This compendium offers manual applications for use with reagents manufactured by DiaSys

Diagnostic Systems on photometric systems. It is as well a guideline and recommendation to

program applications for automated systems.

DiaSys calibrators and controls should be used for calibration and internal quality control.
If the R1/R2 and sample ratio is kept, sample and reagent volumes may be reduced or
increased accordingly. If standards are available, those may be used for calibration
alternatively.

These application proposals represent guidelines only. To avoid misinterpretation, measured

results have to be validated and assessed with caution. Please refer to the corresponding
package inserts for complete information and required calibrators and controls as well. Each
laboratory should establish corrective action in case of deviations in control recovery.

Edition 1, January 2022

Manufacturer:
DiaSys Diagnostic Systems GmbH
Alte Strasse 9, 65558 Holzheim
Germany

www.diasys-diagnostics.com

Seite 2 von 63
ALAT (GPT) FS (IFCC mod.) Product code: 1 2701…
Method
Optimized UV-test according to IFCC (International Federation of Clinical Chemistry and Laboratory
Medicine) [modified]
Assay procedure
Name ALT
Wavelength 340 nm, Hg 365 nm, Hg 334 nm
Optical path 1 cm
Temperature 37°C
Measurement End point, against air
Reagent preparation
Substrate start
Reagents are ready to use
Sample/calibrator
Sample/calibrator 100 µL
Reagent 1 1000 μL
Mix, incubate for 5 min., then add:
Reagent 2 250 μL
Mix, read absorbance after 1 min. and start stopwatch. Read absorbance again after 1, 2 and 3
min.
Sample start (Do not use sample start with P-5-P)
For mono reagent mix 4 parts R1 + 1 part R2
Stability: 4 weeks at 2 -8°C, 5 days at 15 - 25°C
Mono reagent must be protected from light
Sample/calibrator
Sample/calibrator 100 µL
Mono reagent 1000 μL
Mix, read absorbance after 1 min. and start stopwatch. Read absorbance again after 1, 2 and 3
min.
Calculation [U/L]
With factor
Calculate ΔA/min from absorbance readings and multiply by the corresponding factor from table
below:
Δ A/min x factor = ALAT activity [U/L]
Wavelength Factor substrate start Factor sample start
340 nm 2143 1745
334 nm 2184 1780
365 nm 3971 3235
With calibrator
ΔA/min. Sample
ALAT [U/L] = x Conc. Cal [U/L]
ΔA/min. Cal
Conversion factor:
ALAT [U/L] x 0.0167 = ALAT [μkat/L]

Seite 3 von 63
Albumin FS Product code: 1 0220…
Method
Photometric test using bromocresol green. In the presence of bromocresol green at a slightly acid
pH, serum albumin produces a color change of the indicator from yellow-green to green-blue.
Assay procedure
Name ALB
Wavelength Hg 546 nm, 540 – 600 nm
Optical path 1 cm
Temperature 37°C
Measurement End point, against reagent blank
Reagent preparation
Reagent is ready to use
Blank Sample/standard/calibrator
Sample/standard/calibrator - 10 μL
Dist. water 10 μL -
Reagent 1000 μL 1000 μL
Mix, incubate for approx. 10 min. and read the absorbance (A) against reagent blank within 60 min.
Calculation [g/dL]
With standard or calibrator
A Sample
Albumin [g/dL] = x Conc. Std/Cal [g/dL]
A Std/Cal
Conversion factor:
Albumin [g/dL] x 144.9 = Albumin [μmol/L]

Seite 4 von 63
Alkaline phosphatase FS IFCC mod. 37°C Product code: 1 0441…
Method
Kinetic photometric test, according to the International Federation of Clinical Chemistry and
Laboratory Medicine (IFCC) [mod.]
Assay procedure
Name AP
Wavelength Hg 405 nm, 400 – 420 nm
Optical path 1 cm
Temperature 37°C
Measurement Kinetic, against reagent blank
Reagent preparation
Substrate start
Reagents are ready to use
Blank Sample/calibrator
Sample/calibrator - 20 μL
Dist. water 20 μL -
Reagent 1 1000 μL 1000 μL
Mix, incubate for approx. 1 min., then add:
Reagent 2 1000 μL 1000 μL
Mix, read absorbance after 1 min. and start stopwatch. Read absorbance again after 1, 2 and 3
min.

Sample start
For mono reagent mix 4 parts R1 + 1 part R2
Stability: 4 weeks at 2 - 8°C, 5 days at 15 - 25°C
Mono reagent must be protected from light
Blank Sample/calibrator
Sample/calibrator - 20 μL
Dist. water 20 μL -
Mono reagent 1000 μL 1000 μL
Mix, read absorbance after 1 min. and start stopwatch. Read absorbance again after 1, 2 and 3
min.
Calculation [U/L]
With factor
Calculate ΔA/min from absorbance readings and multiply by the corresponding factor from table
below:
Δ A/min x factor = AP activity [U/L]
Factor
Substrate start 405 nm 3433
Sample start 405 nm 2757
Conversion factor:
AP [U/L] x 0.0167 = AP [μkat/L]

Seite 5 von 63
 α-Amylase CC FS Product code: 1 0501…
Method
Enzymatic photometric test, in which the substrate 4,6-ethylidene-(G7)-p-nitrophenyl-(G1)-α-D-
maltoheptaoside (EPS-G7) is cleaved by α-Amylases into various fragments.
These are further hydrolyzed in a second step by α-Glucosidase producing glucose and p-
nitrophenol. The increase in absorbance represents the total (pancreatic and salivary) amylase
activity in the sample.
Assay procedure
Name AMY
Wavelength Hg 405 nm
Optical path 1 cm
Temperature 37°C
Measurement Kinetic, against reagent blank
Reagent preparation
Substrate start
Reagents are ready to use
Serum / plasma Urine
Blank Sample Blank Sample
Sample/calibrator - 20 µL - 10 µL
Dist. water 20 µL - 10 µL -
Reagent 1 1000 μL 1000 μL 1000 μL 1000 μL
Mix, incubate for approx. 1 min., then add:
Reagent 2 250 μL 250 μL 250 μL 250 μL
Mix, read absorbance after 2 min. and start stopwatch. Read absorbance again after 1, 2 and 3
min.
Sample start
For mono reagent mix 4 parts R1 + 1 part R2
Stability: 6 months at 2 - 8°C, 4 weeks at 15 - 25°C
Mono reagent must be protected from light
Serum / plasma Urine
Blank Sample Blank Sample
- 20 µL - 10 µL
Sample/calibrator 20 µL - 10 µL -
Mono reagent 1000 μL 1000 μL 1000 μL 1000 μL
Mix, read absorbance after 2 min. and start stopwatch. Read absorbance again after 1, 2 and 3
min.

Calculation see next page.

Seite 6 von 63
α-Amylase CC FS (calculation) Product code: 1 0501…
Calculation [U/L]
With factor
Calculate ΔA/min from absorbance readings and multiply by the corresponding factor from table
below:
Δ A/min x factor = AP activity [U/L]
Substrate start Sample start
Serum / plasma 5670 4554
Urine 11250 9018
With calibrator
ΔA/min. Sample
α-Amylase [U/L] = x Conc. Cal [U/L]
ΔA/min. Cal
Conversion factor:
α-Amylase [U/L] x 0.0167 = α-Amylase [μkat/L]

Seite 7 von 63
α-HBDH FS Product code: 1 3201…
Method
Optimized UV test according to DGKC (German Society of Clinical Chemistry)
Assay procedure
Name HBDH
Wavelength 340 nm, Hg 334 nm, Hg 365 nm
Optical path 1 cm
Temperature 25°C, 30°C, 37°C
Measurement Kinetic, against air
Reagent preparation
Substrate start
25°C/30 °C 37°C
Sample Sample
Sample 20 µL 10 µL
Reagent 1 1000 μL 1000 μL
Mix, incubate for approx. 1 – 5 min., then add:
Reagent 2 250 μL 250 μL
Mix, read absorbance after 1 min. and start stopwatch. Read absorbance again after 1, 2 and 3
min.
Sample start
For mono reagent mix 4 parts R1 + 1 part R2
Stability: 5 days at 2 - 8°C, 8 hours at 15 - 20°C
Mono reagent must be protected from light
25°C/30 °C 37°C
Sample Sample
Sample 20 µL 10 μL
Mono reagent 1000 μL 1000 µL
Mix, read absorbance after 1 min. and start stopwatch. Read absorbance again after 1, 2 and 3
min.
Calculation [U/L]
Calculate ΔA/min from absorbance readings and multiply by the corresponding factor from table
below:
ΔA/min x factor = α-HBDH activity [U/L]
Substrate start
Wavelength 25°C/30 °C 37°C
340 nm 10080 20000
334 nm 10275 20390
365 18675 37060
Sample start
Wavelength 25°C/30 °C 37°C
340 nm 8095 16030
334 nm 8250 16345
365 15000 29705
Conversion factor:
α-HBDH [U/L] x 0.0167 = α-HBDH [μkat/L]

Seite 8 von 63
Antistreptolysin O FS Product code: 1 7012 …
Method
Particle enhanced immunoturbidimetric test. Determination of the concentration of ASO via
photometric measurement of the antigen-antibody-reaction of latex particles coated with
streptolysin O and antibodies to streptolysin O present in the sample.
Assay procedure
Name ASO
Wavelength 500 – 600 nm
Optical path 1 cm
Temperature 37°C
Measurement Kinetic, against reagent blank
Reagent preparation
Reagent is ready to use
Blank Sample/calibrator
Sample/calibrator - 12 µL
Dist. water 12 µL -
Reagent 1 1000 μL 1000 μL
Mix, incubate for 3 - 5 min., then add:
Reagent 2 200 μL 200 μL
Mix, and read absorbance A1. Incubate for a further 5 min. and read absorbance again A2.
Calculation [mmol/L]
ΔA = [A2 – A1]sample/calibrator

The concentration of antistreptolysin O in unknown samples is derived from a calibration curve

using an appropriate mathematical model such as spline. The calibration curve is obtained with five
calibrators at different levels and NaCl solution (9 g/L) for determination of the zero value.

