0011877771216c501V7.

TRIGB
Triglycerides/Glycerol Blanked
Order information
Analyzer(s) on which cobas c pack(s) can be used
11877771 216 Triglycerides/Glycerol Blanked (12 x 120 tests) System‑ID 07 6868 5 Roche/Hitachi cobas c 311, cobas c 501/502
10166588 130 Precimat Glycerol Code 509
10759350 190 Calibrator f.a.s. (12 x 3 mL) Code 401
10759350 360 Calibrator f.a.s. (12 x 3 mL, for USA) Code 401
12149435 122 Precinorm U plus (10 x 3 mL) Code 300
12149435 160 Precinorm U plus (10 x 3 mL, for USA) Code 300
12149443 122 Precipath U plus (10 x 3 mL) Code 301
12149443 160 Precipath U plus (10 x 3 mL, for USA) Code 301
10781827 122 Precinorm L (4 x 3 mL) Code 304
11285874 122 Precipath L (4 x 3 mL) Code 305
05117003 190 PreciControl ClinChem Multi 1 (20 x 5 mL) Code 391
05947626 190 PreciControl ClinChem Multi 1 (4 x 5 mL) Code 391
05947626 160 PreciControl ClinChem Multi 1 (4 x 5 mL, for USA) Code 391
05117216 190 PreciControl ClinChem Multi 2 (20 x 5 mL) Code 392
05947774 190 PreciControl ClinChem Multi 2 (4 x 5 mL) Code 392
05947774 160 PreciControl ClinChem Multi 2 (4 x 5 mL, for USA) Code 392
04489357 190 Diluent NaCl 9 % (50 mL) System‑ID 07 6869 3
04593138 190 cobas c pack MULTI
On request Open/Close tool

English GK

System information Free glycerol + ATP glycerol‑3‑phosphate + ADP

For cobas c 311/501 analyzers:
TRIGB: ACN 783
For cobas c 502 analyzer: GPO

TRIGB: ACN 8783 Glycerol‑3‑phosphate + O2 dihydroxyacetone phosphate +

Intended use H2O2
In vitro test for the quantitative determination of triglycerides with glycerol
blanking in human serum and plasma on Roche/Hitachi cobas c systems. peroxidase

Summary H2O2 + 4‑chlorophenol oxidation product

Triglycerides measurements are used in the diagnosis and treatment of
patients with diabetes mellitus, nephrosis, liver obstruction, other diseases
involving lipid metabolism, and various endocrine disorders. Before the (The oxidation product formed does not react with 4‑aminophenazone.)
1950’s, triglyceride levels were only estimated from total lipid levels.1 Assay reaction
Triglycerides = Total Lipids - (Cholesterol + Phospholipids) • Addition of R2
In 1957, Van Handel and Zilversmit2 developed a direct manual method
using adsorbent to remove phospholipids from the lipid extract followed by lipase
measuring the concentration of glycerol released by saponification with Triglycerides + 3 H2O glycerol + fatty acids
KOH. In 1966, Kessler and Lederer3 improved the existing semiautomated
method by automating the saponification step.
Further automation of the method was made even simpler and easier with
GK
the introduction of enzymatic methods. Enzymatic methods are based on
the determination of the glycerol part of the triglyceride molecule after Glycerol + ATP glycerol‑3‑phosphate + ADP
hydrolysis of triglycerides into glycerol and fatty acids. Lipase combined
with a protease are the most common enzymes used. These completely
enzymatic systems eliminate the need for caustic reagents, extraction
solvents, high temperature baths and adsorption mixtures for phospholipid GPO
removal thus rendering the method readily available for automation.1 Glycerol‑3‑phosphate + O2 dihydroxyacetone phosphate +
Test principle4 H2O2
Enzymatic colorimetric assay.
Free glycerol is eliminated prior to hydrolysis of triglycerides in a preliminary peroxidase
reaction in which lipase and 4‑aminophenazone are omitted. This reaction
is followed by enzymatic hydrolysis of triglycerides and determination of the H2O2 + 4‑aminophenazone + 4‑(p‑benzoquinone‑monoimino)
liberated glycerol by a fully enzymatic colorimetric assay reaction. 4‑chlorophenol ‑phenazone +
Preliminary reaction 2 H2O + HCI
• Sample and addition of R1

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TRIGB
Triglycerides/Glycerol Blanked

