The influence of temperature and pH on the dissociation curve of oxyhemoglobin of human blood.
The influence of temperature and pH on the dissociation curve of oxyhemoglobin of human blood.
复制标题
温度和pH值对人体血液氧合血红蛋白解离曲线的影响。
DOI：
10.1080/00365516509083359
复制
发表时间：
1965
影响因子：
2.1
通讯作者：
E. Munson
中科院分区：
文献类型：
--
作者：
P. Astrup;K. Engel;J. Severinghaus;E. Munson
方法和程序为了在不同氧分压下平衡血液样品并测量样品的pH，Astrup和Engel使用为pH和Pcoe测定开发的微量平衡装置（Siggaard-Andersen，Engel，JpLrgensen & Astrup 1960）。在每次测量之前，根据美国国家标准局（R. C. Bates 1962），在不同温度下具有已知的pH值。血液pH测量的准确性优于-C 0.005 pH单位。对于氧饱和度的测量，我们使用Siggaard-Andersen，J@ rgensen和Napraa（1962）描述的分光光度显微技术。在毛细管中离心平衡后的血样后，通过冻融使红细胞溶血，并进行分光光度法。通过使用在纯氮气中通过张力测定法脱氧的血液和通过用亚硫酸二钠还原的血液，分别发现完全还原的血液的点。重现性优于 * 1%饱和度。在实验期间，根据货车Slyke测压法对该方法进行了两次检查。将约25 ml新鲜肝素化血液分成3 ml等份。为了避免强酸酸化过程中的溶血，将等分试样离心。向一式两份的上清液血浆中加入50 μ l以下溶液之一：0.15 M NaCl; 0.41 M HCl; 0.82 M HCl; 1.23 M HCl; 0.41 M NaOH; 0.82 M NaOH; 1.23 M NaOH。然后将细胞重悬。平衡后，所得碱过量从+20至-20 mEq/L血液变化，pH值范围为7.0至7.7。然后用含5%CO的氧-氮混合物平衡50 μ l样品。制备的混合物中的氧百分比为：0.95、1.90、3.80、5.70、7.60和9.50，分别对应于以下氧张力（在38 ℃和760 mm Hg的大气压下）：6.8、13.5、27.0、40.5、53.8和67.5 mm Hg。通过使用由Wosthoff oHG，波鸿，西德国制造的专用气体混合泵，由纯氧、氮气和二氧化碳制备气体混合物。用Lloyd气体分析仪（Gallenkamp，英国）检查了泵的准确性，发现浓度优于0.02%。如果没有另外说明，文本和图中给出的张力均指38 C和760 mm Hg。共进行了812次平衡，血样来自52名正常人。每次平衡后，一式两份测量pH和氧饱和度。在4个温度下进行平衡和pH测量，所述温度被选择为13、23、30和40 ° C。
METHODS AND PROCEDURE For equilibration of blood samples at varying oxygen tensions and measurements of the pH of the samples, Astrup and Engel used the micro equilibration apparatus developed for pH and Pcoe determination (Siggaard-Andersen, Engel, JpLrgensen & Astrup 1960). The pH electrode was adjusted before each measurement by using phosphate buffers according to The National Bureau of Standards (R. C. Bates 1962), with known pH values at different temperatures. The accuracy of the blood pH measurements was better than-C 0.005 pH units. For measurements of the oxygen saturation we used the spectrophotometric micro technique described by Siggaard-Andersen, J@ rgensen & Napraa (1962). After centrifugation of the equilibrated blood samples in capillary tubes the red cells were hemolyzed by freezing and thawing, and spectrophotometry was performed. The point for completely reduced blood was found by using blood deoxygenated by tonometry in pure nitrogen and by reduction with disodium thionite, respectively. The reproducibility was better than* 1 per cent saturation. During the experimental period the method was checked twice against the Van Slyke manometric method. About 25 ml of fresh, heparinized blood was divided in 3 ml aliquots. In order to avoid hemolysis during acidification with strong acid, the aliquots were centrifuged. To the supernatant plasma of duplicate aliquots, 50 pl of one of the following solutions were added: 0.15 M NaCl; 0.41 M HCl; 0.82 M HCl; 1.23 M HCl; 0.41 M NaOH; 0.82 M NaOH; 1.23 M NaOH. The cells were then resuspended. The resulting base excess varied from+ 20 to-20 mEq/L blood, and pH ranged from 7.0 to 7.7 after equilibration. A 50 pl sample was then equilibrated with an oxygen-nitrogen mixture containing 5 per cent COs. The oxygen percentages in the prepared mixtures were: 0.95, 1.90, 3.80, 5.70, 7.60, and 9.50, corresponding to the following oxygen tensions (at 38 C and at a barometric pressure of 760 mm Hg): 6.8, 13.5, 27.0, 40.5, 53.8, and 67.5 mm Hg, respectively. The gas mixtures were prepared from pure oxygen, nitrogen and carbon dioxide by using special gas mixing pumps manufactured by Wosthoff oHG, Bochum, Western Germany. The accuracy of the pumps was checked by using a Lloyd gasanalysis apparatus (Gallenkamp, England) and found to be better than 0.02 per cent concentration. The tensions given in the text and figures all refer to 38 C and 760 mm Hg if not otherwise stated. Altogether 812 equilibrations were carried out, with blood samples from 52 normal persons. After each equilibration the pH and oxygen saturation were measured in duplicate. The equilibrations and the pH measurements were made at 4 temperatures, chosen to 13, 23, 30, and