Harmsen E, de Jong J W, Serruys P W
Clin Chim Acta. 1981;115(1):73-84. doi: 10.1016/0009-8981(81)90108-x.
An isocratic high pressure liquid chromatographic system was developed for the estimation of purine nucleosides and oxypurines in blood. Use was made of a reversed-phase column. Nucleotides derived from erythrocytes affected the separation; these compounds were removed with A12O3. The recovery of the whole clean-up procedure exceeded 75%, and the lower detection limit of the assay for blood metabolites was 0.1 mumol/l. In 6 healthy volunteers, non-resting, the following blood concentrations (mean values +/- S.D. in mumol/l) were observed: adenosine (less than 0.1), inosine (0.2 +/- 0.1), hypoxanthine (2.2 +/- 1.3) and xanthine (0.2 +/- 0.1). In plasma and serum the total amount of these compounds was 1.9 and 5.4 times higher, respectively, presumably due to nucleotide breakdown during blood processing. The myocardial arterial-venous differences of blood purine nucleosides, oxypurines and lactate were subsequently measured in blood samples from 13 patients with angiographically documented ischemic heart disease, undergoing an atrial pacing stress test. No significant release of adenosine, inosine and xanthine by the heart was detectable in this study. The myocardial arterial-venous difference of lactate changed from 0.01 +/- 0.03 mmol/l (mean +/- SEM) at rest, to -0.10 +/- 0.04 mmol/l during pacing (p less than 0.002). Relatively larger changes were observed for hypoxanthine: pacing increased the arterial-venous difference from -0.01 +/- 0.05 to -0.51 +/- 0.17 mumol/l (p less than 0.02). We conclude that the high pressure liquid chromatographic assay of blood hypoxanthine is a useful tool in the diagnosis of ischemic heart disease.
开发了一种等度高压液相色谱系统，用于测定血液中的嘌呤核苷和氧嘌呤。使用了反相柱。红细胞衍生的核苷酸会影响分离；这些化合物用Al2O3去除。整个净化过程的回收率超过75%，血液代谢物检测的下限为0.1μmol/l。在6名非静息的健康志愿者中，观察到以下血液浓度（平均值±标准差，单位为μmol/l）：腺苷（小于0.1）、肌苷（0.2±0.1）、次黄嘌呤（2.2±1.3）和黄嘌呤（0.2±0.1）。在血浆和血清中，这些化合物的总量分别高出1.9倍和5.4倍，这可能是由于血液处理过程中核苷酸分解所致。随后，在13名经血管造影证实患有缺血性心脏病且正在接受心房起搏应激试验的患者的血样中，测量了血液嘌呤核苷、氧嘌呤和乳酸的心肌动静脉差值。在本研究中未检测到心脏有明显的腺苷、肌苷和黄嘌呤释放。乳酸的心肌动静脉差值从静息时的0.01±0.03 mmol/l（平均值±标准误）变为起搏时的-0.10±0.04 mmol/l（p<0.002）。次黄嘌呤的变化相对较大：起搏使动静脉差值从-0.01±0.05增加到-0.51±0.17μmol/l（p<0.02）。我们得出结论，血液次黄嘌呤的高压液相色谱测定是诊断缺血性心脏病的一种有用工具。
