JPS6147560A - Organic acid analysis for plant - Google Patents

Abstract

PURPOSE:To reduce the cleaning time of a column in an organic acid analysis of plant by a fast liquid chromatography, by treating green vegetables in heated water to prevent the elution of protein into a sample through the sampling of the heated water. CONSTITUTION:A specified amount of spinach is emmersed into a fixed heated water to elute oxalic acid thereinto. After the heated water is cooled down to the normal temperature, ethyl acetate is added thereto and oil components are removed from the solution through a layer separation to prepare a sample. The sample is injected into a fast liquid chromatograph apparatus employing an intense anion exchange gel column produced by chemically bonding a quaternary ammonium group to a silica gel to detect oxalic acid. A mobile phase is injected at a high flow velocity without absorption of protein into a column filling agent to wash the column. The same method is applicable for vegetables such as Komatsuna, a kind of Chinese cabbage and organic acid contained can be analyzed.

Description

[Detailed description of the invention] B. Fields of application of technology The present invention relates to a method for analyzing organic acids contained in green vegetables.

Conventional technology The analysis of organic acids such as shu-m contained in spinach, Japanese mustard greens, etc. was obtained by adding dilute sulfuric acid to the sample green vegetables, crushing them with a mixer, etc., and centrifuging the resulting mixture. This was done using high-performance liquid chromatography using the supernatant as a sample.

However, since samples are obtained by crushing green vegetables, proteins in the cells are eluted into the sample, and these proteins are adsorbed to the column packing material, which not only takes time to clean the column, but also causes deterioration of the column. There was a problem with inviting people.

C. Objective: In view of these problems, the present invention provides a method for analyzing organic acids in green vegetables that can prevent elution of proteins into a sample, thereby shortening column cleaning time and extending column life. That's true.

20 Structure of the Invention In other words, the feature of the present invention is that organic acids are eluted by treating green vegetables with hot water, and the organic acids are separated and analyzed using high performance liquid chromatography using this hot water as a sample. At the point.

E. Embodiments The details of the present invention will be explained below based on illustrated embodiments.

Soak a certain amount of spinach in a certain amount of hot water,
A sample is prepared by dissolving the oxalic acid in spinach into hot water, cooling it once to room temperature, adding ethyl acetate, and removing the oil in the solution by layer separation.

When the sample thus obtained was injected into a high performance liquid chromatography device using a strong anion exchange gel column made by chemically bonding quaternary ammonium groups to silica gel, oxalic acid was detected with high sensitivity. Moreover, no protein adsorption occurred on the column packing material. Furthermore, after the analysis was completed, the column was washed by injecting the mobile phase at a high flow rate, but no abnormality was observed in the column.

In this example, oxalic acid contained in spinach was explained as an example, but organic acids contained in vegetables such as Komatsuna can also be analyzed using the same method.

[Example] After washing 58.1 g of spinach with water, it was soaked in hot water of 400 mJ1 at a temperature of 90°C for 5 minutes without pulverizing it.
A sample is prepared by immersing the sample to extract oxalic acid, adding 100 m of ethyl acetate thereto, and removing the oil by layer separation.

The column was packed with a strong anion exchanger in which quaternary ammonium groups were chemically bonded to silica gel, and the mobile phase was 0,000, which had a pH of 4.07 with 85% orthophosphoric acid.
Using a high-performance liquid chromatography device that uses a detector that measures ultraviolet absorbance at 202 nm or 210 nm, dilute the above sample 10 times and inject 5 volumes of 1M KH2PO4 into the column. As a result of the analysis, the peak PL of oxalic acid could be detected after approximately 2 minutes and 40 seconds, as shown in FIG.

On the other hand, for comparison, 2.4 mg of standard oxalic acid was
When the solution dissolved in Jl was analyzed using the above-mentioned high-performance liquid chromatography device, as shown in Figure 2, about 2 minutes 4
It was confirmed that peak P2 of oxalic acid appeared after 0 seconds had elapsed.

This revealed that samples prepared by heat-treating green vegetables with hot water contained a large amount of organic acids and no interfering components.

Effects As explained above, according to the present invention, vegetables to be analyzed are directly treated with hot water to elute organic acids into the hot water, and this hot water is used as a sample, making it easy to prepare samples. Not only can it be prepared quickly, but the proteins in the vegetables undergo thermal condensation and do not elute into the sample, so the sample is clean and does not have a negative impact on the column. Since an exchange gel column is used, the mechanical strength of the packing material is high, and the column can be washed by injecting the mobile phase at high speed, thereby shortening the measurement cycle.

[Brief explanation of the drawing]

The t51 diagram is a chromatogram diagram showing an analysis example according to the present invention, and FIG. 2 is a chromatogram diagram for a standard substance. Figure 1 Figure 2

Claims (1)

[Claims] A process of preparing a sample by immersing the plant to be analyzed in hot water and eluting the organic acid of the plant in the hot water, and separating and detecting the sample by high performance liquid chromatography using a strong anion exchange gel column. A method for analyzing organic acids in plants, which comprises the steps of: