GAMMA GT Liquiform Ref.

: 105
Instruction for use

Intended use . Kinetic System for quantitative determination of Reagents

Gamma - Glutamyltransferase (GAMA GT) in serum or plasma (EDTA).

Professional use.

[For in vitro diagnostic use]

1. ( - Reagent 1 - Store at 2-8ºC.
Reagent label bears expiration date. Glycylglycine (197 mmol/L), sodium
azide (£14,6 mmol/L) and surfactant.

Test principle . Gamma-Glutamyltransferase catalyzes the

transference of L-g-Glutamyl-3-carboxi-4-nitroanilide to glycylglycine,
yielding L-g-Glutamylglycylglycine and p-nitroaniline as following the
2. ) - Reagent 2 - Store at 2-8ºC.
Reagent label bears expiration date. Contain buffer ≤ 50 mmol/L, L- ¡-
Glutamyl-3-carboxi-4-g nitroanilide (21 mmol/L), sodium azide (£14,6%)
reactions: and stabilizer.

L-g-Glutamyl-3-carboxi-4-nitroanilide + glycylglycine
L-g-Glutamylglycylglycine + p-nitroaniline
Gamma GT 3. < - Standard - Store at 2-8ºC.
Reagent label bears expiration date. p-nitroaniline (500 mmol/L) and
sodium azide (£14,6 mmol/L).
Do not contain gamma-glutamyltransferase.
The amount of p-nitroaniline produced, which has high absorbance at
The Standard is applicable only in the fixed-time-kinetic method. The p-
405 nm, is proportional to the Gamma GT activity in the sample.
nitroaniline amount is equivalent to 125 U/L Gamma GT's activity.
Summary . Labtest developed Gamma GT Liquiform system based on
the principle of the method of modified Szasz, that propitiates procedure Precautions and warnings
for Gamma GT determination and whose performance is substantially
equivalent to the reference method proposed by the International Avoid the directly sun light exposure.
Federation of Clinical Chemistry and Laboratory Medicine (IFFC)1, 2.
Disposal of all waste material should be in accordance with local
The substances for this reaction are distributed properly in two reagents in guidelines.
order to get more stability in the original liquid form and keep the optimum
conditions of operation conditions, allowing the direct use of the reagents The usual security cares should be applied on the reagent handling.
in automatic systems.
The reagents contain sodium azide as preservative. Avoid ingestion. In
The monoreagent method can be applied by using one Work Reagent, 21 case of eyes contact, immediately flush eyes with plenty of water and get
days stable if stored at 2-8ºC, reaching appropriated performance even in medical assistance.
low requests of the test. The system also allows preparing the volume of Sodium azide may react with lead and copper plumbing to form highly
the Work Reagent needed to one measure of the Gamma GT enzymatic explosive metal azides. On disposal, flush with a large volume of water to
activity. prevent azide accumulation.

The system is linear up to 700 U/L, equivalent to 15 times the high In automatic equipments, the reagents may be contaminated with other
reference threshold, decreasing the dilution necessities and reagents or the air, depending on the equipment's characteristic and the
measurement repetitions in high activities sample. work conditions. These can result in stability reduction and calibration
modifications.
The performed assays show that Labtest products do not suffer
interference by high values of bilirubin, hemoglobin, and triglycerides. As it occurs in all enzymatic activity measurement, the incubation time
and temperature is important for the quality of the results.
The measurements can be done by the fixed-time-kinetic method, in
semi-automatic equipments, using the p-nitroaniline Standard Storage and stability . Unopened reagents, when stored at
(Ref.: 105.3). Also the measurements can be done by the continuous
indicated temperature, are stable up to expiration date shown on the label.
kinetic method, in automatic and semi-automatic equipments. In this
case, Labtest provides the Calibra's series with Gamma GT activity
traceable to the reference material ERMÒ-AD452/IFCC of the Institute for Deterioration . Microbial or chemical contamination may decrease
Reference Materials and Measurements and to the reference method of reagents stability.
the IFCC2. Gamma GT is not suitable for use if Working Reagent has an absorbance
over 1.5 at 405 nm when measured versus water as reference or in case
of contaminations signs or if it develops turbidity.
Methodology . Szasz modified.

