TOX - sc.2030 Rev 02, Effective 2016-09-20 To Present

Harris County Institute of Forensic Sciences
Section: Toxicology Approved by: Chief Toxicologist

Document Type: Non-GC & Non-LC Procedure No.: TOX.SC.2030
Title: Chemistries using Carolina Biolis 24i Rev.:2
1.0 Purpose
1.1 This document is for the operation and maintenance of the Carolina Biolis 24i
analyzer used to measure creatinine, electrolytes (sodium, potassium, and
chloride), glucose, and urea nitrogen (UN) in biological specimens.
1.2 Amounts of creatinine in blood or urine reflect glomerular filtration and are useful
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in the diagnosis and treatment of renal disease. Serum creatinine varies with the
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subject’s age, body weight, and sex. It is sometimes low in subjects with
relatively small muscle mass, cachectic patients, amputees, and in older persons.
A serum creatinine level that would usually be considered normal does not rule
out the presence of impaired renal function.
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1.2.1 Carolina Liquid Chemistries CREA assay uses a coupled enzymatic
procedure. First, creatinine is converted into creatine via creatininase.
Creatine is then converted to sarcosine by creatine amidinohydrolase
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(creatinase), followed by oxidation of sarcosine by sarcosine oxidase
(SOD) which produces hydrogen peroxide. In the presence of peroxidase
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(POD), the hydrogen peroxide produces a colored dye which is quantified
at 546 nm.
Creatinine + H2O Creatinina
 se
→ Creatine
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Creatine + H2O  → Sarcosine + Urea

Creatinase
Sarcosine + H2O + O2  →
SOD
Formaldehyde + Glycine + H2O2
2H2O2 + 4-AA + TOOS  →
POD
4H2O + Quinone dye (λ max 556)
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Any endogenous creatine present in the sample is removed by creatinase

and sarcosine oxidase during pre-incubation.
The Biolis 24i System dispenses sample and reagent into a cuvette where
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the increase in absorbance at 546 nm is monitored. The final absorbance is

directly proportional to the concentration of creatinine in the specimen.
1.3 Electrolytes are involved in most major metabolic functions in the body. Sodium,
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potassium and chloride are among the most important physiological ions found
and as such, are the most often assayed.
1.3.1 The Carolina Biolis 24i ion specific electrode (ISE) module for sodium
(Na+), potassium (K+) and chloride (Cl-) employs crown ether membrane
electrodes for sodium and potassium, and a molecular oriented PVC
membrane for chloride. Each are specific for their respective ion in the
sample. Electrical potentials for each ion are developed according to the
Nernst equation. When compared to the internal reference solution, this
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electrical potential is translated into voltage and then into the ion
concentration in each sample.
1.4 Serum glucose levels may be abnormally high (hyperglycemia) or abnormally

low (hypoglycemia). Glucose measurements are used in the detection of
carbohydrate metabolism disorders including diabetes mellitus, neonatal and
idiopathic hypoglycemia, and of pancreatic islet cell carcinoma.
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1.4.1 Enzymatic method for glucose determination using hexokinase (HK) and
glucose-6-phosphate dehydrogenase (G-6-PDH) is characterized below:
HK
Glucose + ATP G-6-P + ADP
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MG+2
G-6-PDH
G-6-P + NAD 6-phosphogluconate + NADH
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the increase in absorbance at 340 nm is monitored. The final absorbance is
directly proportional to the concentration of glucose in the specimen.
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1.5 Blood urea nitrogen (BUN) makes up approximately 75% of the total non-protein
nitrogen fraction of the blood. It is synthesized in the liver from ammonia
produced by metabolic deamination of proteins. Filtration of urea from the blood
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into the urine by the renal glomeruli is the chief means of eliminating surplus
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nitrogen from the body. Measurements of blood urea nitrogen are used in the
diagnosis and treatment of certain renal and metabolic disorders.
1.5.1 Enzymatic method for blood urea nitrogen (BUN) using urease is
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characterized below:
Urea UREASE
→ NH3 + CO2
NH3 + α-ketoglutarate + NADH GLDH

