Stickle D, Bohrer A, Berger R, Morrissey J, Klahr S, Turk J
Mass Spectrometry Resource, Washington University School of Medicine, St. Louis, Missouri, 63110, USA.
Anal Biochem. 1996 Jul 1;238(2):129-36. doi: 10.1006/abio.1996.0265.
A method is described for detection and quantitation of agmatine [4-(aminobutyl)guanidine] by gas chromatography/negative-ion chemical ionization/mass spectrometry after derivatization with hexafluoroacetylacetone. The lower limit of detection of the derivative was about 25 fmol on-column. For quantitative studies of agmatine content in biological samples, a procedure utilizing an internal standard ([15N4]agmatine prepared from [15N4]arginine) and an extraction step had a lower limit of detection of about 15 pmol for total sample content. Agmatine content was measured in rat tissue samples and normalized to protein content. Kidney and spleen samples exhibited the greatest content of agmatine per unit protein mass but agmatine was also detected in pancreatic islets and brain regions (cerebellum and cerebral cortex). On the basis of these measurements, it is estimated that the pancreatic islet intracellular agmatine concentration may exceed 1 microM. The sensitive and highly specific means of detection and quantitation provided by mass spectrometry may be useful in investigating the physiological role of agmatine in mammalian systems.
本文描述了一种通过气相色谱/负离子化学电离/质谱法检测和定量胍丁胺[4-(氨基丁基)胍]的方法，该方法是在胍丁胺用六氟乙酰丙酮衍生化后进行的。该衍生物的柱上检测下限约为25飞摩尔。对于生物样品中胍丁胺含量的定量研究，使用内标（由[15N4]精氨酸制备的[15N4]胍丁胺）和提取步骤的方法，总样品含量的检测下限约为15皮摩尔。测定大鼠组织样品中的胍丁胺含量，并将其标准化为蛋白质含量。肾脏和脾脏样品每单位蛋白质质量中的胍丁胺含量最高，但在胰岛和脑区（小脑和大脑皮层）也检测到了胍丁胺。根据这些测量结果，估计胰岛细胞内胍丁胺浓度可能超过1微摩尔。质谱提供的灵敏且高度特异的检测和定量方法可能有助于研究胍丁胺在哺乳动物系统中的生理作用。
