USER MANUAL
CS-400 AUTO-CHEMISTRY ANALYZER
Instruction:
Dear user, thanks for purchasing our CS-400 Auto-Chemistry Analyzer.
Please read the user manual carefully in order to operate the instrument correctly. Incorrect operation may affect
the precision and accuracy of the test results, or endanger personal safety.
Please keep the user manual safely for your any time reference.
Note:
● Instrument should be operated by medical inspection specialist, physician, nurse or lab assistant who are
specially trained.
● Instrument should be controlled by special software. Please install the software that is appointed by our
company. Installation of other software/hardware may interferer normal operation. Don’t operate other software
when instrument operating.
● Dust may accumulate on the surface of instrument after long time storage. Soft cloth or gauze can be used for
cleaning work, and a little detergent can be used if necessary. Please cut off the power supply before cleaning.
When instrument is not used, make sure shut the lid down.
● As to the use and storage method of the sample, reagent, Controls, Calibrator, please refer to the relevant
instructions.
● Sample, Controls, Calibrator and waster solution have the potential biochemical infectivity; the detergents are
corrosive that may hurt eyes, skin and mucosa. Operator should refer to the safety regulation for lab operation.
Protective measure should be taken to operator (Such as lab protective clothes and gloves).
● Avoid contact with eyes and skin, in case of skin contact, flush the area with water, rinse immediately with
plenty of water and seek medical advice.
● Operator should comply with the local regulation when draining and dealing with reagent, waste solution, waste
sample, consumable etc. Please dispose the waste solution and instrument consumable according to the regulation
of medical waste, infective waste and industrial waste.
I
Warning:
● Instrument should be operated in a good ground condition, and an independent power supply is a must, the input
power should be conformed to instrument requirement.
● Don’t pull the electrical wire with wet hand, or there is a risk of electrical shock.
● Doesn’t stamp, twist, drag the wire and cable, or it may cause a fire.
● Please don’t open the back and side cover board before cutting the general power supply except DIRUI
Medical special service staff.
● If liquid occurs in instrument interior or there is an internal pipeline leakage, please immediately cut off the
general power supply, and contact DIRUI Medical customer service dept.
● Please don’t touch sample probe, reagent probe and stirring rod, etc. when instrument operating, don’t put your
hand into the opening part, or it may cause body injury or instrument damage.
● Cut off the power supply before replace light source lamp. Don’t touch the lamp before it is cool to avoid
burning.
● Periodic maintenance should be executed strictly according to the user manual. Or it may cause instrument
malfunction, and affect the accuracy and precision of test results.
● Make sure that the Auto-Chemistry Analyzer is operated according to the user manual, or the measuring result is
not a reliable one, and the damage on instrument may endanger human safety.
● Please don’t place combustible material around the instrument.
II
Catalogue
Chapter 1 Brief Introduction...........................................................................................................................1
1.1 Summary ......................................................................................................................................................1
1.2 Main technical index ...................................................................................................................................1
1.3 Composition of instrument .........................................................................................................................3
1.4 Configuration and function ........................................................................................................................8
1.5 Instrument Symbol ....................................................................................................................................21
Chapter 2 Function and Measuring Principle .............................................................................................22
2.1 Mechanism movement principle ..............................................................................................................22
2.2 Assay mode .................................................................................................................................................24
2.3 Check of measure value ............................................................................................................................57
2.4 ISE testing principle ..................................................................................................................................62
Chapter 3 Instrument Installation ................................................................................................................66
3.1 Installation requirement ...........................................................................................................................66
3.2 Open package .............................................................................................................................................67
3.3 Installation procedure ...............................................................................................................................68
Chapter 4 Accessory Device...........................................................................................................................76
4.1 Sample disk barcode reader .....................................................................................................................76
4.2 Reagent barcode reader ............................................................................................................................77
4.3 Purified water equipment .........................................................................................................................78
Chapter 5 CS-400 Software Operation .........................................................................................................79
5.1 Software interface instruction ..................................................................................................................79
5.2 Software Operation ...................................................................................................................................83
5.3 Instrument standard specification ...........................................................................................................86
Chapter 6 Instrument Operation ..................................................................................................................88
6.1 Overview of operation ...............................................................................................................................88
6.2 Detailed operation .....................................................................................................................................89
Chapter 7 Calibration Information ............................................................................................................122
7.1 Colorimetric calibration..........................................................................................................................122
7.2 ISE calibration .........................................................................................................................................127
Chapter 8 Quality Control ...........................................................................................................................129
8.1 QC registration ........................................................................................................................................129
8.2 QC interval ...............................................................................................................................................133
8.3 Monthly quality control ..........................................................................................................................135
Chapter 9 System Setup ...............................................................................................................................137
9.1 Chemistry parameter ..............................................................................................................................137
9.2 Item combination .....................................................................................................................................145
9.3 Calculated item ........................................................................................................................................146
9.4 Cross contamination................................................................................................................................147
9.5 Report sheet format.................................................................................................................................150
9.6 ISE Setup ..................................................................................................................................................153
9.7 Other setup ...............................................................................................................................................154
9.8 Manual item setup ...................................................................................................................................156
9.9 LIS communication setup .......................................................................................................................156
Chapter 10 System management .................................................................................................................158
10.1 User information....................................................................................................................................158
10.2 Hospital information .............................................................................................................................159
10.3 Other information..................................................................................................................................160
10.4 Workload statistics ................................................................................................................................165
10.5 Database maintenance...........................................................................................................................167
10.6 System log ...............................................................................................................................................168
Chapter 11 System Help ...............................................................................................................................169
11.1 System help application.........................................................................................................................169
Chapter 12 System Maintenance.................................................................................................................170
12.1 System maintenance preparation .........................................................................................................170
12.2 The Application of system maintenance menu....................................................................................171
12.3 Maintenance and checkup points and parts........................................................................................180
III
12.4 Maintenance and check up method......................................................................................................183
12.5 ISE device maintenance ........................................................................................................................206
Chapter 13 Alarm Data Processing.............................................................................................................213
13.1 Alarm information type ........................................................................................................................213
13.2 Countermeasure to malfunction do not issue alarm...........................................................................213
13.3 Instrument alarm list.............................................................................................................................214
Chapter 14 Instrument Transportation and Storage .................................................................................244
14.1 Transportation requirement.................................................................................................................244
14.2 Storage requirement ..............................................................................................................................244
14.3 Storage environment .............................................................................................................................244
Addendum A Product Warranty .................................................................................................................245
Addendum B Product Description ..............................................................................................................246
Statement ..........................................................................................................................................................250
IV
Chapter 1 Brief Introduction
1.1 Summary
CS-400 Auto-Chemistry Analyzer is an instrument with discrete system, reagent open function, emergency
priority function as well as an external computer. The instrument is composed of humanized software operation
system, intelligent zed optical unit, complicated mechanism system, precision liquid path and accuracy electrical
system. The instrument could automatically realize sampling, reagent injection, anti-interference, mixture,
pre-temperature, reaction measurement, rinse, calculation, display and print function. The substitution of manual
operation for automatic operation could not only enhance the working efficient but also decrease the test error,
thus greatly enhance the accuracy and precision of test results.
CS-400 Auto-Chemistry Analyzer could carry out the immunology check and biochemical analyze of blood,
urine, ascites, cerebrospinal fluid and other body fluid. The instrument could also carry out clinic test, such as:
myocardium enzymogram, blood sugar, blood fat, liver function, renal function, immunoglobulin, etc.
1.2 Main technical index
Instrument structure:
Throughput:
Discrete system
Constant speed, 400 tests/ hour (800 tests/ hour with ISE)
Simultaneous analysis item No.: At most 88 colorimetric items,
3 ISE items (K, Na, Cl).
Sample volume:
2 to 35μl，
（Stepping 0.1μl）
Reagent volume:
20 to 350μl，（Stepping 1μl）
Reaction solution volume:
150～450μl
Liquid level sensor:
Integration of sample probe and reagent probe with touch sensor and sample probe
block test function.
Stirring:
Independent stirring after reagent injecting.
Sample disk：
115 samples position (50 routine samples, 34 Calibrators, 20 STAS samples, 8
Controls, 3 Detergents)
Reagent disk:
Dual disk
Disk 1: 45 positions for Reagent 1, Reagent 4, diluents and CS anti-bacterial
phosphor-free detergent.
Disk 2: 45 positions for Reagent 2, Reagent 3, CS anti-bacterial phosphor-free
detergent.
Photometer:
Grating spectrophotometry system in a range of 340～750nm, wavelength: 340、
380、405、450、480、505、546、570、600、660、700、750nm
V
Wave length accuracy:
±2nm
Light source:
20W /12V Long life quartz halogen lamp (water cooling)
Measurement range:
0 to 3.3Abs
Reaction disk:
120 pcs of reusable rigid optical plastic reaction cuvette.
Reaction cuvette optical diameter: 6mm
Reaction cuvette rinse:
Automatic
Incubation bath temperature:
37℃±0.1℃
Reaction time:
15 minutes at most (optional for 3, 4,5,10 and 15 minutes)
Analysis method:
Rate assay, end-point assay, 2-point assay.
Calibration method:
1-point linearity, 2-point linearity, multi-point linearity, non-linearity method.
Reagent bottle volume:
20ml, 70ml
Reagent cooling unit:
All reagents keep at 5℃ - 15℃, semiconductor refrigeration.
Barcode scanning：
3 internal barcode scanner ( scan the barcode on the routine sample,
on the R1, R2 disk )
Reagent volume test :
Test and report the reagent remaining volume.
Power supply:
~220/230V
Ambient temperature:
15℃～32℃
Relative humidity:
40% ～ 85%
Appearance dimension:
1060×790×1150mm ( length×width×height)
Fixing power:
2000VA
Weight:
About 300Kg
2
50Hz
Optimum temperature: 18℃ to 25℃
and reagent
1.3 Composition of instrument
1.3.1 Appearance of instrument
1.3.1.1 Front of instrument
① Model symbol
② Cover
③ Left front door
④ Right front door
Figure 1-1 Front of instrument
3
1.3.1.2 Front of opening door
① ISE pipetting syringe
② ISE diluting syringe
③ ISE inner standard solution syringe
④ Alkaline detergent port
⑤ Sample syringe
⑥ R2 syringe
Figure 1-2 Front of opening door
4
⑦ R1 syringe
1.3.1.3 Rear of instrument
①
③
②
① Power supply inlet
② RS-232 interface
⑤ Purified water inlet
⑥ Vacuum tank waste solution outlet
④
③ Cooling fan
⑤
⑥
⑦
⑧
④ Diluted waste solution outlet
⑦ Concentrated waste outlet
⑧ Port of concentrated waste level sensor
Figure 1-3
Rear of instrument
5
1.3.1.4 Top of instrument
①
⑦
⑧
②
③
⑨
④
⑩
⑤
11
○
⑥
12
○
① Sample pipetting mechanism
② Cuvette rinsing mechanism
③ Reaction disk
④ R1 stirring mechanism
⑤ R1 reagent pipetting mechanism
⑥ R1 reagent disk
⑦ Pilot lamp of sample disk rotation
⑩ R2 reagent pipetting mechanism
⑧ cooling lip of inner track of sample disk
11
○
R2 stirring mechanism
Figure 1-4 Top of instrument
6
⑨ Sample disk
12
○
R2 reagent disk
1.3.1.5 Right of instrument
①
②
③
④
① Analytical unit switch (cooling power supply is excluded) ② Pilot lamp of cooling power supply (green)
③ General power supply switch ( breaker)
④ Pilot lamp of power supply ( red)
Figure 1-5 Right of instrument
7
1.3.2 System configuration
Figure 1-6 System configuration
1.4 Configuration and function
CS-400 Auto-Chemistry Analyzer is composed by operating system and analytical system. The two parts is
connected by RS-232 serial wire.
