AgraStrip® Gluten G12

Order #: COKAL0200AS
Gluten
Gluten is the main group of proteins in grains and consists of prolamins (in wheat:
gliadin) and glutelins (in wheat: glutenins) occurring in the same ratio. Due to its
physicochemical characteristics, gluten is used in food products as a binder. Coeliac
disease is an autoimmune disorder of the small intestine. It is caused by a reaction to
gliadin and the only effective treatment is a lifelong gluten-free diet. Due to Codex
Standard 118-1979, “gluten-free” products must comply with gluten levels (including
prolamin fractions from wheat, rye, barley and oats) below 20 mg/kg and “foods specially
processed to reduce gluten content” must comply with levels between 20 and 100
mg/kg.

Short instructions

Perform extraction and dilution steps for finished product, rinse

water or swab.

Shake dilution tube vigorously by hand for 15 seconds.

Place a test strip vertically into the dilution tube and allow liquid
to soak up the strip to the „flow“ level line.

Then place the strip upright into a slot of the Tube Holder and
allow to develop for 10 minutes and read off the result
immediately.

Performance Characteristics:

LOD: 5 – 10 – 20 ppm Gluten* (Finished product)

35 ppb Gluten* (Rinse water)
4 g/25cm2 Gluten** (Swab testing)

* LOD was determined in extraction solution

** LOD was calculated

©2014 by Romer Labs UK Ltd. All Rights Reserved. This document is the property of Romer Labs UK Ltd
PI_COKAL0200AS_LFR_EN_v015 February 2014
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 Sample preparation – Finished Products
1. Homogenize the sample 2. Weigh 0.2g of sample 3. Add the sample to extraction
(i.e. blend, crush, grind). (weigh with Balance or estimate tube
by filling up one of the
extraction tube caps).

4. Fill Extraction Tube with 5. Close tube with tube cap 6. Remove cap from the
Extraction Buffer to level and shake vigorously by hand extraction tube and replace with
shown below (blue arrow). for 1 minute. dropper tip. And transfer 3
drops (100l) to a dilution
tube.

Proceed to the Assay Section (page 3) to complete your test

Sample preparation – Swab Testing

1. Fill extraction tube with 2. Wipe an area of 5cmx5cm using 3. Place the swab into the
Extraction Buffer to level side to side movements, rotating the extraction tube. Carefully break
shown below (blue arrow), swab tip as you go (we recommend the off the end at the pre-scored
take a swab and wet the end “cross-hatch” swabbing technique point.
by dipping into the buffer indicated below)

4. Close the tube with a cap 5. Remove cap from the extraction tube and replace with dropper tip.
and shake vigorously for 1 And transfer 3 drops (100l) to a dilution tube
minute.

Proceed to the Assay Section (page 3) to complete your test

Sample preparation – Rinse water Testing

Add 0.5 mL of rinse water into a dilution tube

Proceed to the Assay Section (page 3) to complete your test

©2014 by Romer Labs UK Ltd. All Rights Reserved. This document is the property of Romer Labs UK Ltd
PI_COKAL0200AS_LFR_EN_v015 February 2014
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 Assay Procedure in detail

1. Transfer various amounts of Dilution Buffer (red label) into the dilution tube already filled with extract

2. Close the dilution tube with the cap and shake vigorously by hand for 15 seconds
3. Take off cap and place a test strip vertically (arrows pointing down) into the dilution tube and allow liquid
to flow up the strip to the „flow“ level line. (this takes about 45 seconds)
4. After the liquid has soaked up to the “flow” level remove the test strip from the dilution tube and place it
upright (arrows pointing down) into a slot of the Tube Holder and allow to develop for 10 minutes and then
read off the result immediately.

Flow level line

15 seconds ~45 seconds 10 minutes

Interpretation of Results

One single blue line (=control line) in the central part of the test = negative
result

One red line (=test line) and one blue line in the result zone = positive result.
The sample contains Gluten higher than the cutoff level and further investigations
should be performed (e.g. quantification
of Gluten using AgraQuant® Gluten G12 ELISA Test Kits).

No control line appears = invalid result, regardless of whether the test line appears.
In the case of an invalid result, please repeat the procedure with a new strip. If the
problem persists, please contact Romer Labs® before continuing further.

Important advice for the proper execution of the test:

It is important to read the results immediately after the 10 minute incubate

step since the AgraStrip® test system has been validated extensively and shows
reliable results after that exact time. Longer incubation times can lead to the
development of false positive results.

©2014 by Romer Labs UK Ltd. All Rights Reserved. This document is the property of Romer Labs UK Ltd
PI_COKAL0200AS_LFR_EN_v015 February 2014
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Performance Characteristics in Detail

Limit of detection: 5 – 10 – 20 ppm Gluten (Finished product)

35 ppb Gluten (Rinse water)
4 g/25cm2 Gluten (Swab testing)

Range of detection: 5 – 10000 ppm Gluten

pH range: Performing the assay in a pH range of 6-8 will lead to reliable results. Highly
acidic samples can lead to false positive results whereas in an alkaline milieu there is the
tendency to false negative results.

Materials Supplied with Kit:

1 Tube containing 10 1 big Dropper Bottle (green 1 small Dropper Bottle

® label) containing 35mL containing 10mL Dilution
AgraStrip Gluten G12 Strips
Extraction Buffer Buffer

10 Extraction Tubes 10 Tube Caps 10 Dilution Tubes with caps

10 Extraction Tube Dropper Tips 10 Breakpoint Swabs 1 Tube Holder

Materials required but not supplied for solid sample preparation

 Blender or Crusher or Blade

©2014 by Romer Labs UK Ltd. All Rights Reserved. This document is the property of Romer Labs UK Ltd
PI_COKAL0200AS_LFR_EN_v015 February 2014
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Technical and Background Information

The AgraStrip® Gluten G12 Test Kit is a lateral flow assay for the detection of Gluten
content in food, rinse waters and environmental swab samples.

