ANALYTICAL EVALUATION OF HIGH-ENERGY PHOSPHATE DETERMINATION BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY IN MYOCARDIAL TISSUE
ANALYTICAL EVALUATION OF HIGH-ENERGY PHOSPHATE DETERMINATION BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY IN MYOCARDIAL TISSUE
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DOI：
10.1006/jmcc.1994.1006
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发表时间：
1994-01-01
影响因子：
5
通讯作者：
NIELSEN, TT
中科院分区：
文献类型：
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作者：
BOTKER, HE;KIMOSE, HH;NIELSEN, TT
高效液相色谱法（HPLC）是测定心肌高能磷酸盐（HEP）的既定方法。通过肌内膜活检技术获得的小组织样本中的HEP化合物的定量需要最大的灵敏度而不损害精确度。采用等度离子对反相高效液相色谱法，高灵敏度和精密度，分别在254和210 nm的检测波长下运行分析腺苷酸和肌酸化合物，分别获得。由于脱蛋白后剩余的高氯酸离子和样品中的pH值对腺苷酸和肌酸化合物有各种影响，因此需要不同的样品制备，这是单独运行的进一步原因。在0.42 mol/l高氯酸中机械均质化20 s，确保心肌HEP提取一致。在活检取样后直接制备样品是优选的，因为HEP化合物在-80 ° C下在几天内在组织中不稳定，即使在液氮中已经诱导了初始代谢抑制。在萃取和中和之后。HEP化合物在-20 ° C下稳定长达3个月。
High performance liquid chromatography (HPLC) is an established method for the determination of myocardial high energy phosphates (HEP). Quantification of HEP compounds in small tissue specimens obtained by endomyocardial biopsy technique requires maximal sensitivity without impairment of precision. Employing isocratic ion-pair reversed-phase HPLC, high sensitivity and precision were obtained by running analyses for adenonucleotides and creatine compounds separately at detection wavelengths of 254 and 210 nm, respectively. Further reasons for separate runs were given by the necessity for different sample preparation as remaining perchloric ion after deproteinizing and pH in the samples had various effects on adenonucleotides and creatine compounds. Mechanical homogenization for 20s in 0.42 mol/l perchloric acid ensured a consistent myocardial HEP extraction. Sample preparation directly following biopsy sampling is preferable since HEP compounds were labile in tissue within days at -80°C even though an initial metabolic inhibition in liquid nitrogen had been induce. Following extraction and neutralization. HEP compounds were stable for up to 3 months at -20°C.