Seite 9 von 63
Apolipoprotein A1 FS Product code: 1 7102 …
Method
Immunoturbidimetric test. Determination of Apo A1 concentration by photometric measurement of
antigen antibody reaction between antibodies to human Apo A1 and Apo A1 present in the sample.
Assay procedure
Name APOA1
Wavelength Hg 580 nm (500 - 700 nm)
Optical path 1 cm
Temperature 37°C
Measurement End point, against reagent blank
Reagent preparation
Reagent is ready to use
Blank Sample/calibrator
Sample/calibrator - 10 µL
Dist. water 10 µL -
Reagent 1 1250 μL 1250 μL
Mix, incubate for 3 - 5 min., read absorbance A1, then add:
Reagent 2 250 μL 250 μL
Mix, incubate for 5 min. and read absorbance A2.
Calculation [mmol/L]
ΔA = [A2 – A1] sample/calibrator

The concentration of apolipoprotein A1 in unknown samples is derived from a calibration curve

using an appropriate mathematical model such as logit/log. The calibration curve is obtained with
five calibrators at different levels and NaCl solution (9 g/L) for determination of the zero value.

Seite 10 von 63
Apolipoprotein B FS Product code: 1 7112 …
Method
Immunoturbidimetric test. Determination of the Apo B concentration via photometric measurement
of antigen-antibody-reaction of antibodies to Apo B with Apo B present in the sample.
Assay procedure
Name APOB
Wavelength Hg 340 nm
Optical path 1 cm
Temperature 37°C
Measurement End point, against reagent blank
Reagent preparation
Reagent is ready to use
Blank Sample/calibrator
Sample/calibrator - 10 µL
Dist. water 10 µL -
Reagent 1 1000 μL 1000 μL
Mix, incubate for 3 - 5 min., read absorbance A1, then add:
Reagent 2 200 μL 200 μL
Incubate for 5 min. and read absorbance A2.
Calculation [mmol/L]
ΔA = [A2 – A1]sample/calibrator

The concentration of apolipoprotein B in unknown samples is derived from a calibration curve using
an appropriate mathematical model such as logit/log. The calibration curve is obtained with five
calibrators at different levels and NaCl solution (9 g/L) for determination of the zero value.

Seite 11 von 63
ASAT (GOT) FS (IFCC mod.) Product code: 1 2601…
Method
Optimized UV-test according to IFCC (International Federation of Clinical Chemistry and Laboratory
Medicine) [modified]
Assay procedure
Name AST
Wavelength 340 nm, Hg 365 nm, Hg 334 nm
Optical path 1 cm
Temperature 37°C
Measurement End point, against air
Reagent preparation
Substrate start
Reagents are ready to use
Sample/calibrator
Sample/calibrator 100 µL
Reagent 1 1000 μL
Mix, incubate for 5 min., then add:
Reagent 2 250 μL
Mix, read absorbance after 1 min. and start stopwatch. Read absorbance again after 1, 2 and 3
min.
Sample start (do not use samle start with P-5-P)
For mono reagent mix 4 parts R1 + 1 part R2
Stability: 4 weeks at 2 - 8°C, 5 days at 15- 25°C
Mono reagent must be protected from light
Sample/calibrator
Sample/calibrator 100 µL
Mono reagent 1000 μL
Mix, read absorbance after 1 min. and start stopwatch. Read absorbance again after 1, 2 and 3
min.
Calculation [U/L]
With factor
Calculate ΔA/min from absorbance readings and multiply by the corresponding factor from table
below:
Δ A/min x factor = ASAT activity [U/L]
Wavelength Factor substrate start Factor sample start
340 nm 2143 1745
334 nm 2184 1780
365 nm 3971 3235
With calibrator
ΔA/min. Sample
ASAT [U/L] = x Conc. Cal. [U/L]
ΔA/min. Cal
Conversion factor:
ASAT [U/L] x 0.0167 = ASAT [μkat/L]

Seite 12 von 63
ATP Hexokinase FS Product code: 1 6201…
Method
Indirect indication for survival rate of erythrocytes by measuring the ATP concentration with the
Hexokinase method
Assay procedure
Name ATP
Wavelength 340 nm, Hg 365 nm, Hg 334 nm
Optical path 1 cm
Temperature 20°C – 25°C
Measurement End Point, against air or water
Sample Preparation
Pipette 1.0 mL blood or erythrocyte concentrate and 1.0 mL trichloroacetic acid 10 – 12% (w/v) into
a centrifuge tube, mix well and put into an ice bath for approx. 5 min. Centrifuge the sample solution
5 – 10 min. at approx. 3000 g. After centrifugation use 250 μL of the clear supernatant directly and
without any waiting time in the assay. Use the ATP standard without sample preparation directly in
the assay. When using ATP standard for calibration, patient results have to be multiplied by 2.
Note: ATP in samples is unstable. The ATP content of blood collected in heparin or EDTA shows a
decrease of 80% within 24 h, if stored at 2 – 8°C. Storage of samples mixed with trichloroacetic
acid at –20°C gives false results, too. Due to this samples clarified with trichloroacetic acid must be
used directly in the assay.
Discard contaminated specimens.
Reagent preparation
The reagent and the standard are ready to use.
Blank Sample/standard
Sample/standard - 250 μL
Dist. water 250 μL -
Reagent 1 2400 μL 2400 μL
Mix and incubate 3 – 5 min. at 25°C. Read absorbance A1, then add:
Reagent 2 600 μL 600 μL
Mix, incubate for approx. 15 min. at 25°C read absorbance A2 within 30 min.
Calculation [µmol/dL]
ΔA = [A2 – A1] sample/standard
With factor
Multiply ∆A by the corresponding factor from the table below in order to calculate the ATP
concentration
Wavelength With sample preparation Without sample preparation
Factor [µmol/dL] Factor [µmol/dL]
340 nm 412.70 206.35
Hg 334 nm 420.71 210.36
Hg 365 764.71 382.35
F = (V x f x 100) / (ε x v x d) [μmol/dL]
V = Total volume in cuvette [μL] = 3250
f = Dilution factor of sample preparation = 2.0
d = Light path [cm] = 1.00
v = Sample volume [μL] = 250
ε = Ext. coefficient NADH [l x cm-1 x mmol-1] = 6.3 at 340 nm
= 3.4 at 365 nm
= 6.18 at 334 nm

Seite 13 von 63
Bicarbonate FS Product code: 1 0950 …
Method
Enzymatic test using phosphoenolpyruvate carboxylase (PEPC) and a stable NADH analog. This
method has been standardized against a primary standard on basis of sodium carbonate.
Assay procedure
Name CO
Wavelength 405 nm, 415 nm
Optical path 1 cm
Temperature 37°C
Measurement Kinetic, against reagent blank
Reagent preparation
The reagent and the standard are ready to use
Sample/standard
Sample/standard 10 µL
Reagent 1000 μL
Mix, incubate and read absorbance A1 after exactly 2 min. and absorbance A2 after exactly 10 min.
against reagent blank.
Calculation [mmol/L]
With standard
ΔA = (A2 – A1) sample/ standard

ΔA Sample
Bicarbonate [mmol/L ] = x Conc. Std. [mmol/L]
ΔA Std.
Conversion factor
Bicarbonate [mmol/L] = Bicarbonate [mEq/L]

Seite 14 von 63
Bilirubin Auto Direct FS Product code: 1 0821 …
Method
Photometric test using 2,4-dichloroaniline (DCA). Direct bilirubin in presence of diazotized 2,4-
dichloroaniline forms a red colored azocompound in acidic solution.
Assay procedure
Name DBIL
Wavelength 546 nm (540 – 560 nm)
Optical path 1 cm
Temperature 20 – 25°C/37°C
Measurement Kinetic, against reagent blank
Reagent preparation
The reagents are ready to use
Blank Sample/calibrator
Sample/calibrator - 50 μL
Dist. water 50 μL -
Reagent 1 1000 μL 1000 μL
Mix, incubate for 3 – 5 min. at 20 – 25°C/37°C, read absorbance A1, then add:
Reagent 2 250 μL 250 μL
Mix, incubate for exactly 5 min. at 37°C or 10 min. at 20 – 25°C, then read absorbance A2.
Calculation [mg/dL]
With calibrator
ΔA = (A2 – A1) sample/calibrator

ΔA Sample
Bilirubin [mg/dL] = x Conc. Cal. [mgl/dL]
ΔA Cal.

Seite 15 von 63
Bilirubin Auto Totale FS Product code: 1 0811…
Method
Photometric test using 2,4-dichloroaniline (DCA). In acidic solution, direct bilirubin forms a red
colored azocompound with diazotized 2,4-dichloroaniline. A specific mixture of detergents enables a
safe determination of the total bilirubin.
Assay procedure
Name TBIL
Wavelength 546 nm (540 – 560 nm)
Optical path 1 cm
Temperature 20 – 25°C/37°C
Measurement Kinetic, against reagent blank
Reagent preparation
The reagents are ready to use
Blank Sample/calibrator
Sample/calibrator - 25 μL
Dist. water 25 μL -
Reagent 1 1000 μL 1000 μL
Mix, incubate for 5 min. at 37°C or 10 min. at 20 – 25°C, read absorbance A1, then add:
Reagent 2 250 μL 250 μL
Mix, incubate for exactly 5 min. at 37°C or 10 min. at 20 – 25°C, then read absorbance A2.
Calculation [mmol/L]
With calibrator
ΔA = (A2 – A1) sample/calibrator
ΔA Sample
Bilirubin [mg/dL] = x Conc. Cal. [mg/dL]
ΔA Cal.
Conversion factor
Bilirubin [mg/dL] x 17.1 = Bilirubin [μmol/L]

Seite 16 von 63
Calcium AS FS Product code: 1 1130…
Method
Photometric test using arsenazo III. Calcium with arsenazo III at neutral pH yields a blue colored
complex, whose intensity is proportional to the calcium concentration. Interference by magnesium is
eliminated by addition of 8-hydroxyquinoline-5-sulfonic acid.
Assay procedure
Name CA_AS
Wavelength 650 nm, Hg 623 nm (630 – 670 nm)
Optical path 1 cm
Temperature 20-25°C/37°C
Measurement End point, against reagent blank
Reagent preparation
The reagent and the standard are ready to use
Blank Sample/standard/calibrator
Sample/standard/calibrator - 10 μL
Dist. water 10 μL -
Reagent 1000 μL 1000 μL
Mix, incubate for 5 min. and read absorbance against reagent blank.
Calculation [mg/dL]
With standard or calibrator
A Sample
Calcium [mg/dL] = x Conc. Std/Cal. [mg/dL]
A Std./Cal.
Conversion factor
Calcium [mg/dL] x 0.2495 = Calcium [mmol/L]
Calcium/U [mg/24 h] x 0.025 = Calcium/U [mmol/24 h]

Seite 17 von 63
Calcium P FS Product code: 1 1181…
Method
Photometric determination with Phosphonazo III. At acidic pH calcium forms a purple-blue colored
complex with phosphonazo III. In a second step calcium is bound to a chelating agent whereby the
specific signal is eliminated. The resulting difference in absorbance is directly proportional to the
calcium concentration in the sample. This guarantees a specific measurement of calcium.
Assay procedure
Name CA_P
Wavelength 660 nm; 700/800 nm bi-chromatic
Optical path 1 cm
Temperature 37°C
Measurement End point, against reagent blank
Reagent preparation
The reagents and the standard are ready to use.
Blank Sample/standard/calibrator
Sample/standard/calibrator - 10 μL
Dist. water 10 μL -
Reagent 1 1000 μL 1000 μL
Mix and incubate for 5 minutes. Read absorbance A1 than add:
Reagent 2 250 μL 250 μL
Mix and read absorbance A2 within 1 minute.
Calculation [mg/dL]
With standard or calibrator
ΔA = (A2 – A1) Sample/standard/calibrator

In case an absorbance of > 1.6 is observed after mixing of R1 and sample, dilute the sample 1 + 1
with NaCl solution (9 g/L), retest and multiply the result by 2.

