Food Allergen Detection

In the business of food preparation, food allergens are a serious concern.
If you produce a product which does not identify an allergen in its ingredient list and/or makes a claim to be free of a specific allergen, it is important to validate those claims.

 


The presence of an undeclared allergen in your food product can have a devastating effect on unsuspecting consumers, in addition to forcing you to recall your product.  It is common that production of your allergen-free product can occur in a manner which could expose it to an allergen because the same facility produces other products which contain allergens.

To ensure your customers’ safety, we offer a type of assay which detects trace amounts of a specific allergen.  We can test your manufacturing equipment to validate your wash down and sanitation procedures taken to prevent the cross-contamination of an allergen have been effective.  Additionally, we can also analyze your final product to validate that any allergen it may have been exposed to is not actually present at current regulatory levels.  This data will provide you with the confidence that your final product meets specified allergen requirements, ensuring your food-sensitive customers’ safety.


The frequency with which you should validate the absence of an undeclared allergen in your product may vary depending on the circumstances for which your product is produced.  In order to assist you more completely in your allergen management plan, please inform us of any and all specific allergen requirements you are striving to meet.


Allergen Testing FAQs

Allergen Testing FAQs

1. What applications are the outline analyses applicable?

The allergen assays we offer are designed to validate the absence of the allergen in question.

They are not appropriate for food products which are known to contain the allergen in question.

In fact, if a sample contains more than 1% (10,000 ppm) of the allergen in question, the assay will be overwhelmed and give a false-negative result.

The assay is applicable on foodstuffs, rinse water, or environmental swab samples.

2. Do you offer environmental swabbing for allergens?

Yes – Your allergen management plan may include validating your cleaning procedure to proof its effectiveness in removing allergens.

Rinse water testing is also an option to ensure the effectiveness of cleaning in terms of allergen contamination.

You may perform this testing on-site yourself, however, if you desire documented third party results, you may schedule us to perform this survey.

Please contact us to discuss the price and availability of this service.

 

3. Do you offer any other alleren assays?

Yes – below is a list of assays available upon request (additional charges may apply):

Allergen Assay Allergen Assay Allergen Assay
Almond Crustacea Peanut
Brazil Nut Hazelnut Sesame
Lactoglobulin Lupin Total Milk
Cashew/Pistachio Macadamia Nut Walnut
Coconut Mustard Whole Egg
The following are exerts from an article published by Romer Labs in their Spotlights Newsletter  Vol 19 and was written by Elisabeth Hammer.
Over the past decade the demand for gluten-free food has soared and, therefore, more and more of these products can be found in the stores. The spectrum of consumers with difficulties in digesting gluten has grown to around 10%.  These individuals show varying degrees of sensitivity towards gluten, but their situation generally improves when following a gluten-free diet. Furthermore, there is a growing perception amongst increasing numbers of consumers that a gluten-free diet is better for you.  But what is gluten?  Why can it be toxic?  And how can gluten be detected in food?
The name “gluten” is derived from the Latin word for glue and it refers to the composite of the proteins called prolamins and glutelins found in wheat, barley rye, oats and their crossbred varieties.  Prolamins are defined as the fraction that can be extracted using 40-70% of ethanol and this fraction is called gliadin, hordein, secalin or avenin respectively, depending on the grain variety.  In general, it can be estimated that prolamins and glutelins occur in the same ratio in gluten.

Worldwide, gluten is an important source of nutritional protein, both in foods prepared directly from sources containing it, and as an additive to foods otherwise low in protein.  Gluten contributes texture and form to food products, due to its physicochemical properties.  Together with water and when kneaded, gluten forms a viscoelastic dough with a special protein network which is responsible for the shape of bakery products.

Following the transition from the hunter-gatherer lifestyle to the beginning of agriculture 10,000 years ago, cereals have been a main pillar of the human diet, which raises the question as to why gluten is causing increasing levels of health problems nowadays.  Approximately 1% of the world’s population is affected by coeliac disease – an immune-mediated enteropathy caused by the ingestion of gluten.  Interestingly, it is more frequent in women than in men.  Symptoms are diverse and not confined to the gastrointestinal tract. Examples are not only diarrhoea, abdominal pain, flatulence, indigestion or weight loss, but also irritability, depression and anxiety.  However, all these symptoms are considered to be unreliable as an indicator for the disease.  Coeliac disease can be diagnosed by a screening for certain antibodies in the serum. Another recommended diagnosis is a biopsy of the mucosa and the small intestine to affirm damage, as the disease leads to the destruction of microscopic, finger-like projections in the small intestine that are called villi.  As intestinal villi are responsible for the absorption of nutrients, malnourishment is a problem that –on a long term basis- may lead to development delays, osteoporosis or nutrient deficiencies, amongst other problems. Coeliac disease is a genetically predisposed auto-immune disorder, in which the immune system responds inappropriately to dietary gluten.  The enzyme called tissue transglutaminase modifies gluten peptides by deamidation in a way that T-cell epitopes are formed.  This stimulates the immune system and cross-reacts with the small intestine tissue, causing an inflammatory reaction that leads to the truncation of the villi.  The majority of proteins responsible for such an immune reaction are prolamins.  The strongest response is directed towards an α2-gliadin fragment that is 33 amino acids long and a principal contributor to gluten immunotoxicity.  This so-called 33-mer is highly resistant to breakdown by digestive enzymes and is, therefore, a suitable molecule for use as an analytical marker.  Homologues have been found in food grains that are toxic for coeliac patients, but are absent in nontoxic grains.

