Authorization of the Ministry of Agriculture as an official laboratory in the field of food and animal feed testing Authorization of the Ministry of Agriculture as an official laboratory in the field of veterinary services

Analysis of pesticide residues

In order to test the health correctness of food and animal feed, a testing of pesticide residues with the multiresidue method is being carried out in our laboratory, and over 400 active substances of pesticides are being quantified.

The multiresidue method is based on multiple mass spectometry related to liquid and gas chromatography (LC-MS/MS and GC-MS/MS), which defines the different groups of chemical pesticides – (organophosphorous, organochlorine, triazine, strobilurine, nicotine, carbamates, avermectins etc.) which also have different uses (insecticides, fungicides, acaricides, herbicides, limacides etc.)

Fungicides from the group of dithiocarbamates (propineb, thiram, ziram, mancozeb, metiram) are determined with the CS2 technique of gas chromatography, with the sampler for volatile samples (headspace autosampler) and the electron-capture detector (HSS-GC-ECD).

Polar pesticides (glyphosate, mepiquat, chlormequat) and their metabolites are determined with a method recommended by the European Community Reference Laboratory (QuPPe method), with the technique LC-MS/MS.

Copper compounds are determined as copper with the atomic absorption spectometry technique (AAS).

Pesticide residues from the group of phenoxycarboxylic acids (clodinafop, haloxyfop, fluazifop and kizalofop) are determined with a method which includes hydrolysis component. Analysis of pesticide residues without hydrolysis can not comply fully the requirements of Regulation (EC) No. 396/2005 on maximum residue levels of pesticides in or on food and feed of plant and animal origin, which prescribes definitions of pesticide residues.

Analytical methods are accredited and comply fully with the legislational requirements, primarily the EU Regulation no. 396/2005.

Through the project IPA2007/HR/16IPO/001-040401 – „Control of the quality and health safety of the food we eat”, our laboratory has improved the possibility of pesticide analysis by acquiry of the instrument GC-MS/MS.

List of pesticides that we currently can determine in our laboratory


Heavy metals are being tested in our laboratory – leadcadmium, mercury and tin as defined in the EU Regulation no. 1881/2006.

Lead – it is being determined in milk, infant formulas, meat, fish, shellfish and seafood, cereals, vegetables, mushrooms, fruits, fats and oils, fruit juices, wine.

Cadmium– it is being determined mainly in all sorts of meat, fish, shellfish and seafood, cephalopods, cereals, vegetables and soybeans.

Mercury – maximum permitted level (MPL) value is defined mainly for fish products.

Tin– it is being determined mainly in canned food (packed in tin packaging).

Apart from heavy metals, we also test the content of other metals and non-metals – ironsodiumpotassiumcalciumlithiumbarium  and others.

The tests are being conducted with the following instruments:

  • Atomic absorption spectrophotometer – Flameless technique (FL-AAS) novAA 350
  • Atomic absorption spectrophotometer – Hydride system (HS-AAS) HS 50
  • Atomic absorption spectrophotometer – Graphite furnace (GF-AAS) Aanalyst 600
  • Microwave Multiwave 3000

aflatoksini 1

Mycotoxins are toxic secondary metabolites produced by molds. They occur in almost all agricultural products, both raw and processed. Apart from posing a risk to human health, their occurance leads to significant economic losses in the field of food and animal feed. About 300 mycotoxins are known, out of which 20 contaminate the agricultural products.

The Act on Contaminants (Official Gazette of the Republic of Croatia “Narodne novine” no. 39/13) and EU Regulation no. 1881/2006 on maximum levels for contaminants in food prescribe the maximum levels of certain mycotoxins which are considered as a potential risk to human health.