Δ A Sample
Calcium [mg/dL] = x Conc. Std./Cal. [mg/dL]
Δ A Std./Cal.
Conversion factor
Calcium [mg/dL] x 0.2495 = Calcium [mmol/L]
Calcium/U [mg/24 h] x 0.025 = Calcium/U [mmol/24 h]

Seite 18 von 63
Chlorid 21 FS Product code: 1 1221…
Method
Photometric test using ferric (III) perchlorate. Chloride forms with ferric ions a yellow colored
complex whose absorption is measured at 340 nm. A discoloring agent in reagent 2 displaces
Chloride out of the complex, thereby discoloring the solution. The difference in absorbance between
the colored and discolored state of the solution is proportional to the concentration of chloride in the
sample.
Assay procedure
Name CL
Wavelength 340/660 nm (bi-chromatic)
Optical path 1 cm
Temperature 37°C
Measurement End Point, against reagent blank
Reagent preparation
The reagents are ready to use
Blank Sample/calibrator
Sample/calibrator - 40 μL
Dist. water 40 μL -
Reagent 1 900 μL 900 μL
Mix and incubate for 5 min. at 37°C, read absorbance A1, then add:
Reagent 2 225 μL 225 μL
Mix, incubate for 1 min. at 37°C, then read absorbance A2.
Calculation [mmol/L]
ΔA = (A2 – A1) sample/calibrator
The concentration of chloride in unknown samples is derived from a linear calibration curve. It is
obtained with the levels 1/2 and 3/4 of the electrolyte calibrator TruCal E.
Conversion factor
Chloride [mmol/L] = Chloride [mEq/L]
Chloride [mmol/L] x 3.545 = Chloride [mg/dL]

Seite 19 von 63
Cholesterol FS Product code: 1 1300…
Method
“CHOD-PAP”: enzymatic photometric test. Determination of cholesterol after enzymatic hydrolysis
and oxidation. The colorimetric indicator is quinoneimine which is generated from 4-aminoantipyrine
and phenol by hydrogen peroxide under the catalytic action of peroxidase (Trinder’s reaction).
Assay procedure
Name CHOL
Wavelength 500 nm, Hg 546 nm
Optical path 1 cm
Temperature 20 – 25°C / 37°C
Measurement End point, against reagent blank
Reagent preparation
The reagent is ready to use
Blank Sample/calibrator/standard
Sample/calibrator/standard - 10 μL
Dist. water 10 μL -
Mono reagent 1000 μL 1000 μL
Mix, incubate for 20 min. at 20 – 25°C or for 10 min. at 37°C. Read absorbance within 60 min
against reagent blank
Calculation [mg/dL]
With standard or calibrator
A Sample
Cholesterol [mg/dL] = x Conc. Std./Cal. [mg/dL]
A Std./Cal.
Conversion factor
Cholesterol [mg/dL] x 0.02586 = Cholesterol [mmol/L]

Seite 20 von 63
Cholinesterase FS Product code: 1 1401…
Method
Kinetic photometric test, optimized method according to the recommendation of the German Society
of Clinical Chemistry (DGKC). Cholinesterase hydrolyses butyrylthiocholine under release of butyric
acid and thiocholine. Thiocholine reduces yellow potassium hexacyanoferrate (III) to colorless
potassium hexacyanoferrate (II). The decrease of absorbance is measured at 405 nm.
Assay procedure
Name CHE
Wavelength Hg 405 nm
Optical path 1 cm
Temperature 37°C
Measurement Kinetic, against reagent blank
Reagent preparation
The reagents are ready to use
Blank Sample/calibrator
Sample/calibrator - 20 μL
Dist. water 20 μL -
Reagent 1 1000 μL 1000 μL
Mix, incubate for approx. 3 min., then add:
Reagent 2 250 μL 250 μL
Mix, read absorbance (A) after 2 min and start stop watch. Read absorbance (A) again after 1, 2
and 3 minutes.
Calculation [U/L]
With factor
ΔA/min x 68500 = CHE activity [U/L]
With calibrator
ΔA/min Sample
CHE [U/L] = x Conc. Cal. [U/L]
ΔA/min Cal.
Conversion factor
Cholinesterase [kU/L] x 16.67 = Cholinesterase [μkat/L]

Seite 21 von 63
CK-MB FS Product code: 1 1641…
Method
Optimized UV test according to DGKC (German Society of Clinical Chemistry) and IFCC
(International Federation of Clinical Chemistry and Laboratory Medicine) for CK with inhibition of
CK-M isoenzymes by monoclonal antibodies. CK-MB consists of the subunits CK-M and CK-B.
Specific antibodies against CK-M inhibit the complete CK-MM activity (main part of the total CK
activity) and the CK-M- subunit of CK-MB. Only CK-B activity is measured, which is half of the CK-
MB activity.
Assay procedure
Name CK_MB
Wavelength 340 nm, Hg 334 nm
Optical path 1 cm
Temperature 37°C
Measurement Kinetic, against reagent blank
Reagent preparation
Substrate start
The reagents are ready to use
Blank Sample/calibrator
Sample/calibrator - 50 μL
Dist. water 50 μL -
Reagent 1 1000 μL 1000 μL
Mix, incubate for approx. 3 min., then add:
Reagent 2 250 μL 250 μL
Mix, read absorbance after 2 min. and start stopwatch. Read absorbance again after 1, 2, 3, 4 and
5 min.
Sample start
For mono reagent mix 4 parts R1 + 1 part R2
Stability: 2 weeks at 2 - 8°C, 24 hours at 15 - 20°C
Mono reagent must be protected from light
Blank Sample/calibrator
Sample/calibrator - 40 μL
Dist. water 40 μL -
Mono reagent 1000 μL 1000 μL
Mix, read absorbance after 5 min. and start stopwatch. Read absorbance again after 1, 2, 3, 4 and
5 min
Calculation [U/L]
With factor
Calculate ΔA/min from absorbance readings and multiply by the corresponding factor from table
below:
Δ A/min x factor = CK-MB activity [U/L]
Wavelength Factor
340 nm 8254
334 nm 8414
With calibrator
ΔA/min Sample
CK MB [U/L] = x Conc. Cal. [U/L]
ΔA/min Cal.
Conversion factor
CKMB [U/L] x 0.0167 = CKMB [μkat/L]

Seite 22 von 63
CK-NAC FS Product code: 1 1601…
Method
Optimized UV-test according to IFCC (International Federation of Clinical Chemistry and Laboratory
Medicine) and DGKC (German Society of Clinical Chemistry).
Assay procedure
Name CK_NAC
Wavelength 340 nm, Hg 365 nm, Hg 334 nm
Optical path 1 cm
Temperature 37°C
Measurement Kinetic, against reagent blank
Reagent preparation
Substrate start
The reagents are ready to use
Blank Sample/calibrator
Sample/calibrator - 50 μL
Dist. water 50 μL -
Reagent 1 1000 μL 1000 μL
Mix, incubate for approx. 3 min., then add:
Reagent 2 250 μL 250 μL
Mix, read absorbance after 2 min. and start stopwatch. Read absorbance again after 1, 2 and 3
min.
Sample start
For mono reagent mix 4 parts R1 + 1 part R2
Stability: 3 weeks at 2 - 8°C, 2 days at 15 - 20°C
Mono reagent must be protected from light
Blank Sample/calibrator
Sample/calibrator - 40 μL
Dist. water 40 μL -
Mono reagent 1000 μL 1000 μL
Mix, read absorbance (A1) after 3 min. and start stopwatch. Read absorbance (A2) again after 1, 2
and 3 min.
Calculation [U/L]
With factor
Calculate ΔA/min from absorbance readings and multiply by the corresponding factor from table
below:
Δ A/min x factor = AP activity [U/L]
Wavelength Factor
340 nm 4127
334 nm 4207
365 nm 7429
With calibrator
ΔA/min Sample
CK [U/L] = x Conc. Cal. [U/L]
ΔA/min Cal.

Seite 23 von 63
Creatinine FS Product code: 1 1711…
Method
Kinetic test without deproteinization according to the Jaffé method Creatinine forms a colored
orange-red complex in an alkaline picrate solution. The difference in absorbance at fixed times
during conversion is proportional to the concentration of creatinine in the sample.
Assay procedure
Name CREA_JAFFE
Wavelength Hg 492 nm, (490 – 510 nm)
Optical path 1 cm
Temperature 20 - 25°C/37°C
Measurement Two point fixed time linear kinetic, against reagent blank
Reagent preparation
The reagents and standard are ready to use.
Blank Sample/standard/calibrator
Sample/standard/calibrator - 50 μL
Dist. water 50 μL -
Reagent 1 1000 μL 1000 μL
Mix, incubate for approx. 0-5 min., then add:
Reagent 2 250 μL 250 μL
Mix and read absorbance A1 after 60 sec, read absorbance A2 after another 120 sec.
Calculation [mg/dL]
With standard or calibrator
ΔA = (A2 – A1) Sample/standard/calibrator

Serum/Plasma:
ΔA Sample
Creatinine [mg/dL] = x Conc. Std./Cal. [mg/dL]
ΔA Std./Cal.
Urine:
ΔA Sample
Creatinine [mg/dL] = x Conc. Std./Cal. [mg/dL] x 50
ΔA Std./Cal.