1.4.1 Operating system
Operating system is composed of host, 17 inch CRT display monitor, keyboard, mouse and printer.
Host computer:
Windows XP system
Special applied software and database.
Computer configuration: Basic frequency ≥ 2.8GHz.
Hard disk≥ 160G
Memory ≥ 1G
With RS-232 serial interface, website interface and USB interface.
CRT display monitor: Display all kinds of form, curve and test data of CS-400 software.
Keyboard:
Operation control and data input.
Mouse:
Carry out software operation
Printer:
8
Print out test data and chart.
1.4.2 Analytical system
Analytical system is composed of sample disk, sample pipetting mechanism, reagent disk, reagent pipetting
mechanism, reaction disk, stirring mechanism, cooling system, rinsing mechanism, optical system etc.
1.4.2.1 Sample disk
! Warning：
△
z When instrument operating, be sure to close the cooling lid on the inner track of sample disk and screw the
knob.
z The sparkling of LED pilot lamp indicates that sample disk is turning or going to turn, do not change sample
or touch sample disk at this time, or it may cause body injury or instrument damage.
① ②
① Outer track
② Middle track
⑤ Handgrip of sample disk inner track
⑧ Handgrip of sample disk outer track
③ ④⑤
⑥
⑦⑧
③ Inner track
⑥ Sample disk guide pin
⑨
④ Sample disk inner track pin
⑦ Locking block of inner/outer disk
⑨ Sample barcode reader
Figure 1-7
Sample disk
(1) Composition and function
Sample disk is composed of outer track, middle track, inner track and LED pilot lamp. The inner track has
cooling function, thus the Controls and Calibrator on inner track of disk can be restored in 5℃～15℃.
Place the containers (standard cup, micro cup, test tube) which contain Calibrator, sample, Controls on the
sample disk, and then the sample disk will send them to the sampling position in the sampling mechanism.
（2）Specifications ( Sample No.)
(Outer track): For routine samples (1-50)………………………………………….50cups
9
(Middle track): For Calibrator (S1-S17)……………………………………………...17cups
For STAT sample (E51-E70)………………………………….…….. 20 cups
For detergent (W1-W3)………………………………………………..3 cups
(Inner track):
For Calibrator (S18-S34)……………………………………………..17 cups
For Controls（C1-C8）………………………………………………..8 cups
（3）Movement
At Power on: Sample disk turns counterclockwise to move routine sample No.1 to the sampling position.
At analysis: At start of analysis, sample disk makes the same movement as ―power on‖. During analysis,
sample disk turns to the direction allowing a quicker access.
At resetting: Make the same movement as at ―power on‖.
(4) Mounting / dismounting
When mounting the sample disk, be sure to set the sample disk matching with the guide pin. Set the latch
on the inner track. Also, be sure to secure the cooling unit lid on the inner track, the outer track can be
demounted without removing the inner track.
Note:When mounting/demounting the routine sample disk (outer track), be sure to hold the hooks with both hands.
In order to change the sample in inner track, be sure that sample probe has stopped sampling or been under the
stand-by status and take out the disk cover first. Before operating, please check the disk position.
(5) Action check
Single-click ― maintenance‖ key, select ― mechanism operation checkup‖, input the check times,
single-click ― Execute ‖ button. Alarm is issued when abnormality exists.
1.4.2.2 Sampling mechanism
! Warning:
△
●
10
Please don’t touch the sampling mechanism when operating, or it may cause body injury or instrument
damage.
①
②
③
④
① Sample probe up and down mechanism
② Sample probe rotation arm
③ Sample probe
④ Rinsing bath of sample probe
Figure 1-8 Sampling mechanism
(1) Function
Assimilates a specified amount of sample from sample container and pipets it into reaction cuvette. The
sample probe possesses the liquid level detect function. Alarm is issued when touch occurs in descending
process, and alarm is issued also when sample probe is blocked.
（2）Specification
Sampling mechanism can assimilate 2.0-35.0 ul sample volume, set in 0.1ul stepping. Sample probe will
assimilate more than specified amount. Minimum sample volume requires more than 100 ul.
（3）Movement
At power on: The sample probe comes over above the reaction cup, and then returns above the sample
probe rinse bath.
At analysis:
The sample probe circulates up and down motions as the following sequence：Sample
container, reaction cuvette, sample rinsing bath. Automatic sample probe rinsing is carried
out when sampling finished. At start of analysis, the sample probe makes the same movement
as at power on. In the rinsing bath, the inner and outer walls of the probe are rinsed. Sample
probe block test is carried out simultaneously.
At resetting: Makes the same movement as at power on.
Assimilate sample: sampling is taken when sample probe descent 1.7mm below liquid level.
(4) Automatic rinsing
Automatic rinsing of probe: After assimilating the sample, the sample probe will return above the rinsing
bath to wash the inner and outer walls of probe。When sampling is finished, the Alkaline detergent is
1
1
assimilated from the position W1 of the sample disk.
（5）Operation check
Single-click the ―Maintenance‖key, select―mechanism operation checkup and input the check times. Click
―Execute ―. If abnormality exists, instrument will issue alarm.
1.4.2.3 Reagent disk
! Warning:
△
● Please don’t touch the lid of the reagent disk when running or it may cause body injury and instrument
damage.
①
②
③
④ ⑤ ⑥
⑦
① Reagent disk cover
② Locking knob of cover
③ Reagent bottle ( on the reagent rack)
④ Track pin of reagent disk
⑤ Handgrip of reagent disk.
⑥ Reagent disk guide pin
⑦ Reagent disk open detector
Figure 1-9 Reagent disk
（1）Function
The reagent disk accommodates reagent bottles and carries the specific reagent, diluents and
Anti-bacterial phosphor-free detergent to the pipetting position in pipetting mechanism. Cooling system can
keep the reagent disk in a lower temperature to store reagent. There is a barcode reader on the wall of
reagent cooling unit, which can scan the barcode on reagent bottle.
（2）Specification
Reagent disk 1 (R1): R1, R4, diluents and detergent, total 45 bottles. Number 45 position is specially used
for Anti-bacterial phosphor-free detergent.
Reagent disk 2 (R2): R2, R3, diluents and detergent, total 45 bottles. Number 45 position is specially used
12
for Anti-bacterial phosphor-free detergent.
Reagent bottle capacity: 70ml、20ml
Single-reagent usage: Single-reagent can be used as Reagent 1 or Reagent 2 that is putted into both two
reagent disk.
（3）Movement
At power on: Turning clockwise, Position 1 of R1 and R2 disks carry each bottle to the reagent pipetting
position.
At analysis: Initial operation is the same as at power on. Subsequently, the disks rotate in the direction
which allows a quicker access.
At resetting: Make sure same as at power on.
（4）Operation check
Single-click the ―Maintenance‖key, select―mechanism operation checkup and input the check times. Click
―Execute―. If abnormality exists, instrument will issue alarm.
（5）Mounting/ Dismounting
Fix the disk with 2 latches at the center of disk. To remove the disk, loose the latches. For mounting, be sure
to set the disk matching with the guide pin and fasten it with the latches. The reagent disk cover must be
attached except for replacement of the disk of reagent. During operation, the reagent probe may moves, so
avoid detaching the reagent disk cover at this time.
Note: If operator open the disk cover during stand-by or running state, alarm occurs. Reagent horizontal scanning
will be automatically carried out when cover the reagent disk lid at stand-by status.
1.4.2.4 Reagent pipetting mechanism
! Warning:
△
● Be sure to close the lid of reagent disk when instrument operating, and fasten the knob.
1
3
①
②
③
④
① Reagent probe up and down mechanism
② Rotating arm of reagent probe
③ Reagent probe
④ Rinsing bath of reagent probe
Figure 1-10 Reagent pipetting mechanism
（1）Function
Assimilate a specified amount of reagent from each reagent bottle and pipets it into a reaction cuvette. The
reagent probe acts as a sensor at the liquid surface level. The remaining amount of reagent in a bottle is
calculated from the descending distance of reagent probe and displayed on the ―reagent info.’ menu.
（2）Specification
Reagent volume: 20 to 350ul set in 1 ul stepping.
（3）Movement
At power on：Move toward the reaction cup side once and then returns above the probe rinsing bath.
At analysis: Reagent probe circulates the motion as the following sequence: reagent bottle--reaction
cuvette—reagent probe rinsing bath.
At resetting: Make the same action as at power on.
（4）Automatic rinsing
Assimilate Anti-bacterial phosphor-free detergent from the position 45 on the reagent disk for three
times and infuse it into reaction cuvette for three times and return to the probe rinsing bath to wash inner
and outer walls of the probe.
（5）Movement check
Single-click the ―Maintenance‖key, select―mechanism operation checkup and input the check times. Click
―Execute ―. If abnormality exists, instrument will issue alarm.
14
1.4.2.5 Reaction disk
! Warning：
△
● Please don’t touch the reaction disks during operating, or it may cause mechanical failure.
①
②
③
④
⑥
⑤
① Reaction cuvette rinsing mechanism
② Reaction disk
③ Fixing screw
④ Fixing knob of reaction disk
⑤ Guide pin and hole
⑥ Reaction cup groupware handgrip
Figure 1-11
Reaction disk
（1）Function
Fasten reaction cuvette by screw, and be sure to keep chemical reaction in constant temperature of 37℃.
Each reaction cuvette also serves as a cell for absorbance measurement.
（2）Specification
Reaction cuvettes No: 20 cuvettes/set * 6 sets ( 120 cuvettes in total)
Optical diameter: 6mm
Material of reaction cuvette: Optical plastic
（3）Operation
Always rotates counterclockwise
At power on：Rotates and stops at the start position. At this time, the first reaction cuvette is located under
the first rinse nozzle.
At analysis: Initially operates the same as at power on, then circulates a cycle of half turn and one pitch
advance (covering 61 reaction cuvettes) followed by temporary stop. Each full turn takes about 18 seconds.
1
5
At resetting: Make the same action as at power on.
（4）Rinsing
At position No. 45 of both reagent disks, set a bottle containing Anti-bacterial phosphor-free detergent and
open its cap. Carry out ― rinsing reaction cuvette‖ in the ―maintenance‖ window, all the reaction
cuvettes will be rinsed. It is usually rinsed by Alkaline detergent in the front-left of instrument, so daily
maintenance for reaction cuvettes is unnecessary.
（5）Operation check
Single-click the ― Maintenance‖ key, select ― mechanism movement checkup‖, and input the check times.
Click ― Execute―. If abnormal exist, instrument will issue alarm.
（6）Demounting
Reaction disk: Remove the rinse mechanism from above the reaction disk and completely loosen the fixing
knob at the center. The reaction disk can be lifted out. For mounting, align the guide pin of instrument with
the guide hole of the reaction disk and tighten the fixing screw.
