GB 17378.6-2007The specification for marine monitoring - Part 6: Organism analysis (English PDF)

4.1 Collection and preparation of samples

4.1.1 Sample types Shellfish, shrimp and fish. Shellfish gathering R.philippinarum general, clams, corners clams, blue mussel, Perna viridis, clam, razor clam, oyster capuchin Oyster and so on.

4.1.2 Reagents

4.1.2.1 deionized or distilled water equivalent, the trace metal content should be below the detection limit of the analytical method, or were not contaminated seawater.

4.1.2.2 synthetic detergent.

4.1.3 instruments and equipment Instruments and equipment are as follows:

--- Plastic freezer. with ice packs. When mussels for storage and transportation, should have a bottom grid, to avoid sample is immersed in water;

---refrigerator;

--- Cryogenic refrigerator;

--- Plates and plastic ruler. for length measurement;

--- Plastic knives;

--- Glass or ceramic dish. Preparation of samples;

--- Tweezers. plastic products or other suitable materials;

--- High-density polyethylene bags and plastic containers. you Save for frozen samples, before loading the sample, the application of synthetic detergent wash with distilled water Wash;

--- Analytical balance. a sense of the amount of 0.1mg;

--- Plastic bottle;

--- Blade. for collecting samples;

4.1.4 Sampling and Transport

4.1.4.1 Preparations Cleaning freezers synthetic detergent (see 4.1.2.2), high density polyethylene bags, plastic sheeting and feet, large metal knives, scrapers, and then steamed Distilled water or seawater (see 4.1.2.1) rinse.

4.1.4.2 mussel samples collected Mussel samples collected by the cleaning blade from its attachments. Select a sufficient number of intact mussels stored in the freezer. For long-distance transport (hot days over 2h), should be filled in plastic sample mussels Barrel, the clean sea water collected at the scene showered on mussels, samples were kept wet state but can not be immersed in water. If the sample shall be carried out after 24h samples can be stored in the mussel-like high-density plastic bag, press out the air bag, tie the bag or Sealing, this bag and the sample labels together into a polyethylene bag and sealed, stored in cryogenic freezer.

4.1.4.3 sized shrimp and fish samples were collected Required to select a sufficient number of intact biological samples into a clean polyethylene bag, the bag should be prevented punctured. Out of the air bag, Tie the bag or heat sealing, this bag and the sample labels together into another polyethylene bag, and sealed, cold storage. If the storage period is not too long When (hot days, no more than 48h), refrigerators or freezers can be used to store samples.

4.1.4.4 Large fish samples were collected Measure and write down the kind of fish fork length, weight and gender. With a lower metal knife clean at least 100g of muscle tissue, the thickness of at least 5cm, the sample handling, removal of stains or visceral part. Deposit In clean polyethylene bag, and seal out air, this bag and the sample labels together into another polyethylene bag, sealed in a deep freezer In storage. If the retention time is not too long (hot days, no more than 48h), available fridge or freezer storage is a sample.

4.1.4.5 Transport of samples After sample collection, if the long-distance transport, should sample into boxes (or plastic bucket), a package of sample should live without clean sea Washout sprinkled on the sample, the sample holding wet state (can not be immersed in water); if the sample should be carried out after 24h samples, the sample can be placed Polyethylene bag, press out the air bag, tie the bag. This together into bags and sample tags another polyethylene bags (or clean wide mouthed glass Glass bottle), the sealing, frozen. Other Executive in accordance with the provisions of GB 17378.3.

4.1.5 Sample preparation

4.1.5.1 Preparations When necessary, frozen samples in a refrigerator (-2 °C ~ 4 °C) to stand overnight, so that part of thaw for slices. Wash with synthetic detergent (see 4.1.2.2) plastic knives, dishes, forceps, plastic sheeting and feet and weighing of plastic film with distilled water or clean seawater (See 4.1.2.1) rinse. Table cover washed plastic film. Synthetic detergent (see 4.1.2.2) carefully wash their hands after using steam Distilled water or clean seawater (see 4.1.2.1) rinse.