Creatinine Clearance [mL/min/1.73 m2]

mg Creatinine/100 mL Urine x mL Urine
=
mg Creatinine/100 mL Serum x min Urine collection time

The calculated creatinine clearance refers to the average body surface of an adult (1.73 m2).
Conversion factor
Creatinine [mg/dL] x 88.4 = Creatinine [µmol/L

Seite 24 von 63
Creatinine PAP FS Product code: 1 1759…
Method
Enzymatic colorimetric test
Assay procedure
Name CREA_PAP
Wavelength Hg 546 nm
Optical path 1 cm
Temperature 37°C
Measurement Two point end, against reagent blank
Reagent preparation
The reagents and standard are ready to use.
Blank Sample/standard/calibrator
Sample/standard/calibrator - 24 μL
Dist. water 24 μL -
Reagent 1 1000 μL 1000 μL
Mix, incubate 5 min. and read absorbance A1, then add:
Reagent 2 500 μL 500 μL
Mix, read absorbance A2 after 5 min.
Calculation [mg/dL]
With standard or calibrator
ΔA = (A2 – 0.672 A1) Sample/standard/calibrator

Serum/Plasma:
ΔA Sample
Creatinine [mg/dL] = x Conc. Std./Cal. [mg/dL]
ΔA Std./Cal.
Urine:
ΔA Sample
Creatinine [mg/dL] = x Conc. Std./Cal. [mg/dL] x 10
ΔA Std./Cal.

Creatinine Clearance [mL/min/1.73 m2]

mg Creatinine/100 mL Urine x mL Urine
=
mg Creatinine/100 mL Serum x min Urine collection time

The calculated creatinine clearance refers to the average body surface of an adult (1.73 m2).
Conversion factor
Creatinine [mg/dL] x 88.4 = Creatinine [μmol/L]

Seite 25 von 63
CRP FS Product code: 1 7002…
Method
Immunoturbidimetric test. Determination of CRP concentration by photometric
measurement of the antigen-antibody reaction of antibodies to human CRP with CRP
present in the sample.
Assay procedure
Name CRP
Wavelength 340 nm, Hg 334 nm
Optical path 1 cm
Temperature 37°C
Measurement End point, against reagent blank
Reagent preparation
The reagents are ready to use.
Blank Sample/calibrator
Sample/calibrator - 60 μL
Dist. water 60 μL -
Reagent 1 1000 μL 1000 μL
Mix, incubate for 5 min. at 37°C and read absorbance (A1), then add:
Reagent 2 200 μL 200 μL
Mix, incubate for 5 min. at 37°C and read absorbance (A2).
Calculation [U/L]
ΔA = (A2 – A1) sample/calibrator

The CRP concentration of unknown samples is derived from a calibration curve using an
appropriate mathematical model such as logit/log. The calibration curve is obtained with five
calibrators at different levels and NaCl solution (9 g/L) for determination of the zero value.

Seite 26 von 63
D-Dimer Product code: 1 7268…
Method
Particle enhanced immunoturbidimetric test. Determination of the D-dimer concentration by
photometric measurement of antigen-antibody-reaction between antibodies against D-dimer bound
to particles and D-dimer present in the sample.
Assay procedure
Name DDIMER
Wavelength 570 nm
Optical path 1 cm
Temperature 37°C
Measurement Two point fixed time linear kinetic, against reagent blank
Reagent preparation
The reagents are ready to use.
Blank Sample/calibrator
Sample/calibrator - 30 μL
Dist. water 30 μL -
Reagent 1 900 μL 900 μL
Mix, incubate for 3-5 min., then add:
Reagent 2 300 μL 300 μL
Mix, read absorbance (A1) within 20 sec., incubate for 5 min, then read absorbance (A2)
Calculation [U/L]
ΔA = (A2 – A1) sample/calibrator

The D-dimer concentration of unknown samples is derived from a calibration curve using an
appropriate mathematical model such as spline. The calibration curve is obtained with 5 calibrators
at different levels and the added diluent for determination of the zero value

Seite 27 von 63
Ethanol FS Product code: 1 0881…
Method
Enzymatic UV test with alcohol dehydrogenase (ADH). In the presence of NAD Ethanol is
converted by alcohol dehydrogenase. The measured absorbance of the produced NADH is
proportional to the ethanol concentration in the sample.
Assay procedure
Name ETH
Wavelength 376 nm (360 – 380 nm)
Optical path 1 cm
Temperature 37°C
Measurement Two point end point, against reagent blank
Reagent preparation
The reagents are ready to use.
Blank Sample/standard
Sample/standard - 10 μL
Dist. water 10 μL -
Reagent 1 1000 μL 1000 μL
Mix and incubate for 5 minutes at 37°C. Read absorbance A1 than add:
Reagent 2 250 μL 250 μL
Mix and incubate 5 min. at 37°C. Read absorbance A2 immediately.
Calculation [mg/mL]
With standard
ΔA = (A2 – A1) sample/standard

A Sample
Ethanol [mg/mL] = x Conc. Std. [mg/dL]
A Std.
Conversion factor
Ethanol [g/L] x 21.7 = Ethanol [mmol/L]
Ethanol [g/L] (serum/plasma) x 0,8 = Ethanol‰ (whole blood)

Seite 28 von 63
Gamma-GT FS (Szasz mod./IFCC stand.) Product code: 1 2801…
Method
Kinetic photometric test according to Szasz/Persijn. Gamma-GT catalyzes the transfer of glutamic
acid to acceptors like glycylglycine in this case. This process releases 5-amino-2-nitrobenzoate
which can be measured at 405 nm. The increase in absorbance at this wavelength is directly
related to the activity of gamma-GT.
Assay procedure
Name GGT
Wavelength 405 nm, (400 – 420 nm)
Optical path 1 cm
Temperature 37°C
Measurement Kinetic against reagent blank
Reagent preparation
Substrate start
The reagents are ready to use.
Blank Sample/calibrator
Sample/calibrator - 100 µL
Dist. water 100 µL -
Reagent 1 1000 μL 1000 μL
Mix, incubate for approx.. 1 min., then add:
Reagent 2 250 μL 250 μL
Mix, read absorbance after 1 min. and start stopwatch. Read absorbance again after 1, 2 and 3
min.
Sample start
For mono reagent mix 4 parts R1 + 1 part R2
Stability: 4 weeks at 2 - 8°C, 5 days at 15 - 20°C
Mono reagent must be protected from light
Blank Sample/calibrator
Sample/calibrator - 100 µL
Dist. water 100 µL -
Mono reagent 1000 μL 1000 μL
Mix, read absorbance after 1 min. and start stopwatch. Read absorbance again after 1, 2 and 3
min.
Calculation [U/L]
With factor
Calculate ΔA/min from absorbance readings and multiply by the corresponding factor from table
below:
Δ A/min x factor = Gamma-GT activity [U/L]
Wavelength Szasz IFCC
Substrat start 405 nm 1421 1606
Sample start 405 nm 1158 1309
With calibrator
ΔA/min Sample
GGT [U/L] = x Conc. Cal. [U/L]
ΔA/min Cal.
Conversion factor:
GGT [U/L] x 0.0167 = GGT [μkat/L]

Seite 29 von 63
GLDH FS (DGKC) Product code: 1 2411…
Method
Optimized UV test, according to recommendations of the DGKC
Assay procedure
Name GLDH
Wavelength 340 nm, Hg 334 nm
Optical path 1 cm
Temperature 37°C
Measurement Kinetic, against air
Reagent preparation
The reagents are ready to use.
Sample/calibrator
Sample/calibrator 150 µL
Reagent 1 1000 μL
Mix, incubate for approx.. 3 min., then add:
Reagent 2 250 μL
Mix, read absorbance after 30 sec. and start stopwatch. Read absorbance again after 1, 2 and 3
min.
Calculation [U/L]
With factor
Calculate ΔA/min from absorbance readings and multiply by the corresponding factor from table
below:
Δ A/min x factor = GLDH activity [U/L]
Wavelength Factor
340 nm -1485
334 nm -1515
With calibrator
ΔA/min Sample
GLDH [U/L] = x Conc. Cal. [U/L]
ΔA/min Cal.
Conversion factor:
GLDH [U/L] x 0.0167 = GLDH [μkat/L]

Seite 30 von 63
Glucose GOD FS Product code: 1 2500…
Method
“GOD-PAP“: enzymatic photometric test. Determination of glucose after enzymatic oxidation by
glucose oxidase. The colorimetric indicator is quinoneimine, which is generated from 4-
aminoantipyrine and phenol by hydrogen peroxide under the catalytic action of peroxidase
(Trinder’s reaction).
Assay procedure
Name GLUC_GOD
Wavelength 500 nm, Hg 546 nm
Optical path 1 cm
Temperature 20°C – 25°C, 37°C
Measurement End Point, against reagent blank
Reagent preparation
The reagent and the standard is ready to use.
Blank Sample/standard/calibrator
Sample/standard/calibrator - 10 μL
Dist. water 10 μL -
Reagent 1000 μL 1000 μL
Mix, incubate 20 min. at 20 - 25°C or 10 min. at 37°C. Read absorbance against the blank within 60
min.
Calculation [mg/dL]
With standard or calibrator
A Sample
Glucose [mg/dL] = x Conc. Std./Cal [mg/dL]
A Std./Cal
Conversion factor
Glucose [mg/dL] x 0.05551= Glucose [mmol/L]

Seite 31 von 63
 Glucose Hexokinase FS Product code: 1 2511…
Method
Enzymatic UV test using hexokinase
Assay procedure
Name GLUC_HK
Wavelength 340 nm, Hg 334 nm, Hg 334 nm
Optical path 1 cm
Temperature 20°C – 25°C, 37°C
Measurement End Point, against reagent blank
Reagent preparation
Substrate start
The reagents are ready to use.
Blank Sample/standard/calibrator
Sample/standard/calibrator - 10 μL
Dist. water 10 μL -
Reagent 1 1000 μL 1000 μL
Mix and incubate 1–5 min. at 20 – 25°C/37°C. Read absorbance A1, then add:
Reagent 2 250 μL 250 μL
Mix, incubate 5 min. at 37°C or 10 min. at 20 – 25°C. Read absorbance A2 against reagent blank
within 30 min.
Sample start
For mono reagent mix 4 parts R1 + 1 part R2
Stability: 3 month at 2 - 8°C, 3 weeks at 15 - 20°C
Mono reagent must be protected from light
Blank Sample/calibrator
Sample/calibrator - 10 µL
Dist. water 10 µL -
Mono reagent 1000 μL 1000 μL
Mix, incubate 5 min. at 37°C or 10 min. at 20 – 25°C. Read absorbance A against reagent blank
within 30 min.

