YY/T 1631.2-2020Compatibility determination of the transfusion sets with blood component - Part 2: Assessment of damage to blood component (English PDF)

3.1 General

This section recommends the use of blood components within the shelf life but near the end of the shelf life, and should meet the requirements of GB 18469. Red blood cell component blood, The collection and storage of platelet concentrate and fresh frozen plasma should be carried out in accordance with 3.2 to 3.4 respectively.

In addition to meeting the requirements of 3.2 to 3.4, the detection indicators of each blood component should also be in the corresponding reference value range or reagent detection range.. In the enclosure.

3.2 Red blood cell component blood

Red blood cell components (RedCelsCompoints, RCC) are stored in the red blood cell maintenance solution. One adult therapeutic dose of red powder Cytoplasmic blood refers to suspended red blood cells separated and prepared from 2 units of whole blood (400mL ± 40mL, without anticoagulant). For blood transfusion The red blood cell component blood in front of the organ usually requires a hemoglobin value > 18g/unit.

3.3 Platelet concentrate

Platelet concentrates (Platelet Concentrates, PCs) were stored at 22°C with shaking. An adult therapeutic dose of platelet concentrate Liquid refers to 1 unit of apheresis platelet or 10 units of mixed platelet concentrate (platelet concentrate prepared from 10 units of whole blood). for The platelet component blood before flowing through the blood transfusion device usually requires the number of platelets >2.4 x 10^11/unit.

3.4 Fresh frozen plasma

Fresh frozen plasma (FreshFrozen Plasma, FFP) was thawed in a 37°C constant temperature water bath. An adult therapeutic dose FFP refers to Fresh frozen plasma prepared from 2 units of whole blood (400mL ± 40mL, without anticoagulant). For the freshness before flowing through the blood transfusion set Frozen plasma usually requires coagulation factor VIII. C >0.7 IU/mL.

Note. "One adult therapeutic dose" is derived from GB 8369, and red blood cell suspension isolated from 1 unit of whole blood (200 mL, without anticoagulant) can also be used Or fresh frozen plasma.

6.1 Sample preparation

Gravity blood transfusion set. Use the infusion stand to adjust the height of the blood transfusion set, so that the blood components flow through the blood transfusion set under a static pressure difference of 1m.

Pressure type blood transfusion set. set the relevant parameters of the pump according to the product manual.

Note. In order to facilitate the analysis of the results, at least 3 sets of tests for each product are tested in parallel.

6.2 Instrument preparation

Initialize the blood cell analyzer, flow cytometer, microplate reader, etc. and perform self-checking for use.

6.3 Preparation of blood components before and after treatment

The stored component blood is taken out, and 10 mL of blood is taken as the component blood before flowing through the blood transfusion set. Follow the instructions in the manufacturer's instructions Methods The puncture of the blood transfusion device is pierced into the blood transfusion socket of the blood bag, and the blood components are caused to flow through the blood transfusion device under gravity or pressure.

Take the emptied unused blood bag and collect the blood components flowing through the blood transfusion set.

7.1 Determination of free hemoglobin

7.1.1 Test principle

Hemoglobin is a colored protein with a molecular weight of 64.458KD. The color develops after the oxidation of o-tolidine, and the color reaction is divided into two phases.

The first-stage oxidation product is blue, the reaction proceeds at about pH 4.6, and the absorption peak is at 630 nm. The second-stage oxidation product is yellow and reacts in The pH is 1.5, and the absorption peak is at 435 nm.

7.1.2 Test equipment and reagents

7.1.2.1 Test equipment

Constant temperature water bath, spectrophotometer, centrifuge, etc.

7.1.2.2 Test reagent

O-tolidine solution (weigh 0.25 g of o-tolidine, dissolve it in 90 mL of glacial acetic acid, and dilute to 100 mL with distilled water.

