GENYSIS SPECTROPHOTOMETRY

ALWAYS Close SPECTROPHOTOMETER door between all sample handling

ALWAYS Pour all liquids AWAY from SPECTROPHOTOMETER, working over bench top

A. TEST SET UP
1. Complete Worksheet with Name, Date, Spect ID, Wavelength, Sample IDs
2. Patient/Unknowns except for QC are ALWAYS performed in duplicate.
3. Check tubes for imperfections, dirt, etc., clean as necessary
4. Label tubes: BLK, STD, QC1, QC2, ID1, ID1, ID2, ID2, etc. Use patient Identification #
rather than names.
5. Most Chemistry methods will use a Regent Blank NOT water.
6. Accurately pipet reagent volume into all tubes; verify reagent color and volume
7. Accurately pipet sample volume into appropriate tubes
8. Mix each tube well
9. Incubate at correct temp and time according to Test Procedure (Package insert), set
timer, log temp/time
10. At incubation time completion, IF not at RT, remove tubes from incubation area (such
as heat block) back into rack at RT.
11. Record current time of end of incubation.
12. Check cuvette for clean, no scratches or cracks, only holding by sides and top. Do not
touch cuvette front and back of the read areas.
13. Proceed to reading all samples in Spectrophotometer within sample stability time as
listed on test procedure.

B. READING Absorbance in SPECT

Close Spect DOOR between all sample handling
Pour all liquids AWAY from Spect
1. Set SPECT to nm according to procedure
2. Beginning with the Reagent BLK, carefully Pour or pipet solution into cuvette
3. Carefully wipe outside of cuvette to remove any moisture; tap gently on counter to
remove any bubbles.
4. Load cuvette into spectrophotometer without touching read surfaces. Cuvette front
indicator should be facing towards you.
5. Close read station door carefully.
6. Press Zero button to set the Reagent BLK to 0.000 A, check ZERO on WS.
7. Remove cuvette from Spect, close the lid carefully.
8. Pour solution back into correct tube.
9. Blot opening of empty cuvette well onto gauze. (tap upside down on gauze)
10. Pour STD into cuvette, load, close door. Do not zero!
11. Record A of STD onto WS to 3 decimal places.
12. PRESS PRINT for each sample when A is displayed before removing cuvette
13. Remove cuvette from Spect, close the lid, and pour solution back into tube.
/conversion/tmp/activity_task_scratch/605951395.docx
Written 2010 rg; Rev 11/14 rg; Rev 12/18 rg
 GENYSIS SPECTROPHOTOMETRY
14. Blot opening of cuvette well onto gauze.
15. Repeat with all tubes, recording A and blotting cuvette between each filling.
16. Close Spect Door, wipe off all surfaces, remove printout, power off if last using.
17. Label printout with your initials and each sample ID

C. GRAPHING
1. Label Graph paper with TEST NAME and Date on center top above graph lines; initial
upper right.
2. Bottom Left line intersection is 0.000 A and 0 C unless otherwise noted in procedures.
The number of C decimals must be the same as the STD concentration
3. Draw line from top to bottom at the Upper Reportable limit concentration.
4. Label axis’ as directed; CONC vs A (X axis C; Y axis A)
5. Plot STD A reading onto labeled graph paper using a small DOT at concentration
6. Using a ruler, draw straight line from 0,0 through STD A DOT up to the upper
reportable range of method. This is your CALIBRATION CURVE.
7. Read QC1,2 from curve:
a. Find QC A on ABSORBANCE axis
b. Follow this line across to CURVE line you drew. Do not write this dot on your
curve line.
c. Follow area down to CONC axis to find the concentration of QC
d. Record QC Concentration onto WS using the same decimal places as STD
8. Repeat with all solutions recording your C reading onto WS

D. Determine Run Acceptability

1. Complete WS with all necessary information.
2. Interpret QC acceptability based on QC material package insert Acceptable Ranges (±2
SD).
3. Patient duplicates must match within 10%
[Reading A-Reading B/Largest Reading X 100]

E. CALCULATION using BEER’S LAW

Cx Cs
Ax
= As

1. Using Beer’s Law formula, insert values for each solution (QC1, QC2, PT1, PT1) into box
on worksheet
2. Calculate showing math set up and log final Concentration result
3. Direct Curve reading vs Calculation final result matching should be within 10%

/conversion/tmp/activity_task_scratch/605951395.docx
Written 2010 rg; Rev 11/14 rg; Rev 12/18 rg