Reaction cuvette: Remove the cuvette setscrew and pull up the reaction cuvette block by holding the
handgrip, and the reaction cuvette can be removed from the reaction disk.
Note:Keep reaction cuvette immersed in purified water. Besides, if the instrument will be not used more than 3
days, reaction cuvette should be removed and immersed in purified water.
1.4.2.6 Incubation bath
! Warning:
△
● Keep the cleanness of purified water in incubation bath, or it may effect the test precision.
● When instrument startup or rinsing incubation bath, make sure there is enough Anti-bacterial
phosphor-free detergent at No.45 position.
（1）Function
Keep the reaction solution in the reaction cuvette at a constant temperature.
（2）Operation
At power on: Automatic exchanges the constant temperature water once, the Anti-bacterial phosphor-free
detergent in position No.45 of both reagent disks is added in incubation bath.
At analysis: Incubation bath water is circulating. Instrument may automatically supply water when water
shortage comes in operation process.
Exchange water: In ―maintenance‖ window, select ―rinsing incubation bath‖ , and then the constant
temperature water may exchange, and then add 6ml CS anti-bacterial phosphor-free detergent in incubation
bath water.
Note: After running for 24 hours, instrument may require ―incubation bath water exchange‖, please carry out
― Rinsing incubation bath‖.
16
1.4.2.7 Stirring mechanism
! Warning:
△
● Please don’t touch stirring mechanism When operate, or it may cause body injury or instrument damage.
(1) Function
Stirring the reaction solution in each reaction cuvette.
(2) Operation
At power on: Move to the side of reaction cuvette and then stops above the rinsing bath, move to the side
of reaction cuvette again, and then stops above the rinsing bath.
At analysis: The mechanism descends, rotates, rises and stops between two locations: reaction cuvette and
stirring rod rinsing bath.
Stirring is carried out after each addition of reagent 1(R1),reagent 2(R2), reagent3(R3), reagent 4(R4). R1
and R4 use Stirring rod mechanism 1. R2 and R3 use Stirring rod mechanism 2.
(3) Automatic rinsing
Automatic rinsing of stirring rod: when stirring rod descends into stirring rod rinsing bath, mechanism may
automatically rotate and washes the stirring rod with purified water.
Sampling finishing: Stirring rod is stirring in reaction cuvette in which detergent is added, thus rinse the
stirring rod.
(4) Operation check
Single-click the ―maintenance‖ key, select ― mechanism operation check‖, and input the check times. Click
― Execute‖. If abnormality exists, instrument will issue alarm.
1.4.2.8 Reaction cuvette rinsing mechanism
! Warning:
△
● Please don’t touch the rinsing mechanism when operate, or it may cause body injury or instrument damage
● Avoid directly contact with body, or it may cause infection. Please adopt protective measure. In case of
skin contact, flush the area with water, rinse immediately with plenty of water and seek medical advice.
（1）Function
Eliminates the reaction solution, rinse the reaction cuvette,
used for test cell blank
injects and eliminates purified water which
(2 ) Composition of rinsing nozzle
1
7
C
D
A B
A
B
B
E
测4 4times
次杯空白（1
次通过）
cell blank次停止，3
measurement.
Rotational
Rotational
direction of
Reaction disk
1 stop, 3 pass.
Figure 12 Arrangement of Rinse Nozzles
Above figure shows that seven steps are needed when rinsing reaction cuvette. (Four times cell blank test is
added) , therefore, to finish rinsing one reaction cuvette, 11 steps are needed.
Step 1: Nozzle 1D assimilates reaction solution, then 1C discharges detergent to reaction cuvette.
Step 2:
Nozzle 2B assimilate detergent from reaction cuvette, then 2A discharges purified water to reaction
cuvette.
Step 3:
Nozzle 3B assimilates the purified water from reaction cuvette, then 3A discharges the purified
water to reaction cuvette.
Step 4:
Nozzle 4B assimilates the purified water from reaction cuvette, and wipes the reaction cuvette at the
same time.
Step 5: Nozzle 5E discharges the purified water to reaction cuvette.
Step 6,7,8,and 9: Carry out the cell blank check measurement at the fifth nozzle. The reaction cuvette with
full purified water allows 4 times cell blank measurement. (1 time static measurement, 3 times
dynamic measurement when reaction cuvette passed by during reaction disk turning )
Step 10: Nozzle 6F assimilates the purified water, which has carried out the cell blank absorbency check.
Step 11:
Nozzle 7 F assimilates the remaining water in reaction cuvette, and wipes the reaction cuvette at
the same time; Nozzle 7G only discharges the purified water in rinsing process before sampling,
rinse the nozzle tip.
Distribution of 11 rinsing nozzles:
A. For discharging purified water ………………………….2 nozzles
B. For assimilating rinse water…………………………….3 nozzles
C. For discharging detergent……..…………………………1 nozzle
D. For assimilating reaction solution ..……………………. 1 nozzle
E. For discharging purified water for cup blank……………1 nozzle
F. For aspirating purified water for cup blank……………….2 nozzles
18
G. For discharging purified water ………………………..…1 nozzle
（3）Operation
At power on: Rises when already down
At analysis:
Rinse the reaction cuvette and carry out the water blank test in the rotational direction of
arrangement of rinse nozzle in Figure.1-13.
（4）Operation check
Single-click the ―maintenance‖ key, select ― mechanism operation check‖, and input the check times. Click
― Execute ‖. If abnormality exist, instrument will issue alarm.
（5）Dismounting
The rinse mechanism can be shifted from the reaction disk by loosening the fixing screw at the head.
1.4.2.9 Reagent cooling system
(1) Composition and function:
Cooling system is composed of reagent cooling system and sample cooling system, which could cool the
reagent, Controls and Calibrator respectively.
（2）Specifications
Temperature: 5 -- 15℃
! Warning:
△
● Even the analyzing system is power off, cooling system is still at working status. The cooling system
only stop working when main power supply is cut off.
● The usage and storage of Reagent should be performed strictly according to user manual.
1.4.2.10 Optical system
Concave
diffraction
grating
Detector (340-750nm)
12 fixed wavelength
Light source
lamp
Reaction
cuvette
Figure 1-13
Photometer
1
9
(1) Function
When the reaction disk rotates, the absorbance of purified water or reaction solution is measured in each
reaction cuvette. As above figure shows.
（2）Specification:
Carry out photometry with dual-wavelength or single-wavelength at wavelengths: 340nm、380nm、405nm、
450nm、480nm、505nm、546nm、570nm、600nm、660nm、700nm、750nm.
Wavelength accuracy: ±2nm
Measuring range: 0 -3.3 Abs
Spectral bandwidth: FHW 8 to 10nm
Detector: Silicon photodiode
Light source: 12V, 20W halogen lamp
20
1.5 Instrument Symbol
Symbol
Meaning
To perform as the instruction under the symbol,
emphasize the important information and special contents.
To perform as the instruction under the mark,
or it may cause biological infection
AC symbol
Only diagnostic use
Storage at
Batch code
Use by
Serial number
Measurement Control
Manufacture by
Grounding terminal
Table 1-2 Instrument symbol
2
1
Chapter 2
Function and Measuring Principle
The function and measuring principle is composed of mechanism movement principle and analyzing assay.
2.1 Mechanism movement principle
CS-400 Auto-Chemistry Analyzer consists primarily of the sample disk, sampling mechanism, reagent
disk, reagent pipeting mechanism, reaction disk, reaction bath, rinsing mechanism and photometer.
Operation of each mechanism is explained according to figure 2-1:
After starting, instrument carries out resetting first, rotates the R1 reagent disk and R2 reagent disk to
Position 1, then rotate them for 360 degrees, the R1 reagent probe, R2 reagent probe, sample probe, R1
stirring rod, R2 stirring rod are all stopped at the upper side of their rinsing bathes respectively.
Upon pressing the start key, the rinse mechanism starts rinsing from reaction cuvette No1. The reaction
disk rotates by 22 patches and stops temporarily, and then rotates by 37 patches and stops, sequently, 2 more
patches and then stops. This sequence is carried out again to cover one full turn plus 2 patches (18 seconds).
Cell blank is measured when the reaction cuvette passes through the photometric unit during rotation of the
reaction disk. The measured value of cell blank becomes the reference value for the subsequent absorbance
measurement. The liquid in the reaction cuvette is assimilated through the nozzle of rinse mechanism.
After rinsing the reaction cuvette for 3 minutes (the reaction disk rotates 10 circles), the sampling
mechanism begins to work when reaction disk rotates the 11th turns, and the sample probe moves above of the
sample cup and decends into the cup. Since the sample probe comprises a liquid level sensor, the probe stops
descending when its tip enters the sample. A set volume of sample is assimilated with the sample pipettor.
Next, the sample probe moves to the top of No.1 reaction cup and descends until its tip reaches the bottom of
the cuvette, where the sample is discharged. The sample probe further moves to the inside of probe rinsing
bath, where its inner and outer walls are washed with purified water.
On the other hand, the reagent pipetting mechanism assimilates reagent 1 (with R1 probe), while the
reaction disk rotates by 22 patches, stops temporarily and the rotates by 37 patches after the fore mentioned
sampling mechanism has discharged the sample into the reaction cuvette. When the reaction disk stops
temporarily after the above rotation, reagent 1 is discharged into the reaction cuvette. And when the reaction
disk stops temporarily after rotating by 2 patches, the R1 stirring mechanism mixes the sample and reagent.
The reagent pipetting mechanism rotates to the top of reagent bottle in the specified channel and descends
while assimilates and discharges reagent. A set volume of reagent is assimilated, then the reagent probe moves
to the top of reaction cuvette and discharges the reagent, followed by returning to the rinsing bath and
washing of its inner and outer walls. Add reagent R1 and start photometry. Measurement is made when each
reaction cuvette passes through the optical path during rotation of the reaction disk. The reaction disk
rotates15 turns plus 22 patches to the reagent 2 pipetting position, where reagent 2 is added with R2 probe.
Reagent 2 is stirred with the R2 stirring mechanism after the reaction disk rotates by 16 turn plus 2 patches
and stops temporarily, and then rotates 22 patches and stops temporarily.
After 26 turns plus 37 patches , reaction disk comes to the reagent 3 sampling position, and reagent
probe 2 assimilates reagent 3. R2 stirring rod begins to work after 2 more patches of reaction cuvette.
After 41 turns plus 37 patches , reaction disk comes to the reagent 4 sampling position, and reagent probe
1 assimilates reagent 4. R1 stirring rod begins to work after 2 more patches of reaction cuvette
After the lapse of about 15 minutes (60 circles), the first cup measurement is over, and the reaction
solution in No. 1 reaction cup is assimilated with the nozzle of cuvette rinsing mechanism, which will
discharge cleaning liquid and water into the reaction cup to rinse the cup. The instrument stops and goes to
22
standby status when the last reaction cuvette has been washed, and goes to the the cell blank status. The
reagent probe and sample probe will be rinsed respectively by their own detergents added by themselves after
every test.
Reagent combination (R1, R2, R3, R4) of each item and reaction time (3~15 mins) is set in the ―Analytical
parameter‖ form in ―System setup‖
2.1.1 Operating Position
Operating principle of 120 reaction cuvette is showed as below:
Reset point
Rinsing mechanism
Position No. of reaction cuvette.