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→ Glutamic acid + NAD+
the change in absorbance at 340 nm is monitored. The final absorbance is
directly proportional to the concentration of UN in the specimen.
2.0 Scope
2.1 This method is applicable to the measurement of creatinine, electrolytes, glucose

and urea nitrogen (UN) in serum or vitreous specimens.
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2.2 Vitreous is the preferred sample for postmortem analysis.
2.3 If serum is used, it should be free from hemolysis and removed from the clot.
At room temperature, glycolysis decreases serum glucose levels approximately
7% in one hour. Therefore, it is important to separate serum from cells as soon
after collection as possible.
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2.4 Assay Interferences
2.4.1 Creatinine
A. There is no significant interference (less than 10% deviation) from
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triglyceride up to 1000 mg/dL, ascorbic acid at 10 mmol/L, bilirubin
up to 40g/dL, bilirubin conjugate up to 30 mg/dL or hemoglobin up to
500 mg/dL with this method.
2.4.2
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Electrolytes
A. Separate serum from the cells as soon as possible. Serum should be
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clear and free of hemolysis. Centrifuge samples that are visibly
contaminated with blood before analysis.
2.4.3 Glucose
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A. There is no significant interference from bilirubin to 16 mg/dL, with

this method. At 1000 mg/dL lipemia exhibits a 12% positive
interference. At 600 mg/dL hemoglobin exhibits a 12% negative
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interference. Drugs and other substances may affect glucose

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determinations.
2.4.4 Blood Urea Nitrogen

A. Hemolysis (2+) and elevated bilirubin up to 7 mg/dL do not cause
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significant interference with this assay. Lipemia will interfere with

this assay.
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3.0 Definitions and Abbreviations
3.1 S (on print out) -- Sample Error

3.2 LH (on print out) -- Instrument performed an automatic dilution (1:2)
3.3 G (on print out) -- Absorbance reading above allowable limit
3.4 L (on print out) -- Linearity error
3.5 B (on print out) -- Absorbance reading below allowable limit
3.6 0A000 (on print out) -- Air detected by flow sensor error
3.7 S0000 (on print out) -- Air detected
3.8 00200 (on print out) -- Potential drift
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3.9 GLUC -- Glucose

3.10 BUN-- Blood Urea Nitrogen
3.11 CREA – Creatinine
3.12 RCT – Reaction Tray
3.13 DI – deionized
3.14 RTM – Reagent Probe Transfer Mechanism
3.15 ISE – ion specific electrode
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4.0 Materials
4.1 Instrument and Equipment
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4.1.1 Carolina Biolis 24i Chemistry Analyzer
4.1.2 16 mm polystyrene sample cup
4.1.3 13 mm nesting sample cups
4.1.4 Alcohol prep pad
4.1.5
4.1.6 d
Cotton swabs
Sample sonicator
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4.1.7 Centrifuge
4.2 Reagents
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4.2.1 Deionized water

4.2.2 Isotonic saline
4.2.3 Fast Detergent 2 ACID wash
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4.2.4 Fast Detergent 1 ALK wash

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4.2.5 ISE Wash solution

4.2.6 Carolina Liquid Chemistry Creatinine Reagent (3 × 100 tests)
4.2.7 Carolina Liquid Chemistry Glucose Reagent (3 × 100 tests)
4.2.8 Carolina Liquid Chemistry BUN Reagent (3 × 150 tests)
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A. The Carolina Liquid Reagents are ready to use and contain

preservatives. Unopened reagents are stable until the expiration date
printed on the label if stored at 2-8°C.
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B. Opened reagent is stable for 30 days when stored in the refrigerated

compartment of the Biolis 24i.
C. Visible signs of microbial growth, turbidity, or precipitate in the
creatinine, glucose, or blood urea nitrogen (BUN) reagent may
indicate degradation and warrant discontinuation of use.
4.2.9 1% acid wash
A. 20 mL Fast Detergent 2 ACID wash + 1980 mL deionized water
B. Stable for 6 months at room temperature.
4.2.10 2% alkaline wash
A. 40 mL Fast Detergent 1 ALK wash + 1960 mL deionized water
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4.2.11 6% alkaline wash
A. 60 mL Fast Detergent 1 ALK wash + 940 mL deionized water
4.3 Standards and controls

4.3.1 ISE Calibrators
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Calibrator 1 contains 420 mL per pouch.
Calibrator 2 contains 20 mL per bottle.
Electrolyte Unit Calibrator 1 Calibrator 2

Sodium mmol/L 140.0 160.0
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Potassium mmol/L 4.00 6.00
Chloride mmol/L 100.0 120.0
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A. ISE calibrators are ready to use and contain preservatives.
B. Unopened reagents are stable until the expiration date printed on the
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label when stored at 18-25 °C. Reagent will be clear as packaged.
Discard if turbid.
C. Stable for 30 days after opening.
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4.3.2 Beckman Coulter SYNCHRON® Systems Multi Calibrator

A. Beckman Coulter SYNCHRON® Systems Multi Calibrator is ready to
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use.
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B. Store at -20 to -15 °C. The calibrator is stable until expiration date on
label.
C. Once opened, bottles are stable for 30 days when stored tightly capped
at -20 to -15 °C.
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D. Immediately prior to use, gently invert vial 5 to 10 times.

