboratory gloves must always be used.

2.2.14.4 Reset of the base line historic

By pressing the base lines historic reset button, an internal


initialisation of the historic in the analyser will occur. The
said historic serves to decide whether a base line is correct
or incorrect. In the event of many base line failure alarms
has, the programme resets certain reference values used
occurring, reset the base line historic by pressing the said
to issue alerts and alarms from the optical system.
button in order for it to generate a new historic.
Bear in mind that the next time you produce a base line, a To access this utility, the work session must be restarted.
log will be generated; to do this it will produce 3 consecutive The lamp must be changed with the analyser in sleeping
base lines. It will take a few minutes longer than normal to mode. If the analyser is on standby mode, the programme
produce these 3 base lines. shuts it down automatically. The lamp must never be
touched with fingers. Once the new lamp has been installed
and the covers of the optics and the rotor have been closed,
2.2.14.5. Fluid system supply
the instructions given by the programme must be followed.
On clicking the Supply button, with the main cover closed, The programme starts up the analyser, checks the light
the analyser fills the fluid system conduits with system intensity of the optical system, shuts down the analyser
liquid. It is possible to supply the dispensing system, the and then requests the user to remove the lamp holder again
washing system or both at the same time. To supply the and replace it again turning it 180º on the axis of the lamp.
dispensing system, the operating arm moves to the washing The programme forces waiting periods to prevent the lamp
station. It is possible to choose the number of supply cycles holder from getting too hot. If the temperature of the lamp
to be performed. When the system liquid container is filled holder is still high, handle it with pincers. The programme
during a work session, the fluid system can be supplied by starts up the analyser again, measures the light intensity
clicking directly on the New System Liquid Container button of the optical system again, compares the light intensity in
on the vertical button bar. both possible positions and chooses the greatest luminosity.
If it is the current position, it tells the user that the test is
complete. If the best position were the previous one, the
2.2.14.6. Cleaning of the dispensing system programme shuts down the analyser and asks the user to
remove the lamp holder and replace it, turning it 180º on
By clicking on the Wash button, with the main cover closed, the axis of the lamp, returning the lamp to its initial position.
the analyser washes the dispensing system internally and
externally. To perform this operation, the operating arm is 2.2.14.8 Change the rotor type
moved to the washing station. The user can choose to
perform the wash with system liquid or with washing solution. In this screen the type of rotor is introduced. Each rotor
In the case of the latter, the analyser asks the user to comes labelled with a letter in its top part. Select in this
position the washing solution container, supplied with the screen the type of rotor to use. For rotors marked with A
analyser, marked in green, in the system liquid position. letter, only select the letter. For the rotors marked with
Once the wash has been performed, the analyser asks other letters, select OTHERS and then introduce the light
him or her to replace the system liquid container and path that will come it within the box of rotors or of the
performs a rinse with the system liquid. distributor.

2.2.14.7. Changing the lamp 2.2.15. Internal quality control


When the user fits a new lamp, this utility must be used to Internal quality control enables the verification of the correct
notify the analyser that the lamp has been changed and functioning of each measurement procedure and the
optimise the luminosity of the photometric system. The validation or rejection of the analytical series. An analytical
programme requests confirmation to know whether or not series is an interval of time or results in which it is
the lamp of the analyser has been physically changed. If it reasonable to suppose that the metrological characteristics