01 English - Ref.: 105

 In order to verify the reaction linearity, it is recommended to measure in 1
Sample
minute as well, and check if the difference of absorbance in each minute is
Use serum or plasma (EDTA). Gamma GT is reportedly stable for about 7 constant.
days at 2-8ºC and 2 months at -20ºC3.
Fixed-time kinetic method . See Calibration and notes 1, 2 and 3.
No known test method can offer complete assurance that human blood This method requires 5% acetic acid solution.
samples will not transmit infectious diseases. Therefore, all blood
derivatives should be considered potentially infectious. In one tube, add 0.8 mL of Reagent 1 and 0.2 mL of Reagent 2. Mix and
transfer 0.5 mL of the Working Reagent to another tube. Label the tubes as
Blank and Test.
Interference
Bilirubin up to 38 mg/dL, hemoglobin up to 180 mg/dL and triglycerides Blank Test
up to 1000 mg/dL do not interfere significantly. Working Reagent 0.5 mL 0.5 mL

False decreased results may occur if heparin is used. Anticoagulants

containing citrate, fluorite, or oxalate inhibit Gamma GT activity. Incubate at 37ºC during 2 minutes. Do not remove the tubes from the
water bath, add:
Gamma GT is lower in women than in men at the same age.
Sample ----- 0.025 mL
Chronic alcoholism increases the Gamma GT activity in serum.
Mix and keep at 37ºC during 10 minutes. Add:
Gamma GT raised levels were observed in patients who used antiepileptic
drugs. 1.0 mL 1.0 mL
5% Acetic acid

Materials required not provided

1. Pipettes to measure reagents and samples Mix and add:
2. Timer Sample 0.025 mL

Continuous kinetic method Mix and measure the Test absorbance at 405 nm or Blue Filter (400 - 420)
1. Photometer capable of measuring absorbance at 400 - 420 nm and against Blank. The color is stable for 60 minutes.
keeping the cuvette at 37ºC.
Calibration . Measure the Standard absorbance in triplicate. The
Fixed-time kinetic method difference between the measures must be less than 2%.
1. A constant temperature water bath (37ºC).
2. 5% Acetic Acid solution. Blank Standard
3. Photometer capable of measuring absorbance at 400 - 420 nm. Water 0.5 mL 0.5 mL
Standard ----- 0.05 mL
Preparing the working reagent . Use one bottle of Reagent 1 5% Acetic acid 1.0 mL 1.0 mL
and Reagent 2 for preparing Working Reagent. Transfer all the contents of
one Reagent 2 bottle to one Reagent 1 bottle and mix gently.
Mix and measure the Standard absorbencies at 405 nm (400 - 420 nm)
The Working Reagent is stable 1 day at 15-25ºC and 21 days at 2-8ºC, against Blank. The color is stable for 60 minutes.
when no chemical or microbial contamination occurs.
Calibration
Procedure Manual calibrations
Continuous kinetic method
Continuous kinetic method . See Calibration and notes 1,
Use Calibrator Calibra H - Labtest. Gamma GT activity is traceably to
2 and 3.
reference material ERM-AD452/IFCC and the reference method of IFCC2.
Reactions conditions: wavelength: 405 nm; cuvette at 37±0.2ºC, 1cm Calibration frequency
light path, pass band £2nm and stray light £0.1. Calibration of 2 points when changing the batch;
Calibration of 2 points when the internal quality control indicates.
1. In a test tube labeled “Test” or “Calibrator”, add 1.0 mL of the Work
Reagent. Fixed-time kinetic method
Use the Standard Ref. 105.3. The Standard is traceable to reference
2. Add 0.05 mL of the sample or enzyme calibrator, homogenize and material ERM-AD452/IFCC and the reference method of IFCC2.
transfer immediately to a cuvette at 37±0.2ºC. Wait one minute.
Perform a new calibration after reagent lot change or when the internal
3. Measure the initial absorbance (A1) at 405 nm (400 - 420 nm), and quality control indicates.
start simultaneously the timer. Measure the absorbance again after 2
minutes (A2).

02 English - Ref.: 105

Automatic Systems Gamma GT (U/L) - 37ºC
Blank of reagents: water or 0.85% NaCl; Women
Use Calibrator Calibra H - Labtest, which are traceable to reference 0 - 6 months 15 - 132
material ERM-AD452/IFCC and the reference method of IFCC2. 6 - 12 months 1 - 39
1 - 12 years 4 - 22
Calibration frequency
13 - 18 years 4 - 24
Calibration of 2 points when changing the batch;
Calibration of 2 points when the internal quality control indicates. Adults 5 - 39

Quality control . The limits and control interval must be adapted to

Gamma GT (U/L) - 37ºC
the laboratory requirements. Each laboratory should establish corrective
measures to be taken if values fall outside the control limits. Men
0 - 6 months 12 - 122
It is recommended to use products from Qualitrol line - Labtest for internal 6 - 12 months 1 - 39
quality control in clinical chemistry trials. 1 - 12 years 3 - 22
13 - 18 years 2 - 42
Calculations Adults 7 - 58

Continuous Kinetic method Conversion: Conventional Unit (U/L) x 16.7 = SI Unit (nKat/L)

A2 - A1 Temperature . The table below allows the conversion of the

DA (test or calibrator) = measured activity at a stated temperature into a value that would be
2 obtained if the measurement was performed in other temperatures.

