EXTRACTION AND MEASUREMENT OF MYOCARDIAL NUCLEOTIDES, NUCLEOSIDES, AND PURINE-BASES BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY
EXTRACTION AND MEASUREMENT OF MYOCARDIAL NUCLEOTIDES, NUCLEOSIDES, AND PURINE-BASES BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY
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DOI：
10.1016/0003-2697(88)90288-6
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发表时间：
1988-03-01
影响因子：
2.9
通讯作者：
ALTSCHULD, RA
中科院分区：
文献类型：
--
作者：
HAMMER, DF;UNVERFERTH, DV;ALTSCHULD, RA
从人肌内膜活检、冷冻夹闭的大鼠心脏和猪冠状窦血浆中提取核苷酸、核苷和嘌呤碱基。高氯酸萃取物用氟里昂-三辛胺中和，并在250 nm处通过反相离子对高效液相色谱法进行分析。为了获得分析小（1-3 mg湿重）组织样品所需的灵敏度，使用小孔径、2.1 mm内径、C18、5 μ m反相柱和0.2ml/min的流速。所有的心肌核苷酸和AMP降解产物在27分钟的总分离时间内与30 mM KH 2 PO 4，7.5 mM磷酸四丁基铵缓冲液，二元pH和乙腈梯度解析。
Nucleotides, nucleosides, and purine bases were extracted from human endomyocardial biopsies, freeze-clamped rat hearts, and porcine coronary sinus plasma. Perchloric acid extracts were neutralized with Freon-trioctylamine and analyzed at 250 nm by reverse-phase ion-pairing high-performance liquid chromatography. To achieve the sensitivity necessary for analyzing small (1-3 mg wet wt) tissue samples, a small-bore, 2.1-mm-internal-diameter, C18, 5-.mu.m reverse-phase column and a flow rate of 0.2 ml/min were used. All of the myocardial nucleotides and AMP degradation products were resolved in a total separation time of 27 min with 30 mM KH2PO4, 7.5 mM tetrabutylammonium phosphate buffers, and binary pH and acetonitrile gradients.