It can be stored in the refrigerator for 8 to 12 weeks. The color will gradually darken during the storage period. If the color is too dark, it should be reconstituted), 1% hydrogen peroxide solution Solution (freshly prepared), 10% acetic acid solution, hemoglobin (Hb) standard solution.

7.1.4 Detection

Measure according to the method in Appendix G of YY 0329-2009.

Note. Depending on the actual situation, it may be necessary to properly dilute the test sample, and other verified equivalent test methods may be used.

7.1.5 Test results

Calculate and record the free hemoglobin content in the upper plasma of the red blood cell components before and after flowing through the blood transfusion device, and evaluate before flowing through the blood transfusion device After the red blood cell damage.

7.2 Determination of potassium ion (K+)

7.2.1 Test principle

The principle of ion selective electrode method (ISE) is an electrochemical technique that uses the relationship between electrode potential and ion activity to determine ion activity.

The core of this technique is the use of a sensitive membrane that selectively responds to the measured ion. The potassium electrode adopts a neutral carrier membrane containing valinomycin, which is suitable for K+ There is a high degree of selectivity. When the selected ion is in contact with the ISE electrode film, the electromotive force in the potentiometer circuit changes immediately, resulting in Health potential difference. The magnitude of the potential difference is proportional to the activity of potassium ions in the solution and also proportional to the ion concentration.

7.2.2 Test equipment and reagents

7.2.2.1 Test equipment

Electrolyte analyzer.

7.2.2.2 Test reagent

The main components of ISE diluent are bis(2-hydroxyethyl)amino(trihydroxymethyl)methane (Bis-Tris), boric acid, and formaldehyde solution.

The main component of ISE reference electrode solution is potassium chloride.

The main components of the ISE internal standard solution are Bis-Tris, boric acid, sodium chloride, potassium dihydrogen phosphate and sodium bicarbonate.

Note. Please refer to the reagent manual of each manufacturer for details.

7.2.3 Sample preparation

Same as 7.1.3.

7.2.4 Detection

7.2.4.1 Turn on the instrument and clean the pipeline.

7.2.4.2 Instrument calibration Use low- and high-value calibration solutions suitable for this instrument to determine the slope value of the potassium and sodium electrode, and then use a blood sample of known concentration The calibration product is calibrated and verified, the compensation value of the standard curve is calculated, and the instrument automatically adds the compensation value (after daily calibration, the slope and compensation of the standard curve The value should be within the allowable range of the instrument).

7.2.4.3 After the calibration is passed, at least 2 quality control products at the concentration level should be made. After the quality control is passed, the samples should be tested.

7.2.4.4 The measurement results are calculated by the microprocessor in the instrument and then printed out.

7.2.4.5 After using up every day, clean the electrodes and pipes before turning off the machine.

Note. According to actual conditions, other proven equivalent test methods can be used.

7.2.5 Test results

Record the potassium ion content in the upper plasma of the red blood cell components before and after the blood transfusion device, and evaluate the red blood cell damage before and after the blood transfusion Injury situation.

8.1 Supernatant lactate dehydrogenase (LDH)

8.1.1 Test principle

Serum lactate dehydrogenase (LDH) catalyzes the oxidation of L-lactic acid to pyruvate and at the same time transfers hydrogen to oxidized nicotinamide adenine dinucleoside Acid (NAD+), to generate reduced nicotinamide adenine dinucleotide (NADH). NADH has strong absorption at the wavelength of 340nm, while NAD+ has no absorption. In the case of excess substrate, the rate of NADH production is proportional to the serum LDH concentration, so it can be monitored NADH rises to determine the serum LDH activity concentration. The reaction formula of LDH activity determination is as follows. L-lactic acid+NAD+ ->+LDH-> Pyruvate+NADH+H+

8.1.2 Test equipment and reagents

8.1.2.1 Test equipment

Constant temperature water bath, spectrophotometer or microplate reader.

8.1.2.2 Test reagent

LDH kit.

8.1.4 Testing

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