Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Kit for measuring oxalic acid content of biological sample by enzyme coupling method and measuring method thereof

A technology of biological samples and oxalic acid, which is applied in the preparation of test samples, measurement of color/spectral characteristics, and analysis through chemical reactions of materials, etc., can solve the problems of wide-scale promotion and application limitations, high cost of oxalate oxidase, etc. Achieve the effect of low cost of reagents and equipment, high sensitivity and high precision

Inactive Publication Date: 2010-07-28
DONGHUA UNIV
View PDF8 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the high cost of oxalate oxidase used in this technology, the wide-scale application of this method has been greatly restricted.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Kit for measuring oxalic acid content of biological sample by enzyme coupling method and measuring method thereof
  • Kit for measuring oxalic acid content of biological sample by enzyme coupling method and measuring method thereof
  • Kit for measuring oxalic acid content of biological sample by enzyme coupling method and measuring method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Pretreatment of oxalic acid samples from different sources

[0061] (1) Sample preparation

[0062] Wash and dry fresh vegetables to remove surface moisture, accurately weigh a certain amount of vegetables, put them into a juicer and crush them into a paste, take out the samples and centrifuge to obtain the supernatant, and use 20% hydrochloric acid or 10M NaOH to adjust the pH value to the range of 4.0 to 6.0, and then put it in the refrigerator for storage;

[0063] A 24-hour urine sample was collected in a container containing 10 mL of concentrated hydrochloric acid, and the volume was measured and stored in a refrigerator;

[0064] (2) Sample pretreatment

[0065] Take the vegetable sample prepared earlier, mix it with 50mM disodium hydrogen phosphate-citric acid buffer solution with a pH value of 7.0 at a volume ratio of 1:3, and measure the pH value with a pH meter. , adjust the pH to 7.0 with hydrochloric acid or NaOH solution, then add activated carbon adsorb...

Embodiment 2

[0068] Since vegetable samples contain enzyme inhibitors or other interfering substances, pretreatment of the samples is necessary. In this experiment, cabbage juice was used as a model to conduct pretreatment research. The experimental results are shown in Table 2.

[0069] Table 2 shows that the oxalic acid content measured in the untreated cabbage juice sample is the smallest, and after being diluted 8 times with water, the measured value rises from 6.1mmol / L to 39.1mmol / L, indicating that the dilution method can reduce the interference factors of the determination. After the sample was treated with activated carbon adsorbent, it was found that the measured value was greatly improved, indicating that the adsorbent could absorb some interfering substances and improve the effect of the enzyme reaction. Combining the above two treatment methods, that is, water dilution method plus adsorbent treatment, the result obtained is much better than using one treatment method alone, an...

Embodiment 3

[0075] Determination of oxalic acid content in different vegetables (compared with the results of the recognized classical HPLC method)

[0076] Take the vegetable juice in embodiment 1 as test sample, measure oxalic acid by the experimental procedure in table 3,

[0077] Table 3 Experimental steps

[0078]

[0079] ΔA was measured according to the experimental procedure in Table 3 340 Finally, the oxalic acid content in the vegetable juice sample was calculated according to the following formula,

[0080] C = V × ΔA ϵ × d × v × 1000 [ mol / L ]

[0081] V——The final volume of the reaction, 3 [mL]

[0082] v——reaction sample volume, 0.1 [mL]

[0083] d——optical path diameter [cm]

[0084] ε——NADH absorption coefficient: 340nm=6.3...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
Ionic strengthaaaaaaaaaa
Ionic strengthaaaaaaaaaa
Ionic strengthaaaaaaaaaa
Login to View More

Abstract

The invention relates to a kit for measuring the oxalic acid content of a biological sample by an enzyme coupling method. The kit comprises a sample pre-treatment kit and a sample reaction kit. The measuring method comprises the following steps of; pretreating the sample to be measured, and removing enzyme reaction inhibitors and interferents; adding oxalate decarboxylase into treated sample solution to perform a reaction at the temperature of between 15 to 16 DEG C for 20 to 60 minutes; adding solution of coenzyme I and deionized water, and measuring a light absorption value A1; adding formate dehydrogenlyase into the mixed solution to perform a reaction for 15 to 60 minutes, and measuring a light absorption value A2 of the reaction solution; and preparing a blank sample for comparison, and finally working out the oxalic acid content according to the measured absorption values. The measuring kit has the advantages of measuring samples from different sources, along with high sensitivity, high stability, convenient operation, low use cost and equipment cost, wide application range, high practicability and precision higher than other kits for measuring the oxalic acid content.

Description

technical field [0001] The invention belongs to the field of detection and analysis of oxalic acid content in biological samples, in particular to a kit for measuring oxalic acid content in biological samples by an enzyme coupling method and a detection method thereof. Background technique [0002] Oxalic acid is an organic carboxylic acid that exists widely in nature, mostly in the form of oxalate, and is a metabolite during the growth of organisms. The metabolic production of oxalic acid is mainly derived from the tricarboxylic acid cycle and the glyoxylate cycle. Oxalic acid can complex with some metal ions in the body to form compounds. [0003] In organisms, the accumulation of oxalate formed by excessive oxalic acid levels will cause certain toxicity to organisms. For example: in some plants (beans, tomato, sunflower, etc.), the accumulation of oxalate can lead to plant nutrient deficiency, growth arrest and other related symptoms. Existing studies have shown that o...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): G01N21/75G01N21/31G01N1/28
Inventor 洪枫曹张军杨光
Owner DONGHUA UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com