Mycotoxins are being tested in our laboratory using the techniques LC-MS/MSHPLC and Elisa:

  • aflatoxin B1 and sum of aflatoxins B1, B2, G1 i G2
  • aflatoxin M1
  • ochratoxin A
  • deoxynivaneol
  • zearalenone
  • fumonisin (sum of fumonisins B1 and B2)
  • T-2 and HT-2 toxin (recommendation)


Acrylamide is a toxicant that occurs via natural processes in the food, during frying or baking at high temperatures. It occurs via the Maillard reactions – a set of chemical reactions between amino acids and sugars such as glucose, fructose and lactose.

The heat launches a series of changes that result in browning of the food and the formation of a characteristic flavor and aroma. The amount of acrylamide that will develop in the food is influenced by several factors such as the type of food, temperature and the time of exposure to high temperatures. In general, foods which contain a large amount of starch and which are prepared at a high temperature for longer contain the most acrylamide. Also, foods rich with the amino acid asparagine may contain high levels of acrylamide after preparation, because it has shown that this amino acid is easily transformed into an acrylamide because of its similar chemical structure.

The maximum permitted level of acrylamides in foods has not been prescribed yet. The Commission Recommendation from November 8th 2013 on testing levels of acrylamide in foods (2013/647/EU) delivers indicative values of acrylamide levels in foods.

Acrylamide is being determined with the LC-MS/MS technique.

3-MCPD (3-monochloropropane-1,2-diol) is a contaminant which occurs as a product of the processing of food and which contains high levels of fat and salt, which are exposed to high temperatures during production. Apart from 3-MCPD, 2-MCPD and glycidyl fatty acids can also develop during production. The EU Regulation no. 1881/2006 defines the maximum permitted level of 3-MCPD in hydrolyzed vegetable proteins and soy sauce.

3-MCPD is being determined in our laboratory with the GC-MS/MS technique.

Nitrates are contaminants whose increased value in food is the result of an intensive use of nitrogen fertilisers and livestock farming. Their increased concentration in the organism can lead to an increased production of nitrites in the digestive tract, as well as ultimately lead to the formation of cancer-causing nitrosamines.

The main source of nitrates in the human diet are leafy vegetables (spinach and lettuce).  The EU Regulation 1881/2006 defines their maximum permitted level in vegetables.

Nitrates are being determined in our laboratory with the HPLC technique.


Microbiological testings of food determine the presence of pathogenic or potentially pathogenic microorganisms and/or their toxins, which can negatively impact on human and animal health.

The provisions of the Act on food hygiene and microbiological criteria for foodstuffs (Official Gazette of the Republic of Croatia “Narodne novine” no. 81/2013) and The EU Regulative no. 2073/2005 prescribe the microbiological criteria for food in order to ensure “safe food”, based on the HACCP system and other hygiene control measures.

The most food-borne agents that cause diseases are:

  • Salmonella sp.
  • Listeria monocytogenes
  • Escherichia coli
  • Staphylococcus aureus
  • Bacillus cereus

Spoilage microorganisms:

  • aerobic mesophilic bacteria
  • yeasts and molds
  • aerobic spore-forming bacteria

Histamine is a compound from the group of biogenous amines that generates via the decarboxylation of the amino acid histidine. It is being created in fish, especially pelagic ones, which is exposed to room temperature for a longer period of time and is considered to be an

indicator for the spoilage of fish due to improper storage. The limit value for histamine is prescribed by the EU Regulative no. 2073/2005 from November 15th 2005 about the microbiological criteria for food, primarily for fish and fish products from the species Scombridae, Clupeidae, Engraulidae, Coryfenidae, Pomatomidae and Scombresosidae.

Histamine is being determined in our laboratory with he HPLC technique, a method which is being prescribed by the EU Regulative no. 2073/2005


Genetically modified organisms are organisms that have an artificially modified genetic structure. Genetic modifications we encounter most commonly are found in corn, soybeans, rice, oilseed rape, potatoes, tomatoes, sugar beet and cotton,

The aim of these modifications is: resistance/tolerance to pests and herbicides (glyphosate, glufosinate), resistance to viral infection…

The most common soy worldwide is Roundup Ready soy, that has gene of an enzyme from bacterium A. tumefaciens. This modification results with glyphosate tolerance.