33
User manual

of series. This parameter is the number of data used to


perform the statistical analysis, i.e. to calculate the
measurement (Xm) and the standard deviation (s). It is
recommended that it is not above 20. A series of results
is accepted if the values of the controls are within the
interval Xm ± ks, where k is the rejection criteria.
· In manual mode, the series of results is accepted if the
results of the controls are between the values of Minimum
(Cmin) and Maximum (Cmax) Concentration, respectively.
These values are programmed by the user on the test
programming screen, for the specific controls, or on the
multiple controls programming, for multiple controls. The
average value and the standard deviation are calculated,
respectively, as Xm = (Cmin+Cmax) / 2 and s = (Cmax-Cmin) /
(2k).
of the measurement procedure are maintained stable. Each The Quality Control screen of the user programme enables
laboratory must establish its own internal quality control access to the quality control results of the analyser. The
programme and the correction procedures to be followed if programme memorises all the control results for each test
the controls do not meet acceptable tolerances. The basic and for each sample type. The user can select a test and a
idea of internal quality control is very simple: one or more sample type and the screen indicates, for each control,
control materials, the controls, are placed between the the name, batch, average value (Xm), standard deviation
patient samples and they are measured together. The result (s), variation quotient CV = 100 s / Xm, and the rejection
from the control material is compared with an expected limits programmed. Whenever a new batch is introduced
value. In this way, it is possible to detect the occasional for one of the controls, the programme automatically creates
existence of anomalies in the measurement procedure. The a new data sheet and the statistical calculations are reset
use of controls is programmed independently for each for this new sheet. All the sheets created can be accessed
measurement procedure from the Test Programming through a dropdown list. A grid displays, for each series
screen. The analyser automatically proposes the carrying performed, the date, concentration, absolute error and
out of controls whenever the work session is reset, in accord relative error, for each control, and an alert for the rejected
with the tests programmed, and the user simply has to series. Absolute error is defined as E abs = C - Xm, where C
activate them. The user can also launch additional controls is the measured control concentration, and the relative error
manually. The use of BioSystems valuated control materials as Erel = Eabs / s. The programme displays the alert for
and calibrators is recommended. rejecting a series when the result of one of the controls is
out of the programmed rejection limits.
From the test programming screen, the user can programme
the number of controls (0, 1 or 2), the rejection criteria With the memorised data, the programme displays the
(between 0.1 and 3 standard deviations), the number of corresponding Levey-Jennings graphs. When any of the
replicates (1, 2 or 3), the control type (multiple or specific) controls of a series exceeds the interval limits Xm ± 2s, the
and the calculation mode (manual or statistical). The programme automatically executes the Westgard algorithm
calculation mode indicates the method used for deciding if and shows the results on the Levey-Jennings graph. This
a series of results is accepted or rejected. algorithm is a set of control rules that are examined
· In statistical mode, the user must introduce the number sequentially. The result of this algorithm is not used by the
programme to accept or reject a series. It is presented as
graphic information only and the user is who must decide

34
to accept or reject the series. The programme only displays 2.2. Alarms and alerts
the rejection alert based on the rejection interval chosen
by the user. The Westgard rules examined are: By clicking on the corresponding button on the horizontal
bar menu, the user can access the Current alarms screen,
· 13s: The result obtained on one of the controls exceeds
which displays the alarms and alerts generated during the
the limits of the interval Xm ± 3s.
current session. Through the main menu, it is also possible
· 22s: The results obtained on two controls exceed, in the to access the Past alarms and alerts, which contains a list
same way, the limits of the interval Xm ± 2s. This can be with the alarms and alerts generated during previous work
the two controls of one series or one single control on sessions.
two consecutive series.
The following is a description of the main alarms and alerts
· R4s: The difference between the results of two controls of
issued by the analyser, requiring user intervention, together
one series or of one control on two consecutive series is
with their possible cause and solution. The alarms that
higher than 4s.
require contact with the technical Assistance Service are
· 41s: Four consecutive results have been obtained, indicated. If any of the alarms persists, it is also necessary
exceeding, in the same way, the limits of the interval to contact the TAS (Technical Assistance Service).
Xm ± 1s. This may occur with two controls on two
consecutive series or with one single control on four
consecutive series. Machine alarms and alerts
· 10Xm: Ten consecutive results have been obtained, all General
greater or all less than the average. This may occur with
two controls on five consecutive series or with one single - Incorrect rack type. Check the configuration of the rack
control on ten consecutive series. tray. If the error persists once the racks have been
correctly positioned, deactivate the sensor of the rack
tray from the corresponding configuration screen and
Buttons contact the TAS.
- Analyser cover open. Close the analyser cover. If the
The Quality Control screen has different buttons that let
error persists, deactivate the cover sensors from the
the user perform different actions:
corresponding configuration screen and contact the
Graphs: This enables the display of the Levey-Jennings TAS. Danger: the main cover of the analyser must
graphs of the test selected for each control. The squares always be closed when it is running.
represent the values obtained in the accepted series. When - Electronics start-up fail. Shut down the analyser and
a Westgard algorithm rule is activated, the values involved start it up again. If the problem persists, contact the
are represented in circles. In the rejected series, the values TAS.
are represented by a triangle. The axis of ordinates indicates
- Error in the Firmware version. Contact the TAS.
the concentration, at standard deviation s. The horizontal
line indicates the number of the series. The horizontal lines Operating arm
indicate the multiples of the standard deviation and the
continual lines the programmed rejection limits. - The operating arm cannot be started. Check that the
arm is not blocked and that there is no object obstructing
Delete: Deletes the series selected on the grid. its movement.
Delete Sheet: Deletes the selected results sheet. - Loss of steps during the functioning of the Z axis. The
Print: Prints a Quality Control report with the data of the arm has collided vertically or is broken. Check that there
selected test and sample type. It is also possible to is no object obstructing the movement of the arm.
print out the Levey-Jennings graph. - Self-elevation safety device broken. Contact the TAS.
Edit: Modifies the rejection criteria and the calculation mode.
The changes affect the selected sheet only. Dispensing system