Calculation see next page.

Seite 32 von 63
Glucose Hexokinase FS (calculation) Product code: 1 2511…
Calculation [mg/dL] substrate start
ΔA = (A2 – A1) Sample/standard/calibrator
Calculation [mg/dL] sample start
∆A = A Sample/Standard
Note:
The pipetting scheme with sample start is recommended only for analyzers with correction of
sample blank (e.g. by bichromatic measurement). Samples often show relatively high absorbances
at the measurement wavelengths which tend to show falsely high glucose values when working with
sample start. The given calculation factors cannot be used for bichromatic measurements
With factor
Calculate ΔA/min from absorbance readings and multiply by the corresponding factor from table
below:
Substrate start Sample start
Wavelength Factor [mg/dL] Factor [mmol/L] Factor [mg/dL] Factor [mmol/L]
340 nm 361 20.0 289 16.0
Hg 334 nm 367 20.5 294 16.4
Hg 365 667 37.1 535 29.7
With standard or calibrator
ΔA Sample
Glucose [mg/dL] = x Conc. Std./Cal. [mg/dL]
ΔA Std./Cal.
Conversion factor
Glucose [mg/dL] x 0.05551= Glucose [mmol/L]

Seite 33 von 63
HDL-c direct FS Product code: 1 3561…
Method
Homogeneous method without centrifugation steps. Block polymer detergents protect LDL, VLDL
and chylomicrons in a way that only HDL-cholesterol is selectively determined by an enzymatic
cholesterol measurement.
Assay procedure
Name HDCL_DIRECT
Wavelength 600/700 nm (bichromatic measurement)
Optical path 1 cm
Temperature 37°C
Measurement End Point, against reagent blank
Reagent preparation
The reagents are ready to use.
Blank Sample/calibrator
Sample/calibrator - 10 μL
Dist. water 10 μL -
Reagent 1 1000 μL 1000 μL
Mix and incubate 5 min. at 37°C. Read absorbance A1, then add:
Reagent 2 250 μL 250 μL
Mix, incubate for 5 min. at 37°C, read absorbance A2.
Calculation [mg/dL]
ΔA = (A2 – A1) sample/calibrator
With calibrator
ΔA Sample
HDL-C [mg/dL] = x Conc. Cal. [mg/dL]
ΔA Cal.

Conversion factor
HDL-C [mg/dL] x 0.02586 = HDL-C [mmol/L]

Seite 34 von 63
Immunoglobulin A FS Product code: 1 7202…
Method
Immunoturbidimetric test. Determination of IgA concentration by photometric measurement of
antigen-antibody-reaction of antibodies to human IgA with IgA present in the sample.
Assay procedure
Name IGA
Wavelength 570 nm
Optical path 1 cm
Temperature 37°C
Measurement End Point, against reagent blank
Reagent preparation
The reagents are ready to use.
Blank Sample/calibrator
Sample/calibrator - 10 μL
Dist. water 10 μL -
Reagent 1 1250 μL 1250 μL
Mix and incubate 3–5 min., read absorbance A1, then add:
Reagent 2 250 μL 250 μL
Mix, incubate 3 min., read absorbance A2
Calculation [mg/dL]
ΔA = (A2 – A1) sample/calibrator
With standard or calibrator
The concentration of IgA in unknown samples is derived from a calibration curve using an
appropriate mathematical model such as logit/log. The calibration curve is obtained with 5
calibrators at different levels and NaCl solution (9 g/L) for determination of the zero value.
Conversion factor
Immunoglobulin A [mg/dL] x 0.0625 = Immunoglobulin A [μmol/L]

Seite 35 von 63
Immunoglobulin G FS Product code: 1 7212…
Method
Immunoturbidimetric test. Determination of the IgG concentration by photometric measurement of
antigen-antibody-reaction between antibodies to human IgG and IgG present in the sample.
Assay procedure
Name IGG
Wavelength 570 nm
Optical path 1 cm
Temperature 37°C
Measurement End Point, against reagent blank
Reagent preparation
The reagents are ready to use.
Blank Sample/calibrator
Sample/calibrator - 10 μL
Dist. water 10 μL -
Reagent 1 1750 μL 1750 μL
Mix and incubate 3–5 min., read absorbance A1, then add:
Reagent 2 350 μL 350 μL
Mix, incubate 3 min., read absorbance A2.
Calculation [mg/dL]
ΔA = (A2 – A1) sample/calibrator
With standard or calibrator
The concentration of IgG in unknown samples is derived from a calibration curve using an
appropriate mathematical model such as logit/log. The calibration curve is obtained with 5
calibrators at different levels and NaCl solution (9 g/L) for determination of the zero value.
Conversion factor:
Immunoglobulin G [mg/dL] x 0.067 = Immunoglobulin G [μmol/L]

Seite 36 von 63
Immunoglobulin M FS Product code: 1 7222…
Method
Immunoturbidimetric test. Determination of the IgM concentration by photometric measurement of
antigen-antibody-reaction of antibodies to human IgM with IgM present in the sample.
Assay procedure
Name IGM
Wavelength 415/700 nm (bi-chromatic)
Optical path 1 cm
Temperature 37°C
Measurement End Point, against reagent blank
Reagent preparation
The reagents are ready to use.
Blank Sample/calibrator
Sample/calibrator - 10 μL
Dist. water 10 μL -
Reagent 1 1250 μL 1250 μL
Mix and incubate 3 min., read absorbance A1, then add:
Reagent 2 250 μL 250 μL
Mix, incubate 3 min., read absorbance A2.
Calculation [mg/dL]
ΔA = (A2 – A1) sample/calibrator
With standard or calibrator
The concentration of IgM in unknown samples is derived from a calibration curve using an
appropriate mathematical model such as logit/log. The calibration curve is obtained with 5
calibrators at different levels and NaCl solution (9 g/L) for determination of the zero value.
Conversion factor::
Immunoglobulin M [mg/dL] x 0.0103 = Immunoglobulin M [μmol/L]

Seite 37 von 63
Iron FS Product code: 1 1911…
Method
Photometric test using Ferene. Iron bound to transferrin is released
Assay procedure
Name FE
Wavelength 595 nm, 600 nm, Hg 623 nm
Optical path 1 cm
Temperature 37°C
Measurement End Point, against reagent blank
Reagent preparation
The reagents are ready to use.
Blank Sample/standard/calibrator
Sample/standard/calibrator - 100 μL
Dist. water 100 μL -
Reagent 1 1000 μL 1000 μL
Mix, read absorbance A1 after 1 – 5 min., then add:
Reagent 2 250 μL 250 μL
Mix, read absorbance A2 after 10 min.
Calculation [µg/dL]
ΔA = (A2 – 0.82 A1) Sample/standard/calibrator

The factor 0.82 compensates the decrease of the absorbance by addition of reagent 2. The factor is
calculated as follows:
(Sample + R1)/Total volume. This compensation is necessary as a high sample volume is used.
With standard or calibrator
ΔA/min Sample
Iron [µg/dL] = x Conc. Std./Cal.[µg/dL]
ΔA/min Std../Cal.
Conversion factor:
Iron [μg/dL] x 0.1791 = [μmol/L]

Seite 38 von 63
Lactate FS Product code: 1 4001…
Method
Enzymatic UV test with lactate dehydrogenase (LDH). In the presence of NAD, lactate is converted
by the lactate dehydrogenase. This procedure releases NADH which is measured at 340 nm. The
absorbance of the produced NADH is proportional to the lactate concentration in the sample.
Assay procedure
Name LACT
Wavelength 340 nm
Optical path 1 cm
Temperature 37°C
Measurement Two point endpoint, against reagent blank
Reagent preparation
The reagents are ready to use.
Blank Sample/calibrator
Sample/calibrator - 15 μL
Dist. water 15 μL -
Reagent 1 1000 μL 1000 μL
Mix and incubate 5 min. at 37°C. Read absorbance A1 then add:
Reagent 2 250 μL 250 μL
Mix and incubate 5 min. at 37°C. Read absorbance A2 within 30 min.
Sample start
For mono reagent mix 4 parts R1 + 1 part R2
Stability: 14 days at 2 - 8°C
Do not use icteric or hemolytic samples with sample start
Blank Sample/calibrator
Sample/calibrator - 10 µL
Dist. water 10 µL -
Mono reagent 1000 μL 1000 μL
Mix and incubate 5 min. at 37°C. Read absorbance A2 within 30 min.
Calculation [mg/dL] substrate start
ΔA = (A2 – A1) sample/calibrator
Calculation [mg/dL] sample start
ΔA = A sample/calibrator
With calibrator
ΔA Sample
Lactate [mg/dL] = x Conc. Cal. [mg/dL]
ΔA Cal.
With factor
ΔA x factor = Lactate concentration [mg/dL]

Wavlength Substrate start factor Sample start factor

340 nm 120.6 144.4
Conversion factor:
Lactate [mg/dL] x 0.1109 = Lactate [mmol/L]

Seite 39 von 63
LDH 21 FS Product code: 1 4251…
Method
Optimized UV-test according to IFCC (International Federation of Clinical Chemistry and Laboratory
Medicine) [modified]
Assay procedure
Name LDH_21_IFCC
Wavelength 340 nm, Hg 365 nm, Hg 334 nm
Optical path 1 cm
Temperature 37°C
Measurement Kinetic, against reagent blank
Reagent preparation
Substrate start
The reagents are ready to use.
Blank Sample
Sample/calibrator - 20 µL
Dist Water 20 µL -
Reagent 1 1000 μL 1000 μL
Mix, incubate for approx. 1 - 5 min., then add:
Reagent 2 250 μL 250 μL
Mix, read absorbance after 1 min. and start stopwatch. Read absorbance again after 1, 2 and 3
min.
Sample start
For mono reagent mix 4 parts R1 + 1 part R2
Stability: 12 hours at 2 - 8°C, 2 hours at 15 - 20°C
Mono reagent must be protected from light
Blank Sample
Sample/calibrator - 20 µL
Dist Water 20 µL -
Mono reagent 1000 μL 1000 µL
Mix, read absorbance after 1 min. and start stopwatch. Read absorbance again after 1, 2 and 3
Calculation [U/L]
With factor
From absorbance readings calculate ΔA/min and multiply by the corresponding factor from table
below:
ΔA/min x factor = LDH activity [U/L]
Wavelength Factor Substrate start Factor Sample start
340 nm 10080 8095
334 nm 10275 8250
365 nm 18675 15000
With calibrator
ΔA/min. Sample
LDH [U/L] = x Conc. Cal. [U/L]
ΔA/min. Cal.
Conversion factor:
LDH [U/L] x 0.0167 = LDH [μkat/L]

Seite 40 von 63
LDL-c direct FS Product code: 1 4131…
Method
Direct immunoinhibition method, endpoint assay. Block polymer detergents protect HDL, VLDL and
chylomicrons in a way that only LDL-cholesterol is selectively determined by an enzymatic
cholesterol measurement.
Assay procedure
Name LDLC_DIRECT
Wavelength 600/700 nm (bichromatic measurement)
Optical path 1 cm
Temperature 37°C
Measurement End Point, against reagent blank
Reagent preparation
Blank Sample/ calibrator
The reagents are ready to use.
Sample/calibrator - 10 μL
Dist. water 10 μL -
Reagent 1 1000 μL 1000 μL
Mix and incubate 5 min. at 37°C. Read absorbance A1, then add:
Reagent 2 250 μL 250 μL
Mix, incubate for 5 min. at 37°C, read absorbance A2.
Calculation [mg/dL]
ΔA = (A2 – A1) sample/calibrator
With calibrator
ΔA Sample
LDL-C [mg/dL] = x Conc. Cal. [mg/dL]
ΔA Cal.

