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Method and system for simultaneously determining contents of various endogenous sugar alcohols in biological sample

A biological sample, endogenous technology, applied in measurement devices, instruments, scientific instruments, etc., can solve the problem of difficulty in detecting multiple endogenous sugar alcohols at the same time, so as to reduce interference and impact, reduce detection costs, and improve detection. The effect of efficiency

Pending Publication Date: 2022-04-29
重庆迪纳利医药科技有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The present invention intends to provide a method for simultaneously determining the content of multiple endogenous sugar alcohols in biological samples, so as to solve the technical problem that the analytical methods in the prior art are difficult to detect multiple endogenous sugar alcohols at the same time

Method used

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  • Method and system for simultaneously determining contents of various endogenous sugar alcohols in biological sample
  • Method and system for simultaneously determining contents of various endogenous sugar alcohols in biological sample
  • Method and system for simultaneously determining contents of various endogenous sugar alcohols in biological sample

Examples

Experimental program
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Effect test

Embodiment 1

[0091] (1) Sample preparation

[0092] Weigh sorbitol, mannitol, erythritol, ribitol and inositol standard respectively, dissolve in 50% dimethylformamide solution (water / dimethylformamide=1 / 1, v / v) Prepare the stock solution of the analyte (the concentration of sorbitol / mannitol / erythritol is 1000000ng / mL, ribitol is 500000ng / mL and inositol is marked as 6000000ng / mL). Prepare two parts of the stock solution of the analyte, which are respectively used to prepare the standard curve working solution and the quality control working solution. The diluent is 50% dimethylformamide solution. The preparation method is shown in Table 1.

[0093] Table 1: Preparation table of five sugar alcohol standard curve working solutions and quality control working solutions

[0094]

[0095] Weigh 6 pieces 13 The C-labeled sorbitol standard substance was dissolved in 50% dimethylformamide solution (water / dimethylformamide=1 / 1, v / v) to prepare an internal standard stock solution (1,000,000 n...

Embodiment 2

[0120] Embodiment 2: Research on the influence of chromatographic column on separation effect

[0121] Using the test method in Example 1, the selection of chromatographic columns was studied, and this example specifically lists the research results of three chromatographic columns. The sample to be detected in this embodiment is a standard curve sample (STD8) containing the isomer components of sorbitol and mannitol. The three columns used are as follows:

[0122] Column 1: Luna Omega SUGAR 100×2.1mm, 3μm

[0123] Column 2: Luna NH2 150×4.6mm, 5μm

[0124] Column 3: Carbo Sep CHO-620 300×6.5mm, 10μm

[0125] For experimental results, see Figure 3-Figure 5 , shows the liquid chromatograms obtained using the three columns. pass image 3 It can be seen that when the chromatographic column 1 is selected, sorbitol and mannitol are completely indistinguishable. From Figure 4 It can be seen that (the peak on the left is mannitol, and the peak on the right is sorbitol), whe...

Embodiment 3

[0126] Embodiment 3: the impact of flow rate on detection

[0127] Adopt the test method of embodiment 1, test successively different flow rates 0.4mL / min, 0.6mL / min, 0.8mL / min to the component separation situation of sugar alcohol under the situation that other conditions are constant, the separation situation of its component is as follows figure 1 , figure 2 , Figure 6 , Figure 7 shown (in figure 2 , Figure 6 , Figure 7 Among them, from top to bottom are the liquid chromatograms containing 5 kinds of sugar alcohols, the liquid chromatograms of ribitol, the liquid chromatograms of erythritol, the liquid chromatograms of inositol, the liquid chromatograms of mannitol and sorbitol liquid chromatogram). It can be seen that the lower the flow rate, the better the separation of the five sugar alcohols, and the longer the separation time. Considering the separation effect and separation time, 0.6mL / min was selected as the optimal flow rate.

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Abstract

The invention relates to the technical field of endogenous substance detection, in particular to a method for simultaneously determining the content of various endogenous sugar alcohols in a biological sample, which sequentially comprises the following steps: pretreating to obtain a to-be-enriched treatment product containing an internal standard; obtaining a sugar alcohol enrichment sample by using the to-be-enriched treatment product containing the internal standard in the UPLC; detecting and / or quantifying sugar alcohols in the sugar alcohol-enriched sample using MS / MS; the sugar alcohol comprises sorbitol, mannitol, erythritol, ribitol and inositol. According to the scheme, the technical problem that an analysis method in the prior art is difficult to detect various endogenous sugar alcohols at the same time is solved, and the technical problem that isomeride sugar alcohols are difficult to completely separate through chromatography is solved. The method is short in detection process time consumption, relatively low in cost, good in specificity, high in sensitivity, capable of saving labor amount and more suitable for being applied to practical operation for evaluating the effect of a medicine or a treatment method.

Description

technical field [0001] The invention relates to the technical field of detection of endogenous substances, in particular to a method and system for simultaneously measuring the contents of various endogenous sugar alcohols in biological samples Background technique [0002] Sugar alcohols are a special class of polyols formed by catalytic hydrogenation of carbohydrates. They are mostly found in specific fruits, vegetables, and fungi in nature, and are usually used as sugar-free in many similar chewing gums, candies, and beverages. sweetener. Under normal conditions, the content of sugar alcohols in the body is low, but after receiving some internal or external stimuli, its endogenous content will change, so sugar alcohols are often used as biomarkers for judging drug response and drug efficacy, etc. things. For example, higher than normal serum erythritol levels predict increased central obesity and cardiovascular disease. Inositol is abundant in the brain and other mamma...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/72
CPCG01N30/02G01N30/06G01N30/72G01N2030/062
Inventor 蒙敏张玉东赵常建王来新
Owner 重庆迪纳利医药科技有限责任公司
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