Laboratory Procedure Manual

Analyte: Fasting Glucose

Matrix: Plasma

Method: Roche Cobas C311 2017—Standard

As performed by: University of Missouri-Columbia

Contact: Dr. Randie Little

Important Information for Users

The University of Missouri periodically refines these laboratory methods. It is the
responsibility of the user to contact the person listed on the title page of each write-up
before using the analytical method to find out whether any changes have been made
and what revisions, if any, have been incorporated.
Fasting Glucose
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Public Release Data Set Information

This document details the Lab Protocol for testing the items listed in the following table:

Variable
File Name SAS Label (and SI units)
Name

LBXGLU Fasting glucose(mg/dL)

LBDGLUSI Fasting glucose (mmol/L)

GLU_I
LBXGLT Two-hour glucose (OGTT) (mg/dL)
OGTT_I
LBDGLTSI Two-hour glucose (OGTT) (mmol/L)

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I. Purpose Statement
a. This SOP is intended for the in vitro test for the quantitative determination of
glucose in human serum and plasma. It is performed on the Roche Cobas C311
system in the Diabetes Diagnostic Laboratory.

G-6-PDH
II. Definitions
a. Not G-6-P + NADP+ gluconate-6-P + NADPH + H+ Applicable

III. Content
a. Principle and Clinical Significance

Glucose is the major carbohydrate present in the peripheral blood. Glucose

derived from dietary sources is either oxidized to provide energy or converted to
glycogen or fatty acids for storage in the liver and tissues. The most frequent
cause of hyperglycemia is diabetes mellitus. Some other factors that contribute
to elevated blood glucose are pancreatitis, pituitary or thyroid dysfunction, renal
failure, and liver disease. Hypoglycemia is less frequently observed, but is found
in conditions such as insulinoma, hypopituitarism, neoplasms, or insulin-induced
hypoglycemia1,2,3,4.

The cobas c311 performs a UV test to detect glucose in blood serum and plasma.

The enzyme hexokinase (HK) catalyzes the reaction between glucose and
adenosine triphosphate (ATP) to form glucose-6-phosphate (G-6-P) and
adenosine diphosphate (ADP). In the presence of nicotinamide adenine
dinucleotide (NAD), G-6-P is oxidized by the enzyme glucose-6-phosphate
dehydrogenase (G-6-PD) to 6-phosphogluconate and reduced nicotinamide
adenine dinucleotide (NADH). The increase in NADH concentration is directly
proportional to the glucose concentration and can be measured
spectrophotometrically at 340 nm.

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The enzymatic reference method is performed with hexokinase.4 Hexokinase

catalyzes the phosphorylation of glucose to glucose-6-phosphate by ATP.

HK
Glucose + ATP G-6-P + ADP
b. Specimen
Collection and
Handling
i. Patient Preparation
1. Patients should be fasting or undergoing an Oral Glucose
Tolerance Test. Patient’s status should be recorded when
specimen is drawn.
ii. Specimen Type
1. Collect blood by venipuncture from individuals using an evacuated
tube system
a. Plasma: Fluoride plasma for NHANES study testing (grey
top tubes)
2. The minimum volume required for analysis directly from
collection tube is 200 µL.
3. Specimens are delivered to the Diabetes Diagnostic Laboratory,
Room M764 by FedEx. Specimens are received frozen on dry ice
and are stored at -70oC. Each specimen must arrive in the
laboratory labeled with two unique accession number generated
by NHANES, and checked against the specimen manifest.

iii. Specimen Stability

1. Stability in fluoride plasma:5 3 days at 15-25 °C
At least 3 months @ -20°C
At least 2 years @-70°C

2. Unacceptable specimen criteria

a. Clotted specimens.

