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Development of highly sensitive analytical system for biologically active hydroxylated fatty acids with a new fluorescent derivatization reagent, AE-OTf

使用新型荧光衍生化试剂 AE-OTf 开发生物活性羟基化脂肪酸的高灵敏度分析系统

基本信息

批准号：
06556018
负责人：
OHRUI Hiroshi
金额：
$ 5.31万
依托单位：
Tohoku University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1994
资助国家：
日本
起止时间：
1994 至 1996
项目状态：
已结题

项目摘要

We have developed a highly sensitive fluorescent derivatization reagent, AE-OTf [2-(2,3-anthracenedicarboximido)ethyl trifluoromethanesulfonate] which will be the most sensitive reagent in the world among the reagents so far prepared, for carboxylic acids. By using AE-OTf, we have studied the development of analytical system for both fatty acids and hydroxylated fatty acids (such as prostagrandines). At first, we have succeeded in development of the subfemto molar HPLC-analydicalsystem for fatty acids which are usually encountered in foods and living bodies by isocratic elution mode. This system has been improved by JASCO group to the system with automatic sampler. Next, a new LC-LC column switching system for the analysis of complex carboxylic acids with hydroxyl groups such as diarrhetic shellfish toxins ; okadaic acid (OA), dinophysistoxin-1 (DTX-I), and dinophysistoxin-3 (DTX-III) has been developed. By the system each toxin can be determined an pico-gram level. This analytical met … More hod is recommended for the official analytical method of WHO by manyl analytical chemists. The successful results with diarrhetic shellfish toxins, complex carboxylic acids with hydroxyl groups, prompted us to develop an analytical system for prostagrandins. We have succeeded in the determination of standard prostagrandins at pico-gram level and are now studying to develop an automatic analytical system of thromboxane B_2 which will be very importand for crinical use. Since there are many biologically important chiral carboxylic acids, our attention was next directed to the development of a highly sensitive chiral fluorescent labeling reagents for chiral carboxylic acids by modifying AE-OTf. We have developed a new chiral fluorescent labeling reagent, (S)-(+)-2-(2,3-anthracenedicarboxido)-1-propyl trifluoromethanesulfonate (AP-OTf) and an HPLC-analytical system for Beraprost sodium in Plasama. All four isomers can be determined at pico-gram level by the system. As summary, we have developed highly sensitive fluorescent labeling reagents for carboxylic acids and pico-gram level determination HPLC-analytical systems using these reagents Less

我们开发了一种高灵敏的荧光衍生化试剂--AE-OTF[2-(2，3-蒽二酰亚胺)乙基三氟甲磺酸盐]，它将是目前世界上对羧酸反应最灵敏的试剂。利用AE-OTF，我们研究了脂肪酸和羟基脂肪酸(如前列腺素)分析系统的开发。首先，我们通过等梯度洗脱的方式成功地建立了食品和生物体中常见的脂肪酸的亚毫微摩尔高效液相色谱分析系统。该系统由JASCO集团改进为带自动取样器的系统。接下来，开发了一种新的LC-LC柱切换系统，用于分析带羟基的复杂羧酸，如腹泻贝类毒素、冈田酸(OA)、甲藻毒素-1(DTX-I)和甲藻毒素-3(DTX-III)。通过该系统，每种毒素都可以被测定到皮克级。这一分析符合…甘露醇分析化学家对WHO的官方分析方法提出了更多的建议。对腹泻贝类毒素的成功结果促使我们开发了一种前列腺素的分析系统。我们已成功地在微微克级测定了标准前列腺素，目前正在研究开发血栓素B_2的自动分析系统，这将是非常重要的临床应用。由于手性羧酸具有许多重要的生物学意义，因此我们的注意力集中在通过修饰AE-OTF来开发高灵敏的手性羧酸荧光标记试剂上。我们开发了一种新的手性荧光标记试剂(S)-(+)-2-(2，3-anthracenedicarboxido)-1-propyl三氟甲磺酸盐(AP-OTF)，并建立了血浆中贝拉前列素钠的高效液相色谱分析系统。该系统可以在皮克级上测定所有四种异构体。综上所述，我们开发了高灵敏的羧酸荧光标记试剂和微微克水平测定高效液相分析系统，使用这些试剂较少