FISHERY PRODUCTS
ANNEX IISupplementary provisions
ANNEX II FISHERY PRODUCTS SECTION I OBLIGATIONS ON FOOD BUSINESS OPERATORS This Section lays down detailed rules relating to visual inspections to detect parasites in fishery products. CHAPTER I DEFINITIONS 1. ‘Visible parasite’ means a parasite or a group of parasites which has a dimension, colour or texture which is clearly distinguishable from fish tissues. 2. ‘Visual inspection’ means non-destructive examination of fish or fishery products with or without optical means of magnifying and under good light conditions for human vision, including, if necessary, candling. 3. ‘Candling’ means, in respect of flat fish or fish fillets, holding up fish to a light in a darkened room to detect parasites. CHAPTER II VISUAL INSPECTION 1. Visual inspection shall be performed on a representative number of samples. The persons in charge of establishments on land and qualified persons on board factory vessels shall determine the scale and frequency of the inspections by reference to the type of fishery products, their geographical origin and their use. During production, visual inspection of eviscerated fish must be carried out by qualified persons on the abdominal cavity and livers and roes intended for human consumption. Depending on the system of gutting used, the visual inspection must be carried out: (a) in the case of manual evisceration, in a continuous manner by the handler at the time of evisceration and washing; (b) in the case of mechanical evisceration, by sampling carried out on a representative number of samples being not less than 10 fish per batch. 2. The visual inspection of fish fillets or fish slices must be carried out by qualified persons during trimming and after filleting or slicing. Where an individual examination is not possible because of the size of the fillets or the filleting operations, a sampling plan must be drawn up and kept available for the competent authority in accordance with Chapter II(4) of Section VIII of Annex III to Regulation (EC) No 853/2004. Where candling of fillets is necessary from a technical viewpoint, it must be included in the sampling plan. SECTION II OBLIGATIONS ON THE COMPETENT AUTHORITIES CHAPTER I TOTAL VOLATILE BASIC NITROGEN (TVB-N) LIMIT VALUES FOR CERTAIN CATEGORIES OF FISHERY PRODUCTS AND ANALYSIS METHODS TO BE USED 1. Unprocessed fishery products belonging to the species categories listed in Chapter II shall be regarded as unfit for human consumption where organoleptic assessment has raised doubts as to their freshness and chemical checks reveal that the following TVB-N limits are exceeded: (a) 25 mg of nitrogen/100 g of flesh for the species referred to in point 1 of Chapter II; (b) 30 mg of nitrogen/100 g of flesh for the species referred to in point 2 of Chapter II (c) 35 mg of nitrogen/100 g of flesh for the species referred to in point 3 of Chapter II. The reference method to be used for checking the TVB-N limit involves distilling an extract deproteinised by perchloric acid as set out in Chapter III. 2. Distillation as referred to in point 1 must be performed using apparatus which complies with the diagram in Chapter IV. 3. The routine methods which may be used to check the TVB-N limit are as follows: — microdiffusion method described by Conway and Byrne (1933), — direct distillation method described by Antonacopoulos (1968), — distillation of an extract deproteinised by trichloracetic acid (Codex Alimentarius Committee on Fish and Fishery Products (1968). 4. The sample must consist of about 100 g of flesh, taken from at least three different points and mixed together by grinding. Member States shall recommend that official laboratories use, as a matter of routine, the reference method referred to above. Where the results are dubious or in the event of dispute regarding the results of analysis performed by one of the routine methods, only the reference method may be used to check the results. CHAPTER II SPECIES CATEGORIES FOR WHICH TVB-N LIMIT VALUES ARE FIXED 1. Sebastes spp., Helicolenus dactylopterus , Sebastichthys capensis . 2. Species belonging to the Pleuronectidae family (with the exception of halibut: Hippoglossus spp.). 3. Salmo salar, species belonging to the Merlucciidae family, species belonging to the Gadidae family. CHAPTER III DETERMINATION OF THE CONCENTRATION OF TVB-N IN FISH AND FISHERY PRODUCTS Reference procedure 1. Purpose and area of application This method describes a reference procedure for identifying the nitrogen concentration of TVB-N in fish and fishery products. This procedure is applicable at TVB-N concentrations of 5 mg/100 g to at least 100 mg/100 g. 2. Definition ‘TVB-N concentration’ means the nitrogen content of volatile nitrogenous bases as determined by the procedure described. The concentration shall be expressed in mg/100 g. 3. Brief description The volatile nitrogenous bases are extracted from a sample using a solution of 0,6 mol perchloric acid. After alkalinisation the extract undergoes steam distillation and the volatile base components are absorbed by an acid receiver. The TVB-N concentration is determined by titration of the absorbed bases. 