4.1.5.2 shellfish sample preparation Plastic shell with a plastic knife or brush to remove all external attachments. With distilled water or clean seawater (see 4.1.2.1) rinsing each individual sample, let the natural flow of dry, pull the foot wire. With the balance, said individual Total weight, and note the weight. With another knife inserted into the plastic foot stretch silk exports, cutting the muscle closed and open shells. With distilled water or clean seawater (see 4.1.2.1) to wash the soft tissue inside the shell, remove the soft tissue with a plastic knife and tweezers, let the water do. Single individual samples. the soft tissue into the plastic container weighed, reweighed, note the fresh weight. Tightly closed and labeled. Measured with a ruler Shell and record length. More individual samples. the steps above to the soft tissue of at least 10 individuals into a known weight of the plastic container, weighing, note the fresh weight. In homogenized homogenized samples, homogenized sample back in a plastic container, and then weigh and record the total weight, calculated homogenized sample weight. Paste sample label. Each individual organism size should be similar, and their individual lengths were measured and the total weight before removing biological tissue.

4.1.5.3 shrimp sample preparation 4.1.5.3.1 single individual samples Long body with a ruler shrimp, shrimp-like film on the said polyethylene, weighing, note length, and fresh weight. The chest and abdomen and head and tail separated by a plastic knife, carefully remove the viscera from the abdomen. Total removal of the leg. Turn down the abdomen with plastic A stripper knife along the outer edge of the abdominal incision, remove the inside of the outer armor with plastic forceps and discarded. Another with the plastic knife loose abdominal muscles, and muscle removed with tweezers. Check the gonads, records identified gender. Tweezers muscle moved plastic container, weighed and the fresh weight. Tightly closed containers, marked with numbers. The several containers together into the same Plastic bag, together with sample registration list, knot it tightly, cold refrigerator. 4.1.5.3.2 more individual samples Samples were prepared according to the method described above, carefully recording each individual length, fresh weight, abdominal muscle weight and gender. Each sample shall comprise six to On the same sex, similar to the size of individual muscles. The samples were homogenized into a homogenizer abdominal muscles, known weight into a plastic container lid Tight, marked with numbers, weighing, note the fresh weight and other data. Several plastic containers in the same plastic bag, along with sample registration list, knot it tightly and stored at low temperature freezer.

4.1.5.4 Sample Preparation sized fish 4.1.5.4.1 single individual samples Fish fork length measurements and said sample in a polyethylene film and weighed. Identification of gonadal sex, write down the fork length and weight. With distilled water or clean seawater (see 4.1.2.1) washing the fish-like, it would be on the bench with a plastic knife cut pectoral and dorsal fin cutting attachment Nearly from head to tail skin. In the vicinity of the gill and tail, across all fish cut back; abdomen, sides and rear of each gill cut back. Only four knives cut in the side of the fish, and not Cut too deep, so as not to cut offal, meat contamination. Tweezers skin and meat separation, the outer skin to guard against contamination of meat. The plastic knife to separate the muscles and spine with another, and tweezers to remove the muscles. Sheng will be organized in a plastic container, weighed and recorded weight. If not satisfy muscle mass analysis of the amount of the side, the other side of the muscle to take supplements. Tightly closed container, label or mark, make a record and stored at low temperature freezer. 4.1.5.4.2 more individual samples Carefully recording each individual body length, fresh weight, muscle weight. Identification of sex. The number of individuals should not be less than 6, and should be the same sex, same size. Homogenized by a homogenizer fish tissue will be homogenized sample into a known weight of the plastic container, tightly closed, labeled and weighed, homogenized sample weight write down And other data. Placed in cold storage in the refrigerator.

4.1.5.5 Large fish sample preparation If necessary, the samples collected at the scene placed -2 °C ~ 4 °C refrigerator overnight, so that part in order to thaw the slice. With distilled water or clean seawater (see 4.1.2.1) and washed fish samples. The fish samples in a clean work surface, remove the remnants of skin and bone, with Plastic knife to the surface, and then the other to repeat a plastic knife, leaving from pollution muscle tissue. The muscle tissue placed in a plastic container Vessel, capped, labeled, weighed, and the data recorded in the record sheet, stored in a deep freezer in the sample.

4.1.5.6 dry sample preparation The portion of the fresh sample drying step according to

4.1.6.1 or 4.1.6.2, the calculation of wet and dry than to correct moisture content. After drying the sample with Agate mortar and pestle, all over 80 mesh and 100 mesh (180µm ~ 154µm) nylon screen for trace element analysis.

