aflatoxin m1 in milk and dairy products screening tools
DESCRIPTION
Aflatoxin M1 is the product of aflatoxin B1 transformation in the digestive apparatus of dairy animals. Once these animals are fed with B1 contaminated feed, liver enzymes transform it into aflatoxin M1. This toxin is found then in milk and dairy products and, even if less toxic than its precursor, is still mutagenic and carcinogenic. Due to aflatoxin M1 toxicity and the likelihood of its presence in milk and dairy products, the level of this toxin is strictly regulated from European and International laws. In the EU, the maximum tolerated level of aflatoxin M1 is 0.05 parts per billion (ppb), and a lower limit of 0.025 ppb is envisaged for so called baby-foods. In the US, the Food and Drug Administration (FDA) established a maximum level for aflatoxin M1 of 0.5 ppb; China –with GB 2761-2011- and the International CODEX ALIMENTARIUS adopted the same limit as well. Tecna presents here its portfolio of products dedicated to aflatoxin M1 screening in milk and dairy products. In order to fulfill international regulations new kits for extra EU markets have been recently introduced.TRANSCRIPT
Tecna port-folio of kits for the determination of aflatoxin M1 in milk and dairy products
Aflatoxin M1
• A"latoxin M1 is a mycotoxin that naturally occurs in milk and dairy products; it is the product of a"latoxin B1 after it is metabolized by dairy animals.
• A"latoxin M1 is less carcinogenic than its parental toxin a"latoxin B1, however it is still highly toxic for liver and kidneys in particular.
• 1-‐3% of the feed a"latoxin consumed is excreted in milk within 24 hours from intake. Clearance time ranges between 1 and 4 days.
• I’screen A"la M1 MILK • I’screen A"la M1
• Celer A"la M1 500 • B ZERO A"la M1 500
Different options for different regulations
The EU limit for aElatoxin M1 in milk is 50 ppt. The Codex Alimentarius and the
FDA Eixed a higher limit at 500 ppt of aElatoxin M1 in milk.
Different requirements can’t be adequately satis6ied by a single product. For this reason Tecna implemented a complete port-‐folio of screening devices to
cover all the requirements.
Product overview • Assay principle: competitive enzyme immunoassay • Applications: raw milk, milk powder
• Assay time: 75 minutes
• Calibration curve: 5 - 250 ppt
• Limit Of Detection
– Raw milk: 0.005 ppb = 5 ppt
– Milk powder: 0.05 ppb = 50 ppt
The leading solution in Italy and Europe for the screening of
a"latoxin M1 in milk.
I’screen AFLA M1 milk is accreditable in compliance with
ISO IEC 14675:2003.
No sample manipulation required
Product overview • Assay principle: competitive enzyme immunoassay • Applications: raw milk, milk powder, cheese, yoghurt
• Assay time: 75 minutes
• Calibration curve: 5 – 250 ppt
• Limit Of Detection
– Raw milk: 0.005 ppb
– Milk powder: 0.05 ppb
– Cheese: 0.037 ppb
– Firm cheese: 0.12 ppb
– Yoghurt: 0.025 ppb
The leading solution in Italy and Europe for the screening of
a"latoxin M1 in milk and dairy products.
I’screen AFLA M1 is accreditable in
compliance with ISO IEC 14675:2003.
Milk validation
• Each prototype developed by Tecna R&D department is subject to internal validation before industrialization and production
• Of 20 samples with < 5 ppt of a"latoxin M1 (HPLC), Tecna kits revealed no positives: 100% speciEicity
• Of 20 samples spiked at 5 ppt of a"latoxin M1, no determination with Tecna kits resulted < 5 ppt: 100% sensitivity
• Kit precision was set onto two Reference milk Materials:
• (21.50 ppt): Intra-‐assay CV 3.7% -‐ Inter-‐assay CV 8.0%.
• (55.19 ppt): Intra-‐assay CV 2.8% -‐ Inter-‐assay CV was 8.5%.
I’screen AFLA M1 performances
• 10 blank low-‐fat and 10 blank whole yoghurt samples were run with theoretical cut off < 25 ppt. Tecna kit revealed no positives: 100% speciEicity
• The same 20 samples were spiked at 40 ppt of a"latoxin M1, no determination with Tecna kit resulted in a false negative: 100% sensibility
• To test the kit accuracy a low fat and a whole yogurt samples were randomly selected and spiked at 50, 100 and 250 ppt of a"latoxin M1. The resulting mean recovery was 81 ± 11%.
• 50 different incurred cheese samples were analysed by HPLC and by I’Screen M1 ELISA kit. The mean recovery value was: 108 ± 15%.
• A speci"ic sample procedure was implemented for Eirm cheese; the mean recovery obtained by analysin incurred samples was 116 ± 15%
• Z-‐scores obtained in two Pro"iciency tests for cheese were -‐0.21 and 0.17.
Yoghurt Cheese
Quality control It is important to monitor the kit performances. As a producer, Tecna performs a strict quality control before releasing each batch of I’screen AFLA M1 and I’screen AFLA M1 milk, by analysing at least three reference milk materials in three sessions, three replicates each.
Acceptance criteria: 68.3% results must must not exceed 1 standard deviation of the reference sample speci6ications
Intra-‐assay CV < 15% Inter-‐assay CV < 20%
Recovery values of the last 10 batches of I’screen AAla M1 milk
Participation to proficiency tests In order to constantly monitor immunoassays performances, Tecna participates every year to various proAiciency tests from different suppliers.
Since 2005, the average z-‐scores is 0.66 (n=35)
Product overview • Assay principle: competitive immunoenzyme assay • Applications: Raw Milk
• Assay time: 20 minutes • Calibration curve: 50 - 1000 ppt (100 - 2000 ppt
optional)
• Limit Of Detection: 50 ppt
No sample manipulation required
A new FAST screening device for the quantitative
detection of a"latoxin M1 in
milk.
Product overview • Assay principle – competitive immunoenzyme assay • Applications: Raw Milk
• Assay time: 20 minutes • Calibration curve: 50 - 1000 ppt (100 - 2000 ppt
optional)
• Limit Of Detection: 50 ppt
No sample manipulation required
A new FAST screening device for the quantitative
detection of a"latoxin M1 in
milk. B ZERO AFLA M1 is the master curve version of Celer
AFLA M1
Only one standard needed for calibration curve: more analysis
with the same kit
Focus on B ZERO AFLA M1
• The B ZERO line of Tecna kits is dedicated to who implements few analysis
• The B ZERO AFLA M1 ELISA kit leads to quantitative results by means of a VIRTUAL MASTER CURVE –provided within each batch-‐ in 20 minutes only.
• The analyst can save the wells dedicated for each calibrator. It is suf"icient to run only the ZERO STANDARD and the SAMPLES!
In a common immunoassay, at least 5 wells have to be used for calibrators + 1 per sample
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In a B ZERO format, only 1 well must be used to run the ZERO standard
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Why TECNA? • Tecna provides more information to check if each assay was good or not
• Tecna organizes theoretical and practical trainings to its customers
• Tecna supports customers on the road to Quality Assurance of screening results (ISO17025)
Visit us at www.tecnalab.com !