E. Slight variations in temperature from brief periods of time from freezer
to room temperature will not affect product performance.
F. Each calibrator lot will have a set value for CREA, GLUC, and BUN
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must be entered into the Carolina Biolis 24i. To update the calibrator
values in the Biolis software perform the following:
1. From the Biolis main screen click the “ITEM” button located
under the “PARAMETER” heading.
2. The item screen lists each analyte by number and name. Double
click the analyte you wish to update:
3. 6 BUN
4. 11 CREA
5. 14 GLUC
6. On the right side of the analyte screen under the “STANDARD”
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heading, enter the value listed on the calibrator specification sheet

into the box.
7. Click “SAVE” and then “RETURN”.
4.3.3 Microgenics Liquid Assayed Chemistry Control (Level 1 and Level 3)

A. The controls are ready to use.
B. Store control at -25 to -15 °C until the expiration date on the box.
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C. Gently invert or rock control when thawing at room temperature.
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D. Once thawed and opened, vials are stable for 14 days when stored
tightly capped at 2-8 °C.
E. Minimize exposure to light and avoid leaving control at room
temperature for long periods of time.
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F. Before the Microgenics controls can be put into service, analyte
concentrations must be verified. A minimum of twenty samples of
each new control lot are assayed over multiple runs to establish a new,
lot -specific acceptable range. The range is calculated as the mean ±
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5% for all analytes and controls except for creatinine QC1 which is
mean ± 10%.
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4.3.4 Pel-Freez Porcine Vitreous Humor Control
A. The control is ready to use.
B. Store vitreous humor control in 10 mL aliquots at -20 °C until
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manufacture’s expiration date.

C. To thaw, refrigerate one aliquot at 2-8 °C for at least 24 hours prior to
analysis.
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D. Once thawed and opened, control is stable for 30 days when tightly
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capped at 2-8 °C.
5.0 Assay Parameters

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5.1 Assay: Creatinine

Sample Volume 4 µL
Reagent 1 Volume 180 µL
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Wavelength 546 nm
Method Enzymatic
Temperature 37º C
5.2 Assay: Electrolytes

Sample Volume 60 µL
Type ISE
Method Direct ISE Undiluted
Temperature 37º C
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5.3 Assay: Glucose

Reagent Volume 300 µL
Sample Volume 3 µL
Wavelength 340 nm
Method UV Hexokinase
Type END
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Temperature 37º C
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5.4 Assay: Blood Urea Nitrogen (BUN)
Sample Volume 3 µL
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Wavelength 340 nm
Method Urease (Rate)
Temperature 37º C
5.5
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A complete list of test parameters and operating procedures can be found in the
Carolina Biolis 24i Operator’s Manual.
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6.0 Procedure
6.1 Start the Biolis 24i.

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6.1.1 If off, turn the computer on. Sign on with the user ID and password. The
start-up procedure will appear on the screen. Click EXIT to clear the
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procedure from the screen so the main software screen can be accessed.
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6.1.2 Turn the system power switch on (front left side of the instrument). A
“Program Downloading” message will pop up followed by “Parameter
Downloading” message. The instrument will be in “Idle” (top right
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corner).
Note: The main power switch on the analyzer (back right) must remain
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turned on at all times to refrigerate the on-board reagents.
6.1.3 Initialize the instrument by clicking on READY.
6.2 Perform day of use maintenance.
6.2.1 Open the front of the instrument and visually check that the sample pump
pistons are moving freely and there are no leaks.
6.2.2 Fill 1% acid and 2% alkaline wash in the reservoirs if necessary.
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6.2.3 Fill the DI water reservoir and empty ISE waste container if necessary.
6.2.4 Clean sample and reagent probes with an alcohol prep pad by rotating the
probes above the sample area. After cleaning, return the probes to the
wash positions.
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6.2.5 Cell Check and Lamp Check.
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A. On the main screen, click on MAINT.
B. Click on CELL CHECK. All 60 cuvettes will be shown. All cuvettes
should be gray or yellow (warning). Red indicates the cuvette needs to
be replaced.
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C. Click on LAMP CHECK. The screen will show you the approximate
life left on the lamp. Below 30% indicates the lamp needs to be
replaced.
6.2.6
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Temperature Check.
A. “TEMP OK” may appear on the screen when the lamp, reaction
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cuvettes and wash are at the proper temperature.
B. Actual temperatures values are found by double clicking just above the
dividing line between ERROR LIST and CANCEL under the MAINT
drop down menu.
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C. When the pop-up window appears for a password, click on OK. Select
TEMP from the drop down menu. The RCT should be approximately
37 °C and the WASH TEMP should be approximately 38 °C.
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D. By opening the front cover of the Biolis 24i, a temperature gauge on

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the left hand side of the instrument will indicate the temperature of the
reagents. This should read 2-8 °C.
6.2.7 Clean the ISE Sample Pot

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A. Open the ISE door on the right side of the instrument.

