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Hazelnut ELISA Kit

Full Name: Hazelnut ELISA Kit
Sample Type: Food (Cookies, Chocolate, Cereals, Ice Cream)
Sensitivity: 0.3 ppm

BACKGROUND

Hazelnut (also called by the name Corylus avellana) is a member of the birch plant and it is a particularly versatile nut due to the many different ways that it can used. The fraction of proteins present is relatively high (approx. 15%), with many of these proteins responsible for causing an allergy (for example Cor a 9 or Cor a 11). They are also useful in a large number of production processes since they are stable due to its high heat resistant capability. Hazelnuts come from the fruit produced by the plant’s male blossoms after they have shed their pollen to be fertilised by a female flower. The seeds are pushed out of the blossom and are very heavy, which aids in their dispersal through rain or animals eating them. The immature fruits are hard with a thin shell while mature fruits have a thick, watery layer on top and a harder shell underneath.

Hazelnut contain between 4-5% oil by weight, but they can be roughly estimated to have 100 calories from fat. They grow wild on hazel trees which are native to Europe and Western Asia. The nut is about 5-12mm in diameter, with a milk chocolate-coloured shell that is hard at first only becoming thin and watery once it has been removed from the tree. A hazelnut’s nutrition profile consists of around 10% oil (about 20 calories), about 1% protein (4 calories), 63% carbohydrates consisting of mostly starch (96%), 18% water as well as a trace amount of fiber and 3% other nutrients. They are rich in plant sterols which can help lower cholesterol by binding to dietary cholesterol and lowering its absorption. The bile acids produced from the digestion of these plant sterols bind to the protein-constructed particles that make up dietary cholesterol, making them difficult for the body to absorb. They also have a high tannin content which acts as an astringent meaning it binds water causing it to be more likely for body tissue damage.

Hazelnut allergies are often quite serious and in some cases can cause life-threatening reactions. Cross-contamination exists as a consequence of a number of different production process (for example during the production of chocolate), this explains the fact that the presence of residues can’t be excluded in foods.

INTENDED USE

Hazelnut ELISA kit is intended for quantitative measurement of presence of hazelnut in foods (such as cookies, chocolate, ice cream and cereals). This assay has a minimum analytical sensitivity limit of 0.3 ppm.

CONTENT

All reagents supplied need to be stored at 2 °C – 8 °C, unopened reagents will retain reactivity until expiration date. Do not use reagents beyond this date.

  • Microtiter Plate: Coated with anti-hazelnut antibodies.
  • Hazelnut Standards 1-5: Concentration 0, 1, 4, 10, 40 ppm.
  • Conjugate (Anti-Hazelnut-Peroxidase).
  • Substrate Solution (TMB).
  • Stop Solution (0.5 M H2SO4).
  • Extraction and sample dilution buffer (Tris)(10x).
  • Washing Solution (PBS + Tween 20)(10x Concentrate).
  • Instruction Manual.

SENSITIVITY

The minimum detection sensitivity level of hazelnut using current hazelnut ELISA kit was 0.3 ppm. The standard range for this assay is 1.0 – 40.0 ppm.

ASSAY CHARACTERISTICS

– Sensitivity: Limit of detection, LOD (0.03ppm), Limit of quantification, LOQ (1.0ppm)
– Cross Reactivity: Walnut (< 0.0022%). No cross reaction was detected for the following: Wheat, Soy, Brazil nut, Barley, Poppy seed, Pistachio, Rye, Sunflower seed, Macadamia nut, Oats, Pumpkin seed, Chestnut, Buckwheat, Pine nuts, Cocoa, Corn, Cashew nut, Dried milk, Rice, Sesame, Gluten, Pea, Peanut, Lecithin, Chickpea, Almond, Gelatine, Bean, Coconut, Apple.
– Intra-Assay Precision: 7 – 12%
– Inter-Assay Precision: 3 – 13%
– Linearity: 87 – 121%
– Recovery: Cookies (101%), Cereals (99%), Ice cream (90%), Chocolate (83%).

REFERENCES

  1. Development of a sandwich ELISA-type system for the detection and quantification of hazelnut_in model chocolates. Food Chem. (2015) 173: 257-65. Costa J., et al.
  2. Detection of hazelnut in foods using ELISA: challenges related to the detectability in processed foodstuffs. J AOAC Int. (2012) 95 (1): 149-56. Cucu T., et al.
  3. Detection of hazelnuts and almonds using commercial ELISA test kits. Anal Bioanal Chem. (2010) 396 (5): 1939-45. Garber E.A. and Perry J.
  4. Development of a highly sensitive and robust Cor a 9 specific enzyme-linked immunosorbent assay for the detection of hazelnut-traces. Anal Chim Acta. (2011) 708 (1-2): 116-22. Trashin S.A., et al.
  5. Hazelnut Allergens: Molecular Characterization, Detection, and Clinical Relevance. Crit Rev Food Sci Nutr. (2016) 56 (15): 2579-2605. Review. Costa J., et al.

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