Gluten Allergy
Gluten is the main group of proteins in grains and consists of prolamins (in wheat:
gliadin) and glutelins (in wheat: glutenins) occurring in the same ratio. Due to its
physicochemical characteristics, gluten is used in food products as a binder. Coeliac
disease is an autoimmune disorder of the small intestine. It is caused by a reaction to
gliadin and the only effective treatment is a lifelong gluten-free diet. Due to Codex
Standard 118-1979, “gluten-free” products must comply with gluten levels (including
prolamin fractions from rye, barley and oats) below 20 mg/kg and “foods specially
processed to reduce gluten content” must comply with levels between 20 and 100
mg/kg.

Assay Principles
The AgraStrip® Gluten G12 Test Kit is an immunochromatographic test for the detection
of gluten in foodstuffs. The test kit uses a new monoclonal antibody called G12 that
specifically recognises the pathogenic fragment of the gliadin protein present in gluten.
This fragment is called 33-mer and triggers the auto-immune reaction in coeliac patients.
During the test, the sample reacts with a coloured conjugate (anti-gliadin 33mer
monoclonal antibody – red-coloured microsphere) which forms a complex with the
reagent on the strip. This complex spreads along the membrane by capillary action. The
AgraStrip® Gluten G12 is easy to use, fast and reliable.

Approvals
This test kit’s performance was reviewed by AOAC Research Institute and was found to
perform to the manufacturer’s specifications. The following matrices have been validated
according to AOAC Performance Tested Methods (PTM) protocols: rice flour, bread,
cookies, ice cream and dark chocolate. Stainless steel has been validated per AOAC PTM
protocol for environmental surface swab testing.

The rinse water test method has been validated internally by Romer Labs ®.

©2014 by Romer Labs UK Ltd. All Rights Reserved. This document is the property of Romer Labs UK Ltd
PI_COKAL0200AS_LFR_EN_v015 February 2014
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 Precautions

1. The product must be stored in its original package, between 15 and 25°C (room
temperature). Do not use components beyond the expiration date indicated on the
kit labels. Do not open the product until needed.
2. Test strips must be kept inside their original packaging, closed as tightly as
possible. Do not freeze.
3. Adhere to the instructions for test procedures.
4. The components in this test kit have been quality control tested as a standard
batch unit. Do not mix components from different lot numbers.

Sampling:
Consideration must be taken that the food may contain an uneven distribution of Gluten
(spot contamination). It is important to test a representative portion of food as only a
small amount of material is tested with the AgraStrip® Gluten test.

Detection:
The detection limit of the AgraStrip® Gluten G12 test is at the low ppm level but will vary
depending on the food matrix being tested. To give reliable results each individual matrix
should be validated before the kit is used routinely. Since the assay is for screening
purpose a positive result might require confirmation or further testing.
For further information regarding validation please contact Romer Labs.

Note:

! !
Chocolate and flour samples may block the filter tip of the extraction tube. This can be
avoided by transferring the extract directly from the extraction tube to the reaction vial
using a pipette or by hand to a level just under the 0.5ml graduation of the reaction vial

©2014 by Romer Labs UK Ltd. All Rights Reserved. This document is the property of Romer Labs UK Ltd
PI_COKAL0200AS_LFR_EN_v015 February 2014
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 Handling of the new improved AgraStrip® extraction & dilution buffer bottles

To open the bottle just hold the neck of the upper part (indicated
by the asterix) with your thumb and index finger and twist the
corrugated screw top counterclockwise .

To close the bottle twist the screw top clockwise up to the stop.

For further information please contact:

COMPETENCE CENTER ALLERGENS:

Romer Labs UK Ltd. Tel:+44 (0) 845 519 50 10

Block 5, The Heath Business & Email: enquiry@romerlabs.com
Technical Park,
Runcorn, Cheshire, WA7 4QX

GLOBAL HEAD QUARTERS:

Romer Labs Division Holding GmbH Tel: +43 (0) 2272 615 33
Technopark 1 Email : office-europe@romerlabs.com
3430 Tulln, Austria

For customer service contact details in your country please visit:

www.romerlabs.com

Warranty
The user assumes all risk in using Romer Labs UK Ltd products and services. Romer
Labs UK Ltd will warrant that its products and services meet all quality control standards
set by Romer Labs UK Ltd, and Romer Labs UK Ltd will, at its option, repair or replace
any product, components, or repeat services which prove to be defective in workmanship
or material within product specific warranty periods or expiration dates and which our
examination shall disclose to our satisfaction to be defective as such. This warranty is
expressly in lieu of all other warranties, expressed or implied, as to description, quality,
merchantability, fitness for any particular purpose, productiveness, or any other matter.
Romer Labs UK Ltd shall be in no way responsible for the proper use of its products.
Romer Labs UK Ltd hereby disclaims all other remedies, warranties, guarantees or
liabilities, expressed or implied, arising by law or otherwise, and it shall have no liability
for any lost profits or damage, direct, indirect or otherwise, to person or property, in
connection with the use of any of its products or services. This warranty shall not be
extended, altered or varied except by a written instrument signed by an authorized
representative of Romer Labs UK Ltd

©2014 by Romer Labs UK Ltd. All Rights Reserved. This document is the property of Romer Labs UK Ltd
PI_COKAL0200AS_LFR_EN_v015 February 2014
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 FOR NOTES

©2014 by Romer Labs UK Ltd. All Rights Reserved. This document is the property of Romer Labs UK Ltd
PI_COKAL0200AS_LFR_EN_v015 February 2014
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