B. Click on MAINT on the main screen and select ISE.
C. Click on MANUAL SEND then <MANT> and continue after
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“ISE!” is displayed.
D. Moisten a swab with DI water.
E. Insert the swab tip into the sample pot and rotate to clean.
F. When finished, click on <PURG>. After “ISE!” is displayed a second
time, close the ISE door. Click on EXIT twice.
6.2.8 ISE Wash

A. Place a sample cup of ISE Wash solution onto the yellow Calibration
rack labeled “Wash”.
B. On the main screen, click “READY”, click on ISE WASH, and then
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click OK.
6.2.9 Chemistry Reagent Check

A. On the main screen, click on BOTTLE.
B. Confirm enough reagent is on board and the expiration date has not
passed.
C. If a reagent is expired or needs replacing, highlight the reagent row
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that will be deleted.
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D. Click on LINE CLEAR. Remove the reagent bottle from its position in
the refrigerated reagent carousel. Press OK.
E. Press UPDATE, OK, and EXIT.
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Note: It is extremely important that items are cleared and
updated before other reagents can be loaded in the same position.
F. Load new reagent(s). On the main screen, press BOTTLE.
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G. Click BOTTLE READ so the instrument can read the reagent
barcode(s).
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1. If the reagent barcode does not read, manually input the reagent
bottle number by highlighting the line where the reagent was
placed, right clicking and inputting the reagent bottle number in
the pop up window.
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H. Enter the number of tests per assays (4.2.6 – 4.2.8) into R1 and/or R2
as appropriate.
I. Press UPDATE and EXIT.
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J. To fill the DIL2 (deionized water) and Ultra Bottles on reagent rack,
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press BOTTLE. Type “100” in the corresponding reagent’s R1

column. Press UPDATE and EXIT.
6.2.10 Electrolyte Reagent Check

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A. On the main screen, click on REAGENT (bottom of screen).

B. Verify enough ISE Standard 1 is on board (test counts) and expiration
date has not passed.
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C. If the standard is expired or needs replacement, click on MAINT (main

screen).
D. Select ISE (double click).
E. Click on the CAL 1 tab.
F. Update the standard count to 2100.
G. Enter the change date and time on the calendar.
H. Select UPDATE SET and EXIT.
6.3 Calibration
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6.3.1 Calibration is performed on the day of use or whenever any of the

following conditions exist:
A. A reagent or calibrator lot number has changed or there is an observed
shift in control values.
B. A new bottle of reagent is opened and used.
C. Major preventative maintenance was performed on the analyzer.
D. A critical part was replaced.
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6.3.2 To request a calibration and control testing
A. On the main screen, press CALIB.
B. When the calibration procedure box pops up, click EXIT.
C. A screen will appear with the test name, lot number, and position of
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calibrators on the yellow rack (including blank, and number of reps for
each sample, example: S1-1 is position S1, 1 rep).
D. Dispense approximately 500 µL of the Multi Calibrator into a sample
cup and place in position “S1”.
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E. Pipette approximately 500 µL of DI water into a sample cup and place
it in position “B1”.
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F. Dispense approximately 500 µL of the ISE Standard 2 into a sample
cup and place it in the “CAL” position under ISE.
G. Select the “CH ODR” box for each assay except ULTRA. For ISE
CAL select “serum”.
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H. Select UPDATE and OK, then click EXIT.

I. Order the controls by selecting ORDER on the Main screen.
J. Select NEW (at the bottom of the screen).
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K. Enter in the position screen “C1”.

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L. Type in QC1 in the PATIENT ID field. Under the CONTROL field,

make sure the selection MAS LEVEL 1 appears.
M. Click on CHEMISTRY under “Profile”.
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Note: To order individual test(s), click on CREA, GLU, or BUN (will

highlight blue). Na, K, and CL cannot be ordered separately.
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N. Click on ORDER to proceed to the next screen and it will advance to

the next position number “C2”.
O. Type in QC3 in the PATIENT ID and select MAS LEVEL 3 from the
drop down menu under CONTROL.
P. Click the desired testing (single or CHEMISTRY panel).
Q. Click on ORDER and then EXIT.
R. Pipette approximately 500 µL of each control into sample cups and
place them in their programmed positions on the yellow rack (C1-
QC1, C2-QC3 respectively). Press START.
S. To view the calibration data, click CAL DATA on the bottom left of
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the main menu screen.