Conversion factor:
LDL-C [mg/dL] x 0.02586 = LDL-C [mmol/L]

Seite 41 von 63
Lipase DC FS Product code: 1 4321…
Method
Direct colorimetric method. Asynthetically produced lipase substrate (1,2-o-dilauryl-rac-glycero-3-
glutaric acid-(6-methylresorufin) ester) is added to a micro-emulsion which is specifically split by
lipase in the presence of colipase and bile acids. The combination of lipase and bile acids make this
specific and reliable for pancreatic lipase without any reaction due to lipolytic enzymes or esterases.
The reagent composition has been thoroughly optimized to avoid serum matrix effects. The
generated methylresorufin ester is spontaneously degraded to methylresorufin. The absorbance by
this red dye is directly proportional to the lipase activity in the sample
Assay procedure
Name LIPASE
Wavelength 580 nm, Hg 578 nm
Optical path 1 cm
Temperature 37°C
Measurement Kinetic, against reagent blank
Reagent preparation
The reagents are ready to use. Do not shake!
A slight apparent red precipitate may occur in reagent 2, which does not affect the performance of
the test. Please do not resuspend before use
Blank Sample/calibrator
Sample/calibrator - 20 μL
Dist. water 20 μL -
Reagent 1 1000 μL 1000 μL
Mix carefully (do not shake), incubate 1 to 5 min.. Start reaction by adding reagent 2
Reagent 2 250 μL 250 μL
Mix, incubate 2 min at 37°C, read absorbance and start stopwatch. After exactly 1 and 2 min. read
absorbance again and then calculate ΔA/min.
Calculation [U/L]
With calibrator
ΔA/min. Sample
Lipase [U/L] = x Conc. Cal. [U/L]
ΔA/min. Calibrator
Conversion factor:
Lipase [U/L] x 0.0167 = Lipase [μkat/L]

Seite 42 von 63
Lp(a) 21 FS Product code: 1 7139…
Method
Particle enhanced immunoturbidimetric test. Determination of the Lp(a) concentration by
photometric measurement of antigen-antibody-reaction between antibodies against Lp(a) bound to
particles and Lp(a) present in the sample.
Assay procedure
Name LPA
Wavelength 700 nm
Optical path 1 cm
Temperature 37°C
Measurement Two point kinetic, against reagent blank
Reagent preparation
The reagents are ready to use.

Blank Sample/calibrator
Sample/calibrator - 15 μL
Dist. water 15 μL -
Reagent 1 600 μL 600 μL
Mix, incubate for 3 – 5 min., then add:
Reagent 2 300 μL 300 μL
Mix, read absorbance A1 within 30 sec., incubate for 5 min., then read absorbance A2.
Calculation [mg/dL]
ΔA = (A2 – A1) sample/calibrator
With calibrator
The Lp(a) concentration of unknown samples is derived from a calibration curve using an
appropriate mathematical model such as spline. The calibration curve is obtained with 5 calibrators
at different levels and NaCl solution (9 g/L) for determination of the zero value.

Seite 43 von 63
Magnesium XL FS Product code: 1 4610…
Method
Photometric test using xylidyl blue. Magnesium ions form a purple colored complex with xylidyl blue
in alkaline solution. In presence of GEDTA, which complexes calcium ions, the reaction is specific.
The intensity of the purple color is proportional to the magnesium concentration.
Assay procedure
Name MG
Wavelength 520 nm, Hg 546 nm, 500 - 550 nm (Increase of absorbance)
628 nm, Hg 623 nm, 570 - 650 nm (Decrease of absorbance)
Optical path 1 cm
Temperature 20 – 25°C/37°C
Measurement End point, against reagent blank
Reagent preparation
The reagent is ready to use.
Blank Sample/standard/calibrator
Sample/standard/calibrator - 10 μL
Dist. water 10 μL -
Reagent 1000 μL 1000 μL
Mix and read absorbance against blank after 5-60 min. at 20-25°C/37°C.
Calculation [mg/dL]
With calibrator
ΔA Sample
Magnesium [mg/dL] = x Conc. Std./Cal. [mg/dL]
ΔA Std./Cal.
Conversion factor:
Magnesium [mg/dL] x 0.4114 = Magnesium [mmol/L]

Seite 44 von 63
Myoglobin FS Product code: 1 7098…
Method
Particle enhanced immunoturbidimetric test. Fixed time determination of the concentration of
myoglobin through photometric measurement of antigen-antibody-reaction among antibodies to
human myoglobin coated to latex particles and myoglobin present in the sample.
Assay procedure
Name MYO
Wavelength 580 nm
Optical path 1 cm
Temperature 37°C
Measurement Two point kinetic, against reagent blank
Reagent preparation
The reagents are ready to use.
Blank Sample/calibrator
Sample/calibrator - 20 μL
Dist. water 20 μL -
Reagent 1 600 μL 600 μL
Mix, incubate for 3 – 5 min., then add:
Reagent 2 200 μL 200 μL
Mix and read absorbance A1 within 30 sec. Incubate for 5 min. and read absorbance again A2.
Calculation [µg/L]
ΔA = (A2 – A1) sample/calibrator
With calibrator
The myoglobin concentration of unknown samples is derived from a calibration curve using an
appropriate mathematical model such as spline. The calibration curve is obtained with four
calibrators at different levels and NaCl solution (9 g/L) for determination of the zero value.
Conversion factor:
Myoglobin [μg/L] x 0.059 = Myoglobin [nmol/L]

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NEFA FS Product code: 1 5781…
Method
Enzymatic endpoint method. Non-esterified fatty acids and coenzyme A react in the presence of
acyl coenzym A synthetase (ACS) to acylated coenzyme A. Acylated coenzyme A is oxidized by
acyl coenzyme A oxidase under development of H2O2. H2O2 is converted to a coloured product by
the use of Trinder substances in the presence of peroxidase (POD).
Assay procedure
Name NEFA
Wavelength 546 nm/600 nm (bichromatic)
Optical path 1 cm
Temperature 37°C
Measurement End point, against reagent blank
Reagent preparation
The reagents are ready to use.
Blank Sample/standard/calibrator
Sample/standard/calibrator - 20 μL
Dist. water 20 μL -
Reagent 1 1000 μL 1000 μL
Mix, incubate for 5 min. Read absorbance A1, then add:
Reagent 2 250 μL 250 μL
Mix, incubate 10 min. and read absorbance A2 within 20 min.
Calculation [µg/L]
ΔA = (A2 – A1) Sample/standard/calibrator
With standard
ΔA Sample
NEFA [mg/dL] = x Conc. Std./Cal. [mg/dL]
ΔA Std./Cal.
Conversion factor:
Non-esterified fatty acids [mg/dL] x 0.0354 = Non-esterified fatty acids [mmol/L]

Seite 46 von 63
Pancreatic amylase CC FS Product code: 1 0551…
Method
Enzymatic photometric test. Enzymatic photometric test, in which the substrate 4,6-ethylidene-(G7)-
p-nitrophenyl-(G1)-α-D-maltoheptaoside (EPS-G7) is cleaved by α-amylases into various
fragments. These are further hydrolyzed in a second step by α-glucosidase producing glucose and
p-nitrophenol [1,2]. As the salivary isoenzyme is inhibited selectively by a combination of two
monoclonal antibodies during the preincubation phase, the increase in absorbance represents the
pancreatic amylase activity in the sample.
Assay procedure
Name PAMY
Wavelength Hg 405 nm
Optical path 1 cm
Temperature 37°C
Measurement Kinetic, against reagent blank
Reagent preparation
The reagents are ready to use.
Blank Serum/plasma Urine
Sample/calibrator - 20 µL 10 µL
Reagent 1 1000 μL 1000 μL 1000 μL
Mix, incubate for approx. 3 min., then add:
Reagent 2 250 μL 250 μL 250 μL
Mix, read absorbance after 2 min. and start stopwatch. Read absorbance again after 1, 2 and 3 min.
Calculation [U/L]
ΔA = (A2 – A1) sample/calibrator
With factor
Calculate ΔA/min from absorbance readings and multiply by the corresponding factor:
Δ A/min x 5670 = Pancreatic amylase activity [U/L]
With calibrator
ΔA/min Sample
P-Amyl. [U/L] = x Conc. Cal. [U/L]
ΔA/min Cal.
Conversion factor::
Pancreatic amylase [U/L] x 0.0167 = Pancreatic amylase [μkat/L]