Reagents - working solutions 6. Unscrew the screw cap of the bottle in position C on the right side of the
cobas c pack MULTI using the Open/Close tool.
R1 TRIS buffer: 0.15 mol/L, pH 7.6; magnesium sulfate: 17.5 mmol/L;
7. Pour exactly 15 mL of the content of bottle 2 (R2) into the opened bottle
EDTA, disodium salt: 10 mmol/L; 4‑chlorophenol: 3.5 mmol/L; of the cobas c pack (position C).
potassium hexacyanoferrate (II): 6 µmol/L; sodium cholate: 0.15 %;
hydroxypolyethoxy‑n‑alkanes: 0.12 %; ATP: ≥ 1 mmol/L; glycerol 8. Close the bottle tightly using the Open/Close tool.
kinase (Geobacillus stearothermophilus): ≥ 0.4 U/mL; glycerol 9. Leave position A empty.
phosphate oxidase (microbial): ≥ 5 U/mL; (horseradish): The TRIGB cobas c pack is now ready for use.
peroxidase ≥ 0.3 U/mL; preservative Note
R2 TRIS buffer: 0.15 mol/L, pH 7.6; magnesium sulfate: 17.5 mmol/L; Use only the cobas c pack MULTI. Always use a new cobas c pack MULTI
when preparing fresh reagent. Never reuse accessories designed for single
EDTA, disodium salt: 10 mmol/L; 4‑chlorophenol: 3.5 mmol/L; use, as this may result in reagent contamination and could affect test
potassium hexacyanoferrate (II): 6 µmol/L; sodium cholate: 0.15 %; results. If the cobas c pack MULTI bottles are not filled correctly, this may
hydroxypolyethoxy‑n‑alkanes: 0.12 %; lipase (Pseudomonas result in faulty reagent pipetting and could cause erroneous results.
species): ≥ 6 U/mL; 4‑aminophenazone: 0.7 mmol/L; preservative Storage and stability
Precautions and warnings TRIGB
For in vitro diagnostic use.
Exercise the normal precautions required for handling all laboratory Shelf life at 2‑8 °C: See expiration date
reagents. on cobas c pack
Disposal of all waste material should be in accordance with local guidelines. label.
Safety data sheet available for professional user on request.
On‑board in use and refrigerated on the analyzer: 2 weeks
For USA: Caution: Federal law restricts this device to sale by or on the
order of a physician. Diluent NaCl 9 %
This kit contains components classified as follows in accordance with the Shelf life at 2‑8 °C: See expiration date
Regulation (EC) No. 1272/2008: on cobas c pack
Peroxidase label
EUH 208 May produce an allergic reaction. On‑board in use and refrigerated on the analyzer: 12 weeks
Product safety labeling follows EU GHS guidance. Specimen collection and preparation
Reagent preparation and cobas c pack MULTI assembly For specimen collection and preparation only use suitable tubes or
Reagent handling collection containers.
Only the specimens listed below were tested and found acceptable.
R1 Connect one Bottle 1a (Enzymes) to one Bottle 1 (Buffer) using Serum
one of the enclosed adapters. Mix by gentle inversion. Plasma: Li-heparin and K2‑EDTA plasma.
R2 Connect one Bottle 2a (Lipase/4‑Aminophenazone) to one Bottle 2 The sample types listed were tested with a selection of sample collection
tubes that were commercially available at the time of testing, i.e. not all
(Buffer) using one of the enclosed adapters. Mix by gentle available tubes of all manufacturers were tested. Sample collection systems
inversion. from various manufacturers may contain differing materials which could
One set of Roche/Hitachi bottles yields two cobas c packs. affect the test results in some cases. When processing samples in primary
tubes (sample collection systems), follow the instructions of the tube
Labeling the cobas c pack MULTI manufacturer.
Turn the barcode labeled side of a new cobas c pack MULTI toward you. Centrifuge samples containing precipitates before performing the assay.
Affix the supplied TRIGB barcode label directly over the existing barcode See the limitations and interferences section for details about possible
label. sample interferences.
Sample stability claims were established by experimental data by the
manufacturer or based on reference literature and only for the
temperatures/time frames as stated in the method sheet. It is the
responsibility of the individual laboratory to use all available references
and/or its own studies to determine specific stability criteria for its
laboratory.
Stability in serum: 2 days at 20‑25 °C5
10 days at 4 °C6
3 months at -20 °C7
several years at -70 °C7
Stability in plasma: 2 days at 20‑25 °C5
15 days at 4 °C8
Filling the cobas c pack MULTI
3 months at -20 °C7
1. Turn the cobas c pack MULTI toward you as shown previously. 
several years at -70 °C7
2. Position A of the cobas c pack is now in the center, position B on the
left side, position C on the right side of the cobas c pack. Materials provided
3. Unscrew the screw cap of the bottle in position B on the left side of the See “Reagents – working solutions” section for reagents.
cobas c pack MULTI using the Open/Close tool.
Materials required (but not provided)
4. Pour exactly 15 mL of the content of bottle 1 (R1) into the opened bottle
of the cobas c pack (position B). ▪ See “Order information” section
5. Close the bottle tightly using the Open/Close tool. ▪ General laboratory equipment