The only effective treatment for coeliac disease up to now has been a lifelong gluten-free diet.  This is challenging to maintain as gluten is very common in food.  “Hidden” gluten that is used as a protein filler can be found in unexpected products such as pharmaceuticals, sausages, sauces and desserts.  In addition, gluten-free products may contain gluten due to cross contamination during milling, storage and production.  Gluten-free food is usually based on rice, maize or buckwheat, as well as purified starch that still contains low levels of gluten.  It is very difficult to set limits because sensitivity varies from individual to individual.  According to scientific studies, the ingestion of gluten should be maintained at below 50 mg per day.

The Codex Alimentarius Committee started to discuss recommendations for limits in the late 1970s, cumulating in the 2008 CODEX Standard for Foods for Special Dietary Use for Persons Intolerant to Gluten (CODEX STA N 118 – 1979).  This recommendation was taken into European legislation through Commission Regulation (EC) No 41/2009 of 20 January 2009, concerning the composition and labelling of foodstuffs suitable for people intolerant to gluten.  In contrast to other food allergens, thresholds have been defined.  Food labeled as gluten-free must not exceed 20 ppm, whereas food containing low levels of gluten has to be lower than 100 ppm.  A proposed rule for gluten-free labeling of foods is in preparation in the U.S.
As there are regulations in place, there is a need for appropriate detection methods for gluten in food. Several technologies such as specific antibody based tests e.g. enzyme linked immunosorbent assays (ELISA) or lateral flow assays, polymerase chain reaction (PCR) methods and newer concepts like mass spectrometry are available – all with varying degrees of commercialisation – giving both qualitative and quantitative results.  An analytical test system should preferably be able to detect epitopes that are involved in coeliac disease.

The fact that gluten is a complex mixture of proteins and that it occurs in a wide range of unprocessed as well as processed matrices creates a huge challenge in terms of correct quantification and makes it difficult to find a suitable reference material. In 1985, the Prolamin Working Group (PWG) was founded in Europe.  Its first task was to establish a recognized gluten – respectively gliadin – standard.  By extracting gliadins from a selection of the most common wheat varieties, they managed to get a reference material. The IRMM (Institute for Reference Material and Measurements) initially accepted the PWG gliadin as a certified reference material, but later withdrew its acceptance.  However, it is still the only reference that has some acceptance and has been widely used for calibration of test systems.

Enzyme linked immunosorbent assays (ELISA) are the recommended method for the detection of gluten in food and a large number of test kits are available commercially.  In immunological methods, antibodies are applied that have been raised against different prolamin fractions or specific sequences that are harmful.  Different test kits do not necessarily give similar results for several reasons. These include different specificities of the polyclonal and monoclonal antibodies used, different extraction methods, and different materials for the calibration of the assays.

Numerous monoclonal and polyclonal antibodies have been developed for gluten testing, but only some of them are accepted on a broader basis.  In the late 1980s, the Skerritt antibody was developed.  This monoclonal antibody was raised against wheat gliadin from an Australian wheat variety and recognizes HMW (high molecular weight) glutenin subunits and the heat stable subfraction called ω-gliadins, which makes the Skerritt antibody suitable for gluten analysis in processed foods.  Even so, as the quantitation is based on the amount of ω-gliadins, which differ among cereals species, this can cause considerable differences in results.  Moreover, the Skerritt antibody only has a weak response to hordein.

Another monoclonal antibody for the detection of gluten is the R5 antibody, which was developed by Professor Mendez in Spain. The R5 antibody was raised against rye secalin, but showed strong cross reactivity to wheat gliadin.  However, it also cross-reacts with proteins from soy and lupin that are not harmful prolamins.  The change in direction to detecting immunotoxic peptides that play a role in the pathogenesis of coeliac disease from the detection of prolamins, led to the development of a next generation of antibodies.  The G12 antibody employed in the AgraQuant® G12 ELISA and AgraStrip® Gluten G12 Lateral Flow Test belongs to this next generation.