Introduce Series: Enables manual introduction of series • Needle


up to a maximum of 30 series per sheet. - No needle detected. Check that the needle is correctly
New sheet: Enables the creation of a new sheet when the installed.
30 series of the current sheet are exceeded. If a new - Needle is too bent. The needle has been detected to
sheet is not created, the following series overwrite the be outside its safety margins. Check the straightness
oldest series. A new sheet is automatically created when of the needle. If necessary, replace it with a new needle.
a test parameter is changed.
- Needle level detection error. Contact the TAS.
• Needle thermostatisation system
- Needle thermostatisation error. Contact the TAS. The
alarm gives supplementary information to be used by
35
User manual

the TAS to diagnose the breakdown. correctly in position.


• Dispensing pump - Reagent volume run out. Add more reagent and indicate
it to the analyser from the Positions screen for it to
- The dispensing pump cannot be started. Contact the
perform the analyses. Check the reagent bottle is
TAS.
correctly in position.
• Containers
Results
- There is no system liquid container. Place the contai-
ner correctly. see the “guide to messages” document directly in the user
- There is no waste container. Place the container program in the help menu.
correctly.
- Waste container full. Empty the waste container.
- System liquid container empty. Fill the system liquid
container.
- Error in the container level control system. Deactivate
the level control scales from the corresponding
configuration screen and contact the TAS.

Reactions rotor and readings


- The rotor cannot be started. Check that the reactions
rotor is not blocked.
- No rotor. Place a rotor in the analyser.
- End of rotor. 120 wells of the current rotor have been
used. Place a new empty rotor in the analyser.
- No rotor cover. Position the cover correctly. If the error
persists, deactivate the cover sensors from the
corresponding configuration screen and contact the
TAS. Danger: the main cover of the analyser must
always be closed when it is running.
• Rotor thermostatisation system
- Rotor thermostatisation error. Contact the TAS. The
alarm gives supplementary information to be used by
the TAS to diagnose the breakdown.

Optical system
- Base line error: reduced light intensity. Place a new
rotor. If the alert persists, replace the lamp.
- Lamp’s life. Information referring to the insufficient
luminosity of the lamp. This message will appear during
the Warming up or during the change of rotor. Replace
the lamp.
- Reduced light intensity in x filter. Replace x filter
recommended / necessary. Replace corresponding filter.
- Filter wheel cannot be started. Contact the TAS.
- Error in photometric system. Contact the TAS.
- Lamp lifetime exceeded. Replace the lamp.

Work session alarms and alerts

Preparations
- Sample volume run out. Add more sample and indicate
it to the analyser from the Positions screen for it to
perform the analyses. Check the sample tube is

36
3. Calculation and measurement procedures
This chapter describes the different analysis modes of the are indicated. The analysis modes are the same for blanks,
analyser and the calculations made to obtain the analytical calibrators, controls and samples of patients. The controls
results, i.e. the concentration values of the different analytes are treated in the same way as the patient samples in all
of the samples. In each case, the different formulas used the calculations.

Symbols used in the formulas

Asample Sample absorbance (patient or control)


Acalibrator Calibrator absorbance
Ablank Blank absorbance
λmain
[...] Absorbance value at main wavelength
[...]λreference Absorbance value at reference wavelength
TR Factor that depends on the type of reaction programmed. Its value is +1 for increasing reactions
and -1 for decreasing reactions.
F Calibration factor
Csample Sample concentration (patient or control)
Ccalibrator Programmed calibrator concentration
Func[...] Calibration curve or function (with multipoint calibrators)
nsample Number of sample replicates (patient or control)
ncalibrator Number of calibrator replicates
nblank Number of blank replicates
i Replicate index
[...] R1 Absorbance value with the first reagent in a bireagent differential method.
R1+R2
[...] Absorbance value after adding the second reagent in a bireagent differential method.
[...] T1 Absorbance value in the first reading in a fixed time method.
T2
[...] Absorbance value in the second reading in a fixed time method.
∆A Variation of absorbance of a sample per unit of time (patient or control)
∆t
sample