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b. Unlabeled samples
c. Specimens not collected in sodium fluoride.
d. Whole blood specimens
e. Refer to limitations section for additional details
f. The below must be followed for unaccepted specimen
collections:
i. Record all unacceptable samples on sample
manifest
ii. Contact NHANES regarding unacceptable specimen

iv. Handling Conditions

1. For specimen collection and preparation, only use suitable tubes
or collection containers. Only the specimens listed below were
tested and found acceptable: Plasma: fluoride plasma for NHANES
study testing. (Grey-top tubes)
2. To minimize glycolysis, one should place the sample tube
immediately in an ice-water slurry, and the plasma should be
separated from the cells within 30 min. Blood for FPG analysis
should be drawn in the morning after the individual has fasted
overnight (at least 8 hours).
3. If testing is not performed immediately, samples are to be
maintained under frozen conditions (-70°C).
4. For NHANES samples: Laboratory services are requested through
the Westat system operations via an email notification containing
a unique manifest list of the samples and sample analysis type
(e.g. Glucose), which confirms that specimens have been shipped
to DDL. Transport under frozen conditions.
5. Each Manifest Form should include and verified against each
sample received:
a. Patient Sample ID #
b. Test Name
c. Date Collected
d. Shipment ID #
e. Shipment Date
f. Lab Name
g. Lab ID
h. Survey Year
6. Once received and prepared for analysis, specimens are to be
immediately returned to -70°C storage. When ready to be
analyzed, specimens are to be thawed at room temperature.

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Following testing, specimens are to be returned to -70°C for long-

term storage.

c. Equipment and Materials

i. Equipment
1. Roche Cobas C311 analyzer consists of the following main
components:
a. Analyzer Unit
i. Sample disk: 110 position disk for loading of up to
108 samples
ii. Sample detectors and barcode readers: on inner
and outer rings
iii. Sample Pipetting System: clot detecting by
pressure measurements, liquid level detection by
capacitance, and abnormal descent detection.
1. Metal shield pipe
2. Rinse station
3. Drying cylinder
4. Basic (sample cleaner 1) and acid wash
(sample cleaner 2) solutions for special
probe washes and maintenance functions.
iv. Reagent Compartment
v. Reagent Pipetting System
vi. Reaction Disk Area
vii. ISE System (not in use)
viii. Water supply tank
ix. Vacuum System
x. Sound Volume Knob
xi. Cell Wash Solutions
xii. Waste Container
xiii. Inlet Water Line
xiv. Dilute Waste Line
b. Control Unit
i. Computer
1. Hard Disk Drive
2. 3-1/2 in floppy disk drive and a DVD-RW
rewrite device
ii. Keyboard
iii. Mouse
iv. Color Monitor: touch screen
v. Printer

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ii. Materials
1. Reagents and other items
a. R1: MES buffer: 5.0 mmol/L, pH 6.0; Mg2+: 24 mmol/L;
ATP: ≥ 4.5 mmol/L; NADP: ≥ 7.0 mmol/L; preservative
b. R2: HEPES buffer: 200 mmol/L, pH 8.0; Mg2+: 4 mmol/L; HK
(yeast): ≥ 300 µkat/L; G-6-PDH (E. coli): ≥ 300 µkat/L;
preservative
c. Sample vials and false bottom tubes
d. Printer paper.
e. Toner cartridge.
f. Refer to MSDS located in the Operator’s Manual for
reagent description and composition
2. Other materials
a. Powder free hypoallergenic latex examination gloves.
b. Biohazardous waste storage bags and boxes (Jefferson
Smufit Corporation, Highland, IL).
c. Viro Research Envirocide Disinfectant Decontaminant
Cleaner (Fisher Scientific, St. Louis, MO)
d. Transfer pipettes (Fisher Scientific, St. Louis, MO)
e. Single fold paper towels (Ft. Howard Corp. Co, Green Bay,
WI).
f. Kim Wipe lintless tissues (Kimberly Clark Corp., Roswell,
GA).
g. Low and high in-house plasma controls (low and high,
respectively).
h. Bio-Rad controls
i. Volumetric pipettes for measuring different volumes.

iii. Equipment Maintenance

1. Roche Cobas c311– Routine maintenance
a. Perform daily maintenance as outlined in Cobas c311
manual every time machine is used
b. Perform weekly maintenance as outlined in Cobas c311
manual once a week (generally whenever machine is used)
c. Perform monthly, quarterly and bi-annually maintenance
when assigned
2. Record all daily, weekly, monthly, and periodical maintenance in
the “C311 Maintenance” binder. This paperwork is approved by
the supervisor monthly and filed in “C311 Maintenance” binder.
3. Roche – Periodic/Preventative maintenance – Preventative
maintenance inspections will be done at least yearly.
4. Pipettes are to be calibrated once a year.