T. Click on PRINT for BUN, CREA, and GLUC calibration values.
U. Click on ISE CAL DATA and PRINT for the ISE slope values.
V. To print QC data, click on QC at the top of the main menu screen.
W. Select “Current QC” and PRINT.
X. Record QC and ISE slopes in the maintenance binder.
Y. To repeat a QC, click on QC then click CURRENT QC. Mark the
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analyte to be repeated under “Re-run”. Click SAVE then click EXIT.
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Z. Click START.
Note: QCs can only be repeated using “Re-run” (item Y and Z) twice.
If a QC requires a 3rd repeat it will have to be ordered as a specimen
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and run on the gray sample rack.
6.3.3 Acceptance Criteria

A. Ensure that Microgenics control results are within established ranges
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and document in ToxLab and in TOXF.039 Carolina Chemistry QC and
Calibration Log and TOXF.040 Carolina ISE QC and Calibration Log.
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B. If a control is outside its respective ranges, corrective action must be
taken before case samples are run. Document all corrective action in
the Carolina Biolis 24i log book.
C. Corrective action may include but is not limited to:
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1. Make sure all daily maintenance was performed correctly and

documented (fill DI water, acid/base containers, clean ISE cup,
sufficient reagent etc.). If new reagents have been added, make
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sure there are no bubbles and additional reagents are matched lot
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numbers (i.e. two glucose reagents on board).

D. Check to make sure all calibrations have been ordered and passed.
Note: The instrument should be programmed to calibrate and run
controls. If the calibration fails, the controls will still run. If the
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calibration is not ordered, it will print the last calibration and still run
the controls.
E. Check open expiration dates of standards and quality controls.
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F. Check all standard/QC sample cups for proper levels and make sure
there are no bubbles.
G. If A-F are OK, repeat control.
H. If QC fails, recalibrate analyte in question and rerun QC. If QC is OK,
proceed with run. If QC fails, consult Tox I or higher. This may
require opening a new QC, calibrator or reagent one at a time to
resolve the issue. Successful QCs are necessary before starting the run.
I. Consult Tox I or higher before placing a service call.
6.4 Prepare Samples
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6.4.1 Pull samples and place in order of the worklist.

6.4.2 If a sample is not already aliquotted in a 12 x 75 mm culture tube, transfer
at least 0.5 mL (~1 mL is preferred) of sample to a labelled 12 x 75 mm
culture tube. If less than 0.5 mL is available, report as “Quantity
insufficient for analysis”.
6.4.3 Sonicate pig vitreous control and samples for several seconds.
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6.4.4 Allow samples to sit for several minutes to allow any froth to settle.
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6.4.5 If necessary, centrifuge samples at 3500 RMP for 5 minutes to remove
remaining particulate.
6.5 Requisition Samples
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6.5.1 On the Main screen, click on ORDER.
6.5.2 Select NEW at the bottom of the screen.
6.5.3 Enter the position number (Tray 1 contains positions 1-30, Tray 2 contains
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positions 41-60….). Sample trays are gray.
Type in the case number in PATIENT ID along with the sample type
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(example ML14-0001 VIT).
6.5.5 Order the test(s) by clicking on CHEM.
6.5.6 Click on ORDER at the bottom of the screen and it will advance to the
next position number.
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6.5.7 Repeat steps 5.5.3 to 5.5.7 for the remaining samples.

6.5.8 Print the work list by clicking on WORKLIST (bottom screen).
Select “Patient” under the Sample box. Click on SEARCH and the tests
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and sample positions will be listed. Click PRINT.

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6.5.9 Pipette approximately 500 µL of sample into a 1 mL sample cup and place
it into a labeled 12x75mm culture tube. Load the sample into its
respective position on the sample tray.
6.5.10 Click on EXIT to go to the main menu screen and then START.
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6.5.11 After the analysis is complete, print the completed worklist by clicking on
WORKLIST (bottom screen). Select “Patient” and “Control” under the
sample box. Click on SEARCH and the instrument will print the worklist
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with results. Click on PRINT. Refer to TOX07.1006 for post-run