63,光电检测位
Photoelectric
detection
62,R1 stirring
62,R1 stirring
rod
position
搅拌探针
63,R4 stirring
position
Sop and wipe
Sam. probe
Sample probe
60 R1 Pipetting
R1, R4 Probe
61 R4 Pipetting
Reference position
number
Reagent probe 2,3
Reagent probe
Stirring probe
Stirring
Figure 2-1 Each mechanism operation position
2.1.2 Analytical flow
The analytical flow of CS-400 Auto-Chemistry Analyzer is show in figure 2-2:
2
3
Table 2-1 Analytical flow
2.1.3 Photometric Features
This instrument adopts the whole reaction monitoring system, which intermittently measures the absorbance of
reaction solution for a reaction time of 15 minutes.
The reaction disk rotates 1 turn plus 2 pitches in about 18 seconds and during this time the absorbance is
measured for all of 120 reaction cuvettes which go across the optical axis of the photometer. For each reaction
cuvette, measurement is made10 times in a reaction time of about 3 minutes. 13 times measurement is made
during 4 minutes (13 photo metric point.). 16 times measurement is made during 5 minutes (16 photo metric
point33 times measurement is made during 10 minutes (33 photo metric point.) .49 times measurement is made
during 15 minutes (49 photo metric point.)
CS-400 multi-wavelength photometer condenses the white light emitted from the light source lamp through the
lens, passes the condensed light through the reaction cuvette and separates the light with the concave diffraction
grating. The separated respective wavelength components are simultaneously received on the 12 fixed detectors
and amplified by 12 amplifiers, then logarithmically converted to obtain the absorbance or difference in
absorbance. In 2 wavelengths photometry, concentration is measured by the value of the difference of dominant
wavelength and complementary wavelength. This means that the photometer features a correcting effect for
lipemia, hemolysis and icterus of sample and has a compensating effect for fluctuation in source voltage, thus
realizing stable measurement.
2.2 Assay mode
The assay mode of Auto-Chemistry Analyzer is based on the Beer-Lambert law that the material selective
absorption light
The main principle is: When monochromatic light with specific wavelength passes through the cuvette with
sample, the monochromatic light absorbency and sample liquid concentration are varies directly as the distance
24
which is passed through sample liquid by light:
I
A = lg（1/T）= lg（ 0 ）= ε b c
It
A － Absorbency of the light when passing through liquid
T － Transmitted intensity and incident intensity ratio: transmittance It/I0；
I0 － Incident intensity
It － Transmitted intensity
ε － Molar absorption coefficient of solution（ml×mmol 1×cm 1）
；
－
－
c － Mol concentration of the solution（mmol/ml）；
b － Solution layer thickness（cm）；
Solution layer thickness (b): Optical path, which is fixed by instrument. Molar absorption coefficient (ε) is the
correlation coefficient of the wavelength, solution and solution temperature. Linear relationship is displayed
between solution thickness and absorbency when in stable temperature and single wavelength（ε value is given
on the reagent bottle by factory）
If the sample liquid adequate distribution, interaction between liquid and incidence monochromatic light only
happens during absorbing process. No fluorescence, disperse and photochemical appear. No interaction between
substances in the solution while absorbing process. The absorbency possess conducts nature, and this condition
conforms to the Beer-Lambert law
2.2.1 Assay mode variety
As to how to set the assay parameter and standard liquid parameter, please refer to user manual. Assay mode is
showed as table 2-2:
2
5
Method
Item
Photometry point
L– 0– 0– 0
1＜L≤49
1-point
Assay
Cell blank
Formula
B1 + B 2 + B 3
3
AL + AL−1
2
Note
t ：time
B1 + B 2 + B 3
L– M– 0- 0
1＜L＜M≤49
2-point
Rate Assay
3
( AM + AM −1 ) − k ( AL + AL −1 )
2
(minute)
between
photometry
point L、m
1-point &
Rate Assay
Second
half
Second
half
M–N–0–0
1＜L≤M＜N≤49
L– M – 0 - 0
3≤L＜M＜N＜P≤49
L +2＜M
Second
half
B1 + B 2 + B 3
△（AQ- P）-k△（AN -M）
3
3
AL + AL−1
2
B1 + B 2 + B 3
3
( AN + AN − 1 ) − k ( AM + AM −1 )
2
B1 + B 2 + B 3
3
N–P–0–0
3≤L＜M＜N＜P≤49
N+2＜P
First half
AL + AL−1
2
B1 + B 2 + B 3
3
B1 + B 2 + B 3
L– 0 – 0 – 0
1＜L≤M＜N≤49
First half
△A（M-L）
3
L– 0 – 0 – 0
1＜M＜N≤L＜P＜Q≤49
Rate A
Assay with
Serum
Index
Measurem
ent.
AM + AM −1 AL + AL −1
−
2
2
t
B1 + B 2 + B 3
M–N–P–Q
1＜M＜N≤L＜P＜Q≤49
M+2＜N，P+2＜Q
Rate A
Assay
Rate B
Assay
(mode 2 )
3
L– M – 0 - 0
1＜L＜M≤49
L +2＜M
First half
Rate B
Assay
(mode 1 )
B1 + B 2 + B 3
L– M– 0– 0
1＜L＜M≤49
2-point
assay
P-N （ different wavelength from the
Half）
Two
△ A(P-N)–k △ A(M-L) （ same
conditions
wavelength as the second half）
B1 + B 2 + B 3
L– M – 0 – 0
3≤L＜M＜N＜P＜Q＜R≤49
L +2＜M
B1 + B 2 + B 3
3
Table 2-2 Assay mode table
Explanation of symbols:
26
△A（M-L）
3
L,m,n,p,q,r
: Photometric points
Rn
: Volume of nth reagent ,n=1 to 4
△A(R-Q)– k△A(P-N)
SB
: Stopped cell blank
B1,B2,B3
: Passed cell blanks
Ax
: Absorbance at photometric point x
△A(m-L)
: Change in absorbance per minute between photometric points L and M
k
: Liquid volume correction factor
a
k=
S + ∑ Rj
j =1
b
S + ∑ Ri
i =1
S
Rj 、Ri
: Sample volume
a: No. of reagents with correction (at Al measurement)
B: No. of reagents without correction (at Am measurement)
Note 1: The 5 th Photometric point won’t be Stirred after adding reagent 2. Stirring when the reaction disk
pauses after rotates one circle plus 2 pitches
Note 2: liquid in the reaction cuvette should be more than or equal to 150 ul, less than or equal to 450ul.
Note 3: Do input 0 if the photometric point is not used.
(1) 1-point Assay
Absorbance
Endpoint assay in which absorbance is measured at a designated photometric point (specific time point when
reaction reach balance) after addition of reagents. Figure 2-2 explains the 1-point assay.
Cell blanks
（B1+B2+B3）/3
R2～R4
R1
S
AL + AL − 1
2
SB B1 B2 B3
Time
Figure 2-2 1-point Assay
(a) Photometric point : 【L】-【0】-【0】-【0】 （1< L ≤ 49）
(b) Calculation of absorbance
2
7
The average of absorbance at measurement points Land L-1 is used.
AX = AL + AL − 1
2
(c) Calculation of concentration
C X = {K × ( AX − B ) + C1 }× IFA + IFB
SB: Stopped cup blank R1~R3:
Passed cup blank
R1~R4: Reagent adding position
Cx: Concentration of standby sample
C1: Concentration of standard 1 solution(reagent blank)
K: Factor
B: Absorbance of blank
IFA and IFB: Instrument constants, representing slope and intercept
(d) Analytical items
TP, ALB, etc.
(2) 2-point Assay
Absorbance
Endpoint assay in which measurement is made twice at different points to obtain the difference in absorbance.
One point is measured as the action initial, the other point is measured when the action reach endpoint or
balance. The difference between the absorbance of two photometric points is used for calculation sample
concentration. Figure 2-3 explains the 2-point assay:
Cell blanks
（B1+B2+B3）/3
R2～R4
Am + Am −1
R1
S
AL + AL −1
2
2
SB B1 B2 B3
Time
Figure 2-3 2-point Assay
(a) Photometric point : 【L】-【M】-【0】-【0】 （1≤ L ≤ 49）
28
(b) Calculation of absorbance
The difference between the average of absorbance at measurement point m and m-1 and that at
measurement point l and l-1 is used.
( Am + Am − 1) − k ( AL + AL − 1)
AX=
2
a
S + ∑ Rj
k=
j −1
b
S + ∑ Ri
i −1
a: No. of reagents at Al measurement
b: No. of reagents at Am measurement
(c) Calculation of concentration
C X = {K × ( AX − B) + C1 }× IFA + IFB
SB: Stopped cup blank R1~R3:
Passed cup blank
R1~R4: Reagent adding position
Ax: the deference between the photometric point M and L
Cx concentration of standby sample
C1: Concentration of standard 1 solution(reagent blank)
K: Factor
B: Absorbance of blank
IFA and IFB: Instrument constants, representing slope and intercept
(d) Analytical items
CRE, etc.
(3) 2-point Rate Assay
Measurement is made twice at different measurement points (The two point are neither measured initial nor
endpoint) to determine the change in absorbance per minute in order to calculate sample concentration. For
check of reaction limit level, refer to Figure 2-4:
Absorbance
Reaction limit level
Am-1
Cell blanks
R2～R4
Am
29
Figure 2-4 2-point Rate Assay
(a) Photometric point : 【L】-【M】-【0】-【0】 （1< L <M≤ 49）
(b) Calculation of absorbance
The difference between the average of absorbance at measurement points M and M-1 and that at
measurement points L and L-1, then divide the result by time.
( Am + Am − 1) ( AL + AL − 1)
−
AX=
2
2
t
t: Time (minute) between measurement points L and M
(c) Calculation of concentration
C X = {K × ( AX − B ) + C1 }× IFA + IFB
SB: Stopped cup blank B1~B3:
Passed cup blank R1~R4:
Reagent adding position
Ax: Average change in absorbance per minute between measurement points L and M
Cx: Concentration of standby sample
C1: Concentration of standard solution 1(reagent blank)
K: Factor
B: Absorbance of blank
IFA and IFB: Instrument constants, representing slope and intercept.
(d) Analytical items
30
BUN, CRE etc.
(4) Rate A Assay
Ordinary Rate Assay. The concentration or activity level is obtained from the change in absorbance between the
specified measurement points. Figure 2-5 explains the Rate A Assay.
Absorbance
S
R1
R2～R4
Cell blanks
（B1+B2+B3）/3
AL
Am
Reaction limit level
SB B1 B2 B3
Time
Figure 2-5 Rate A Assay
(a) Photometric point : 【L】-【M】-【0】-【0】 （1<L <M≤ 49） L+2<m)
(b) Calculation of absorbance
The change in absorbance per minute between measurement point L and M is obtained by the least
squares method
AX=△A(L-m)
(c) Calculation of concentration
C X = {K × ( AX − B) + C1 }× IFA + IFB
SB: Stopped cup blank
B1~B3: Passed cup blank
R1~R4: Reagent adding position
△A(L-m): Change in absorbance per minute between measurement point L and M
Cx: Concentration of standby sample
C1: Concentration of standard solution 1(reagent blank)
31
K: Factor
B: Absorbance of blank
IFA and IFB: Instrument constants, representing slope and intercept.