∆A Variation of absorbance of a calibrator per unit of time


∆t
calibrator

∆A Variation of absorbance of a blank per unit of time


∆t
blank

37
User manual

3.1. End point Asample-Ablank


Csample= Ccalibrator
Acalibrator-Ablank
3.1.1. Absorbance

The absorbance of the reaction is measured once only on


Considering that TR2=1, this formula is equivalent to the
a base line of distilled water. In this procedure, one or two
previous one with a factor
reagents may be used and the absorbance may be
measured at one or two wavelengths. The calibration can
Ccalibrator
be based on the use of multiple or specific calibrators (one F=TR·
or several), or on a programmed factor. For each test, a Acalibrator-Ablank
blank is prepared using distilled water instead of the sample
or with reagent only. The absorbance of the blank is also
measured on the base line of distilled water. 3.1.2.3. Several calibrators

If a multipoint calibrator is used, the concentration is


3.1.1.1. Monoreagent / Bireagent calculated using a calibration function or curve. This curve
is obtained from the programmed concentration values of
The procedure is different for the tests using one or two the calibrators and the absorbance values measured for
reagents. For the monoreagents, the analyser dispenses each one with regard to the base line Acalibrator, using an
the reagent and the sample in cycle 1 and measures the interpolation method (polygonal or spline) or a regression
absorbance of the reaction during a later cycle, in accord method (linear or quadratic) and linear or logarithmic axes,
with how it is programmed in the test. For bireagents, the in accord with how it is programmed fro each test. With
analyser dispenses reagent 1 and the sample in cycle 1. this curve, the analyser calculates the concentration of the
In a later cycle, it dispenses reagent 2 in the same well sample in accord with its absorbance with regard to the
and then measures the absorbance of the reaction in another base line
later cycle, in accord with how it is programmed in the
test. Csample=Func[Asample]

3.1.1.2. Monochromatic / Bichromatic 3.1.3. Replicates

The absorbance can be measured at one or two Up to 50 replicates can be programmed for each sample
wavelengths. In the case of bichromatic readings, the and up to 3 for each blank, calibrator or control.
difference between the absorbance at the main wavelength
and the absorbance at the reference wavelength is taken 3.1.3.1. Blank
as the absorbance value.
The average of the absorbance values measured is taken
Amuestra=[Asample]λmain-[Asample]λrefefence as the absorbance of the blank.
Acalibrator=[Acalibrator]λmain-[Acalibrator]λreference n blank

ΣA
1 i
Ablank=
Ablank=[Ablank]λmain-[Ablank]λreference nblank
blank

i=1

3.1.2. Concentration 3.1.3.2. Calibrator


Based on the absorbance value obtained, the analytical The average of the absorbance values measured is taken
concentration in the sample can be calculated. as the absorbance of the calibrator.
ncalibrator

ΣA
3.1.2.1. Factor 1 i
Acalibrator= calibrator
The concentration is calculated using a programmed fac- ncalibrador
i=1
tor with the formula
3.1.3.3. Sample
Csample=TR·F·(Asample-Ablank)
The average values calculated for the absorbance of the
blank and the calibrator or calibrators are used in the
3.1.2.2. Single calibrator calculations set forth in the previous section to obtain the
If a single point calibrator is used, the concentration is concentration of each replicate of the sample. The average
calculated with the formula value of the calculated concentrations is taken as the
concentration of the sample.

38
n sample 3.2.2.3. Several calibrators

Σ
1
Csample= Cisample
nsample If a multipoint calibrator is used, the concentration is
i=1 calculated using a calibration function or curve. This curve
is obtained from the programmed concentration values of
3.2. Bireagent differential the calibrators and the absorbance values measured for
each one with regard to the base line Acalibrator, using an
interpolation method (polygonal or spline) or a regression
3.2.1. Absorbance method (linear or quadratic) and linear or logarithmic axes,
The analyser dispenses reagent 1 and the sample in cycle in accord with how it is programmed fro each test. With
1. In a later cycle, it measures the absorbance of the this curve, the analyser calculates the concentration of the
mixture on a base line of distilled water. later, it dispenses sample in accord with its absorbance with regard to the
reagent 2 in the same well and then measures the base line
absorbance of the reaction in a later cycle, in accord with
how it is programmed in the test. The absorbances are Csample=Func[Asample]
measured at one single wavelength. For each test, a blank
is prepared using distilled water instead of the sample or 3.2.3. Replicates
with reagents only. The absorbances of said blank, with
the first reagent and with both reagents, are also measured Up to 50 replicates can be programmed for each sample
on the base line of distilled water. The calibration can be and up to 3 for each blank, calibrator or control.
based on the use of multiple or specific calibrators (one or
several), or on a programmed factor. The difference between
3.2.3.1. Blank
the absorbance measured with the two reagents and the
absorbance measured with the first reagent only is taken The average of the absorbance values measured is taken
as the absorbance value. as the absorbance of the blank.
n blank