verification.
6.5.12 Samples that require repeated analysis can be tested immediately
following the completion of the sample run. Perform the following steps:
A. Place sample(s) in the next available position or at the beginning of the
second tray (hand write repeats at the end of the work list).
B. Order the repeat test using the sample case number followed by a
unique identifier (ML14-0001 VIT RPT, VIT RPT 1, etc.).
C. Order the appropriate test(s), then click EXIT. Apply the appropriate
treatment to the sample if required (dilution).
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D. Click START.
7.0 Data Analysis/Interpretation/Documentation
7.1 Calculations are not required, except in cases where a specific test required
manual dilution. In these instances, multiply the printed value by the dilution
factor on the result printout.
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7.2 All maintenance items are documented on TOXF.087 Carolina Biolis
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Maintenance Record.
8.0 Reporting
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8.1 Creatinine
8.1.1 The dynamic range is 0.2 – 38.8 mg/dL.
8.1.2 If less than 0.2 mg/dL, report as “Less than 0.2 mg/dL”.
8.1.3 If a result is greater than 38.8 mg/dL, the instrument automatically will
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perform a dilution (1:2). If the sample requires further dilution, it must be
performed manually with isotonic saline as the diluent. The results shall
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be multiplied by the dilution factor.
8.2 Electrolytes
8.2.1 The dynamic ranges are:
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Sodium 90 – 240 mEq/L

Potassium 1.0 - 20.0 mEq/L
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Chloride 60 – 200 mEq/L

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8.2.2 If less than the dynamic range, the result will be reported as “Less than
90/1.0/60 mEq/L”.
8.2.3 If a result is greater than linearity of the assay, the sample must be diluted
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manually with deionized water. The results shall be multiplied by the

dilution factor.
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8.3 Glucose
8.3.1 The dynamic range is 10 – 740 mg/dL.
8.3.2 If less than 10 mg/dL, report as “none detected.”
8.3.3 If a result is greater than 740 mg/dL, the instrument automatically will
performed manually using isotonic saline as the diluent. The results shall
8.4 Blood Urea Nitrogen (BUN)

8.4.1 The dynamic range is 2 – 91 mg/dL.
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8.4.2 If less than 2 mg/dL, report as “Less than 2 mg/dL”.

8.4.3 If a result is greater than 91 mg/dL, the instrument will automatically
performed manually using isotonic saline as the diluent. The results will
8.5 Abnormal results
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8.5.1 Abnormal results are flagged by the Carolina Biolis 24i according to the
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normal values entered by the user into the instrument parameters.
9.0 Instrument Maintenance
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9.1 Weekly
9.1.1 Check Air Filter

A. Remove front cover of the Biolis 24i.
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B. Locate the filter (lower left side of instrument).
C. Remove the foam filter from the black plastic frame.
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D. Rinse the filter with DI water and dry thoroughly. Re-install filter.
9.2 Bi-Weekly
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9.2.1 Ultra Wash (6% alkaline detergent solution)

A. Fill a brown 60 mL reagent bottle with 6% alkaline detergent solution,
and place it into position 1 on the reagent carousel.
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B. Fill 6 sample cups with 6% alkaline solution.

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C. On the main screen click the ORDER button.

D. Enter the correct position number and type the cup ID.
EX: Ultra 1 for cup 1, ULTRA 2 for cup 2, etc.
E. Enter 10 for the number of replicates and select ULTRA for the test.
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F. Click ORDER at the bottom of the screen (the screen will advance to
the next position number).
G. Repeat for cups 2 through 6.
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H. Select EXIT then START.
9.3 Monthly
9.3.1 Clean acid and alkaline wash reservoirs

A. Discard remaining acid and alkaline washes.
B. Rinse well with DI water.
C. Wipe inside the container.
D. Refill with reservoirs with acid and alkaline wash.
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9.4 As Needed
9.4.1 Changing the Water Reservoir Filter

A. Loosen the water reservoir cap and pull both the filter and float switch
unit up.
B. Wipe all DI water off the old filter with a Kimwipe.
C. Pull down on the old filter to remove. Insert the new filter and push up
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until it hits the stopper.
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D. Confirm the float switch moves up and down freely.
E. Return the filter and float switch unit into the reservoir and tighten the
cap.
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9.4.2 Changing the Cleaning Reservoir Solution
A. Loosen the cleaning solution reservoir cap and pull both the filter and
float switch unit up.
B. Wipe all liquid off the old filter with a Kimwipe.
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C. Pull down on the old filter to remove. Insert the new filter and push up
until it hits the stopper.
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D. Confirm the float switch moves up and down freely.
E. Return the filter and float switch unit into the reservoir and tighten the
cap.
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9.4.3 Changing the Light Source

A. For safety, ensure the Biolis 24i and workstation are turned off and
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that the Biolis has cooled down for at least 10 minutes.