4.1.6 Determination of the dry weight

4.1.6.1 Drying The weighing bottle placed in an oven at 105 °C, 2h, remove the weighing bottle and placed in the dryer to cool 30min. Cover caps, weighed with an analytical balance, note the weight. Take 5g ~ 10g of the biometric sample preparation in the weighing bottle, cover caps, and then said Weight (± 0.5mg) and note the weight. The weighing bottle containing the sample into the semi-open lid 105 °C oven, removed after 24h, placed in the dryer to cool 30min. After the cover cap Weigh and record the weight alleged. Repeat the drying operation, the drying to the weight before and after the two is less than 0.5% of the total weight. Wet and dry weight ratio calculation.

4.1.6.2 lyophilized High levels of lipids in biological samples, can not be dried to constant weight, the application freeze-dried. Weigh accurately 1g ~ 2g the biometric system Preparation of the sample in a clean container freeze-dried samples, weighed after the first freeze-drying 24h. Again lyophilized 24h, reweighed. twice Weigh weight difference should be less than 0.5% by weight of the total. Otherwise, it should continue to be dried to meet the requirements. Wet and dry weight ratio calculation.

4.1.7 Precautions The provisions should note the following.

--- Mussels collected near the laboratory, there is no special transport and storage problems. When transported to the laboratory, samples should be ventilated with mussels Seawater remains moist. Inter collected from intertidal mussels can survive in air for 24h. Mussels should not be placed in water transport;

--- After hand washing, not touching anatomy, should wear gloves; if conditions permit, preparation and sample preparation shall be in a clean strip Element is carried out;

--- When multiple individual sample preparation, should take the same sex, similar to the size of individual organisms. Before removing the soft tissue should be measured separately for each Body length and weight of the body;

--- Sample before digestion, the vessel containing the total weight of the sample and the weight of the storage time of the comparison, can be found in the samples during storage if weightlessness;

--- Trace metal content in different parts of the muscle may differ, so the actual sample information should be recorded in detail as far as possible;

--- A biological sample for organochlorine pesticides and petroleum hydrocarbons measured, sample collection and pre-treatment equipment and reagents make the appropriate corresponding changes Change, should avoid the use of plastic containers containing halogenated reagents;

--- Determination of total mercury and organisms harmful organisms, not dried sample application, the application of wet sample measurement, the results are still dry sample analyte Content to represent.

4.2 regulations and requirements

4.2.1 Analysis of samples drying. drying temperature and time are not specified, refer to 105 °C ± 1 °C, drying 2h.

4.2.2 Standard Solution, all pipettes and flasks test should be performed or capacity correction.

4.2.3 Unless otherwise stated, the purification of the container are to (1 + 3) soaked with a solution of nitric acid 2d ~ 3d after carefully with deionized water shower Wash, dry spare.

4.3 Description

4.3.1 various chemical density (rho) refers to the 20 °C, mass divided by volume, in units of g/mL.

4.3.2 desiccant when not indicate a specific name, refer to color gel.

4.3.3 concentration of the standard elements are formulated refers to the concentration of the element.

4.3.4 does not specify solvent solution is an aqueous solution.

5.1 Atomic Fluorescence Spectrometry

5.1.1 Scope and Application This method is applicable to the determination of total mercury in marine organisms. This method is a method of arbitration.

5.1.2 The principle of the method In nitric acid - perchloric acid digestion system, the total amount of mercury in the organism into mercury ions into solution. With potassium borohydride as a reducing agent Solution of mercury ions to mercury vapor. Argon as a carrier gas of atoms of mercury vapor into the atomic fluorescence spectrometer of atomic reactor, with special Species mercury hollow cathode lamp as the excitation source, the determination of mercury atomic fluorescence intensity.

5.1.3 Reagents and preparation

5.1.3.1 nitric acid (HNO3). rho = 1.42g/mL, pure class distinctions.

5.1.3.2 perchloric acid (HClO4). pure class distinctions.

5.1.3.3 hydrochloric acid (HCl). rho = 1.19g/mL, pure class distinctions.

5.1.3.4 oxalic acid (H2C2O4 · 2H2O).

5.1.3.5 potassium borohydride (KBH4).

5.1.3.6 potassium hydroxide (KOH). pure class distinctions.

5.1.3.7 nitric acid solution (1 + 19).

1 The volume ...