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Phosphate FS Product code: 1 5211…
Method
Photometric UV test with endpoint determination
Assay procedure
Name PHOS
Wavelength 340 nm, Hg 334 nm, Hg 365 nm
660 nm bichromatic
Optical path 1 cm
Temperature 20 – 25°C/37°C
Measurement End point, against reagent blank
Reagent preparation
The reagents and the standard are ready to use.
Blank Sample/standard/calibrator
Sample/standard/calibrator - 10 μL
Dist. water 10 μL -
Reagent 1 800 μL 800 μL
Mix, incubate for 5 min., read absorbance A1, then add:
Reagent 2 200 μL 200 μL
Mix and read absorbance A2 within 5 – 60 min.
Sample start
For mono reagent mix 4 parts R1 + 1 part R2
Stability: 1 year at 2 - 8°C
Blank Sample
Sample/calibrator - 10 µL
Dist Water 10 µL -
Mono reagent 1000 μL 1000 µL
Mix and incubate for 5 min. Read absorbance against reagent blank within 60 min.
Calculation [mg/dL] substrate start
ΔA = (A2 – A1) Sample/standard/calibrator
Calculation [mg/dL] sample start
ΔA = A Sample/standard/calibrator
With standard or calibrator
ΔA Sample
Phosphorus [mg/dL] = x Conc. Std./Cal. [mg/dL]
ΔA Std./Cal.
Conversion factor:
Phosphate [mmol/L] = Phosphorus [mmol/L]
Phosphorus [mg/dL] x 0.3229 = Phosphorus [mmol/L]
Phosphorus [mg/dL] x 3.06619 = Phosphate [mg/dL]

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Phospholipids FS Product code: 1 5741…
Method
Enzymatic colorimetric test
Assay procedure
Name PHOSPHOLIPIDS
Wavelength 570 nm
Optical path 1 cm
Temperature 37°C
Measurement Fixed time, against reagent blank
Reagent preparation
The reagents are ready to use.
Blank Sample/standard/calibrator
Sample/standard/calibrator - 10 μL
Dist. water 10 μL -
Reagent 1 1200 μL 1200 μL
Mix, incubate for 5 min., read absorbance A1, then add:
Reagent 2 300 μL 300 μL
Mix and read absorbance A2 after exactly 5 min.
Calculation [mg/dL]
ΔA = (A2 – A1) Sample/standard/calibrator
With standard or calibrator
ΔA Sample
Phospholipids [mg/dL] = x Conc. Std./Cal [mg/dL]
ΔA Std./Cal.
Conversion factor:
Phospholipids [mg/dL] x 0.0129 = Phospholipids [mmol/L]

Seite 49 von 63
Potassium FS Product code: 1 5221…
Method
Enzymatic photometric test. Pyruvate kinase is activated by K+ ions in the sample and
subsequently catalyzes the dephosphorylation of phosphoenolpyruvate to pyruvate. In a second
step pyruvate is transformed to lactate under consumption of a NADH analogue. The signal
decrease measured at 340 nm is proportional to the amount of potassium in the sample.
Assay procedure
Name K
Wavelength 340 nm
Optical path 1 cm
Temperature 37°C
Measurement Linear kinetic, against reagent blank
Reagent preparation
The reagents are ready to use.
Blank Sample/calibrator
Sample/calibrator - 100 μL
Dist. water 100 μL -
Reagent 1 1000 μL 1000 μL
Mix, incubate for 5 min. at 37°C., then add:
Reagent 2 250 μL 250 μL
Mix, incubate at 37°C, read absorbance A1 after 2 min. and start stopwatch. Read absorbance A2
after 1 min., absorbance A3 after 2 min. and absorbance A4 after 3 min. at 37°C and calculate
ΔA/min.
Calculation [mg/dL]
ΔA/min Sample
Potassium [mmol/L] = x Conc. Cal. [mmol/L]
ΔA/min Cal.
With calibrator
The concentration of potassium in unknown samples is derived from a calibration curve using an
appropriate mathematical model such as cubic spline. The calibration curve is obtained with the
levels 1 - 4 of the electrolyte calibrator TruCal E
Conversion factor:
Potassium [mmol/L] = Potassium [mEq/L]
Potassium [mmol/L] x 3.91 = Potassium [mg/dL]

Seite 50 von 63
Prealbumin FS Product code: 1 0292…
Method
Immunoturbidimetric test. Fixed time determination of the prealbumin concentration by
photometric measurement of antigen-antibody-reaction between antibodies against prealbumin and
prealbumin present in the sample.
Assay procedure
Name PALB
Wavelength 415 nm
Optical path 1 cm
Temperature 37°C
Measurement End point, against reagent blank
Reagent preparation
Blank Sample/calibrator
Sample/calibrator - 12 μL
Dist. water 12 μL -
Reagent 1 1000 μL 1000 μL
Mix, incubate for 5 min., read absorbance A1, then add:
Reagent 2 200 μL 200 μL
Mix, incubate for 5 min. and read absorbance A2.
Calculation [g/L]
ΔA = (A2 – A1) sample/calibrator
The prealbumin concentration of unknown samples is derived from the calibration curve using an
appropriate mathematical model such as 4-parameter Logit-log. The calibration curve is obtained
with five calibrators at different levels and NaCl solution (9 g/L) for determination of the zero value.

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Rheumatoid factor FS Product code: 1 7022…
Method
Immunoturbidimetric test. Determination of the RF concentration by photometric measurement of
antigen antibody reaction among heat aggregated IgG and rheumatoid factors present in the
sample.
Assay procedure
Name RF
Wavelength 340 nm, Hg 334 nm
Optical path 1 cm
Temperature 37°C
Measurement Fixed time kinetic, against reagent blank
Reagent preparation
Blank Sample/calibrator
Sample/calibrator - 15 μL
Dist. water 15 μL -
Reagent 1 250 μL 250 μL
Mix, incubate for 3 - 5 min., read absorbance A1, then add:
Reagent 2 50 μL 50 μL
Mix, incubate for 5 min. and read absorbance A2.
Calculation [g/L]
ΔA = (A2 – A1) sample/calibrator
The concentration of RF in unknown samples is derived from a calibration curve using an
appropriate mathematical model such as logit/log. The calibration curve is obtained with 5
calibrators at different levels and NaCl solution (9 g/L) for determination of the zero value.

Seite 52 von 63
Sodium FS Product code: 1 4808…
Method
Enzymatic photometric test. β-galactosidase catalyzes the conversion of o-nitrophenyl-β-
Dgalactopyranoside (ONPG) to o-nitrophenol and galactose. The activity of β-galactosidase
depends on the sodium concentration in the sample. The absorbance increase at 405 nm is
proportional to the sodium concentration in the sample.
Assay procedure
Name NA
Wavelength 405/660 nm (bichromatic)
Optical path 1 cm
Temperature 37°C
Measurement Kinetic, against reagent blank
Reagent preparation
Blank Sample/calibrator
Sample/calibrator - 40 μL
Dist. water 40 μL -
Reagent 1 900 μL 900 μL
Mix, incubate for 5 min. at 37°C, then add:
Reagent 2 300 μL 300 μL
Mix, incubate at 37°C, read absorbance A1 after 1 min. and start stopwatch. Read absorbance A2
after 1 min. and absorbance A3 after 2 min. at 37°C and calculate ΔA/min.
Calculation [µg/L]
ΔA/min. Sample
Sodium [mmol/L] = x Conc. Cal. [mmol/L]
ΔA/min. Cal.
With calibrator
The concentration of sodium in unknown samples is derived from a linear calibration curve. It is
obtained with the levels 1/2 and 3/4 of the electrolyte calibrator TruCal E.
Conversion factor:
Sodium [mmol/L] = Sodium [mEq/L]
Sodium [mmol/L] x 2.30 = Sodium [mg/dL]

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Total bile acids 21 FS Product code: 1 2238…
Method
Enzymatic cycling method. Two reactions are combined in the new generation enzymatic cycling
method. In the presence of Thio-NAD, the enzyme 3-α-hydroxysteroid dehydrogenase (3-α-HSD)
converts bile acids to 3-ketosteroids and Thio-NADH. The reaction is reversible and 3-α-HSD can
convert 3-ketosteroids and NADH to bile acids and NAD. In the presence of excess NADH, the
enzyme cycling occurs efficiently and the rate of formation of Thio-NADH is determined by
measuring the specific change of absorbance at 405 nm. This cycling reaction leads to significant
signal amplification.
Assay procedure
Name TBA_21
Wavelength 405 nm / 600 nm (bichromatic)
Optical path 1 cm
Temperature 37°C
Measurement Kinetic, against reagent blank
Reagent preparation
Reagents are ready to use
Blank Sample/calibrator
Sample/calibrator - 10 μL
Dist. water 10 μL -
Reagent 1 675 μL 675 μL
Mix, incubate for 5 min. at 37°C. Then add:
Reagent 2 225 μL 225 μL
Mix and read absorbance after 1 minute and start stop watch. Read absorbance again after 1 and
2 minutes.
Calculation [mg/dL]
With calibrator
ΔA/min. = ΔA/min. sample/calibrator

ΔA/min Sample
Bile acids [µmol/L] = x Conc. Cal. [µmol/L]
ΔA/min Cal.

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Total protein FS Product code: 1 2311…
Method
Photometric test according to biuret method. Proteins form a violet blue color complex with copper
ions in alkaline solution. The absorbance of the color is directly proportional to the concentration.
Assay procedure
Name TP
Wavelength 540 nm, Hg 546 nm
Optical path 1 cm
Temperature 20 – 25°C/37°C
Measurement End point, against reagent blank
Reagent preparation
Reagents are ready to use
Blank Sample/standard/calibrator
Sample/standard/calibrator - 20 μL
Dist. water 20 μL -
Reagent 1 1000 μL 1000 μL
Mix, read absorbance A1 after 1 – 5 min. at 20 – 25°C/ 37°C, then add:
Reagent 2 250 μL 250 μL
Mix, incubate for 5 min. at 20 – 25°C/37°C and read absorbance A2 within 60 min.
Calculation [g/dL]
With standard or calibrator
ΔA = (A2 – A1) Sample/standard/calibrator

ΔA Sample
Total protein [g/dL] = x Conc. Std./Cal. [g/dL]
ΔA Std./Cal.

Seite 55 von 63
Total protein UC FS Product code: 1 0210…
Method
Photometric test using pyrogallol red. Proteins form a red complex with pyrogallol red/ molybdate.
The absorbance is directly proportional to the protein concentration.
Assay procedure
Name TPU
Wavelength 600 nm
Optical path 1 cm
Temperature 37°C
Measurement End point, against reagent blank
Reagent preparation
Reagents are ready to use
Blank Sample/standard
Sample/standard - 20 μL
Dist. water 20 μL -
Reagent 1000 μL 1000 μL
Mix and read absorbance against reagent blank exactly after 10 min.
Calculation [g/dL]
With standard
A Sample
Total protein [mg/L] = x Conc. Std. [mg/L]
A Std.