ΣA
Asample=[Asample]R1+R2-[Asample]R1 1 i
Ablank= blank
Acalibrator=[Acalibrator] R1+R2
-[Acalibrator] R1 nblank
i=1
3.2.3.2. Calibrator
R1+R2 R1
Ablank=[Ablank] -[Ablank]
The average of the absorbance values measured is taken
as the absorbance of the calibrator.
3.2.2. Concentration

Based on the absorbance value obtained, the analytical ncalibrator

ΣA
concentration in the sample can be calculated. 1 i
Acalibrator= calibrator
ncalibrator
i=1
3.2.2.1. Factor 3.2.3.3. Sample
The concentration is calculated using a programmed fac- The average values calculated for the absorbance of the
tor with the formula blank and the calibrator or calibrators are used in the
calculations set forth in the previous section to obtain the
Csample=TR·F·(Asample-Ablank) concentration of each replicate of the sample. The average
value of the calculated concentrations is taken as the
3.2.2.2. Single calibrator concentration of the sample.

If a single point calibrator is used, the concentration is nsample

ΣC
1 i
calculated with the formula Csample= sample
nsample
i=1
Asample-Ablank
Csample= Ccalibrator
Acalibrator-Ablank 3.3. Fixed time
Considering that TR2=1, this formula is equivalent to the
previous one with a factor 3.3.1. Absorbance

The absorbance of the reaction is read at two particular


Ccalibrator times with regard to a base line of distilled water at one
F=TR· single wavelength. The calibration can be based on the
Acalibrator-Ablank

39
User manual

use of multiple or specific calibrators (one or several), or on is obtained from the programmed concentration values of
a programmed factor. For each test, a blank is prepared the calibrators and the absorbance values measured for
using distilled water instead of the sample or with reagent each one with regard to the base line Acalibrator, using an
only. The absorbances of this blank at the two specific interpolation method (polygonal or spline) or a regression
times are also measured on the base line of distilled water. method (linear or quadratic) and linear or logarithmic axes,
in accord with how it is programmed fro each test. With
this curve, the analyser calculates the concentration of the
3.3.1.1. Monoreagent / Bireagent sample in accord with its absorbance with regard to the
The procedure is different for the tests using one or two base line
reagents. For the monoreagents, the analyser dispenses Csample=Func[Asample]
the reagent and the sample in cycle 1 and measures the 3.3.3. Replicates
absorbance of the reaction in two later cycles, at the
moments in time T1 and T2, in accord with how it is Up to 50 replicates can be programmed for each sample
programmed in the test. For bireagents, the analyser dis- and up to 3 for each blank, calibrator or control.
penses reagent 1 and the sample in cycle 1. In a later
cycle, it dispenses reagent 2 in the same well and then
measures the absorbances of the reaction at the moments 3.3.3.1. Blank
in time T1 and T2, in accord with how it is programmed in The average of the absorbance values measured is taken
the test. The difference between the absorbance measured as the absorbance of the blank.
at time T1 and the absorbance measured at time T2 is
taken as the absorbance value. n blank

ΣA
1
i
Ablank= blank
T2
Asample=[Asample] -[Asample] T1 nblank
i=1

Acalibrator=[Acalibrator]T2-[Acalibrator]T1
3.3.3.2. Calibrator
Ablank=[Ablank]T2-[Ablank]T1
The average of the absorbance values measured is taken
as the absorbance of the calibrator.
3.3.2. Concentration
ncalibrator

ΣA
Based on the absorbance value obtained, the analytical 1 i
concentration in the sample can be calculated. Acalibrator= calibrator
ncalibrator
i=1

3.3.2.1. Factor 3.3.3.3. Sample


The concentration is calculated using a programmed fac- The average values calculated for the absorbance of the
tor with the formula blank and the calibrator or calibrators are used in the
Csample=TR·F·(Asample-Ablank) calculations set forth in the previous section to obtain the
concentration of each replicate of the sample. The average
value of the calculated concentrations is taken as the
3.3.2.2. Single calibrator concentration of the sample.
nsample