The  EU Regulation no. 1829/2003  defines GMO legislative for food and feed. Products need to be labeled if they contain more than 0.9% of GMO.

In the Department for Molecular Diagnostics, presence of GMOs is being conducted using Real Time PCR method on Stratagene Mx3005P. This procedure involves:

  1. DNA extraction
  1. Qualitative method – screening – detection of specific transgene elements: p35S (promoter sequence of Cauliflower mosaic virus), tNOS (terminator of nopaline synthase from A. tumefaciens), pFMV (promoter from Figwort mosaic virus), TE9 (terminator of the pea ribulose-1,5-bisphosphate carboxylase small subunit (rbcS) E9 gene), MON87701 (MON-87701-2)
  1. If screening is positive for certain elements, identification (GTS 40-3-2, MON89788, A2704-12, A5547-127) and quantification of GMO events have to be conducted (expressed as %).


Tests of basic food ingredients are being conducted in our laboratory, such as the testing of contents of proteins,fatscarbohydratesfibersashesmoisturesalts etc. The abovementioned tests are being conducted for the purpose of declaring the nutritional value of food products and the creation of a nutritive declaration, which complies with the EU Regulative no. 1169/2011 about informing consumers about food, to secure informed food choices and to use and utilize food in a safe manner.



Basic content:

  • Energy value (kJ/kcal)
  • Fats / of which saturated fats
  • Carbohydrates / of which sugars
  • Proteins
  • Salt


  • mono- and polyunsaturated fatty acids
  • starch
  • fibers
  • vitamins and minerals

TESTING OF CARBOHYDRATES Carbohydrates are, besides proteins and fats, the most important nutritients in food. They are divided into:

  • sugars (mainly monosaccharides and disaccharides)
  • starch and other digestible polysaccharides
  • dietary fibers


Allergens are substances that cause the reaction of the immune system of the organism, which can cause swelling and itching in the mouth, skin changes and even a life threatening anaphylactic shock. The most common allergens are – peanutsnuts (almonds, hazelnuts, walnuts, Brazil nuts, cashews, pistachios, pine nuts, chestnuts), soybeansmilkeggscrustaceans (shrimps, prawns, lobster, crayfish), fishwheatshellfish and other molluscs.

The tests on allergens are being conducted in our laboratory by using the ELISA test. The allergens must be pointed out on the declaration (a different type font).


According to the EU Regulation no. 1333/2008 and the Act on the implementation of the Regulations of the European Union in the field of food additives, food enzymes and flavorings (Official Gazette of the Republic of Croatia “Narodne novine” no. 39/13), a “food additive” is any substance that is usually by itself not enjoyed as food and that is usually not used as a typical food ingredient, regardless of whether it has food value or not, and the intentional adding of which to food for technological reasons in the production, processing, preparation, treatment, packaging, transportation or storing of such food results in, or could reasonably be expected to have, the effect that it or its byproducts become, directly or indirectly, a component of such foods.

The content of following additives is being determined in our laboratory:

preservatives                              E200-E203    sorbic acid and its salts

                                                       E210-E213    benzoic acid and its salts

                                                       E220-E228   sulfur dioxide and sulphites expressed as SO2

                                                       E235             natamycin

                                                       E249-E250   nitrites

                                                       E251-E252   nitrates


antioxidants                                    E310             propylgallate



sweeteners                                    E950               acesulfame potassium

                                                      E951               aspartame

                                                      E954               saccharin and its salts

colors                                            E102               tartrazine

                                                     E104               quinoline yellow

                                                     E110               sunset yellow

                                                     E120               carmine

                                                     E124               ponceau 4R

                                                     E129               allura red

                                                     E132               indigotine

                                                     E133               brilliant blue FCF

                                                     E142               sulphur green

                                                     E151               brilliant black BN

humectants   E452               polyphosphates