13.1.1
• The enzymatic methods are adjusted to the test protocols of the kits listed below and can only be used with these kits.
• A cell changer is needed for the analyses (6- or 8-cell changer or cell carousel).
Overview of the enzymatic methods for foodstuffs analysis 13.1.2
Method Test kit Catalog number Method name
Acetate R-Biopharm Art. Nr 10 148 261 test combination für 3 x 11 Determinations
Acet
Cholesterol R-Biopharm Art. Nr. 10 139 050 035 Test combination for 31 Determinations
Chol
Citric acid R-Biopharm Art. Nr. 10 139 076 035 Test combination for 3 x 12 Determinations
Cit
D-/L-Lactic acid R-Biopharm Art. Nr. 11 112 821 035 Test combination for 30 determinations
DLLacticacid
D-Glucose R-Biopharm Enzytec Fluid Line Art. Nr. E5140 Test combination for 4x10 determinations
Gluc
Ethanol R-Biopharm Enzytec Fluid Line Art. Nr. E5340
Test combination for 4x10 determinations
Eth
Fructose R-Biopharm Enzytec Fluid Line Art. Nr. E5120
Test combination for 4 x 9 determinations
Fruc
Galactose R-Biopharm Art. Nr. 10 176 303 035 Test combination for 32 determinations
Galac
Glutaminsäure R-Biopharm Enzytec Generica Line Art. Nr. E1269
Test combination for 3x12 determinations
Glut
Hydroxy butyric
acid R-Biopharm Art. Nr. 10 907 979 035 Test combination for 3 x ca.12 determinations
D3hba
Lactose (Gluc) R-Biopharm Enzytec Fluid Line Art. Nr. E5600
Test combination for 4 x 10 determinations
LactGluc
Lactose (Galac) R-Biopharm Art. Nr. 10 176 303 035 Test combination for 32 determinations
LactGalac
L-Malate R-Biopharm Art. Nr. 10 139 068 035 Test combination for 30 determinations
LMal
L-Lactic acid R-Biopharm Art. Nr. 10 139 084 035 Test combination for 30 determinations
LLacticacid
Nitrate R-Biopharm Art. Nr. 10 905 658 035 Test combination for 3 x 13 determinations
Nitr
Oxalic acid R-Biopharm Art. Nr. 10 755 699 035 Test combination for 11 determinations
Oxal
Saccharose R-Biopharm Enzytec Fluid Line Art. Nr. E5180
Test combination for 4x10 determinations
Sacc
Sorbit R-Biopharm Art. Nr. 10 670 057 035 Test combination for 3 × ca.12 determinations
Sorb
Starch R-Biopharm Art. Nr. 10 207 748 035 Test combination for 27 determinations
Star
Sulfite R-Biopharm Art. Nr. 10 725 854 035 Test combination for 31 determinations
Sulf
Xylit R-Biopharm Art. Nr. 10 670 057 035 Test combination for 3 × ca.12 determinations
Xylit
Overview of the enzymatic methods for the medical field 13.1.3
Method Test kit Catalog number Method
name Urea/ Ammoniac Megazyme K-URAMR
100 assays per kit Urea Free fatty acid Wako ACS-ACOD-MEHA –
method NEFA-HR(2) R1 Set 434-91795
(4x for 50ml or R1a 4x 50ml) Fat
Porphyrine RECIPE 17100 ClinRep
Complete kit for Porphyrine total in urea
for 50 determinations
Por
delta ALA (5- amino-
levulin acid) RECIPE 17200ClinRep
complete kit for 5-ALA and PPG in urea
for 50 determinations
Ala
PBG (Porpho-bilinogen) RECIPE 17200ClinRep
complete kit for 5-ALA and PPG in urea
for 50 determinations
Pbg
Bile acid
Note: For the proof microcells for small sample volumes are needed.