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5. Temperatures for the refrigerators and freezers where specimens,

controls and calibrators are stored are to be recorded. Any
readings that fall outside acceptable ranges are to be reported to
the supervisor for corrective action. These areas are monitored
by 7 day 24 hour temperature recorders for minimum and
maximum temperatures and are checked daily (working day) by a
technician to make sure the temperatures are within acceptable
ranges. The charts are to be checked and initialed by the
supervisor at least weekly.

iv. Storage Requirements – Reagent Use and Storage:

1. GLUC3
a. Shelf life at 2-8 °C: see expiration date on cobas c pack
label
b. On-board, in use and refrigerated on the analyzer: 8 weeks
2. Diluent NaCl 9 %
a. Shelf life at 2-8 °C: see expiration date on cobas c pack
label
b. On-board, in use and refrigerated on the analyzer: 12
weeks
3. All regents are ready for use; to load packs onto machine, follow
instructions in Cobas c311 analyzer Basic Operator Training Guide.

v. Reagent Labeling: Reagents, calibrators, controls, and solutions should be

traceably identified to indicate the following:
1. Content and quantity, concentration or titer
2. Storage requirements.
3. Date
4. The below should be followed for reagents used daily
a. Preparation date or opened and the identity of the
preparer;
b. Tech’s initials
5. When new reagents are received they must be initialed and dated
by the technician who receives them. When the reagents are
opened they are initialed and dated with open date by the
technician.
a. Regent packs loaded onto the machine are logged by the
machine with the load date and will flag when expired.

d. Calibration:
i. Preparation and Stability

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1. Material: C.f.a.s (Calibrator for automated systems) manufactured

by Roche Cobas. Human serum with chemical additives and
material of biological origin as specified on package insert.
2. Calibrator values are given in the electronically available value
sheets, organized by lot. Determinations were performed under
strictly standardized conditions on multiple Roche analyzers using
Roche system reagents and C.f.a.s. master calibrator or reference
materials.
a. Barcodes are given with each package of calibrator. They
may be used to scan the calibrator lot into the c311
3. Carefully open a bottle avoiding the loss of lyophilizate. Using a 3
mL volumetric pipette, add in exactly 3.0 mL of distilled/deionized
water. Carefully close the bottle and dissolve the contents
completely by occasional swirling within 30 minutes. Avoid
formation of foam.
4. Storage and Stability
a. Store unopened C.f.a.s at 2-8oC
i. Stability of lyophilized calibrator at 2-8oC: see
expiration date
b. Stability of glucose in the reconstituted calibrator
i. 15-25 oC: 8 hours
ii. 2-8 oC: 2 days
iii. (-15)—(-25 C) :
o 4 weeks (when frozen once)

ii. Frequency of Calibration:

1. Calibration verification is performed when a system is first placed
in service and at least every six months (or as specified by the
manufacturer) thereafter for all quantitative tests. Recalibration is
required (regardless of the length of time since last performed)
immediately if any of the following occurs;
a. A change of reagent lots for chemically or physically active
or critical components, unless the laboratory can
demonstrate that the use of different lots does not affect
the accuracy of patient/client test results or the range used
to report patient/client test data
b. If QC materials reflect an unusual trend or shift or are
outside of the laboratory's acceptable limits, and other
means of assessing and correcting unacceptable control
values fail to identify and correct the problem
c. After major maintenance or service. The Laboratory
Director must determine what constitutes major
maintenance or service.

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d. When recommended by the manufacturer.

iii. Calibration Procedure: To calibrate the c311 for glucose

1. Calibrators: S1: H20
S2: C.f.a.s
2. Calibration Mode: Linear 2-point calibration
3. Choose the “Calibration” tab
4. Select the glucose test
5. Select “2 point” under the method window
6. Insert calibrators and controls into the correct vial positions.
These positions can be found in the Calibration load list report
and QC load list report
7. Select “Save”
8. Select the “Start” button
9. Traceability: This method has been standardized against ID/MS.
10. Calibration Acceptability Criteria
a. When QC results fail to meet the acceptable criteria, check
the sample cup containing the QC specimen for bubbles
and reanalyze the QC specimen.
b. If the QC results meet the acceptable criteria, accept the
run and report the results.
c. If steps above do not result in correction of the "out-of-
control" values for QC materials, troubleshoot the
instruments and reagents until the system is back “in
control”
d. Recalibrate the system using a new vial of glucose
standard.
e. Reanalyze the calibrator, controls, and specimens.
Specimens are stable at 4-8 °C overnight. If the system
requires more than 24 hours before it can be restored to
functionality, use new aliquots of standard, controls, and
specimens for analysis.
f. Refer to Operator’s Manual for additional troubleshooting
advice.
g. If the calibration is still unsuccessful speak with your
supervisor for further help. Do not perform glucose
analysis until the system is declared "in-control" again.