4. Chemicals Unless otherwise indicated, reagent-grade chemicals should be used. The water used must be either distilled or demineralised and of at least the same purity. Unless otherwise indicated, ‘solution’ means an aqueous solution as follows: (a) perchloric acid solution = 6 g/100 ml; (b) sodium hydroxide solution = 20 g/100 ml; (c) hydrochloric acid standard solution 0,05 mol/l ((0,05 N), Note: When using an automatic distillation apparatus, titration should take place with a hydrochloric acid standard solution of 0,01 mol/l ((0,01 N); (d) boric acid solution = 3 g/100 ml; (e) silicone anti-foaming agent; (f) phenolphtalein solution = 1 g/100 ml 95 % ethanol; (g) indicator solution (Tashiro Mixed Indicator) 2 g methyl-red and 1 g methylene-blue are dissolved in 1 000 ml 95 % ethanol. 5. Instruments and accessories (a) A meat grinder to produce a sufficiently homogenous fish mince. (b) High-speed blender with a speed of between 8 000 and 45 000 revolutions/min. (c) Fluted filter, diameter 150 mm, quick-filtering. (d) Burette, 5 ml, graduated to 0,01 ml. (e) Apparatus for steam distillation. The apparatus must be able to regulate various amounts of steam and produce a constant amount of steam over a given period of time. It must ensure that during the addition of alkalising substances the resulting free bases cannot escape. 6. Execution Warning: When working with perchloric acid, which is strongly corrosive, necessary caution and preventive measures should be taken. The samples should, if at all possible, be prepared as soon as possible after their arrival, in accordance with the following instructions: (a) Preparing the sample The sample to be analysed should be ground carefully using a meat grinder as described in point 5(a). Exactly 10 g + 0,1 g of the ground sample is weighed out into a suitable container. This is mixed with 90,0 ml perchloric acid solution as specified in point 4(a), homogenised for two minutes with a blender as described in point 5(b), and then filtered. The extract thereby obtained can be kept for at least seven days at a temperature of between approximately 2 o C and 6 o C; (b) Steam distillation 50,0 ml of the extract obtained in accordance with point (a) is put into an apparatus for steam distillation as described in point 5(e). For a later check on the extract's alkalinisation, several drops of phenolphtalein as specified in point 4(f) are added. After adding a few drops of silicone anti-foaming agent, 6,5 ml of sodium hydroxide solution as specified in point 4(b) is added to the extract and steam distillation begins immediately. The steam distillation is regulated so that around 100 ml of distillate is produced in 10 minutes. The distillation outflow tube is submerged in a receiver with 100 ml boric acid solution as specified in point 4(d), to which three to five drops of the indicator solution as described in point 4(g) have been added. After exactly 10 minutes, distillation is ended. The distillation outflow tube is removed from the receiver and washed out with water. The volatile bases contained in the receiver solution are determined by titration with standard hydrochloric solution as specified in point 4(c). The pH of the end point should be 5,0 + 0,1. (c) Titration Duplicate analyses are required. The applied method is correct if the difference between the duplicates is not greater than 2 mg/100 g. (d) Blank A blind test is carried out as described in point (b). Instead of the extract, 50,0 ml perchloric acid solution as specified in point 4(a) is used. 7. Calculation of TVB-N By titration of the receiver solution with hydrochloric acid as in point 4(c), the TVB-N concentration is calculated using the following equation: V 1 = Volume of 0,01 mol hydrochloric acid solution in ml for sample V 0 = Volume of 0,01 mol hydrochloric acid solution in ml for blank M = Weight of sample in g. Remarks 1. Duplicate analyses are required. The applied method is correct if the difference between duplicates is not greater than 2 mg/100 g. 2. Check the equipment by distilling solutions of NH 4 Cl equivalent to 50 mg TVB-N/100 g. 3. Standard deviation of reproducibility S r = 1,20 mg/100 g. Standard deviation of comparability S R = 2,50mg/100 g. CHAPTER IV TVB-N STEAM DISTILLATION APPARATUS Text of image Steam generator Distillation tube Steam injection tube Sample extract Cooler Cool water End of condenser Flask or beaker (Boric acid)