DiaSys Diagnostic Systems
1 2212 99 90 313 (5x10ml R1;
1x10ml R2; 1x2ml standard) Bile
Pyruvat Megazyme K-PYRUV
100 assays per kit Pyru
DiaSys http://www.diasys-diagnostics.com/
Making available enzymatic methods in WinASPECT PLUS 13.1.4
1. Call the Method / Edit method list / Add method menu command.
The Open standard window is opened.
2. Select the // WinASPECT PLUS/Methods Enzymology/Method Name subfolder.
For details on the name of the Method Name macro folder, refer to the lists above.
3. Select the Method Name.mac file there and confirm your selection with [OK].
The analytical method is now available in the Method menu.
Note
At first the reference measuring is carried out after method start. After this the request for the addition of the reagent follows.
The 1st position into all cell changers remains free, even if the samples are measured in several ranks. The null measurement takes place at this position.
Two 8-cell changer are used always in offset mode. In this mode both cell changers are moved consecutively through the beam paths so that both cell changers are used for sample measurement.
Executing the macro for enzymatic analyses 13.1.5
1. Turn on the printer if you want to print the analytical results. The results are printed automatically after analysis.
2. Open the Method menu and click on the macro in the list.
3. Later in the analysis follow the instructions on screen.
The analytical data is automatically stored in the following folders.
//Enzymology /Method Name
Saved files
/export The analytical results are automatically exported into the Excel-legible ".csv" format and saved as a text file.
The file name is made up as follows:
Method Name_Date_Time.csv or Method Name_Date_Time.txt
e.g., Acet_01.03.2011_17_18_21.csv
/profile Contains the stored sample tables. Sample tables can be saved and loaded during the course of analysis on demand. The extension of the sample table is ".prl".
The name of the sample table file can be freely selected.
/print Contains the saved print protocols of the analyses. The file name is made up as follows:
Operator-Date-Time.pr
Operator is the entry made during the course of the analysis in the Operator field.
e.g., NeleNyx_01.03.2011_17_18_21.pr
/prot The files in this folder were only required internally for the process.
All files can be renamed as required. However, the extensions must be kept.
Subsequent printout of analytical results
Analytical results can be printed subsequently.
1. Call the Method / Print protocol menu command.
The Open standard window is opened.
2. In the //Methods/Enzymology/Method Name/Print folder select the desired protocol file.
This starts the printout of this file.
Method / Print protocol
Determination of glucose as an example for enzymatic 13.1.6
measurement
1. Select the method "Gluc.mac" from Method menu.
2. Create a new sample table.
Note:
If you load a saved sample table, you can also edit the parameters. But it is not possible to change the number of samples.
3. Enter the number of samples.
4. Activate the parameters dilution and net weight, if it is necessary.
5. Enter the name of the operator. The concentration unit is a default value, but it is possible to edit this parameter.
6. Enter the sample name, the dilution factor and the weight, if it is necessary.
7. Save the sample table, if you want to use it later again.
8. Place empty cells in the cell changer according the displayed scheme:
1st pos. – empty, used for baseline determination,
2nd pos. – blank value 3rd pos. – standard further pos. – samples.
Click [OK]. The cell changer moves to the 1st position for baseline determination.
9. Pipette reagent 1, water, standard sample and sample into the cells
according to the pipetting scheme of test kit.
Click on [OK].
10. After the 1st incubation time has elapsed the absorbance value E1 is measured automatically.
11. Pipette reagent 2 into the cells according to the pipetting scheme of test kit.
12. Click on [OK].
13. After the 2nd incubation time has elapsed the absorbance value E2 is measured automatically.
14. The values of calculated glucose concentration, the differences of absorbance (E1-E2) and the difference corrected by blank value (dE) are displayed in the sample table.
15. Power on the printer. The print out starts automatically with [OK].
Note:
The analysis data are saved automatically. So the data can be printed out at a later date (
section "Executing the macro for enzymatic analyses" p. 176).
The method starts again with step (2).