e. Quality Control:
i. Two types of quality control (QC) systems are used in this analytical
method: 1) "sample QC" and 2) "batch QC." For sample QC, 2% of

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specimens are randomly selected and analyzed either within-assay or

between-assay for quality assurance purposes. If the difference between
duplicates is greater than 10%, the specimen is reanalyzed. Batch QC
specimens are placed in the rack at the end of the entire run. The
following criteria are for “batch QC.”
ii. Control Materials and Stability:
1. Lyphocheck Assayed Chemistry Control Levels 1 and 2
a. Prepared from human serum with added chemicals,
purified biochemical material (tissue extracts of human
and animal origin), therapeutic drugs, stabilizers and
preservatives. This product is provided in lyophilized form
for increased stability
b. Stability
i. Store unopened controls at 2-8oC
ii. Stability of lyophilized calibrator at 2-8oC: see
expiration date
iii. Stability of Glucose in reconstituted controls
1. 2-8 oC: 7 days
2. Frozen : 30 days (when frozen once)

2. In-house controls IHL and IHH

a. Controls were prepared by collecting 450 mL (one unit) of
whole blood from two diabetic (IHH) and two non-diabetic
(IHL) subjects. The blood was collected in blood bags
without an anticoagulant. The serum was allowed to clot
for 1 hour and then separated from the red blood cells by
centrifugation in a refrigerated centrifuge (4oC) for 25 min
at 1500 g. The serum was then removed from the red
blood cells, aliquoted in 0.5-mL portions and stored at -70
°C or colder in Nalgene cryogenic vials.
b. If the stock of these controls becomes low, another batch
is prepared in time to analyze it concurrently with the
current QC materials. The new controls are used only
after their means and the ranges have been established by
performing 20 characterization runs.
c. Stability
i. In liquid nitrogen: 5 years
ii. At -70 oC: 1 year
iii. 2-8 oC: 72 hours
iv. 20-25 oC: 24 hours

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v. Current stability studies can be found in the Quality

Control Binder located in room M771.

iii. Preparation for analysis

1. For each assay a single use aliquot is prepared in the same
manner as an unknown specimen as described in Procedure.
2. Bio-Rad Controls: Using a volumetric pipet or equivalent,
reconstitute each vial with 5 mL of distilled or deionized water.
Replace the stopper and allow this product to stand for
approximately 20 minutes swirling occasionally. Before sampling,
gently swirl the vial several times to ensure homogeneity. After
each use, promptly replace the stopper and return to the
appropriate storage condition.
3. In-House Controls: One vial of each control is thawed and used in
each assay. Reconstitution is not required for these controls.

iv. Frequency for control materials: All four batch QC controls are analyzed
at the beginning and end of each assay.

v. Mean and Ranges:

1. Daily means and ranges of the controls are calculated from at
least 20 inter-assay determinations. The bias ranges of the daily
means are set at ±1 SD or the 67% confidence interval (CI); the
warning limits (WL) are the ±2 SD or the 95% CI and the control
limits (CL) are the ±3 SD or the 99% CI. For the daily ranges, the
bias limit is the mean + 1 SD with warning and control limits set at
the mean +2 SD and the mean + 3 SD, respectively.
a. The means and ranges for the current controls are posted
near the c311 instrument
b. Current control value assignments and limits can be found
in the Quality Control Binder located in room M771.

2. After each assay run, all control data are recorded on the Daily
Diary Log Sheet. The results of the analysis are accepted or
rejected according to the guidelines established.

