Bossant M J, Ninio E, Delautier D, Bessou G, Trouvin J H, Benveniste J
INSERM U200, Clamart, France.
Anal Biochem. 1989 Nov 1;182(2):419-23. doi: 10.1016/0003-2697(89)90617-9.
A new method to quantitate paf-acether (paf) was developed. It is based on the measurement of serotonin released from washed rabbit platelets challenged with paf. Platelets (1 X 10(8)/ml) were exposed with or without stirring to various concentrations of paf (26-130 pM) at 37 degrees C or at room temperature. Supernatants were submitted to a 4-min liquid chromatography run and serotonin was measured by electrochemical detection. We quantitated paf from three different biological sources, human neutrophils, mouse peritoneal macrophages, and cultured mast cells, comparing a classical method, i.e., platelet aggregation with the electrochemical detection of endogenous serotonin. We found similar results since, when compared with the aggregation method, the results differed by 12 to 47%. The sensitivity of both methods was 26 pM. The between-day variation coefficient was 23 and 14% (n = 12) for the aggregation method and the serotonin release, respectively, whereas the within-day variation coefficient for serotonin quantitation was less than 5% (n = 12). The superiority of the new method lies in its simplicity, the economy of platelets, and its possibility of automation. It can be applied to any agonist or any mechanism capable of releasing serotonin from platelets and more generally when a simple and fast method for measuring serotonin is desirable.
开发了一种定量血小板活化因子（PAF）的新方法。该方法基于对用PAF刺激的洗涤兔血小板释放的血清素的测量。血小板（1×10⁸/ml）在37℃或室温下，在搅拌或不搅拌的情况下，暴露于不同浓度的PAF（26 - 130 pM）。将上清液进行4分钟的液相色谱分析，并通过电化学检测测量血清素。我们对来自三种不同生物来源的PAF进行了定量，即人中性粒细胞、小鼠腹腔巨噬细胞和培养的肥大细胞，将一种经典方法（即血小板聚集结合内源性血清素的电化学检测）与之进行比较。我们发现结果相似，因为与聚集法相比，结果相差12%至47%。两种方法的灵敏度均为26 pM。聚集法和血清素释放法的日间变异系数分别为23%和14%（n = 12），而血清素定量的日内变异系数小于5%（n = 12）。新方法的优势在于其简单性、血小板用量少以及可自动化的可能性。它可应用于任何能从血小板释放血清素的激动剂或任何机制，更普遍地说，当需要一种简单快速的血清素测量方法时均可应用。