Calibrator activity
Factor = Factors for temperature correction
Work Temperature
DA Calibrator 25°C 30°C 37°C
25°C ----- 1.37 1.79
Gamma GT activity = DA Test x Factor 30°C 0.73 ----- 1.30
37°C 0.56 0.77 -----
When the optimum reaction conditions are obtained the 2550 factor can
be applied.
Example: the enzymatic activity obtained at 37ºC must be multiplied to the
Fixed-time kinetic method: factor 0.77 in order to obtain the activity at 30ºC or to the factor 0.56 in
order to obtain the activity at 25ºC.
Standard = 125 U/L
Performance characteristics9
Test absorbance
Gamma GT (U/L) = x 125
Standard absorbance R ecover y studies . I n t w o s a m p l e s w i t h G a m m a -
Glutamyltransferase activities of 295 and 485 U/L were added different
125 quantities of the enzyme, obtaining the follow results:
Calibration factor =
Standard absorbance
Initial Activity (U/L) 295 485
Added Activity (U/L) 101 101
Gamma GT (U/L) = Test absorbance X Factor
Expected Activity (U/L) 396 586
Obtained Activity (U/L) 391 585
Linearity
Recovery (%) 98.7 99.8
Up to 700 U/L.

If Gamma GT concentration exceeds 700 U/L, the sample must be diluted The mean proportional systematic error at 59 U/L decision level was
with 0.85% NaCl. Multiply the result by the appropriate dilution factor. 0.4 U/L and at 181 U/L decision level was 1.3 U/L.

Dilute the sample so that the obtained value is around 50 and 400 U/L. Method comparison . The proposed method was compared to the
reference method, obtaining the follow results:
Expected values7,2 . Each laboratory should evaluate the
transferability of the expected values to its own patient population and, if
necessary, estimate its own reference interval.

03 English - Ref.: 105

 Comparison Labtest References
Method Method
n 80 80 1. Szasz G. Clin Chem 15:124, 1969.
Range (U/L) 14.7 - 244.0 11.4 - 241.4
Mean (U/L) 119.9 118.8 2. IFCC Reference Procedure for the Measurement of Catalytic
Labtest method = 1.015 x Concentration of g-Glutamyltransferase. Clin Chem Lab Med 2002;
Regression Analysis
comparison method - 2.97 40:734-38.
Correlation Coefficient 0.999
3. Kaplan LA, Pesce AJ.: Methods in Clinical Chemistry, St. Louis: The C.
V. Mosby Co. 1122, 1987.
The estimated total error is -3.5 % in a decision level of 59 U/L and -0.14 %
in a decision level of 181 U/L. 4. Otimal Biological Variation Database specifications. Available at:
<https://www.westgard.com/optimal-biodatabase1htm.htm>
(access in 02/2022).
Imprecision Within Run
N Mean (U/L) SD (U/L) %CV 5. Basques JC. Especificações da Qualidade Analítica. Labtest
Diagnóstica 2005.
Sample 1 20 59 0.7 1.13
Sample 2 20 181 1.0 0.57 6. Westgard JO, Barry PL, Hunt MR, Groth T. Clin Chem. 1981, 27:493-
501.

Imprecision - Run-to-Run 7. Soldin SJ, Brugnara C, Wong EC: Pediatric Reference Intervals, 5.ed.
%CV Washington: AACC Press, 2005. p.98-99.
N Mean (U/L) SD (U/L)
Sample 1 20 59 1.5 2.58 8. Burtis CA, Ashwood ER. Tietz Textbook of Clinical Chemistry,
Sample 2 20 181 3.2 1.80 Philadelphia: Saunders Company 1994;980-86.

