Derivatizing reagent of polyhydroxy compound

Abstract

The invention discloses a derivatizing reagent of polyhydroxy compound. The reagent is composed of DMSO, 1-methylimidazole and acetic anhydride. The reagent of the invention can carry out acetylation derivatization reaction on the basis of maintaining native conformation of simple sugar, single derivatization product can be obtained after derivatization reaction, and qualitative or quantitative analysis can be accurately carried out on various polyhydroxy compounds, thereby avoiding the interference of various derivatization products in analysis.

Description

technical field [0001] The invention relates to a polyhydroxy compound derivatization reagent, in particular to a sugar acetylation derivatization reagent. Background technique [0002] The monosaccharides existing in nature can be divided into neutral monosaccharides (aldose and ketose), acidic monosaccharides (uronic acid), sugar alcohols, sugar amines and thiosugars, etc., which are analyzed by gas chromatography Occasionally, derivatization is required due to the presence of its hydroxyl groups. At present, the methods commonly used for the derivatization of monosaccharides mainly include methyl silylation (trimethyl silylation), acylation (sugar oxime acetylation and sugar alcohol acetylation) and methylation. Among the currently commonly used monosaccharide derivatization methods, the silanization method requires a strict anhydrous operating environment to prevent experimental failure because the derivatization reagent and derivatization product are easily decomposed ...

AI Technical Summary

Problems solved by technology

This method can be better used for the derivatization analysis of aldose and sugar alcohol, but for ketose, fructose, etc., it cannot be effectively oximated and derivatized, because hydroxylamine hydrochloride cannot completely oximate the ketone group, resulting in Not a single but multiple product coexistence phenomenon (such as figure 2 ) makes this method unable to detect ketose

[0006] 2. There are problems in the mechanism of acetylation derivatization of sugar alcohols

[0007] This method has a good acetylation derivatization effect on sugar alcohols, but for aldose and ketose, it needs to be reduced to the corresponding sugar alcohol with sodium borohydride before derivatization, resulting in the inaccurate analysis and determination of ketose , aldose and alcohol sugars exist at the same time

[0008] (1) Aldose problem: use sodium borohydride to reduce the aldehyde group or ketone group of the sugar to a hydroxyl group, and then acetylate it

Since the aldose-reduced sugar alcohols cover up the sugar alcohols that actually exist in nature and have important physiological functions, it not only makes it impossible to accurately determine the natural aldose content, but also directly leads to the inability to detect these naturally occurring sugar alcohols

[0009] (2) The problem of ketose --- this method cannot directly detect ketose

[0012] 2. Commonly used derivatization methods have problems in derivatization reaction conditions --- high temperature requirements, long time, and inappropriate reaction solvents

[0017] 2. Commonly used derivatization methods require reaction time

[0020] 3) In the sugar alcohol acetylation method, the sodium borohydride reduction reaction needs at least 60-90 minutes, and there are many reaction steps and chemical reagents, wasting valuable time [Blakeney A.B., Harris P.J., Henry R.J.and Stone B.A. (1983) A simple and rapid preparation of alditol acetates for monosaccharide analysis. Carbohydrate Research 113, 291-299.〕

[0021] 3. The problem of solvents used in common derivatization methods

[0022] Need to use anhydrous pyridine as reaction solvent in the silanization method, and isomerization phenomenon (such as Figure 7 ), a phenomenon of multiple chromatographic peaks of monosaccharides appears during chromatographic separation, which affects the qualitative and quantitative analysis of monosaccharides

[0023] 3. Problems in the stability of commonly used derivatization reagents and derivatization products

This method is greatly affected by moisture, and needs to be operated under strict anhydrous conditions, otherwise the methyl silylation reagent and its derivative products are easily degraded by water, which will affect the authenticity of the analysis results [Marie-christineet. (1997) Determination of Carbohydrates in two ferrlitic soil: analysis by capiliary gas chromatography after derivatization by silylation.SoilBiol.Biochem.Vol.29, No.9 / 10, pp.1585-1589〕

[0026] 2) In the sugar alcohol acetylation method, sodium borohydride is easy to absorb moisture and deteriorate, and the storage conditions are relatively high

[0027] 4. Problems in the qualitative and quantitative analysis of monosaccharides by common derivatization methods

This will inevitably lead to isomers of monosaccharides naturally present in plant samples due to the use of pyridine as a solvent when detecting plant samples, which not only affects the accurate qualitative and quantitative analysis of the monosaccharides, but also affects the chromatogram produced by isomers. The peak occupies precious time in chromatographic separation and affects the separation of other monosaccharide chromatographic peaks [Marie-christine et. (1997) Determination of Carbohydrates in two ferrlitic soil: analysis by capiliary gas chromatography after derivatization by silylation.SoilBiol.Biochem.Vol.29 , No.9 / 10, pp.1585-1589]

[0029] 2. In the sugar alcohol acetylation method, since all aldoses and ketoses are reduced to sugar alcohols for detection, it is impossible to qualitatively quantify sugar alcohols with important physiological significance, which reduces the detection range of monosaccharides and their derivatives

[0030] 3. In the sugar oxime acetylation method, hydroxylamine hydrochloride cannot oximate the ketose group of ketose, which makes the method unable to effectively detect ketose [Ye Fangting, Yan Xiaojun, etc. (2005) used differential derivatization to simultaneously analyze aldose and ketone Sugar, Analytical Chemistry, Issue 11, Volume 33, 1569~1572]

[0032] 1) At present, acidic monosaccharides such as uronic acid and sugar amines such as glucosamine can only be derivatized by silanization method, but due to the harsh conditions of silanization reaction, silanization products are easy to decompose when exposed to water, resulting in reliable qualitative and quantitative results. discounted sex

[0033] 2) Sugar alcohol acetylation and sugar oxime acetylation are relatively easy compared to the conditions of silylation, and the acetylated products are relatively stable under dry conditions and can be stored for a long time, but these two methods cannot convert acidic monosaccharides Effective derivatization of sugar amines, limiting the scope of use of these methods [Marie-christine et. (1997) Determination of Carbohydrates in two ferrlitic soil: analysis by capiliary gas chromatography afterderivatization by silylation. Soil Biol. Biochem. Vol.29, No.9 / 10, pp. 1585-1589]

[0034] 5. Commonly used derivatization methods have a problem with the demand for testing materials

[0035] Each analysis of traditional monosaccharide analysis technology usually requires 0.5-1.0g of biological sample material, which will undoubtedly cause great damage to the study of trace materials or endangered organisms

[0036] 6. The impact of commonly used derivatization methods on the environment

In case of high heat, the internal pressure of the container will increase, and there is a danger of cracking and explosion

[0038] 2. The acetylation of sugar oxime uses hydroxylamine hydrochloride as the oximation reagent, but hydroxylamine hydrochloride is highly toxic and irritating to the skin

[0040] From the above analysis, it can be seen that commonly used derivatization solvents, oximation reagents, and reducing reagents will cause serious harm to the environment and human health. Chemical reagents have very important application value

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