DD298348B5 - Method for controlling the quality of stored erythocytes and blood transfusions - Google Patents

Description

For direct control of the blood supply, localized P31 spectroscopy in the high field tomograph (B> 1.5T) is most appropriate.

P-31 NMR spectroscopy in no way affects the life and functioning of red blood cells. The invention will be explained in more detail below with reference to two exemplary embodiments:

Embodiment 1

A blood bank was processed as usual. The determination of the 2.3-DPG content for the quality control of preserved blood was carried out 1 to 6 hours after blood sampling and up to 14 days after storage at 4 ° C. In addition, some blood preparations have been temporarily warmed to room temperatures for several hours to determine the effects of a cold chain interruption during storage. The 31 P NMR spectra were recorded with a PFT (Pulse Fourier Transform) spectrometer using quadrature detection at a resonant frequency of 162 MHz without proton broadband coupling (typical spectral acquisition parameters: sweep width: 36ppm, time domain: 16K data points, number of spectra accumulations: 400, Repetition time per scan: 5s for Ρί / 2 excitation pulses). The erythrocyte preserves to be examined were introduced into the spectrometer. In addition, each erythrocyte preserve was given a double capillary. The outer capillary contained D2O for sample blocking and the inner capillary 85% H3PO4, whose NMR signal was used both as standard for chemical shift (δ = O) and as integration standard for quantitative evaluation. Depending on the medium and the pH value, the 2 phosphorus signals of the 2.3-DPG were found at δ (3Ρ) = 3.1 ± 0.5 and δ (2Ρ) = 2.9 ± 0.5 and both signals became quantitative difference, the difference δ (3Ρ) - δ (2Ρ) was between 0.4 to 0.6 ppm. The measurements were carried out at room temperature

 The results are shown in Table 1.

Embodiment 2

To assess the therapeutic effect, patient blood (blood group A) was fractionated by differential agglutination after transfusion of 4-6 blood groups of 0 blood group at intervals of 10 min, 3,6,12,18,24,48 post-transfusion and in vivo regeneration determined by 2.3 DPG using P-31 NMR spectroscopy. A spectrometer was used as described in Example 1. The prepared blood samples were filled into NMR tubes (outer diameter 10 mm, minimum filling quantity: 3 ml) into which a double capillary was also introduced in each measurement, the same as described in Example 1, D ₂ O for Probenlock and 85% H ₃ PO ₄ as standard and integration standard contained. The examination and evaluation were carried out as in the embodiment 1. The results are shown in Table 2.

Table 1: P-31 NMR measurements of 2.3-DPG content of fresh and preserved red blood cells in blood conserves (dilution factor 2.0)

sample material Hemoglobin- Rel. NMR signal 2.3-DPG 2.3-DPG (Mmol / molHB) salary intensity salary salary (Mmol / l) (Integral)* (Mmol / l) (Mmol / molHb) fresh blood ACD-A-stabilized 8.4 2.02 1.58 188.1 bottle blood ACD-A-stabilized Storage time at 4 ° C 2 days 11.0 2.12 1.65 150.0 5 days 11.4 1.64 1.28 112.3 14 days 11.0 0.00 0.00 0.0 warming bisauf25 ° C for 10 hours (Vorkonservierung 2 days at 4 ⁰ C) 10.0 0.60 0.47 46.5

* Integral (85% H ₃ PO ₄ ) = 1 set (corresponds to 0.39 mmol glycerate ^ .S diphosphate / l)

Table 2: P-31 NMR measurements for in vivo regeneration of 2.3-DPG after transfusion of DPG-depleted blood (dilution factor for the hemolysate 11.93)

time hemoglobin Rel. NMR signal 2.3-DPG 2.3-DPG the measurement salary intensity salary salary (Mmol / l) (Integral)· (Mmol / l) (Mmol / lmolHb) bottle blood 14 days storage 11.0 0.00 0.00 0.00 at 4 ⁰ C Patient's blood, praetransfusionem 4.2 0.20 0.93 221.4 posttransfusionem 10 min 10.6 0.08 0.37 34.9 3 hours. 10.0 0.17 0.79 79.0 6h. 9.6 0.16 0.74 77.1 12h. 8.4 0.19 0.88 104.8 8h. 9.2 0.22 1.02 110.9 24h. 9.1 0.22 1.02 112.1 48h. 4.0 0.12 0.56 140.0

* Integral (85% H ₃ PO ₄ ) = 1 set (corresponds to 0.39mmol glycerate ^ .S diphosphate / l)

Claims (1)

Method for quality control of packed red blood cells and blood transfusions, characterized in that - The RBCs are examined in the appropriate conservation ratio by means of P-31-NMR spectroscopy at the beginning, during and after storage as in-process quality control on their content of intracellular metabolites, in particular on the concentration of 2,3-diphosphoglycerate and adenine nucleotides, and / or After post-transfusion, the patient's own blood after differential agglutination is separated from the circulating preserved red blood cells in the circulation and the in vivo regeneration of 2,3-diphosphoglycerate and the adenine nucleotides are also determined by P-31 NMR spectroscopy. Field of application of the invention The application of the invention relates to the non-invasive quality control of erythrocyte-containing blood conserves and to the evaluation of the therapeutic effect of a transfusion. Characteristic of the known state of the art The currently known technical solutions for the quality assessment and control of erythrocyte-containing blood conserves relate to the characterization of preservation-related damage to the red blood cell. For this purpose, samples must always be taken from the preservation container in order to carry out the investigations. These concern the in vito studies of the life and functioning of the red blood cells, such as. Biochemical parameters (ATP and 2.3-DPG content), pH, extracellular hemoglobin and extracellular potassium ion content), parameters of membrane function (deformability, morphological changes, osmotic resistance, lipid peroxidation, lipid and cholesterol loss, vesiculation, sialic acid cleavage, etc.). ) or the in vivo determination of the actual function of the transfused red blood cells, the measurement of the oxygen transport function and the survival time of these blood cells in the recipient organism. Object of the invention The aim of the invention is to carry out both a noninvasive quality control of packed red blood cells in order to be able to make statements on the life and functionality of the erythrates at any time of preservation, as well as to make an assessment of the therapeutic effect of a transfusion. Explanation of the essence of the invention The object of the invention is to develop a method for the noninvasive determination of the content of 2,3-diphosphoglycerate and the adenine nucleotides in packed red blood cells and for the determination of the quality of blood transfusions. According to the invention this object is achieved in that the erythrocyte preserves in the preservation container (blood bank or blood bag) by P-31-NMR spectroscopy at the beginning, during storage as process-accompanying quality control or immediately before a planned transfusion are tested or transfusion the patient's own blood after differential agglutination is separated from the circulating preserved red blood cells in the circulation and also subjected to the testing by P-31 NMR spectroscopy. Thus, the transfusion effect of any red blood cell-containing blood transfusion can be accurately predicted and controlled, and is particularly significant in patients in need of transfusion, whose cardiac and cerebral compensation capabilities are limited or who require massive transfusions. In these cases, knowledge of the current level of 2.3-DPG of the blood conserved, which may already be less than 10% of baseline after 14 days of storage, is particularly important since the in vivo regeneration of this metabolite is 3-6 hours post-transfusion (pt). only about 40% and hours pt about 80%. It is furthermore advantageous if blood preserves are examined noninvasively by means of P-31 NMR spectroscopy, because they can be used to detect cavitation-related interruptions of the cold chain or other storage or preservation-related damage to the red blood cells without intervention in the sterile preservation container.