The G12 antibody specifically recognizes the 33-mer of the gliadin protein present in gluten.  This toxic fragment was identified by the University of Stanford and published in 2002 in a paper in Science.  The G12 antibody was raised against this 33-mer peptide using knowledge gained from this publication, and recognizes the hexapeptide sequence QPQLPY and similar peptides found in barley, rye and oats.  In contrast, the R5 antibody was raised against a secalin extract and later the epitope it reacts with was identified as the QQPFP pentapeptide.  The distinction between the two antibodies relates to the fact that the G12 antibody specifically targets the toxic fragment that triggers the auto-immune reaction in coeliac patients, rather than a peptide sequence unrelated to clinical outcomes.  It was confirmed during validation studies that G12 does not give any false positive signals with soy and is, therefore, suitable for measuring gluten in products containing soy.  There is also no cross reactivity to maize or rice.

There is an on-going debate whether oats are safe.  Several publications conclude that certain oat varieties may cause an auto-immune response in coeliac patients.  During the validation of AgraQuant® Gluten G12 ELISA test and AgraStrip® Gluten G12 Lateral Flow Test, positive and negative responses to oat varieties were observed.  The positive results appear to be a specific reaction of the antibody with the toxic fragment, rather than a non-specific response.  Therefore, the G12 antibody may shed new light on this debate by recognizing oat varieties that trigger a response in coeliac patients.  The Spanish Coeliac Association has recently awarded the 6th National Prize for Research on coeliac disease to a scientific team that used the G12 antibody to identify oat varieties containing low levels of gluten, in this regard.

Coeliac patients depend on the correct labelling of glutenfree food in order to maintain their health.  Ensuring the safety of food is a demanding task and, therefore, new developments in the field of detection methods for gluten are on-going.  The results obtained from new immunochemical test systems based on the G12 antibody should be considered to be closer to the ideal of a food safety test as they establish the important link between coeliac disease and detection of the immunotoxic peptides.

FREQUENTLY ASKED QUESTIONS


General Sample Submission FAQs

General Sample Submission FAQs

How do I get my results?
MBL delivers results via E mail in the form of a PDF Certificate of Analysis to the recipients designated in the Laboratory Deliverables section of the New Customer Package.
MBL will only report to a third-party at the consent of Customer.
Upon request, results may also be delivered via fax or USPS mail, if this is your preferred method of delivery.
For multiple recipients of results, we recommend establishing an internal distribution list whereby you can define and maintain such recipients (i.e. labresults@yourcompany.com).
How long do you keep my sample & can I have them back?
Sample Retention Times (after reporting results) are as listed in the table below.
Any repeat, additional, and/or confirmation analysis must be requested by the Customer within the retention time frame.
Repeat analysis performed at the Customers' request will be charged to the Customer if the original results are confirmed.
To limit liability, samples submitted to MBL will not be returned to Customer because they have been stored in a facility which is known to have live pathogens.

Sample Retention Times (After Reporting Results) are as Follows:


Sample Type Retention Time
Qualitative Microbiology Negative Enrichments / Samples – Not Retained
Positive Enrichments / Samples – 24 Hours
Quantitative Microbiology 2 Business Days
Chemistry Analyses 2 Business Days
Other Analyses 2 Business Days 

How long does it take to get results?

Specific turnaround times for each test can be found in our Test Library.

Our turnaround times are approximate, and do not include weekends or holidays for chemical and some microbiology analyses.

We will always try to begin analysis on the day we receive your samples.  However, samples submitted after 3pm may be held until the next day.

A 50% surcharge will be assessed for samples submitted after 3 pm requiring same day set up for microbiological analysis.

E. coli O157:H7 results are typically reported by 6:00am Tuesday through Friday, 7:00am on Saturday, 2:00pm on Sunday, and 10:00am on holidays.

Alternative terms regarding turnaround times may be arranged on a case-by-case basis.

What is the Sample Description?

This is assigned by You, the Customer, to identify the name of samples submitted to MBL, as indicated on the Sample Submission Form.

You should describe your sample in a way which makes sense to you and is meaningful now and later when referring to your records.

Describing samples in a like and similar manner each time you submit the same type of sample can help you later in comparing data from one lot to another.

General Methodology FAQs

Methodology FAQs

How much sample do you need for a test?

Typically, larger sample sizes equate to more representative results.

In general, most analyses require 100 grams to 500 grams of your product for an accurate test.

Each analysis is different, so please visit our Test Library for specific sample size requirements.

What Certifications do you maintain and where can I download them?

We maintain two Certifications (click on the image to download).

Please see our Accreditations Page for more information on our Certifications and our Commitment to Quality.