(d) Analytical items
AST, ALT, etc.
(5) 1-point & Rate Assay
Two tests are analyzed by the endpoint assay and rate assay in a single channel. Test A is analyzed by the
endpoint assay in the first half of the specified reaction time and test B is analyzed by the rate assay in the
second half. Figure 2-6 explains the 1-point & rate assay.
Figure 2-6 1-point& Rate Assay
(a)Input of ―analytical parameter‖
Correct input of test A and test B is required respectively
(Text A): Photometric point : 【L】-【0】-【0】-【0】
Dual tests assay: designate B
（Text B） :Photometric point: 【M】-【N】-【P】-【Q】
1<m＜n≤L＜P＜Q≤49
M+2＜N，P+2＜Q
(b) Calculation of absorbance
For test A, the average of absorbance at measurement points L and L-1 is used, and used for the test B is
the difference obtained by subtracting the change in absorbance per minute between measurement points
32
L and M from that between measurement points N and P
AA=
AL + AL − 1
2
AB=△Ap×q-k△Am×n
a
S + ∑ Rj
K=
j −1
b
S + ∑ Ri
i −1
a：△A(N-M): No. of reagents.
b：△A(Q-P) : No. of reagents.
(c) Calculation of concentration.
For each of tests A and B, calculation is made according to the following formula.
C X = {K × ( AX − B) + C1 }× IFA + IFB
SB: Stopped cup blank
B1~B3: Passed cup blank
Rn: Reagent adding position
AA and AB : Each calculated absorbance of tests A and B
k: Liquid volume correction factor
S: Sample volume
Rj、Ri: Volume of each reagent
Cx: concentration of standby sample
C1: Concentration of standard solution 1(reagent blank)
K: Factor
AX: Calculated absorbance
B: absorbance of standard solution 1(reagent blank)
IFA and IFB: Instrument constants, representing slope and intercept.
In the rate assay, the change in absorbance per minute is obtained by the least squares method
(6) 3-point dual assay
Endpoint assay and endpoint assay without sample blank are carried out simultaneously in the same cup. The
first half reaction time is used for test A, and the left half is used for test B. Figure 2-7 explains the rate A assay
33
Absorbance
with serum index measurement.
Cell blank
Time
Figure 2-7 3-point dual assay
(a)Input of analytical Parameters
Correct input of test A and test B is required respectively
(Test A)Photometric point : 【L】-【0】-【0】-【0】
Dual tests assay: designate B
(Test B) Photometric point： 【M】-【N】-【0】-【0】
1< L＜M<N≤49
(b) Calculation of absorbance
The average value of absorbance at photometric points L and L-1 is for test A, and test B, the difference
between the average absorbance at photometric points N and N-1 and that of photometric points Mind
M-1.
AA = AL + AL − 1
2
AB =
( AN + AN −1 ) − k × ( AM + AM −1 )
2
a
k=
S + ∑ Rj
j =1
b
S + ∑ Ri
i =1
a：△ AM : No. of reagent when tested
b：△ AN : No. of reagent when tested
(c) Calculation of concentration
34
C X = {K × ( A X − B) + C1 }× IFA + IFB
SB: Stopped cup blank
B1~B3: Passed cup blank
Rn: Reagent adding position
AA and AB : Each calculated absorbance of tests A and B
k: Liquid volume correction factor
S: Sample volume
Rj、Ri: Volume of each reagent
Cx: concentration of standby sample
C1: Concentration of standard solution 1(reagent blank)
K: Factor
AX: Calculated absorbance
B: absorbance of standard solution 1(reagent blank)
IFA and IFB: Instrument constants, representing slope and intercept.
(7) Rate B Assay (mode 1)
Two tests are analyzed by the rate assay in a single channel. In the first half of reaction time, test A is measured,
and test B is measured in the second half. In the rate B assay, correction is possible with sample blank and for
an endogenous reaction. However, the method of correction of such reaction varies with measuring wavelength.
So this assay is categorized into modes 1 and 2 for easier explanation. Mode 1 is subdivided depending on
whether or not measuring wavelength is the same between test A and B. Figure 2-8 explains the rate B assay
(mode 1).
35
Figure 2-8 Rate B Assay (mode 1)
When wavelength differs between tests A and B
(a) Input of analytical parameters
Respective entry is required for each of tests A and B.
(Test A) photometric point: 【L】-【M】-【0】-【0】
(Test B ) photometric point:
【N】-【P】-【0】-【0】
3≦L ＜m＜n＜p≦49。L+2＜m、n+2＜p
(b) Calculation of absorbance
Used for test A is the change in absorbance per minute between measurement point l and m, which is
obtained by least squares method. Used for test B is the change in absorbance per minute between
measurement points n and p, which is obtained by the same method, but do not carry out blank
correction.
△AA=△A(M-L)
△AB=△A(P-N)
(c) Calculation of concentration.
For each of tests A and B, calculation is made according to the following formula.
C X = {K × ( A X − B) + C1 }× IFA + IFB
SB: Stopped cup blank
B1~B3: Passed cup blank
AA and AB : Each calculated absorbance of tests A and B
k: Liquid volume correction factor
S: Sample volume
Rj、Ri: Volume of each reagent
Cx: concentration of standby sample
C1: Concentration of standard solution 1(reagent blank)
K: Factor
AX: Calculated absorbance
B: absorbance of standard solution 1(reagent blank)
IFA and IFB: Instrument constants, representing slope and intercept.
36
When wavelength is the same between tests A and B
(a) Input of analytical parameters
Respective entry is required for each of tests A and B.
(Test A) photometric point: 【L】-【M】-【0】-【0】
(Test B ) photometric point: 【N】-【P】-【0】-【0】
3≦L＜m＜n＜p≦49。L+2＜m、n+2＜p
(b) Calculation of absorbance
Used for test A is the change in absorbance per minute between measurement points l and m, which is
obtained by least squares method. Used for test B is the difference obtained by subtracting the above
change for test A from the change in absorbance per minute between measurement points n and p, which
is obtained by the same method
△ AA =△A（L- M）
△ AB =△A（P- N）-k×△A（L- M）
a
k=
S + ∑ Rj
j =1
b
S + ∑ Ri
i =1
a: △A（M - L）No. of reagents with correction
b: △A（P- Q）No. of reagents without correction
(c) Calculation of concentration
For each of tests A and B, calculation is made according to the following formula.
C X = {K × ( A X − B) + C1 }× IFA + IFB
SB: Stopped cup blank
B1~B3: Passed cup blank
AA and AB : Each calculated absorbance of tests A and B
k: Liquid volume correction factor
S: Sample volume
Rj、Ri: Volume of each reagent
Cx: concentration of standby sample
C1: Concentration of standard solution 1(reagent blank)
37
K: Factor
AX: Calculated absorbance
B: absorbance of standard solution 1(reagent blank)
IFA and IFB: Instrument constants, representing slope and intercept.
(8) Rate B Assay (mode 2)
Two tests are analyzed by the rate assay in a single channel. In the first half of reaction time, test A is measured,
and test B is measured in the second half. Correction of endogenous reaction is carried out by using the ratio of
change in absorbance within a time period after measurement of the first test in the first half of reaction time.
Figure 2-9 explains the rate B assay (mode 2).
s
Figure 2-9
Rate B Assay (mode 2)
(a) Input of analytical parameters
Respective entry is required for each of tests A and B.
(Test A) photometric point: 【L】-【M】-【0】-【0】
Dual tests assay: designate B
(Test B ) photometric point:
【N】-【P】-【Q】-【R】
3≦L＜m＜n＜p≦49。L+2＜m、n+2＜R
(b) Calculation of absorbance
Used for test A is the change in absorbance per minute between measurement points l and m, which is
obtained by the least squares method. Used for test B is the difference obtained by subtracting the change
in absorbance per minute between measurement points n and p from that between measurement points q
and r, which is obtained by the same method
△ AA =△A（L- M）
△ AB =△A（P- Q）-k×△A（P- N）
38
a
S + ∑ Rj
k=
j =1
b
S + ∑ Ri
i =1
a: △A（P- N）No. of reagents
b: △A（R- Q）No. of reagents
(c) Calculation of concentration
For each of tests A and B, calculation is made according to the following formula.
C X = {K × ( A X − B) + C1 }× IFA + IFB
SB: Stopped cup blank
B1~B3: Passed cup blank
AA and AB : Each calculated absorbance of tests A and B
k: Liquid volume correction factor
S: Sample volume
Rj、Ri: Volume of each reagent
Cx: Concentration of standby sample
C1: Concentration of standard solution 1(reagent blank)
K: Factor
AX: Calculated absorbance
B: absorbance of standard solution 1(reagent blank)
IFA and IFB: Instrument constants, representing slope and intercept.
2.2.2 Calibration Method
(1) Linearity Method (1-point linearity)
The absorbance and input K value of blank (or standard 1) is measured to prepare a working curve. Figure 2-10
explains the linear method.
39
Absorbance
Concentration
Figure 2-10 1-point linearity
(a) Calibration Parameter input
Calibration type: 【1-point linear】
Calibration point: 【1】 (number of standard sample )
Span point: 【0】
(b) Input K factor in ―calibration result‖ menu
Input K factor in the ―calibration result ‖
(c) Calculation of parameters for working curve
S1ABS (B): Change in absorbance per minute of blank (standard 1)
K: Input value.
C1: Concentration of standard 1（reagent blank ), input value.
(d) Calculation of concentration
C X = {K × ( A X − B) + C1 }× IFA + IFB
Cx: Concentration of standby sample
AX: Calculated absorbance or change of absorbance per minute.
IFA and IFB: Instrument constants, representing slope and intercept.
(e) Applicable assay
1-point assay, 2-point rate assay, 2-point assay, 3-point assay, 1-point&rate assay, rate A assay and rate B
assay
(2) Linearity Method ( 2-point linearity )
40
Blank (or standard 1) and standard sample (standard 2) are measured to prepare a linear working curve Figure
Absorbance
2-11 explains the linear method.
Concentration
Figure 2-11 2-point linearity
(a) Calibration Parameter input
Calibration type: 【2-point linearity】
Calibration point: 【2】 (number of standard sample )
Span point: 【2~6】
(b) Calculation of parameters for working curve
S1ABS (B): Absorbance or change in absorbance per minute of blank (standard 1)
K: Calculated from measured values and input values of blank (standard 1) and standard sample (standard
2)
C1: Concentration of standard 1（reagent blank)
C2: Concentration of standard 2
A2: Absorbance or change in absorbance per minute of standard 2.
K=
C 2 − C1
A2 − B
(c) Calculation of concentration
C X = {K × ( A X − B) + C1 }× IFA + IFB
Cx: Concentration of standby sample
AX: Absorbance or change in absorbance per minute
IFA and IFB: Instrument constants, representing slope and intercept.
41
(d) Applicable assay
1-point assay, 2-point rate assay, 2-point assay, 3-point assay, 1-point&rate assay, rate A assay and rate B
assay
(3) Linearity Method (Multi-point linearity)
Absorbance
Blank (or standard 1) and standard samples (standard 2 and standards 6) are measured and linear working curve.
Figure 2-12 explains the linear method.