9. Labtest: data on file.

Total error . The estimated total error (random + systematic) in a
decision level of 59 U/L is 7.8 % and in a decision level of 181 U/L is 3.1 %. 10. Desirable Biological Variation Database specifications. Available at:
<https://www.westgard.com/biodatabase1.htm> (access in 02/2022).
Analytical sensitivity . Detection limit: 2.48 U/L. The detection limit
represents the lowest measurable Gamma-Glutamyltransferase activity Presentation
that can be distinguished from zero.
Product Reference Contents
Effects of matrix dilution . Samples with values equal 710 and 1 2 X 24 mL
603 U/L were used for evaluating the system response on dilution with 105-2/30 2 2 X 6 mL
150 mmol/L NaCl (0.85%). Using dilution factor from 2 to 16, the 1 X 3 mL
recovery was 99.0 %. Gama GT Liquiform
1 2 X 40 mL
105-2/50 2 2 X 10 mL
Notes 1 X 3 mL
1 2 X 41 mL
1. The material cleaning and drying are fundamental factors to the Gama GT Liquiform 105-2/55 2 2 X 14 mL
reagent stability and to obtain correct results. Labmax 560/400
1 X 3 mL
1 1 X 40 mL
2. The water in the laboratory to prepare reagents and use in the Gama GT Liquiform
105-1/53 2 1 X 13 mL
measurements, must have resistivity ³1 megaohm.cm, or conductivity Linha Audmax i
1 X 3 mL
£1 microsiems/cm and silicates concentration must be £0.1mg/L (Type II
Gama GT Liquiform 1 2 X 40 mL
reagent water). The water for washing must be Type III, having resistivity
CS400/800 105-2/53 2 2 X 13 mL
³0.1 megaohms or conductivity £10 microsiemens. For the final
Audmax 400/600 1 X 3 mL
washing, use Type II reagent water.

3. It is suggested to consult http:/www.fxol.org in order to review For information about other commercial presentations, consult the
physiopathological source and drugs interference in results and website www.labtest.com.br or contact SAC.
methodology.
The number of tests in automated instruments depends on the
programmed parameters.

04 English - Ref.: 105

Application procedures using Gamma-Glutamyltransferase are available Labtest Diagnóstica S.A.
for various automated instruments. CNPJ: 16.516.296/0001-38
Av. Paulo Ferreira da Costa, 600 - Vista Alegre - CEP: 33240-152
Customer information Lagoa Santa, Minas Gerais - Brasil - www.labtest.com.br
Customer Service email: [email protected]
[Warranty conditions]

Labtest Diagnóstica warrants the performance of this product under the Edition: May, 1994 Copyright by Labtest Diagnóstica S.A.
specifications until the expiration date shown in the label since the Revision: March, 2024 Reproduction under previous autorization
Ref.: 040324(03)
application procedures and storage conditions, indicated on the label and
in this insert, have been followed correctly.

05 English - Ref.: 105

 Símbolos utilizados com produtos diagnósticos in vitro
Símbolos usados con productos diagnósticos in vitro
Symbols used with IVD devices

Conteúdo suciente para < n > testes Risco biológico

Contenido suciente para < n > tests Riesgo biológico
Contains sufcient for < n > tests Biological risk

Data limite de utilização (aaaa-mm-dd ou mm/aaaa) Corrosivo

Estable hasta (aaaa-mm-dd o mm/aaaa) Corrosivo
Use by (yyyy-mm-dd or mm/yyyy) Corrosive

Limite de temperatura (conservar a) Tóxico

Temperatura limite (conservar a) Tóxico
Temperature limitation (store at) Poison

Representante Autorizado na Comunidade Europeia Marca CE

Representante autorizado en la Comunidad Europea Marcado CE
Authorized Representative in the European Community CE Mark

Carcinogênico/mutagênicoe/ou sensibilizante à respiração Atenção

Carcinogénico/mutagénico y/o sensibilizante respiratorio Atención
Carcinogenic/mutagenic and/orsensitizingto breathing Attention

Tóxico para os organismos aquáticos Fabricado em

Tóxico para los organismos acuáticos Elaborado en
Toxic for aquatic organisms Manufactured on

Gases/líquidos comburentes Fabricado por

Gases/líquidos oxidantes Elaborado por
Oxidizing gases/liquids Manufactured by

Substância inamável Uso veterinário

Sustancia inamable Uso veterinario
Flammable substance Veterinary use

Período após abertura Liofilizado

Período post-abertura Liofilizado
Period after-opening Lyophilized

Produto de uso único Reagente

Producto de un solo uso Reactivo
Single use product Reagent

Consultar instruções de uso Número do lote

Consultar instrucciones de uso Denominación de lote
Consult instructions for use Batch code

Instalar até Número do catálogo

Instalar hasta Número de catálogo
Install before Catalog Number

Material Calibrador/Padrão Controle

Material Calibrador/Estándar Control
Calibrator/Standard Material Control

Produto diagnóstico in vitro Controle negativo

Dispositivo de diagnóstico in vitro Control negativo
In vitro diagnostic device Negative control

Reagente Controle positivo

Reactivo Control positivo
Reagent Positive control

Reagente contendo micropartículas

Reactivo con micropartículas
Reagent with microparticles

06 English - Ref.: 105