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B. Pull the RTM probe up and rotate to the left so it is between the
reagent and reaction trays.
C. Loosen the two lamp lead terminal screws at the terminal block and
remove the lamp leads.
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Note: Only loosen the screws enough to allow the leads to be removed.
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D. Loosen the fixing screw knobs of the lamp holding plate and pull the
plate upwards.
E. Using a hex wrench, remove the lamp from the lamp holding plate. Set
the new lamp with the cut part of the lamp to the pin.
Note: Do not touch the glass surface of the lamp.
F. Position the lamp holding plate so the holes on the plate fit the pins of
the lamp housing. Tighten the two fixing screw knobs and set the
lamp leads to the terminals of the thermal block.
Page 15 of 1
G. Turn on the Biolis 24i and workstation and initialize instrument.

H. Click on MAINT then LAMP in the sub-menu. Click CHANGE and
then OK. The lamp percentage will show 100%.
I. Perform a gain adjustment.
9.4.4 Changing Cuvettes

A. Ensure that the Biolis 24i is turned off. Pull the RTM probe up and
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rotate to the left so it is between the reagent and reaction trays.
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B. Remove the black cover that sits over the cuvette wheel. Cuvette
groups are composed of 5 cuvettes, so rotate the wheel until you find
the cuvette group that contains the cuvette you want to replace.
C. Loosen, but do not remove, the screws holding the plastic cell holder
C
with a 2.5 mm Allen key and lift the cell holder up. There are 3 cell
holders, with each holder housing 20 reaction cuvettes.
D. Remove the old cuvette and replace it with a new one. When
replacing, wear gloves and avoid touching the bottom of the new
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cuvette.
E. Return the plastic cell holder and tighten the fixing screws. After
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replacing the cell holder, turn the cuvette wheel manually to ensure it
moves freely.
F. Replace the black cover, move the RTM probe back into position, and
turn on both the Biolis 24i and workstation.
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G. Initialize the instrument and perform a gain adjustment.
9.4.5 Gain Adjustment

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A. From the main menu select MAINTENANCE then USER

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MAINTENANCE.
B. Click AUTO GAIN. You will be prompted to remove cuvette number
50.
C. Refer to 7.4.5 B-D to remove the number 50 cuvette. Once removed,
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put the cuvette off to the side on a Kimwipe and retrieve an unused
cuvette. Fill the cuvette with 500µL of DI water.
D. Refer to 7.4.5 D-F to return the DI water-filled cuvette to position 50.
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Click OK on the workstation.

E. When the auto gain completes a “Save Data?” prompt will show on the
screen. Click OK.
F. Repeat C and D to remove the DI water-filled cuvette and return the
original number 50 cuvette back to the instrument. Empty the cuvette
used for the auto gain and store with other new cuvettes.
9.4.6 Replacing Expired ULTRA and DIL2 Barcodes

A. On the main screen click BOTTLE. Select the ULTRA (position 1)
that is currently listed.
Page 16 of 1
B. Click LINE CLEAR then click UPDATE. Exit the BOTTLE screen
and then re-open it.
C. Under the POSITION column, place the mouse cursor on number 1
and right click. When prompted “Please input bottle ID”, enter one of
the barcodes listed on the updated barcode sheet.
D. The position 1 line will autofill with new information. Click UPDATE
and then exit the BOTTLE screen.
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Note: When the last of the barcodes has been used, contact Carolina
for a new list of ULTRA and DIL2 barcodes.
9.5 Every 6 months or as needed
C
9.5.1 Replace ISE electrodes.
9.5.2 Replace pump windings.
A. Open the front cover of the Biolis 24i.
d
B. The waste and calibrator pump windings are located on the bottom
right hand side of the instrument.
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C. For the waste pump winding, release motor tension by pushing down
on the silver metal release block.
D. Remove the pump winding from W1 and W2 tubing attachments. The
W1 and W2 connections are labeled on the back wall of the
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instrument.
E. Replace with a new pump winding by attaching one end to the pump
winding mount and connecting to the W2 tubing; and then attaching
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the other end to the pump winding mount and connecting to the W1
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tubing.
F. Repeat steps B through E for the calibrator pump winding. The tubing
connections for these windings are C1 and C2.
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9.6 Annually
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9.6.1 Preventative maintenance by manufacturer

9.6.2 Replace reference electrode.
10.0 References
10.1 Tietz, N.W. (Ed), Fundamentals of Clinical Chemistry, 3rd Edition, WB

Saunders. p. 991 (1976).
10.2 Biolis 24 Operators Manual, Version 1.84, Tokyo Boeki Machinery Ltd.
10.3 Carolina Chemistry QC and Calibration Log – TOXF.039
10.4 Carolina ISE QC and Calibration log – TOXF.040
Page 17 of 1
10.5 ISE Calibration and Slope log – TOXF.085
11.0 Revision History
Revision Description of Change Reviewed By Date