Concentration
Figure 2-12 Multi-point linearity
(a) Calibration Parameter input
Calibration type:【 multi-point linearity】
Calibration point: 【3-6 】(number of standard sample)
Span point: 【3-6】
(b)Calculation of parameters for working curve
S1ABS (B):Linear primary regression intercept for absorbance or change in absorbance per minute of
blank (standard)
K: Inverse number of working curve slope in the result of linear primary regression.
S1ABS and K values can be calculated by the formulas below:
S1ABS (B) = A −
K =
42
X × Cr
Y
Y ×10 4 ×10 a
X
n
(
)(
X ： ∑ Cri − Cr × Ai − A
)
i =1
n
(
Y ： ∑ Cri − Cr
)
2
i =1
⎛ n
⎞
A : ⎜ ∑ Ai ⎜ / n
⎝ i=1 ⎟
⎛ n
⎞
Cr : ⎜ ∑ Cri ⎜ / n
⎝ i=1
⎟
A1,A2: Each measured absorbance in duplicate measurement of standard(1)
n: No. of standards (N) ×2
Cri: Concentration of standard (i)
(c)Calculation of concentration
C X = {K × ( AX − B) + C1 }× IFA + IFB
Cx: Concentration of standby sample
AX: Absorbance of sample or its change per minute.
IFA and IFB: Instrument constants, representing slope and intercept.
(d) Applicable assay
1-point assay, 2-point rate assay, 2-point assay, 3-point assay, 1-point&rate assay, rate A assay and rate B
assay
(4) Logit-log 3P (Non-linearity Method)
This is applied to a working curve in which the absorbance converges as the concentration increase. Figure 2-13
explains the non-linearity method.
43
Figure 2-13 Logit-log3P
(a) Calibration Parameter input
Calibration type: 【Logit-log3P】
Calibration point: 【3-6】(number of standard sample)
Span point: 【0】span calibration invalid
(b) Calculation of parameters for working curve
B: the absorbance or approximate value measure of the absorbance change per minute
approaches ∞.
when CX
K: blank (standard 1) absorbance or value calculate by the approximation formula of the absorbance
change per minute subtraction B
a: Constants in approximation formula. Automatically calculated.
B,K,a are displayed on the Calibration List screen.
(c) Calculation of concentration.
C X = (C + C1 ) × IFA + IFB
AX = B +
C=
K
1 + aC)
1 ⎧ K − ( AX − B)⎫
×⎨
⎬
a ⎩ AX − B ⎭
Cx: Concentration of standby sample
C1: Blank concentration.
AX: Absorbance of sample or its change per minute.
K: Constants in approximation formula. The more Cx approaches ∞, AX approaches B
When K＜0， AX≤B+K or K＞0，When AX≥B+K，C=C1
IFA,IFB Instrument constants, representing slope and intercept.
44
(d) Calculation of SD value
∑ ∑( A
N
SD =
2
IJ
i =1 j =1
)
, 2
− A1
2N − 3
（N=3~6、j=1or 2）
(Aij-Ai’): Difference between approximate absorbance Ai’ and measured value Aij or A12. Each standard
sample is measured in duplicate so the number as measurement points Aij is 12 at maximum
(e) Applicable assay
1-point assay,
2-point rate assay, 2-point assay,
Rate A assay.
(5) Logit-log4P (Non-linearity method 2)
Absorbance
It is applied to a working curve in which the absorbance converges as the concentration increases. Figure 2-14
explains the non-linearity method.
Concentration
Figure 2-14 Logit-log4P
(a) ―Calibration Parameter‖ input
Calibration type: 【Logit-log4P】
Calibration point: 【4-6】 (number of standard sample)
Span point: 【0】
(b) Calculation of parameters for working curve
B: approximation for the absorbance or its change per minute when CX approaches ∞.
K: blank (standard 1) absorbance or value calculate by the approximation of the absorbance change per
minute subtraction B
45
a ,b : Constants in approximation formula. Automatically calculated.
B,K,a are displayed on the Calibration List screen.
(c) Calculation of concentration.
C X = (C + C1 ) × IFA + IFB
K
1 + aC b )
AX = B +
C = b×
1 ⎧ K − ( AX − B)⎫
×⎨
⎬
a ⎩ AX − B ⎭
Cx: Concentration of standby sample
C1: Blank concentration.
AX: Absorbance of sample or its change per minute.
K: Constants in approximation formula. The more Cx approaches ∞, AX approaches B
When K＜0， AX≤B+K or when K＞0，AX≥B+K C1=0
IFA,IFB Instrument constants, representing slope and intercept.
(d) Calculation of SD value
∑ ∑( A
N
SD =
2
i =1 j =1
IJ
)
, 2
− A1
2N − 4
（N=4~6、j=1or 2）
（Aij-Ai’）: Difference between approximate absorbance Ai’ and measured value Aij or A12. Each
standard sample is measured in duplicate so the number as measurement points Aij is 12 at maximum
(e) Applicable assay
1-point assay, 2-point rate assay, 2-point assay, Rate A assay.
(6) Logit-log5P (Non-linear method 3)
There is no distinct difference between the working curves prepared by the non-linear method 2 and 3. However,
in some cases, the non-linear method 3 allows more accurate approximation because this method has one more
calculation because this method has one more calculation parameter than the non-linear method 2. Figure 2-15
explain the non-linear method 3.
46
Absorbance
Concentration
Figure 2-15 Logit-log5P
(a) ―Calibration Parameter‖ input
Calibration type : 【Logit-log5P】
Calibration point: 【5-6】( number of standard sample )
Span point: 【0】Span point calibration invalid.
(b) Calculation of parameters for working curve
B: approximation for the absorbance or its change per minute when CX approaches ∞.
K,a,b,c: Constants in approximation formula. Automatically calculated.
B,K,a ,b,c :are displayed as S1ABS,K,A,B,C on the Calibration List screen.
(c) Calculation of concentration.
a+b·lnC+c·C-ln{
AX − B
}=0
K − ( A − BX )
Calculate C according to the Newton approximation formula.
C X = (C + C1 ) × IFA + IFB
K
1 + exp× (− a − b × ln C − c × C )
AX=B+
Cx: Concentration of standby sample
C1: Blank concentration.
AX: Absorbance of sample or its change per minute.
K: Constants in approximation formula. The more Cx approaches ∞, AX approaches B
47
When K＜0， AX≤B or when K＞0，AX≥B，C=0
IFA,IFB Instrument constants, representing slope and intercept.
(d) Calculation of SD value
∑ ∑( A
N
SD =
2
IJ
i =1 j =1
)
, 2
− A1
2N − 4
（N=5~6、j=1 or 2）
（Aij-Ai’）: Difference between approximate absorbance Ai’ and measured value Aij or A12. Each
standard sample is measured in duplicate so the number as measurement points Aij is 12 at maximum
(e) Applicable assay
1-point assay, 2-point rate assay, 2-point assay, Rate A assay.
(7) Exponential function method (Non-linear method)
Absorbance
Unlike non-linear methods 1,2 and 3.Exponetial function method prepares a working curve in which the
absorbance disperses as the concentration increases. Figure 2-16 explains the exponential function method.
Concentration
Figure 2-16 Exponential function method
(a) Calibration Parameter input
Calibration type: 【exponential function】
Calibration point: 【5-6】(number of standard sample)
Span point: 【0】Span point calibration invalid.
(b) Calculation of parameters for working curve
B: approximation formula for the absorbance or its change per minute of blank (standard 1)
K,a,b,c: Constants in approximation formula. Automatically calculated.
48
B,K,a ,b,c :are displayed as S1ABS,K,A,B,C on the Calibration List screen.
(c) Calculation of concentration.
AX=B+K × exp{a × (ln C ) + b ×(ln C ) + c × (ln C )
2
3
}
⎛ A − B⎞
2
3
a × (ln C ) + b × (ln C ) + c × (ln C ) -ln ⎜ X
⎜=0
⎝ K ⎟
Calculate C according to the Newton approximation formula.
C X = (C + C1 ) × IFA + IFB
Cx: Concentration of standby sample
C1, C2~CN: Blank and standard concentration.
AX: Absorbance of sample or its change per minute.
When K>0， AX≤B or when K<0，AX≥B，C=0
IFA,IFB Instrument constants, representing slope and intercept.
(d) Calculation of SD value
∑ ∑( A
N
SD =
2
i =1 j =1
IJ
)
, 2
− A1
2N − 5
（N=5~6、j=1 or 2）
（Aij-Ai’）: Difference between approximate absorbance Ai’ and measured value Aij or A12. Each
standard sample is measured in duplicate so the number as measurement points Aij is 12 at maximum
(e) Applicable assay
1-point Assay, 2-point Rate assay, 2-point Assay, Rate A assay.
(8) Spline function method (Non-linear method)
In this method, line is connected in each section so as to form a curve as a whole. Since each section is
smoothed including the error in measured value, more accurate approximation is possible than the polygonal
line approximation. Figure 2-17 explains this method.
49
Absorbance
Figure 2-17 Spline function method
(a) ―Calibration Parameter‖ input
Calibration type: 【Spline function】
Calibration point: 【5-6】(number of standard sample)
Span point: 【0】Span point calibration invalid.
(b) Calculation of parameters for working curve
A(1),b(1),c(1),d(1): Constants in approximation formula, l=1~N
In the ―calibration‖ menu, S1ABS show as a (1) (intercept of absorbance axis ).
(c) Calculation of concentration.
(
AX=a(I)+b(I) × (C X − C (1) + c(I)) × CX − C(1)
2
) + d (I) × (C
X
- C(1) )
3
f × (C X − C(I )) = a × (I ) + b × (I ) × (C X − C (I )) + d × (I ) × (C X − C (I )) 2 + d (I ) × (C X
Calculate C according to the Newton approximation formula.
C X = (C + C1 ) × IFA + IFB
Cx: Concentration of standby sample
C1~CN: Blank and standard concentration.
AX, A2~AN: Absorbance of sample and standard or its change per minute.
IFA, IFB Instrument constants, representing slope and intercept.
(d) Calculation of SD value
50
− C (I )) 3 − AX
∑ ∑( A
N
SD =
)
2
i =1 j =1
, 2
− A1
IJ
2N − 4
（N=5~6、j=1 or 2）
（Aij-Ai’）: Difference between approximate absorbance Ai’ and measured value Aij or A12. Each
standard sample is measured in duplicate so the number as measurement points Aij is 12 at maximum
(e) Applicable assay
1-Point Assay, 2-Point Rate Assay, 2-Point Assay, Rate A Assay.
(9) Polygon method (Non-linear method)
The range between standard samples 1 to 6 is subject to approximation in consideration of measured values
across them and line is connected in each section so as to form a curve as a whole. Since each section is
smoothed including the error in measured value, more accurate approximation is possible than the polygonal
line approximation. Figure 2-18 explains the polygonal line method
Absorbance
Concentration
Figure 2-18 Polygon method
(a) ―Calibration Parameter‖ input
Calibration type: 【polygon】
Calibration point: 【3-6】(number of standard sample)
Span point: 【0】Span point calibration invalid.
(b) Calculation of parameters for working curve
S1ABS is the two measure value (absorbance or it’s change) of STD (1)
K=
C 2 − C1
A2 − B
B: Absorbance or it’s change of standard (1)
51
A2: Absorbance or it’s change of standard (2)
C1: standard (1) concentration (input value)
C2: standard (2) concentration (input value)
(c) Calculation of concentration.