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Assay Calibration
For optimum performance of the assay follow the directions given in this
document for the analyzer concerned. Refer to the appropriate operator’s Calibrators S1: Precimat Glycerol
manual for analyzer‑specific assay instructions. S2: C.f.a.s.
The performance of applications not validated by Roche is not warranted Calibration mode Linear
and must be defined by the user.
Calibration frequency 2‑point calibration
Application for serum and plasma
- after 24 hours on board
cobas c 311 test definition - after cobas c pack change
Assay type 1‑Point - after reagent lot change
- as required following quality control
Reaction time / Assay points 10 / 57
procedures
Wavelength (sub/main) 700/505 nm
Calibration interval may be extended based on acceptable verification of
Reaction direction Increase calibration by the laboratory.
Units mmol/L (mg/dL, g/L) Traceability: This method has been standardized against SRM 909b and
triolein standards.
Reagent pipetting Diluent (H2O)
Quality control
R1 100 µL – For quality control, use control materials as listed in the "Order information"
R2 100 µL – section.
Sample volumes Sample Sample dilution In addition, other suitable control material can be used.
The control intervals and limits should be adapted to each laboratory’s
Sample Diluent (NaCl) individual requirements. Values obtained should fall within the defined
Normal 2 µL – – limits. Each laboratory should establish corrective measures to be taken if
values fall outside the defined limits.
Decreased 4 µL 15 µL 150 µL
Follow the applicable government regulations and local guidelines for
Increased 2 µL – – quality control.
cobas c 501 test definition Calculation
Roche/Hitachi cobas c systems automatically calculate the analyte
Assay type 1‑Point concentration of each sample.
Reaction time / Assay points 10 / 70
Conversion factors:
Wavelength (sub/main) 700/505 nm
mmol/L x 88.5 = mg/dL mmol/L x 0.885 = g/L
Reaction direction Increase
mg/dL x 0.0113 = mmol/L mg/dL x 0.01 = g/L
Units mmol/L (mg/dL, g/L)
Limitations - interference
Reagent pipetting Diluent (H2O)
Criterion: Recovery within ± 10 % of initial values at triglyceride levels of
R1 100 µL – 2.3 mmol/L (203 mg/dL).
R2 100 µL – Icterus:9 No significant interference up to an I index of 60 for conjugated
and 50 for unconjugated bilirubin (approximate conjugated bilirubin
Sample volumes Sample Sample dilution concentration: 1026 µmol/L or 60 mg/dL; approximate unconjugated
Sample Diluent (NaCl) bilirubin concentration: 855 µmol/L or 50 mg/dL).
Normal 2 µL – – Hemolysis:9 No significant interference up to an H index of 400
(approximate hemoglobin concentration: 248 µmol/L or 400 mg/dL).
Decreased 4 µL 15 µL 150 µL Lipemia:9 The L index correlates with sample turbidity but not with
Increased 2 µL – – triglycerides level. Extremely lipemic samples (triglycerides greater than
3000 mg/dL) can produce a normal result.10
cobas c 502 test definition Prozone Check: The flag  > Kin is an indicator for extremely high
triglyceride concentrations in the sample. False normal results are due to
Assay type 1‑Point oxygen depletion during assay reaction.
Reaction time / Assay points 10 / 70 Drugs: No interference was found at therapeutic concentrations using
Wavelength (sub/main) 700/505 nm common drug panels.11,12
Exception: Intralipid is directly measured as analyte in this assay and leads
Reaction direction Increase to high triglyceride results.
Units mmol/L (mg/dL, g/L) Acetaminophen intoxications are frequently treated with N‑Acetylcysteine.
Reagent pipetting Diluent (H2O) N‑Acetylcysteine at a plasma concentration above 499 mg/L and the
Acetaminophen metabolite N‑acetyl‑p‑benzoquinone imine (NAPQI)
R1 100 µL – independently may cause falsely low results.
R2 100 µL – Venipuncture should be performed prior to the administration of
Metamizole. Venipuncture immediately after or during the administration of
Sample volumes Sample Sample dilution Metamizole may lead to falsely low results. A significant interference may
Sample Diluent (NaCl) occur at plasma Metamizole concentrations above 0.1 mg/mL.
Normal 2 µL – – In very rare cases, gammopathy, in particular type IgM (Waldenström’s
macroglobulinemia), may cause unreliable results.13
Decreased 4 µL 15 µL 150 µL For diagnostic purposes, the results should always be assessed in
Increased 4 µL – – conjunction with the patient’s medical history, clinical examination and other
findings.