ΣC
If a single point calibrator is used, the concentration is 1 i
calculated with the formula Csample= sample
nsample
i=1
Asample-Ablank
Csample= Ccalibrator
Acalibrator-Ablank 3.4. Kinetics
Considering that TR2=1, this formula is equivalent to the
previous one with a factor 3.4.1. Variation of the absorbance per unit of time
Ccalibrator The kinetic mode is used to measure the catalytic activity
F=TR· of an enzyme. The absorbance of the reaction on a base
Acalibrator-Ablank
line of distilled water is measured periodically during several
cycles, between the times Ti and Tf programmed in the
test. The readings are taken at one single wavelength.
3.3.2.3. Several calibrators
Based on these absorbance measurements, the analyser
If a multipoint calibrator is used, the concentration is calculates the variation of the absorbance of the reaction
calculated using a calibration function or curve. This curve per unit of time. The calibration can be based on the use of

40
multiple or specific calibrators (one or several), or on a Considering that , this formula is equivalent to the previous
programmed factor. For each test, a blank is prepared using one with a factor
distilled water instead of the sample or with reagent only. Ccalibrator
The absorbances of this blank are also measured on the Csample=TR
base line of distilled water. ∆A ∆A
3.4.1.1. Monoreagent / Bireagent ∆t ∆t
calibrator blank

The procedure is different for the tests using one or two 3.4.2.3. Several calibrators
reagents. For the monoreagents, the analyser dispenses If a multipoint calibrator is used, the concentration is
the reagent and the sample in cycle 1 and measures the calculated using a calibration function or curve. This curve
absorbance of the reaction in several later cycles, between is obtained from the programmed concentration values of
the times Ti (start) and Tf (end), in accord with how it is the calibrators and the absorbance gradient values measured
programmed in the test. For bireagents, the analyser dis- for each one with regard to the base line ,
penses reagent 1 and the sample in cycle 1. In a later
cycle, it dispenses reagent 2 in the same well and then ∆A using an interpolation method (polygonal or
measures the absorbances of the reaction at the moments ∆t spline) or a regression method (linear or
calibrator
in several later cycles, between the times Ti and Tf, in quadratic) and linear or logarithmic axes, in
accord with how it is programmed in the test. accord with how it is programmed fro each test. With this
curve, the analyser calculates the concentration of the
3.4.1.2. Checking linearity sample in accord with the absorbance gradient with regard
Catalytic activity is measured by the speed of the reaction, to the base line
which is proportional to the gradient of the absorbance- ∆A
time curve. This ∆A/∆t gradient is calculated using the li-
Csample=Func [ ∆t ]
sample
near method over the set of absorbances measured between
the times Ti and Tf. The most common units for measuring 3.4.3. Replicates
the gradient are ∆A/min. Depending on the test, the period
of measurements can be different. In general, for the more Up to 50 replicates can be programmed for each sample
common tests, around 13 readings are taken at regular 15- and up to 3 for each blank, calibrator or control.
second intervals. The analyser automatically calculates the
best line of regression by the squared minimums method
3.4.3.1. Blank
and checks the linearity of the measurements base don
the correlation quotient. If the linearity is low, the analyser The average of the absorbance gradient values measured
issues the corresponding alert together with the result of is taken as the absorbance gradient of the blank.
the test.
n blank

Σ
∆A 1 i
3.4.2. Concentration = ∆A
∆t nblank ∆t
blank i=1
Based on the absorbance gradient obtained, the analytical blank

concentration in the sample can be calculated.


3.4.3.2. Calibrator
3.4.2.1. Factor
The average of the absorbance gradient values measured
The concentration is calculated using a programmed fac- is taken as the absorbance gradient of the calibrator.
tor with the formula
ncalibrator
∆A
Σ
∆A i
∆A 1
∆A
Csample=TR·F ( ∆t ∆t
) ∆t =
ncalibrator ∆t
sample blank calibrator i=1 calibrator
3.4.2.2. Single calibrator
3.4.3.3. Sample
If a single point calibrator is used, the concentration is
calculated with the formula The average values calculated for the absorbance gradient
of the blank and the calibrator or calibrators are used in the
∆A ∆A calculations set forth in the previous section to obtain the
∆t ∆t concentration of each replicate of the sample. The average
sample blank value of the calculated concentrations is taken as the
Csample= Ccalibrator concentration of the sample.
∆A ∆A nsample

ΣC
∆t ∆t 1 i
Csample= sample
calibrator blank
nsample
i=1

41