C X = {K N × ( AX − AN ) + C N }× IFA + IFB
(e) Applicable assay
1-Point Assay, 2-Point Rate Assay, 2-Point Assay, Rate A assay.
(10) Isozyme Method
A2
STD(2)
A3
STD(3)
AF
Sample
STD(4)
B’
STD(1)
C1
STD(3)
A4’
STD(4)
B
A3’
AM’
Sample
A4
absorbance
absorbance
In a sample in which 2 different isozymes coexist, a reagent containing inhibitor may fail to completely
suppress the activity of either isozyme alone. In this case, isozyme activity is determined from total activity and
activity residual rate. Each working curve for total activity and isozyme activity are prepared by using 2
channels. If the activity of a specific isozyme of the coexistent two can be suppressed completely by using
monoclonal antibody, etc. this calibration method is unnecessary. As figure 2-19, 2-20 shows:
CF
C2
concentration
Figure 2-19 Isozyme Method
STD(1)
C1
CM’
concentration
Figure2-20 Isozyme Method
(a) calibration principle
Isozymes method uses 2 reagent position. The total activity Cf is supposed to be calculated first with a
certain reagent in the isozyme P position. Calculate the activity Cm or Cn of isozymes M or N with a
reagent which can suppress N or M substance.
The isozymes M inhibitor testing isozymes M.
Generally speaking isozymes N inhibitor cannot suppress the activity of isozymes N completely, and the
activity of isozymes M is suppressed in a degree at the meantime.
The isozymes Method uses 2 channels to test the total activity and standard cost of isozymes M,N, K
value is tested by total activity channel, both two channels are used.
52
M and N activity residual rate of M and N is calculated by inhibitor (ratio between absorbance of
standard 3 and standard 4 in two channel.).Calculate the total activity and isozymes M activity upon
above method. The activity of isozymes N is observed by calculation.
(b) Input of parameters:
Reagent and standard samples.
Reagent: Reagent for measurement of total activity, reagent for measurement of isozyme activity.
Standard sample: Standard sample F (containing both isozymes M and N), standard sample M
(containing isozyme M), standard sample N (containing isozyme N)
Reagent position: isozymes M and N are placed in different positions
(d) Entry on Chemistry parameters screen.
Make entry for each of the isozyme P and Q channels as shown in below Table:
Con. and Pos.of F Activity
(Isozyme P) (Isozyme Q)
Calibrator
(concentration)
Calibrator（1）
Calibrator（2）
Calibrator（3）
Calibrator（4）
Con.and Pos. of M isozyme
(Isozyme Q)
(position)
(concentration)
(position)
Blank concentration …………………….. [S1]
Blank concentration……………..[S1]
Concentration value of calibrator F………[S2]
0…………………………………….0
0 …………………[S3] (Isozyme M Calibrator)
0………..[S3](Isozyme M Calibrator)
0………………….[S4] (Isozyme N Calibrator)
0………. [S4](Isozyme N Calibrator)
Table 2-3
S1 to S4 are calibrator code numbers of calibrator 1 to calibrator 4 respectively. Enter the same calibrator
code number in both channels for each of calibrator 1,3,4. Set standard sample of isozyme M at position
Calibrator 3 and that of isozyme N at position Calibrator (4). It is unnecessary to enter the concentrations
of calibrator 3,4,2 for the isozyme Q channel. The channel number M of isozyme M is specified in the
isozyme P channel. But it need not be specified on the isozyme Q side. Unless M is entered, Isozyme
parameter error alarm occurs to disable analysis
(c) Working curve for total activity measurement channel (isozyme P) K factor is calculated by the
following formula:
K=
C 2 − C1
A2 − B
B: Absorbance of blank (standard 1) or its change per minute
A2: Absorbance of standard sample F (standard 2) or its change per minute
53
C1: Activity of blank (standard 1 )
C2:Activity of standard sample F(standard 2 )
(d) Calculation of activity for total activity measurement channel (isozyme P)
CF={K·（AF-B）-C1}·IFA-IFB
C3={K·（A3-B）-C1}·IFA-IFB
C4={K·（A4-B）-C1}·IFA-IFB
CF 、C3 、C4: is sample, activity of standard 3, standard4;
Ap 、A3 、A4: is absorbance or change of the
absorbance per minute of standard 3 , standard 4;IFA,IFB Instrument constants, representing slope and
intercept.
(e) Calculation of activity for isozyme measurement channel (Q)
CM’={K·（AM’-B’）-C1}·IFA-IFB
C3’’={K·（A3’- B’）-C1}·IFA-IFB
C4’={K·（A4’- B’）-C1}·IFA-IFB
CM’: Isozyme M activity of sample；AM’: Isozyme absorbance of sample or its change per minute；C3’ 、
C4’:each inhibited activity of standard 3 and 4；A3’、A4’分 Each inhibited absorbance of standard 3 and 4
or its change per minute；B: Absorbance of blank or its change per minute IFA,IFB Instrument constants,
representing slope and intercept.
(f) Calculation of activity residual rate
α=
{K × (A 3 '-B) + C1}× IFA + IFB
{K × (A 3 - B ) + C1}× IFA + IFB
β=
{K × (A 4 '-B) + C1}× IFA + IFB
{K × (A 4 - B ) + C1}× IFA + IFB
when C1=0, IFA=1，IFB=0
α=
A3, -B,
A3-B
β=
A4，
-B，
A4-B
(g) Calculation of isozyme M activity Cm
CM’=α×CM+β×CN
54
CM= CF-CN= CF - C M − α × C M
β
β×CM=β×CF -CM’+α×CM
（α-β）×CM= CM-β×CF
CM=
C M '− β × CF
(α − β )
(h) Applicable assay
1-Point Assay, 2-Point Rate Assay, 2-Point Assay, Rate A Assay.
Note: Suggest operator use 6 calibrators in Logit-log5P, exponential, spline non-linear calibration.
2.2.3 Calibration points
According to the number of calibration points, calibration can be effected in different ways. Blank calibration:
Only reagent blank (standard 1 ) is calibrated. Span calibration: Only one standard solution other than the
reagent blank is calibrated. 2-point calibration: Reagent blank and a single standard solution are calibrated.
Full-point calibration: All standard solutions specified on the Chemistry Parameters screen are calibrated. These
are selectively usable so as to meet your analytical purpose. Each calibration is explained below.
(1) Blank Calibration
Only reagent blank (standard 1) is calibrated. The previously measured working curve is corrected for a change
in absorbance or absorbance change rate (through calculation of S1ABS). Table 2-4 lists the calculation method
for each calibration type. (a) S1ABS calculation
Calibration type
SIABS calculation
2 point linearity
（A11+ A12）/2
Multi-point linearity
{（AU+ A12）/2-（AU’+ A12’）/2}+SIABS’
Isozyme P
（A11+ A12）/2
Isozyme Q
（A11+ A12）/2
Logit-Log 3P
{（A11+ A12）/2-（A11’+ A12’）/2}+SIABS’
Logit-Log 4P
{（A11+ A12）/2-（A11’+ A12’）/2}+SIABS’
Logit-Log 5P
（A11+ A12）/2
Exponential function
（A11+ A12）/2
Spline function
{（A11+ A12）/2- SIABS’}+a（I）
Polygon method
（A11+ A12）/2
Table 2-4 S1ABS Calculation
A11、 A12：1st and 2nd absorbance values of standard (1) measured presently.
55
A11’, A12’：1 st and 2 nd absorbance values of standard (1) measured previously.
SIABS’：Previous SIABS value
a（I）：I=1~N，N representing the number of standard solution and factor of the curve（refer to 5.3.3（8）
Logit-Log 5P method）
This calibration method needs be specified for each test on the calibration test selection screen of Routine
job Menu.
Note*：Only blank calibration can be carried out in case 1 is entered for the number of standard solutions (K
factor method)
(b)Applicable calibration types
Linear (2-point), Linear (multi-point), Linear (1-point: K factor), Isozyme P, Isozyme Q, Logit-log 3P,
Logit-log4P, Logit-log 5P, Exponential, Spline
(2) Span Calibration
Only one standard solution other than reagent blank is calibrated. The standard solution which corresponds to
the span point entered on the Chemistry Parameters screen is measured and the previously measured working
curve is corrected for its slope. Table 2-5 lists the calculation method for each calibration type. This calibration
method need be specified for each test on the Calibration Test Selection screen.
(a) K factor and S1ABS calculation
Calibration type
K factor calculation
S1ABS
Two-point calibration
（C2-C1）/（A2-S1ABS）
Previous value
Full-point calibration
（C2-C1）/（AN-S1ABS）
Previous value
Table 2-5 S1ABS, K value calculation
C2：Concentration of standard (2)
C1：Concentration of standard (1)
CN：Concentration of standard N (N represents span point)
A2：Average of measured absorbance values of standard (2)
AN：Average of measured absorbance values of standard (N)
(b) Applicable assay
Linear (2-point), Linear (multi-point),
(3) 2-point calibration
Reagent blank and a single standard solution are calibrated. The standard solution and reagent blank, which
correspond to the span points entered on the Chemistry Parameters screen, are measured and the previously
measured working curve is corrected for S1ABS and slope. Table 2-6 explains the calculation method.
(a) K factor and S1ABS calculation
56
Calibration type
S1ABS Calculation
K factor calculation
Two-point linearity
（A11+ A12）/2
（C2-C1）/（A2-S1ABS）
Full-point linearity
（AU+ A12）/2
（CN-C1）/（AN-S1ABS）
Table 2-6 S1ABS, K value calculation
st
A11、 A12：1 and 2nd absorbance values of standard (1) measured presently.
C2：Concentration of standard (2)
C1：Concentration of standard (1)
CN：Concentration of standard N (N represents span point)
A2：Average of measured absorbance values of standard (2)
AN：Average of measured absorbance values of standard (N)
A1: Average of measured absorbance values of standard(1)
(b) Applicable assay
Linear (2-point), Linear (multi-point),
(4) Full-point Calibration
All standard solutions (including reagent blank) specified on the ―Chemistry Parameters‖ screen are calibrated.
After this calibration, all working curve parameters S1ABS,K,a,b and c displayed on the Calibration List screen
are updated.
(a) Calculation formula
The calculation varies from the calibration methods.
(b)Applicable calibration types
Linearity (multi-point),Isozyme P, Isozyme Q, Logit-log 3P, Logit-log 4P, Exponential, Spline
2.3 Check of measure value
Various checks are performed to enhance the reliability of measured results. Below are check description
2.3.1 Calibration check
(1) Blank level check
In calibration, a warning –level alarm is issued if the measured absorbance of blank is not within the input
range of standard 1 absorbance. In this case, the result of measurement and alarm (S1ABS) are printed out. To
avoid check, enter-“ -3.3～3.3” .
(2) Discrete check
A warning level alarm is issued if the difference of the two times measured absorbance value is larger than the
set value. In calibration, each Calibrator (include reagent blank: Calibrator 1) is tested twice.
57
To
avoid
enter“ 3.3”
the
check,
.
Discrete check is performed by the below formula:
≤ Absorbance Discrete Check（ABS）
(3) Sensitivity check
If the difference of standard absorbance (average between 2 times measure) from Max. concentration standard
absorbance (sensitivity) exceeds the permissible absorbance sensitivity value (Sensitivity Limit), a warninglevel alarm is issued. In this case, alarm mark is printed out together with the result of measurement.
