原料全血30℃保存24 h后制备的悬浮红细胞和血浆质量变化情况的初步研究

Pilot study on the changes of suspended red blood cells and plasma quality prepared from material whole blood stored at 30℃ for 24 hours

目的将全血分别在冷藏(4±2)℃与室温(30±1)℃条件下保存24 h,比较2种温度条件下由全血分离所得的悬浮红细胞及血浆在保存期内的质量变化。方法本次研究纳入20名健康无偿献血者,每人献400 mL全血,将其分成2份,每份200 mL。对照组在冷藏条件下保存;实验组在室温条件下保存,于24 h后将2组全血离心,分离血浆入转移袋,加入50 mL MAP红细胞保存液,制备得到悬浮红细胞。2组悬浮红细胞均放置在冷藏条件下保存,于保存d 1、7、14、21、35分别无菌取样检测Hb、Hct、MCV、溶血率、pH、K^+、葡萄糖、ATP、2,3-DPG、红细胞渗透脆性值等指标;2组血浆分别于全血采集1 h后及全血保存24 h后进行无菌取样,检测Ⅷ因子含量。结果分离所得血浆在全血采集1 h后与保存24 h后Ⅷ因子含量无明显变化。在35 d保存期内,悬浮红细胞的Hb、Hct和MCV值无显著性差异;d 35,研究发现实验组红细胞渗透脆性高于对照组(4.9±0.2 vs 4.7±0.2;P<0.05);2组溶血率、K+值随保存时间延长而增加,分别于d 7、14、21、35;d 1、7、14、21组间有显著性差异(0.29±0.14 vs 0.11±0.09、0.39±0.22 vs 0.18±0.08、0.80±0.22 vs 0.24±0.07、0.93±0.22 vs 0.38±0.11;10.5±1.2 vs 5.4±0.7、21.7±2.0 vs 18.6±2.6、24.8±1.6 vs 22.7±2.7、29.2±2.5 vs 27.1±2.8;P均<0. 05);2组ATP、2,3-DPG、葡萄糖、pH值随保存时间延长而下降,分别于d 14、21、35;d 1、7、14;d 1、7、14、21、35;d 1、7、21组间有显著性差异(4.2±0.2 vs 4.7±0.2、3.6±0.2 vs 4.5±0.2、3.1±0.3 vs 3.9±0.2;1.96±0.44 vs 10.67±0.87、1.33±0.34 vs 3.27±0.88、0.82±0.29 vs 1.49±0.45;23.2±1.1 vs 29.4±1.5、22.0±1.3 vs 27.4±1.1、21.5±0.8 vs 25.1±1.1、18.7±1.5 vs 22.8±1.2、16.1±1.8 vs 19.1±1.7;6.8±0.1 vs 7.0±0.1、6.7±0.1 vs 6.8±0.1、6.4±0.1 vs 6.5±0;P均<0.05)。结论与冷藏(4±2)℃保存条件下相比,全血在室温(30±1)℃保存24 h后进行离心制备,35 d保存期内的溶血率增加,2,3-DPG值下降;而分离所得血浆在全血采集1 h后与保存24 h后Ⅷ因子含量无明显变化。本研究为相关标准或规程的制定提供参考依据或以此来指导临床用血。

Objective The quality changes of suspended red blood cells and plasma, prepared from whole blood(WB) stored at cold storage(4±2)°C or room temperature(30±1)°C for 24 hours, were compared. Methods Each bag of WB(400 mL/bag) collected from 20 healthy voluntary blood donors in Shanghai Blood Center was split into halves(200 ml each) and stored overnight at(4±2)°C(control group) or(30±1)°C(test group) for 24 h. The split WB units of two groups were centrifuged after 24 h holding, and separated into packed red blood cells(RBCs) and plasma. RBCs prepared in both groups were suspended in additive solution MAP(50 mL), thereafter stored at(4±2)℃ and sampled aseptically on day 1, 7, 14, 21, and 35 to evaluate regarding storage parameters, including hemoglobin, Hct, MCV, hemolysis ratio, pH, electrolyte, glucose, ATP, 2,3-DPG, and erythrocyte osmotic fragility. The Factor VIII content of plasma in both groups were determined 1 h after collection and repeated 24 h after storage. Results There was no significant difference in the content of factor VIII of plasma. Hb, Hct and MCV of erythrocyte suspension had little changes during storage between the two groups. The erythrocyte osmotic fragility of test group was higher than control group(4.9±0.2 vs 4.7±0.2;P<0.05). The hemolysis rate and K+ value of two groups was increasing with the prolongation of storage time, and there was significant difference between the two groups,d 7、14、21、35;d 1、7、14、21(0.29±0.14 vs 0.11±0.09、0.39±0.22 vs 0.18±0.08、0.80±0.22 vs 0.24±0.07、0.93±0.22 vs 0.38±0.11;10.5±1.2 vs 5.4±0.7、21.7±2.0 vs 18.6±2.6、24.8±1.6 vs 22.7±2.7、29.2±2.5 vs 27.1±2.8;P<0.05). In addition, the ATP, 2,3-DPG, glucose and pH values of the two groups was decreasing with the prolongation of storage time. The difference between the two groups was obvious,d 14、21、35;d 1、7、14;d 1、7、14、21、35;d 1、7、21(4.2±0.2 vs 4.7±0.2、3.6±0.2 vs 4.5±0.2、3.1±0.3 vs 3.9±0.2;1.96±0.44 vs 10.67±0.87、1.33±0.34 vs 3.27±0.88、0.82±0.29 vs 1.49±0.45;23.2±1.1 vs 29.4±1.5、22.0±1.3 vs 27.4±1.1、21.5±0.8 vs 25.1±1.1、18.7±1.5 vs 22.8±1.2、16.1±1.8 vs 19.1±1.7;6.8±0.1 vs 7.0±0.1、6.7±0.1 vs 6.8±0.1、6.4±0.1 vs 6.5±0;P<0.05). Conclusion Compared with RBCs suspension stored at(4±2)°C for 35 days, substantial decrease in 2,3-DPG and increase in hemolysis rate of erythrocyte were observed in RBCs suspension prepared from(30±1)°C overnight held WB. As to the content of Factor VIII of plasma, no significant change was found between 1 h after collection and 24 h holding. This study provides a reference for the development of relevant standards or protocols and guidance for the clinical practice of blood use.