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ACTION REQUIRED Intermediate precision Mean SD CV

Special Wash Programming: The use of special wash steps is mandatory
when certain test combinations are run together on Roche/Hitachi mmol/L (mg/dL) mmol/L (mg/dL) %
cobas c systems. The latest version of the carry‑over evasion list can be Precinorm U 0.53 (46.9) 0.01 (0.9) 2.7
found with the NaOHD-SMS-SmpCln1+2-SCCS Method Sheets. For further
instructions refer to the operator’s manual. cobas c 502 analyzer: All Precipath U 0.62 (54.9) 0.02 (1.8) 2.8
special wash programming necessary for avoiding carry‑over is available Human serum 3 0.64 (56.6) 0.02 (1.8) 2.7
via the cobas link, manual input is required in certain cases.
Where required, special wash/carry‑over evasion programming must Human serum 4 5.82 (515) 0.07 (6) 1.1
be implemented prior to reporting results with this test. Method comparison
Limits and ranges Triglycerides values for human serum and plasma samples obtained on a
Measuring range Roche/Hitachi cobas c 501 analyzer (y) were compared with those
determined using the same reagent on a Roche/Hitachi 917 analyzer (x).
0.1‑10.0 mmol/L (8.85‑885 mg/dL)
Sample size (n) = 72
Determine samples having higher concentrations via the rerun function.
Dilution of samples via the rerun function is a 1:5.5 dilution. Results from Passing/Bablok16 Linear regression
samples diluted using the rerun function are automatically multiplied by a
factor of 5.5. y = 1.019x - 0.012 mmol/L y = 1.030x - 0.027 mmol/L
Low recovery of triglycerides values may be due to low free glycerol values τ = 0.979 r = 0.999
(< 0.0113 mmol/L or < 1 mg/dL). Samples with a value of < 1.13 mmol/L (or
< 100 mg/dL) triglycerides must be diluted with Precimat Glycerol as The sample concentrations were between 0.372 and 9.33 mmol/L (32.9
follows: 20 parts sample plus 1 part Precimat Glycerol. Rerun the diluted and 826 mg/dL).
sample. Multiply the rerun result by the dilution factor of 1.05.
References
Lower limits of measurement 1 Kaplan LA, Pesce AJ. Clinical Chemistry: Theory, Analysis and
Lower detection limit of the test Correlation. Ladig D, Kasper R (ed), St Louis, CV Mosby Co
0.1 mmol/L (8.85 mg/dL) 1984;1221-1229.
The lower detection limit represents the lowest measurable analyte level 2 Van Handel E, Zilversmit DB. Micromethod for the direct determination
that can be distinguished from zero. It is calculated as the value lying of serum triglycerides. J Lab Clin Med 1957;50(1):152-157.
3 standard deviations above that of the lowest standard (standard 1 + 3 SD, 3 Kessler G, Lederer H. Fluorometric measurement of triglycerides. In:
repeatability, n = 21). Skeggs LT Jr, et al; eds: Automation in Analytical Chemistry. Technicon
Expected values according to NCEP14 Symposia, Tarrytown, NY 1965;341-344.
Normal range: < 1.70 mmol/L (< 150 mg/dL). 4 Kohlmeier M. Direct enzymic measurement of glycerides in serum and
Clinical interpretation according to the recommendations of the European in lipoprotein fractions. Clin Chem 1986;32(1):63-66.
Atherosclerosis Society:15 5 WHO Publication: Use of anticoagulants in diagnostic laboratory
mmol/L mg/dL Lipid metabolism disorder investigations, WHO/DIL/LAB/99.1 Rev.2:Jan 2002.
6 Evans K, Mitcheson J, Laker M. Effect of Storage at 4 °C and -20 °C on
Cholesterol < 5.18 < 200 Lipid, Lipoprotein, and Apolipoprotein Concentrations. Clin Chem.
Triglycerides No 1995;41:392-396.
< 2.26 < 200
7 Tietz NW, ed. Clinical Guide to Laboratory Tests, 3rd ed. Philadelphia,
Yes PA: WB Saunders Company 1995;610-611.
Cholesterol 5.18‑7.77 200‑300 if HDL‑cholesterol 8 Kronenberg F, Lobentanz EM, König P, et al. Effect of sample storage
< 0.9 mmol/L (< 35 mg/dL) on the measurement of lipoprotein[a], apolipoproteins B and A-IV, total
Cholesterol > 7.77 > 300 and high density lipoprotein cholesterol and triglycerides. J Lipid Res.
1994 Jul;35(7):1318-28.
Triglycerides Yes
> 2.26 > 200 9 Glick MR, Ryder KW, Jackson SA. Graphical Comparisons of
Interferences in Clinical Chemistry Instrumentation. Clin Chem
Each laboratory should investigate the transferability of the expected values 1986;32:470-475.
to its own patient population and if necessary determine its own reference
ranges. 10 Shephard MDS, Whiting MJ. Falsely low estimation of triglycerides in
lipemic plasma by the enzymatic triglyceride method with modified
Specific performance data Trinder’s chromogen. Clin Chem 1990;36(2):325-329.
Representative performance data on the analyzers are given below.
Results obtained in individual laboratories may differ. 11 Breuer J. Report on the Symposium “Drug effects in Clinical Chemistry
Methods”. Eur J Clin Chem Clin Biochem 1996;34:385-386.
Precision 12 Sonntag O, Scholer A. Drug interference in clinical chemistry:
Precision was determined using human samples and controls in an internal recommendation of drugs and their concentrations to be used in drug
protocol with repeatability (n = 21) and intermediate precision (3 aliquots interference studies. Ann Clin Biochem 2001;38:376-385.
per run, 1 run per day, 21 days). The following results were obtained:
13 Bakker AJ, Mücke M. Gammopathy interference in clinical chemistry
Repeatability Mean SD CV assays: mechanisms, detection and prevention.
Clin Chem Lab Med 2007;45(9):1240-1243.
mmol/L (mg/dL) mmol/L (mg/dL) %
14 U.S. DEPARTMENT OF HEALTH AND HUMAN SERVICES Public
Precinorm L 1.11 (98.2) 0.02 (1.8) 1.6 Health Service, NIH Publication No. 01-3305, May 2001.
Precipath L 2.90 (257) 0.02 (1.8) 0.6 15 Study Group, European Atherosclerosis Society. Strategies for the
prevention of coronary heart disease: A policy statement of the
Human serum 1 0.45 (39.8) 0.00 (0.0) 0.9 European Atherosclerosis Society. European Heart Journal 1987;8:77.
Human serum 2 8.44 (747) 0.08 (7) 0.9 16 Bablok W, Passing H, Bender R, et al. A general regression procedure
for method transformation. Application of linear regression procedures
for method comparison studies in clinical chemistry, Part III. J Clin
Chem Clin Biochem 1988 Nov;26(11):783-790.

