JPS59176672A - Reagent for forming fluorescent derivative for detecting hydroxyl group compound to be used for high-speed liquid chromatograph - Google Patents

Abstract

PURPOSE:To enable sepn. and detection of a hydroxyl group contg. compd. which is separated by a high speed liquid chromatograph with high specificity and high sensitivity by using chloride of coumarinic acid as a reagent for forming a fluorescent deriv. for detecting a hydroxyl group contg. compd. CONSTITUTION:The compd. of the formula obtd. by bringing SOCl2 into reaction with a coumarinic acid is mixed with a sample to be analyzed to convert the hydroxyl group contg. compd. to coumarinate which is separated by each component by using a high speed liquid chromatograph. The respective hydroxyl group contg. compds. are detected with the fluorescence generated by the same. The sepn. and detection of higher alcohol having approximate C atoms such as, for example, C14H29OH, C16H33OH, C18B37OH or thelike are accomplished with high sensitivity without generation of side peaks, etc.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a fluorescent derivatization reagent for detecting hydroxyl compounds, which is represented by the chemical formula [11] and is suitable for high performance liquid chromatography.

Conventionally, dinitrobenzyl chloride has been used as a reagent to detect hydroxyl compounds such as alcohols and steroids separated by high-performance liquid chromatography.
Since the detection must be carried out by ultraviolet absorption, there are problems in that not only the specificity is low but also the sensitivity is low.

In order to solve these problems, various fluorescent derivatization reagents have been proposed in the past, but they have had the problem of detection errors occurring in samples that contain many impurities and have a low concentration of the target component.

In view of these problems, an object of the present invention is to provide a fluorescent derivatization reagent that is not only easy to obtain with high purity but also has high detection sensitivity and specificity.

That is, the present invention is a compound obtained by reacting coumaric acid with thionyl chloride as shown in Formula 1. to convert the target component into a fluorescent substance. When the sample to be analyzed mixed with this derivatization reagent is separated into components by high performance liquid chromatography and detected by a fluorescence detector, fluorescence is specifically generated from the hydroxyl group compounds, and the hydroxyl group compounds are selectively detected. can be detected.

[Experiment example] The above-mentioned reagent itself and the reaction of this reagent with each of three types of higher alcohols C, Hiro H290H, Cab Hg30 H, and C, Hg H3-OH were injected into a high performance liquid chromatograph. What? When these were detected using a fluorescence detector, the results shown in the figure were obtained.

That is, it was found that although the fluorescence of the reagent itself was present, no sub-peaks were generated (a). It was found that the elution time increased at a certain rate in relation to the number of carbon atoms in the higher alcohol, and that the reagent was able to combine with the target substance having a hydroxyl group without producing any by-products, resulting in no sub-peaks ( b), (
c), (d).

As described above, according to the present invention, it is possible to specifically detect with high sensitivity without being affected by sub-peaks such as hydroxyl compounds separated by compound high performance liquid chromatography.

[Brief explanation of the drawing]

The figure shows the results of analysis of hydroxyl compounds using the derivatization reagent of the present invention. Procedural amendment (method) September 1981? 91] 1. Indication of Tenant 1981 Patent Original F1 No. 52258-2 Title of Invention Hydroxylic Acid Used in High Performance Liquid Chroma I Graph (Fluorescence Derivatization Reagent for Compound Detection 3 Compensator 21 Relationship with Matter A Applicant 378 Funaomachi, Kawaramachi-dori Nijo-shita Ru, Nakagyo-ku, Kyoto City (199) Horotsu Manufacturing Co., Ltd. Representative Setsuo Yokochi 4, Agent 〒112 @
03 (815) 810 (1 2-1 Koishikawa, Bunkyo-ku, Tokyo
No. 2 No. 5 Amendment Order 1” dated 11/3 June 8, 1980 (June 2, 1982)
(Delivered on the 8th) 6. Subject details of supplementary IF; 17. Brief description of drawings and drawing 7. Contents of amendments (1) Page 4 of the specification, lines 16 to 17 [Figures]
is...a thing. ” shall be corrected as follows. [Fig. 1 a, b, c, and d are all characteristic diagrams showing the analysis results of water-resistant group compounds using the derivatization reagent of the present invention. (2) Amend the drawing as shown in the attached sheet.

Claims (1)

[Claims] A fluorescent derivatization reagent for detecting hydroxyl compounds used in high performance liquid chromatography, represented by a chemical formula.