Merged TOX.SC.2023, TOX.SC.2024,
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TOX.SC.2025, and TOX.SC.2026 into one procedure
TOX.SC.2030. Added calibrator update steps for
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SYNCHRON Multi-Cal. Edited items 2.0, 3.0, 4.2,
and 4.6.7. Added matrix-match vitreous humor QC
information to 4.4.5 and 4.5.3. Added items 9.3 and J.
9.4 (QC logs). Added maintenance information (set. Hollowell/A.
C
0 7.0). Salazar 1214
Added ISE form reference (11.5) A Salazar 0415
Added Carolina ISE calibrator to 4.4 and removed
1 Diamond Diagnostic
d
Reordered sections, removed plasma from scope,
A Salazar 0615
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added auto-dilute feature, added appropriate diluents
to each test, added corrective action steps for failed
controls, added acceptable ranges for Microgenics P.Small/T.
2 control Gray 0816
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12.0 Attachment(s) or Appendices

Not Applicable.
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Page 18 of 1

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TOX - sc.2030 Rev 02, Effective 2016-09-20 To Present

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Harris County Institute of Forensic Sciences

Section: Toxicology Approved by: Chief Toxicologist

Creatine + H2O  → Sarcosine + Urea

Any endogenous creatine present in the sample is removed by creatinase

the increase in absorbance at 546 nm is monitored. The final absorbance is

1.4 Serum glucose levels may be abnormally high (hyperglycemia) or abnormally

NH3 + α-ketoglutarate + NADH GLDH

→ Glutamic acid + NAD+

2.1 This method is applicable to the measurement of creatinine, electrolytes, glucose

2.2 Vitreous is the preferred sample for postmortem analysis.

A. There is no significant interference from bilirubin to 16 mg/dL, with

interference. Drugs and other substances may affect glucose

2.4.4 Blood Urea Nitrogen

significant interference with this assay. Lipemia will interfere with

3.0 Definitions and Abbreviations

3.1 S (on print out) -- Sample Error

3.9 GLUC -- Glucose

4.1 Instrument and Equipment

4.2.1 Deionized water

4.2.4 Fast Detergent 1 ALK wash

4.2.5 ISE Wash solution

A. The Carolina Liquid Reagents are ready to use and contain

B. Opened reagent is stable for 30 days when stored in the refrigerated

B. Stable for 6 months at room temperature.

4.3 Standards and controls

Electrolyte Unit Calibrator 1 Calibrator 2

4.3.2 Beckman Coulter SYNCHRON® Systems Multi Calibrator

D. Immediately prior to use, gently invert vial 5 to 10 times.

heading, enter the value listed on the calibrator specification sheet

4.3.3 Microgenics Liquid Assayed Chemistry Control (Level 1 and Level 3)

manufacture’s expiration date.

capped at 2-8 °C.

5.0 Assay Parameters

5.1 Assay: Creatinine

5.2 Assay: Electrolytes

5.3 Assay: Glucose

6.1 Start the Biolis 24i.

turned on at all times to refrigerate the on-board reagents.

6.1.3 Initialize the instrument by clicking on READY.

6.2 Perform day of use maintenance.

6.2.2 Fill 1% acid and 2% alkaline wash in the reservoirs if necessary.

D. By opening the front cover of the Biolis 24i, a temperature gauge on

6.2.7 Clean the ISE Sample Pot

A. Open the ISE door on the right side of the instrument.

6.2.8 ISE Wash

6.2.9 Chemistry Reagent Check

F. Load new reagent(s). On the main screen, press BOTTLE.

press BOTTLE. Type “100” in the corresponding reagent’s R1

6.2.10 Electrolyte Reagent Check

A. On the main screen, click on REAGENT (bottom of screen).

C. If the standard is expired or needs replacement, click on MAINT (main

6.3.1 Calibration is performed on the day of use or whenever any of the

H. Select UPDATE and OK, then click EXIT.

K. Enter in the position screen “C1”.

L. Type in QC1 in the PATIENT ID field. Under the CONTROL field,

Note: To order individual test(s), click on CREA, GLU, or BUN (will

N. Click on ORDER to proceed to the next screen and it will advance to

the main menu screen.

6.3.3 Acceptance Criteria

1. Make sure all daily maintenance was performed correctly and

numbers (i.e. two glucose reagents on board).

6.4 Prepare Samples

6.4.1 Pull samples and place in order of the worklist.