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A point (period/stop) is always used in this Method Sheet as the decimal

separator to mark the border between the integral and the fractional parts of
a decimal numeral. Separators for thousands are not used.
Symbols
Roche Diagnostics uses the following symbols and signs in addition to
those listed in the ISO 15223‑1 standard (for USA: see
https://usdiagnostics.roche.com for definition of symbols used):
Contents of kit
Volume after reconstitution or mixing
GTIN Global Trade Item Number

FOR US CUSTOMERS ONLY: LIMITED WARRANTY

Roche Diagnostics warrants that this product will meet the specifications
stated in the labeling when used in accordance with such labeling and will
be free from defects in material and workmanship until the expiration date
printed on the label. THIS LIMITED WARRANTY IS IN LIEU OF ANY
OTHER WARRANTY, EXPRESS OR IMPLIED, INCLUDING ANY IMPLIED
WARRANTY OF MERCHANTABILITY OR FITNESS FOR PARTICULAR
PURPOSE. IN NO EVENT SHALL ROCHE DIAGNOSTICS BE LIABLE
FOR INCIDENTAL, INDIRECT, SPECIAL OR CONSEQUENTIAL
DAMAGES.
COBAS, COBAS C, PRECICONTROL, PRECIMAT, PRECINORM and PRECIPATH are trademarks of Roche.
All other product names and trademarks are the property of their respective owners.
Additions, deletions or changes are indicated by a change bar in the margin.
© 2018, Roche Diagnostics

Roche Diagnostics GmbH, Sandhofer Strasse 116, D-68305 Mannheim

www.roche.com
Distribution in USA by:
Roche Diagnostics, Indianapolis, IN
US Customer Technical Support 1-800-428-2336

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