3. Two types of QC charts are used in assessing the quality of an

assay.
a. The first chart plots the mean of all the replicate
determinations in a run and compares it with the
established target mean, which is the overall mean
established by the 20 pr more characteristic runs. The

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guideline declares a system as "out-of-control" if any of

the following events occur:
i. The mean for one control from a single run falls
outside the 99% confidence limits.
ii. The means for two controls from a single run fall
outside the 95% confidence limits.
iii. The daily means for one control from eight
successive runs (excluding the runs in which the
mean is within ±1 SD or the bias range) fall either
all above or all below the center line.
b. The second type of QC chart plots the range of the
replicates (the difference between the highest and the
lowest value of a single control within a run) and compares
it with the established target range, which is the overall
mean of daily ranges established by the 20 or more
characteristic runs. The QC guidelines for DDL declare a
system as "out-of-control" if any of the following events
occur:
i. The daily range for one control exceeds the 99%
confidence limit.
ii. The daily ranges for two controls exceed the 95%
confidence limits.
iii. The daily ranges for one control from eight
successive runs (excluding the runs in which the
mean is within1 SD or bias range) are all above the
mean line (trend rule).
c. If a run is declared out of control, investigate the system
(instrument, standards, controls etc.) to determine the
cause of the problem. Do not perform any analysis until
the problem has been resolved.

vi. Remedial Action if QC Systems Fail to Meet Acceptable Criteria

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1. When QC results fail to meet the acceptable criteria, check the

sample cup containing the QC specimen for bubbles and reanalyze
the QC specimen.
2. If the QC results meet the acceptable criteria, accept the run and
report the results.
3. If steps above do not result in correction of the "out-of-control"
values for QC materials, troubleshoot the instruments and
reagents until the system is back “in control”
4. Recalibrate the system using a new vial of glucose standard.
5. Reanalyze the calibrator, controls, and specimens. Specimens are
stable at 4-8 °C overnight. If the system requires more than 24
hours before it can be restored to functionality, use new aliquots
of standard, controls, and specimens for analysis.
6. If the above steps do not correct the "out of control" condition,
consult with the supervisor for further corrective action. Do not
perform glucose analysis until the system is declared "in-control"
again.

f. Procedure – Stepwise:
i. Special safety precautions:
1. Gloves and lab coat are required for handling all human blood
specimens.
2. All plastic tips, sample vials, gloves, etc. that contact blood are
considered contaminated and are to be placed in an approved
waste container.
3. Work surfaces are protected by absorbent pads. The pads are
discarded into biohazardous waste container weekly or whenever
blood contamination occurs. All work surfaces are wiped down
with Envirocide weekly.

ii. Initial processing of specimens - Clinical trial specimens are checked

against the clinical trial data sheet to insure that the patient information
matches. The specimens are placed at -70oC until ready to be tested.
Barcodes are printed by the DDL that correspond with the NHANEs
accession numbers on the samples. False bottom tubes are labeled with
these barcodes.

iii. To run the Roche Cobas c311:

1. Before turning on the analyzer, perform the maintenance checks
as outlined on the cobas c311 analyzer Maintenance Log under
the “Check” and “Hands-on” sections.

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2. All healthcare personnel shall routinely use appropriate barrier

precautions to prevent skin and mucous membrane exposure
when contact with blood or other body fluids of any patient is
anticipated. All products or objects that come in contact with
human or animal body fluids should be handled, before and after
cleaning, as if capable of transmitting infectious diseases. Wear
appropriate Personal Protective Equipment (PPE), including facial
protection, gloves, and protective clothing. Dispose of all
biological samples and diluted specimens in a biohazard waste
container at the end of analysis. Dispose of all liquid hazardous
waste in properly labeled hazardous waste container.
3. Turn on the power to the analyzer and control unit. The Analytical
Unit must be turned on before the control unit.
4. Log-in to the control unit using name: adm and password: adm
5. The system will perform the “Power On Pipe” automatically.
6. Ensure all “Push-button” maintenance functions were performed
by checking the utility screen.
7. Perform any maintenance functions that have expired
8. Put machine into maintenance mode by flipping switch on the
analyzer. Perform any necessary maintenance outlined in the
Maintenance Log that has not already been done.
9. Return machine to operation mode
10. Go to “System Overview” screen. Ensure that temperature of the
incubator is within 37oC ± 0.1oC by clicking “AU” button and
observing the core temperature.
11. Check the “Work Flow Guide” area at the top of the screen. If any
of the first five buttons is highlighted, perform that action (Daily
Maintenance, Sample Data Clear, Regent Preparing, Calibration
and QC Select, Parameter Download). These actions should be
automatically completed by the analyzer.
12. Calibrate if necessary following the calibration procedures.
13. Enter controls to run as samples
a. Go to Utility menu
b. Click on sample information
c. Type in control name
d. Hit enter
e. Select “Gluc” test button
f. Select “Save” test button
g. Enter “Barcode Read Error” mode
h. Enter name of control and position on machine
i. Repeat for all 8 controls (beginning and end)