ISO/IEC 17025:2005 Certificate

Murray-Brown Laboratories ISO 17025 Certificate Exp 08-31-18

CDPHE Drinking Water Certificate

MBL State of Colorado Drinking Water Certificate 010117 to 123117

What matrices can you analyze?

We can handle a variety of matrices, including nearly all foods and food products, environmental samples, and dietary supplements.  Please inform us if you have any matrix concerns.

What methods will you use to analyze my sample?

Murray-Brown Labs’ ISO/IEC 17025:2005 Certificate itemizes all analyses/methods for which we are accredited.

Unless specified by the Customer in writing, we will select the method believed to be the most appropriate for analyzing samples based upon the information you provided.

We will notify you when a method you specify is believed to be inappropriate or out of date, and reserve the right to decline the use of such a method.

Cost & Payment FAQs

Cost & Payment FAQs

What are your Payment Terms and what forms of payment do you accept?

Payment Terms and Conditions

Samples are as submitted by Customer; MBL will not invoice a third party unless agreed upon by MBL, at MBLs’ sole discretion.
Customer must have a valid credit card on file with MBL; this card will be charged immediately upon completion of analyses (per project or weekly invoice as dictated by MBL), unless credit terms have been granted.  Customers scheduling $500 or more per week in MBL tests and services must apply for credit terms to pay by check or be must be approved by MBL to pay via credit card on file.
MBL may extend credit to Customer based upon the findings of a credit or other reference check, and/or the history with Customer and/or its representatives.  Customer granted credit agrees: payment terms are Net 15 days; MBL will charge credit card on file for overdue balances; a late charge of 1.5% per month will be added to all unpaid balances; all current and/or ongoing work will be halted for accounts overdue by 30 days and no additional work will be accepted until the balance is paid in full; accounts delinquent 90 days will be scheduled for collections; Customer is responsible for collection and/or legal fees MBL incurs as a result of securing payments for invoices which are in default.
MBL reserves the right to collect prepayment from Customer; costs of analyses are as defined via our Pricelist and/or Quotations.  Additional charges, as assessed by MBL and agreed upon by Customer, may be applied for: sample preparation; rush and/or weekend analyses; sample collection and/or pick-up; consulting; time required to complete above and beyond Customer requests.

All Major Credit Cards Accepted

Credit Cards

What are your prices? Are there discounts for volume samples?

Please contact us for pricing including large volume discounts.

Sample Collection & Transportation FAQs

Sample Collection & Transportation FAQs

Do you offer sample pick up and how much does it cost?

Yes – Below you can find our sample pick up pricing, schedule, and fees.

Sample Pick Up is Free if:

 

 

 

Otherwise, Pick Up Fees are:

  Cost of testing is greater than or equal to $100; 24 hours notice is given;

  Pick up location is within 20 miles of MBL;

  Wait time for samples upon arriving at facility is less than 5 minutes.

 

$25 per pick up

(within 20 mile radius; plus $1 per mile over 20 miles; plus $1 per minute past 5 minute wait; plus $25 if less than $100)

When the Customer employs MBL to courier samples, MBL agree to do so in a manner which preserves the integrity of the sample(s).

Do you perform on-site sample collection?

Yes – Our trained staff is available to perform sampling at your or a third-party site.

Please contact us to discuss the price and availability of this service.

Sample collection by MBL on behalf of the Customer will take place according to an appropriate Sampling Plan, selected and/or designed by MBL, which will be available to the Customer for reference.  MBL is not responsible for alterations made by the Customer to the larger portion from which MBL collected samples after such sample collection occurred; such alterations may invalidate reported results.

When the Customer employs MBL to collect and/or courier samples, MBL agree to do so in a manner which preserves the integrity of the sample(s) and the larger portion from which it was collected.

We can also, if needed, assist in training your staff in proper sample collection techniques and procedures.

How do I collect a representative sample?

There is no short answer to this question and it is best to discuss this with any regulatory body that you are dealing with and any associated regulatory program by which your are governed.

We can provide guidance and assistance with most of the common regulatory requirements and associated sampling protocols.

MBL will provide a result which is representative of the sample submitted; it is the responsibility of the Customer to ensure that the sample submitted is representative of the larger portion from which it was collected.

How do I ship my sample to the laboratory?

Usually via overnight courier if refrigerated or frozen with enough coolant to maintain the proper temperature until receipt at our laboratory.

If you are uncertain about the details, please call us to discuss them.

How should I handle my sample before submitting it to the laboratory?

The integrity and representativeness of each sample must be maintained through delivery to the laboratory.

What type of container should I collect my sample in?

The appropriate container for each type of sample is included in our list of tests.  Please call to discuss any non-typical situations.