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Transferrin FS Product code: 1 7252…
Method
Immunoturbidimetric test. Determination of the transferrin concentration through photometric
measurement of antigen-antibody-reaction among antibodies to transferrin and transferrin present
in the sample.
Assay procedure
Name TRF
Wavelength 570 nm
Optical path 1 cm
Temperature 37°C
Measurement End point, against reagent blank
Reagent preparation
Reagents are ready to use
Blank Sample/calibrator
Sample/calibrator - 10 μL
Dist. water 10 μL -
Reagent 1 1250 μL 1250 μL
Mix, incubate for 3 – 5 min., read absorbance (A1), then add:
Reagent 2 250 μL 250 μL
Mix, incubate for 5 min., read absorbance A2.
Calculation [mg/dL]
With calibrator
ΔA = (A2 – A1) sample/calibrator
The concentration of transferrin in unknown samples is derived from a calibration curve using an
appropriate mathematical model such as logit/log. The calibration curve is obtained with 5
calibrators at different levels and NaCl solution (9 g/L) for determination of the zero value.

Iron [µg/dl] x 79570

TRF - Saturation [%] =
Tfr [mg/dL] x 2 x 56 x 10
Conversion factor:
Transferrin [mg/dL] x 0.126 = Transferrin [µmol/L]

Seite 57 von 63
Triglycerides FS Product code: 1 5710…
Method
Colorimetric enzymatic test using glycerol-3-phosphate-oxidase (GPO). Determination of
triglycerides after enzymatic splitting with lipoprotein lipase. Indicator is quinoneimine which is
generated from 4-aminoantipyrine and 4-chlorophenol by hydrogen peroxide under the catalytic
action of peroxidase.
Assay procedure
Name TRIG
Wavelength 500 nm, Hg 546 nm
Optical path 1 cm
Temperature 20 – 25°C/37°C
Measurement End point, against reagent blank
Reagent preparation
Reagent is ready to use
Blank Sample/standard/calibrator
Sample/standard/calibrator - 10 μL
Dist. water 10 μL -
Reagent 1000 μL 1000 μL
Mix, incubate for 20 min. at 20 – 25°C or 10 min. at 37°C. Read the absorbance against reagent
blank within 60 min
Calculation [g/dL]
With standard or calibrator
A Sample
Triglycerides [mg/dL] = x Conc. Std./Cal. [mg/dL]
A Std./Cal.

To correct for free glycerol, subtract 10 mg/dL (0.11 mmol/L) from the triglycerides value calculated
above.
Conversion factor:
Triglycerides [mg/dL] x 0.01126 = Triglycerides [mmol/L]

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UIBC FS Product code: 1 1921…
Method
Photometric test using Ferene. A known ferrous ion concentration incubated with sample, binds
specifically with transferrin at unsaturated iron binding sites. Remaining unbound ferrous ions are
measured with the ferene
reaction. The difference between the amount of excess iron and the total amount added to the
serum is equivalent to the quantity bound to transferrin. This is the UIBC (unsaturated iron binding
capacity) of the sample.
Assay procedure
Name UIBC
Wavelength 600 – 620 nm, Hg 578 nm, 623 nm
Optical path 1 cm
Temperature 37°C
Measurement End point, against reagent blank
Reagent preparation
Reagents are ready to use
Blank Sample/calibrator
Sample/calibrator - 75 μL
Dist. water 75 μL -
Reagent 1 1000 μL 1000 μL
Mix, read absorbance A1 after 5 min., then add:
Reagent 2 250 μL 250 μL
Mix, read absorbance A2 after exactly 5 min.
Calculation [µg/dL]
With calibrator
ΔA = (A2 – 0.81 A1) sample/calibrator

The factor 0.81 compensates the decrease of the absorbance by addition of reagent 2. The factor is
calculated as follows: (Sample + R1)/Total volume. This compensation is necessary as a high
sample volume is used.

ΔA Sample
UIBC [µg/dL] = x Conc. Cal. [µg/dL]
ΔA Cal.

Conversion factor:
UIBC [μg/dL] x 0.1791 = UIBC [μmol/L]
TIBC [μg/dL] = UIBC [μg/dL] + Iron [μg/dL]
Transferrin [mg/dL] = 0.7 x TIBC [μg/dL]

Seite 59 von 63
UREA CT FS Product code: 1 3115…
Method
Colorimetric test. Urea is hydrolyzed in the presence of water and urease to produce ammonia and
carbon dioxide. Ammonium ions react with hypochlorite and salicylate to give a green dye. The
increase in absorbance at 578 nm is proportional to the urea concentration in the sample.
Assay procedure
Name UREA_CT
Wavelength 578 nm, 560 – 600 nm
Optical path 1 cm
Temperature 20 – 25°C/37°C
Measurement End point, against reagent blank
Reagent preparation
Mix R1 + R3 in the ratio 100 + 1 e.g. 20 mL R1 + 0.2 mL R3 = R1A
Stability of R1A: 2 weeks at 2 – 8°C, 2 days at 15 – 25°C
R1A and R2 must be protected from light!
Reagent 2 and standard are ready for use.
Blank Sample/standard/calibrator
Sample/standard/calibrator - 10 μL
Dist. water 10 μL -
Reagent 1 A 1000 μL 1000 μL
Mix, incubate 10 min. at 20 – 25°C or 5 min. at 37° C, then add:
Reagent 2 1000 μL 1000 μL
Mix, incubate 10 min. at 20 – 25°C or 5 min. at 37° C. In case of 20 – 25°C read the absorbance
against reagent blank within 30 min.; in case of 37°C within 5 min.
Calculation [mg/dL]
With standard or calibrator
ΔA Sample
Urea [mg/dL] = x Conc. Std./Cal. [mg/dL]
ΔA Std./Cal.
Conversion factor:
Urea [mg/dL] x 0.1665 = Urea [mmol/L]
Urea [mg/dL] x 0.467 = BUN [mg/dL]
BUN [mg/dL] x 2.14 = Urea [mg/dL]
(BUN: Blood urea nitrogen = Urea-N in blood)

Seite 60 von 63
Urea FS Product code: 1 3101…
Method
Enzymatic UV test
Assay procedure
Name UREA
Wavelength 340 nm, Hg 334 nm, Hg 365 nm
Optical path 1 cm
Temperature 25°C/30°C/37°C
Measurement Two point kinetic, against reagent blank
Reagent preparation
Reagents are ready to use
Blank Sample/standard/calibrator
Sample/standard/calibrator - 10 μL
Reagent 1 1000 μL 1000 μL
Mix, incubate 0 - 5 min., then add:
Reagent 2 250 μL 250 μL
Mix, incubate for approx. 60 sec. at 25°C/30°C or approx. 30 – 40 sec. at 37°C, then read
absorbance A1. Read absorbance A2 exactly after another 60 seconds.
Calculation [mg/dL]
ΔA = (A2 – A1) Sample/standard/calibrator
Notes
1. The method is optimized for 2-point kinetic measurement. It is recommended to perform the
method only on mechanized equipment because it is difficult to incubate all samples and the
reagent blank strictly for the same time intervals. The assay scheme may be used for adaptation
purposes for instruments with no specific adaptation sheet. The volumes may be proportionally
smaller.
2. The statement “approx. 60 sec. or approx. 30 - 40 sec“ means that the time period chosen does
not need to be exactly 60 resp. 30 – 40 sec. A time period once chosen (e.g. 55 sec.) has to be
respected exactly for all samples, standards and the reagent blanc.
With standard or calibrator
ΔA Sample
Urea [mg/dL] = x Conc. Std./Cal. [mg/dL]
ΔA Std./Cal.
Conversion factor:
Urea [mg/dL] x 0.1665 = Urea [mmol/L]
Urea [mg/dL] x 0.467 = BUN [mg/dL]
BUN [mg/dL] x 2.14 = Urea [mg/dL]
(BUN: Blood urea nitrogen = Urea-N in blood)

Seite 61 von 63
Uric acid FS TBHBA Product code: 1 3021…
Method
Enzymatic photometric test using TBHBA. Uric acid is oxidized to allantoin by uricase. The
generated hydrogen peroxide reacts with 4-aminoantipyrine and 2,4,6-tribromo-3-hydroxybenzoic
acid (TBHBA) to quinoneimine.
Assay procedure
Name UA_TBHBA
Wavelength 520 nm, Hg 546 nm, 500 - 550 nm
Optical path 1 cm
Temperature 20 – 25°C/37°C
Measurement End point, against reagent blank
Reagent preparation
Reagents are ready to use
Blank Sample/standard/calibrator
Sample/standard/calibrator - 20 μL
Dist. water 20 μL -
Reagent 1 A 1000 μL 1000 μL
Mix, incubate 5 min., then add:
Reagent 2 250 μL 250 μL
Mix, incubate 30 min. at 20 – 25°C or 10 min. at 37°C. Read the absorbance against the reagent
blank within 60 min.
Calculation [mg/dL]
With standard or calibrator

ΔA Sample
Uric acid [mg/dL] = x Conc. Std./Cal. [mg/dL]
ΔA Std./Cal.
Conversion factor:
Uric acid [mg/dL] x 59.48= Uric acid [μmol/L]

Seite 62 von 63
Uric acid FS TOOS Product code: 1 3001…
Method
Enzymatic photometric test using TOOS (N-ethyl-N-(hydroxy-3-sulfopropyl)-m-toluidin) Uric acid is
oxidized to allantoin by uricase. The generated hydrogen peroxide reacts with 4-aminoantipyrine
and N-ethyl-N-(hydroxy-3-sulfopropyl)-m-toluidin (TOOS) to a blue violet dye. Ascorbate oxidase
avoids interference by ascorbic acid.
Assay procedure
Name UA_TOOS
Wavelength 550 nm, Hg 546 nm
Optical path 1 cm
Temperature 20 – 25°C/37°C
Measurement End point, against reagent blank
Reagent preparation
Blank Sample/standard/calibrator
Sample/standard/calibrator - 20 μL
Reagent 1 1000 μL 1000 μL
Mix, incubate 5 min., then add: read the absorbance 1 and then add
Reagent 2 250 μL 250 μL
Mix, incubate 5 min. at 37°C or 10 min. at 20 – 25°C. Read the absorbance 2 within 30 min. Pay
attention to apply exactly the same incubation time for standard/calibrator, blank and sample
Calculation [mg/dL]
ΔA = (A2 – A1) Sample/standard/calibrator
With standard or calibrator
ΔA Sample
Uric acid [mg/dL] = x Conc. Std./Cal. [mg/dL]
ΔA Std./Cal.
Conversion factor:
Uric acid [mg/dL] x 59.48= Uric acid [μmol/L]

Seite 63 von 63