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14. Enter samples and tests to run

a. Go to Utility menu
b. Click on sample information
c. Scan barcode for sample
d. Hit enter
e. Select “Gluc” test button
f. Select “Save” test button
g. Repeat for all samples
15. To run samples, choose “Start” button at lower right of the screen
16. Hit “Start” when prompted
17. To manually shut down the instrument, choose the “Deactivate
system” option.
18. Preliminaries:
a. All reagents should be at room temperature before assay.
b. Allow frozen reference standards, QC specimens, and any
frozen blood samples to thaw. Mix all samples at least ten
minutes before putting on the machine
19. Sample preparation: Using printed labels, label the appropriate
sample vials with the corresponding sample identification, and
place the whole, uncapped original tube into the sample vials.
20. After run is completed, print hard copy of the results and proceed
to reporting results.

iv. Calculations:
1. Roche cobas c systems automatically calculate the analyte
concentration of each sample.
a. Conversion factors: mmol/L x 18.02 = mg/dL
mmol/L x 0.1802 = g/L
mg/dL x 0.0555 = mmol/L

v. Reporting results:
1. All replicate values of QC data plus all pertinent assay information
(date of analysis, reagent lot number, technician ID, samples ID
etc.) are recorded in the Microsoft Access Glucose Daily Diary Log
database located on the network drive. The calibrator value is also
recorded. Enter the data under the form “Diary Sheet Entry Form”.
The Microsoft Access program will automatically calculate the daily
mean and range for each control and determine if a run is accepted
or rejected. The current above or below the mean trend is also
calculated. The program will print out a diary sheet for each run
and the information is checked and signed by a supervisor.

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2. The clinical trial sample values are automatically uploaded into the
computer database in mg/dL units. Data check sheets are printed.
3. Any comments associated with the specimen are entered in the
comment field. If a result is below the assay detection limit, or a
sample is missing, or if the sample volume is less than 200 µL, or
the sample is otherwise unacceptable, the result field is left blank
or a –1 is entered and an appropriate comment is entered in the
assay comment field.
4. Recorded results are then to be checked against the original
printout by the supervisor or, in the event the supervisor is not
present, the designated senior technician. Record the date
checked and initials on the c311 Diary Sheet. A copy of the data
check sheet is kept in the appropriate data book at the Diabetes
Diagnostic Laboratory at the University of Missouri.

g. Reference Ranges:
i. ADA reference ranges for fasting individuals3
1. Normal: < 100 mg/dL
2. Pre-diabetic (impaired fasting glucose): 100-125 mg/dL
3. Diabetic: ≥ 126 mg/dL

ii. ADA reference ranges for 2-hour postprandial individuals3

1. Normal: < 140 mg/dL
2. Pre-diabetic (impaired glucose tolerance): 140-199 mg/dL
3. Diabetic: ≥ 200 mg/dL

h. Procedures for Abnormal Results:

i. Critical Value Adults and Children: < 40mg/dL or > 400mg/dL
ii. Critical Value Infants < 1 month : < 40mg/dL or > 300 mg/dL
iii. Critical results must be repeated and verified.
1. Glucose results do not have STAT turnaround. We typically
receive these samples at least 24 hours after collection.
Therefore there is no “panic situation”.

i. Reporting Format:
i. Results are expressed on the report as mg/dL
ii. Measuring range for Plasma: 2-600 mg/dL without dilution
1. Reanalyze samples containing more than 600 mg/dL by diluting
the specimen two-fold (1+1) with distilled water. The result
output must then be multiplied by 2 to account for the dilution.
2. Lower detection limit: 2 mg/dL

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a. The lower detection limit represents the lowest

measurable analyte level that can be distinguished from
zero. It is calculated as the value lying three standard
deviations above that of the lowest standard (standard 1 +
3 SD).