The working curve of the alarmed analytical item will be renewed, and the K value won’t be renewed. To
avoid the check, enter-―0‖.
Check the permissible sensitivity by the below formula:
A
lower
STD（N）lim it ≤
− ASTD（1）
≤ upper lim it
C STD（N）− C STD（1）
(4) K factor check
If the fluctuation in factor K value between previous calibration and current calibration is 20% ,a warning
level alarm is issued. The working curve and K factor will be renewed and testing can be carried out. Make
sure check the reason of alarm.
Check the K factor by the below formula:
K this − K last
× 100% ≤ 20%
(K this + K last ) / 2
(5) Drift rate check
In calibration, a warning –level alarm is issued if the difference between the calculated absorbance and tested
absorbance has exceeded the drift rate set value. The working curve and K factor will be renewed and testing
can be carried out. Make sure to check the reason of alarm. To avoid the check, enter―3.3‖.
2.3.2 Absorbance of reaction limited check
As to the Rate assay, correct data won’t be obtained when concentration or activity exceeds the quantitative
span. Thus, set the upper limit value and lower limit value of the absorbance, print the alarm sign. Input the
calibration value on the screen. To avoid the check, enter-0 (decrease) or 3.3 (increase).
When 4 or more than 4 tested absorbance value is not accord with the set value of reaction limit absorbance,
alarm is issued as figure 2-21 shows:
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ABS
Reaction limit level
Time
Input photometry range
Figure 2-21
2.3.3 Linearity Abnormal Check
In the rate assay, relation between absorbance change and time should be Linearity. Thus, check on the
linearity is a necessity.
Select ―Linearity check‖ in ―Alarm Info.‖, and input the limit check value in corresponding textbox as figure
2-22 shows:
Figure 2-22
If not selected, even if input value in textbox, linearity check is not carried out.
(1) When number of measurement points (N) more than 9 (N>9)
Linearity is checked by dividing the difference in absorbance change between the first and last 6
measurement points by the average absorbance change for all. If the value thus obtained is beyond the limit
linearity value, alarm is printed out together with the result of measurement as figure 2-23 shows:
Δ Af
− Δ Ab
×100＞Limit linearity value（%）
ΔA
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Photometry point
Time
Figure 2-23
Linearity check N≥ 9
(2) When number of measurement points (N) between 4 and 8 （4≤N≤8）
Linearity is checked by dividing the difference in absorbance change between the first and last 6 measurement
points by the average absorbance change for all. If the value thus obtained is beyond the limit linearity value,
alarm is issued as figure 2-24 shows:
Δ Af ，− Δ Ab，
Δ A，
×100＞Limit linearity value（%）
Photometry point
Time
Figure 2-24 Linearity check（4≤ N≤ 8）
2.3.4 Prozone check
In immunoreaction, working curve descends if antigen concentration is abnormally high beyond the suitable
range (prozone area). This is called prozone or zone phenomenon.
This instrument can check whether concentration is in the absorbance decreasing range (post zone). For prozone
check, the following 2 methods are available: antigen readdition method in 1-point assay with prozone check
and reaction rate method in 2-point assay. To Avoid check, input ―-3.3 lower limit‖ in ―checkup value‖ function
box in ―analyze parameter‖ menu.
(1) Antigen supplement method:
Take 1-point essay for example, measure reagent 1, and take it’s value as reference value. Replace reagent with
serum diluents, which contain antigen, add 20ul. Compare the prozone limit value with absorbance difference
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(before add reagent 2 and after add reagent 2).
Input method:
Prozone check value (PC value): 【 】
Upper /lower limit: 【】
Analytical method: 【】
Photometric point: 【Q1】 【Q2】 【0】 【0】
1≤Q1≤Q2≤49
PC =
Aq 2 + A( q 2 − 1)
Aq1 + A( q1 − 1)
− k×
2
2
K=total liquid volume when test q1/total liquid volume when test q2
When 1≤Q1≤Q2≤4 or 5≤Q1≤Q2≤16, 17≤Q1≤Q2≤31, 32≤Q1≤Q2≤49 K=1.
Absorbance
AQ1、AQ2：absorbance of photometric point:Q1,Q2
Time
Figure 2-25 Antigen supplement method
(2) Reaction speed ratio method:
Take 2-point assay for example, the ratio between average reaction speed and reaction speed with serum
(prozone check value: PC value), then compare the PC value with prozone limit value.
Prozone checkup value(PC value) 【 】
Upper/lower limit: 【】
Analyze method: 【】
photometric point: 【q1】 【q2】 【q3】 【q4】
5≤q1≤q2≤49、5≤q3≤q4≤49
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PC =
( Aq 4 − Aq 3) /( q 4 − q 3)
×100
( Aq1 − Aq 2 ) /( q1 − q 2)
photometric point: Aq1、Aq2、Aq3、Aq4：absorbance: q1、q2、q3、q4
photometric point q1、q2、q3、q4 test time point:q1、q2、q3、q4
Do not carry out prozone check when following condition occur:
①
test point: q1=q2=q3=q4。
②
test for No.1 standard liquid (reagent blank)
Absorbance
Figure 2-26 explains antigen supplement method:
Time
Figure 2-26 Reaction speed ratio method
2.4 ISE testing principle
2.4.1 Operation principle
Once analysis starts, SIP injection pump aspirates 65UL reference liquid and infuses it into the reference
electrode liquid line---sample probe aspirates15uL sample and infuse it into diluting cup---DIL injection pump
infuses 450uL diluent---SIP injection pump aspirates the diluted sample and infuses it into electrode liquid line,
and test its potential, and the rest of diluted sample will be assimilated by vacuum nozzle---IS injection pump
infuses 600uL internal standard liquid into the diluting cup for rinsing it, and the internal standard liquid will be
sucked by vacuum nozzle sequently---IS injection pump infuses 450uL internal standard liquid into the diluting
cup---SIP injection pump aspirates 65uL reference liquid and infuses it into the reference electrode liquid line
first, and then SIP injection pump aspirates it and infuses it into the electrode liquid line, and test its
potential---the vacuum nozzle assimilates the redundant internal standard liquid in order to empty the diluting
cup for next turn test.
2.4.2 Electric potential generation principle
The electric potential is calculate by ―Nernst‖ formula.
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E = E0 + 2.303×
RT
× l og(ai) …………………………………………………………(1)
nF
ai = f × Ci ………………………………………………………………………………. (2)
E 0 ：Standard electrode potential of tested system
R：Gas constant（8.314510 J*mol-1*K-1）
T : Absolute temperature（t℃+273.15）(K)
F : Faraday constant（9.6485309×104C* mol-1）
ai ：Ion（i）activity
f ：Activity coefficient
Ci：Concentration
n ：electric charge of given icon（Cation is positive, Anion is negative）
2.4.3 Test method
The following procedure explains the preparation of working curve, the test of internal standard liquid
concentration, calculation of concentration, and result revision.
2.4.3.1 Working curve preparation
Test low concentration slope liquid (S1) and high concentration slope liquid (S2), set the slope (sensitivity) of
K,NA, Cl electrode.
SL =
E(H ) − E(L)
……………………………(3)
C (H )
log
C (L)
SL：Slope
E(H): Electrode potential of high concentration slope liquid
E(L)：Electrode potential of low concentration slope liquid
C(H):Concentration value of high concentration slope liquid （input value 0）
C(L): Concentration value of low concentration slope liquid （input value 0）
2.4.3.2 Internal standard liquid concentration test
Test the concentration of internal standard liquid when working curve prepared.
C (IS ) = C (L) ×10
E ( IS )− E ( L )
SL
………………………………(4)
C(IS)：Concentration of internal standard liquid
E(IS): Electrode potential of internal liquid.
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2.4.3.3 Concentration calculation.
Make internal standard liquid concentration as reference value, calculate basic sample, emergency sample, QC
product concentration. The internal standard liquid is tested according to different sample.
C (S ) = C (IS ) ×10
E ( S )− E ( IS )
SL
……………………………………….(5)
C(S)：Sample concentration
E(S): Sample electrode
2.4.3.4 Result revision
Test Calibrator (S3) after calibration, and the calculate the concentration, make the difference between test
concentration and input value as compensation value, and add/ subtract the sample concentration of standby
sample.
C (VALUE) = C (C ) − C ( X ) ……………………………..(6)
C(VALUE): Compensate value
C(C): Input value of serum Calibrator concentration
C(X):Measure value of serum Calibrator concentration
C ' (S ) = IF {C (S ) + C (VALUE )} …………………(7)
C’(S):Sample concentration after revision
IF: Instrument constant（1.0）
2.4.3.5 Standard specification of ISE
Item
Specification
Sample syringe
15ul
Diluent liquid volume
450ul
Capacity
80 sample/hour（only for electrolyte test）
Range
Reagent consumption
volume
Na 20~200mmol/L（only for serum test）
10~400mmol/L（for urine test）
+
K 1.0~15.0mmol/L（only for serum test）
1~200mmol/L（for urine test）
Cl 20~200mmol/L（only for serum test）
10~400mmol/L（for urine test）
Internal standard liquid 1050ul/sample (only for electrolyte continual test)
Diluent
450 ul/sample
Reference electrode liquid 130 ul/sample
Table 2-7 ISE standard specification
64
Note: During operation, if ISE analysis is not taken more than 10 minutes, the internal standard liquid will be
pipetted once in order to activate electrode.
65
Chapter 3 Instrument Installation
Analyzer has passed all tests before transportation. Packing it gently in order to avoid damage while
transportation. To make sure normal operation, install or initialize the CS-400 only by authorized staffs from
DIRUI Medical Company.
3.1 Installation requirement
Before installation, operator should check the space, power and environment requirement.
3.1.1 Space Requirement
To make sure the space of maintenance, please follow the instruction as below:
● Space between left (right) side of analyzer and the wall should ≥ 50cm
● Space between rear panel of analyzer and the wall should ≥ 50cm
● Space in front of analyzer should≥ 100cm
● Make sure there is enough space for waste device and purified water equipment.
3.1.2 Environment requirement
Operate or store the analyzer according to the following requirement:
● Working environment: 1 5 ℃～32 ℃
● Relative humidity: 4 0 ％ ～85 ％
● Atmospheric pressure: 7 6 k P a ～106kP a
● Environment should with no dust, no mechanical vibration, no noise source and power interference
● Do not put the analyzer in the vicinity of brush motor, flicker fluorescent tube and other constant on-off
electrical equipment.
● Avoid direct sunlight, do not put the analyzer in front of heat source and wind source
3.1.3 Power requirement:
● Power supply: ~2 20 V, 50 Hz
● Power: 2000 VA
● Circuit breaker: 250V, 20A
●A well grounded power supply socket is a must. (Socket at least with one 15 A and three 5A). Large electrical
appliance such as air condition, refrigerator, even cannot use the same electrical wire as analyzer. Instrument
is equipped with a three core electrical wire, red wire is live line, blue wire is zero line, and yellow green wire
is ground lead. As figure 3-1 shows:
66
HOT
Figure 3-1
Note: The unqualified environment may cause test value inaccuracy, analyzer damage and it is also
harmful to human body.
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