iii. Linearity data is found in the c311 Maintenance Binder under the
Linearity tab. Linearity studies are performed at least twice per year.

j. Supervisor Responsibility:
i. The supervisor ensures quality control passes within the acceptable
ranges prior to releasing patient results.

k. Limitations of the Procedure/ Interfering Substances:

i. Criterion: Recovery within ± 10 % of initial value at a glucose
concentration of 70.3 mg/dL.
ii. Serum/plasma
1. Icterus: No significant interference up to an I index of 60
(approximate conjugated and unconjugated bilirubin
concentration: 60 mg/dL.)
2. Hemolysis: No significant interference up to an H index of 1000
(approximate hemoglobin concentration: 1000 mg/dL.
3. Lipemia (Intralipid): No significant interference up to an L index of
1000. There is poor correlation between the L index (corresponds
to turbidity) and triglycerides concentration.
4. Drugs: No interference was found at therapeutic concentrations
using common drug panels.7,8
5. In very rare cases gammopathy, in particular type IgM
(Waldenström's macroglobulinemia), may cause unreliable
results.

iii. Actions Required

1. Special Wash Programming: The use of special wash steps is
mandatory when certain test combinations are run together on
Roche/Hitachi cobas c systems. Refer to the latest version of the
Carry over evasion list found with the
NaOHD/SMS/Multiclean/SCCS Method Sheet and the operator
manual for further instructions.
2. Where required a special wash or carry over evasion
programming must be implemented prior to reporting results
with this test.

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IV. SUMMARY STATISTICS AND QC STATISTICS

See next page

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2015-2016 Summary Statistics and QC Chart for Fasting Glucose (mg/dL)

Start End Standard Coefficient of

Lot N Date Date Mean Deviation Variation
16742 18 15JAN15 29APR15 296.8 3.8 1.3
16741 18 15JAN15 29APR15 84.8 1.4 1.6
IHH8 82 07MAY15 02FEB17 324.8 5.3 1.6
172732 24 07MAY15 29OCT15 235.0 2.9 1.2
IHL8 82 07MAY15 02FEB17 93.1 1.5 1.6
173070 8 07MAY15 18JUN15 90.4 0.7 0.8
176407 16 24JUN15 29OCT15 91.6 1.6 1.7
BR14492 58 04NOV15 02FEB17 298.5 5.3 1.8
BR14491 58 04NOV15 02FEB17 88.2 1.3 1.5

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Fasting Glucose
NHANES 2015-2016

2015-2016 Summary Statistics and QC Chart for Two Hour Glucose (OGTT) (mg/dL)

Start End Standard Coefficient of

Lot N Date Date Mean Deviation Variation
16742 16 15JAN15 29APR15 296.9688 3.9894 1.3
16741 16 15JAN15 29APR15 84.8438 1.4109 1.7
IHH8 83 06MAY15 02FEB17 324.7831 5.2737 1.6
172732 25 06MAY15 29OCT15 234.8067 2.9359 1.3
IHL8 83 06MAY15 02FEB17 93.1084 1.4976 1.6
173070 9 06MAY15 18JUN15 90.5000 0.6972 0.8
176407 16 24JUN15 29OCT15 91.5625 1.5586 1.7
BR14492 58 04NOV15 02FEB17 298.5388 5.2833 1.8
BR14491 58 04NOV15 02FEB17 88.2098 1.3137 1.5

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Fasting Glucose
NHANES 2015-2016

V. References, Regulatory References, Related Documents, or Links

1. Sacks DB. Carbohydrates. In: Tietz NW, ed. Fundamentals of Clinical Chemistry.
4th ed. Philadelphia: WB Saunders 1996:351-374.
2. Knudson PE, Weinstock RS. Carbohydrates. In: Henry JB, ed. Clinical Diagnosis
and Management by Laboratory Methods. 20th ed. Philadelphia: WB Saunders
2001:211-223.
3. Sacks DB. Carbohydrates. In: Burtis CA, Ashwood ER, eds. Tietz Textbook of
Clinical Chemistry. 3rd ed. Philadelphia: WB Saunders 1999:750-785.
4. Kunst A, Draeger B, Ziegenhorn J. In: Bergmeyer. Methods of Enzymatic Analysis,
3rd ed. Volume VI, Metabolites 1: Carbohydrates. 1984:163-172.

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