阅读说明：本技术 一种果汁中22种糖、糖醇及醇类同时检测的方法 (A kind of method that 22 kinds of sugar, sugar alcohol and alcohols detect simultaneously in fruit juice ) 是由 宋卫得 高尧华 许美玲 叶佳宇 李林杰 林令海 于 2019-09-25 设计创作，主要内容包括：本发明公开了一种果汁中22种糖、糖醇及醇类同时检测的方法,属于分析化学领域,本发明采用积分脉冲安培-离子色谱法,通过优化前处理方法,研究影响组分分离的实验因素,建立了一次性同时检测果汁中22种糖类、糖醇、醇类(2,3-丁二醇、丙二醇、甲醇、丙三醇、赤藓糖醇、木糖醇、鼠李糖、阿拉伯糖醇、山梨醇、海藻糖、半乳糖醇、甘露醇、2-脱氧-D-葡萄糖、阿拉伯糖、蜜二糖、葡萄糖、半乳糖、果糖、核糖、蔗糖、棉子糖、麦芽糖)的方法,解决了对果汁中糖类、糖醇及醇类多组分准确分析的技术难题,该方法快速、准确、灵敏,实现了多类别多组分的同时分析,对果汁成份检测做出重要的贡献。(The invention discloses 22 kinds of sugar in a kind of fruit juice, the method that sugar alcohol and alcohols detect simultaneously, belong to analytical chemistry field, the present invention uses integrated pulsed amperometric-chromatography of ions, by optimizing pre-treating method, research influences the empirical factor of component separation, it establishes disposable while detecting 22 kinds of carbohydrates in fruit juice, sugar alcohol, alcohols (2, 3- butanediol, propylene glycol, methanol, glycerine, antierythrite, xylitol, rhamnose, arabite, sorbierite, trehalose, galactitol, mannitol, 2-deoxy-D-glucose, arabinose, melibiose, glucose, galactolipin, fructose, ribose, sucrose, gossypose, maltose) method, it solves to carbohydrate in fruit juice, the technical problem that sugar alcohol and alcohols multicomponent are accurately analyzed, this method is quick, accurately, it is sensitive, it realizes multi-class more It is analyzed while component, important contribution is made to fruit juice detection.)

1. a kind of method that 22 kinds of sugar, sugar alcohol and alcohols detect simultaneously in fruit juice, it is characterised in that the following steps are included:

Step 1: the configuration of mixed standard solution:

By 2,3- butanediol, propylene glycol, methanol, glycerine, antierythrite, xylitol, rhamnose, arabite, sorbierite, Trehalose, galactitol, mannitol, 2-deoxy-D-glucose, arabinose, melibiose, glucose, galactolipin, fructose, core Sugar, sucrose, gossypose, maltose standard solution configure mixed standard solution as needed；

Step 2: sample to be tested pre-treatment:

It takes 0.100 g of fruit juice measuring samples in 100 mL beakers, about 80 mL of ultrapure water is added, mix and stand 10 min, use 0.10 mol/L sodium hydroxide solution adjusts dilution pH value to 6.5~8.6, is fully transferred in 100 mL volumetric flasks, Scale is settled to using ultrapure water, shakes up, stand 15 min, takes 10.0 mL solution, first crosses 0.22 μm of filter membrane, after in advance The IC-RP10 column of activation discards preceding 3 mL, collects back segment clear liquid, to be detected；

Step 3: ion chromatography condition and multistage gradient elute condition

Analytical column: CarboPac MA1 (4 × 250 mm)；

Guard column: CarboPac MA1 (4 × 50 mm)；

Working electrode: Gold；

Reference electrode: AgCl electrode；

Mobile phase: A:1000 mmol/L NaOH solution, B: ultrapure water；

Flow velocity: 0.40 mL/min；

Column temperature: 30 DEG C；

Sample volume: 10 μ L；

Multistage gradient elutes condition:

Multistage gradient rinse procedures are as follows: when 0.0-80.0 min, leacheate NaOH concentration is 500.0 mmol/L；80.0-85.0 Min, leacheate NaOH concentration are 500.0-600.0 mmol/L；85.0-105.0 min, leacheate NaOH concentration is 600.0 mmol/L；105.0-110.0 min, leacheate NaOH concentration is 600.0-500.0 mmol/L；110.0-120.0 min, leaching Washing lotion NaOH concentration is 500.0 mmol/L；

Step 4: calibration curve is drawn:

The chromatographiccondition and multistage gradient that the mixed standard solution configured in step 1 is provided according to step 3 elute item Part is detected using ion chromatograph, using constituent mass concentration to be measured as abscissa, using component peak area to be measured as ordinate, To draw calibration curve, quantified by external standard method；

Step 5: the detection of sample solution:

Chromatographiccondition and multistage gradient elution condition that gained clear liquid in step 2 is provided according to step 3 are used into ion Chromatograph is detected, and calculated result；

Step 6: result calculates:

By result measured by step 4 mixed standard solution come calculated result measured by aligning step five, to calculate institute Survey each component content in fruit juice.

2. the method that 22 kinds of sugar, sugar alcohol and alcohols detect simultaneously in a kind of fruit juice according to claim 1, feature exist In above-mentioned solution uses the ultrapure water that resistivity is 18.2 M Ω cm to prepare.

Technical field

The invention belongs to analytical chemistry fields, and in particular to 22 kinds of sugar, sugar alcohol and alcohols detect simultaneously in a kind of fruit juice Method.

Background technique

Fruit juice is the important beverage in people's daily life, contains protein, amino acid, carbohydrate, glycitols etc. in fruit juice Ingredient.Wherein saccharide compound is that people sustain life the main source of energy needed for activity.Sugar alcohol is the aldehyde radical or ketone of carbohydrate The substance that base is reduced into hydroxyl and is formed will not cause blood glucose rise mainly by the sweetener of corresponding sugar reaction generation, Therefore have the function of preventing diabetes and obesity.Alcohols is in molecule containing combined with the carbon on alkyl or benzene ring side chain The compound of hydroxyl, many alcohols are beneficial to human health, but some alcohols have certain toxicity.Carbohydrate, glycitols component Detection be both analytical chemistry field difficult point, and analysis testing field research hotspot, at present China's food safety country Regulation in standard " prepackaged food nutritional labeling general rule ", it is necessary in the sugared content of food labelling mark food.

The content size of sugar, sugar alcohol and alcohols directly affects the quality of fruit juice in fruit juice.Current carbohydrate, sugar alcohol, alcohols master The detection method wanted has gas chromatography, liquid chromatography, Enzyme Electrode, chromatography of ions etc..Gas chromatography has fast Fast, sensitive advantage, but pre-treatment step is complicated, it is quantitative not accurate enough.Liquid chromatography has spy accurate, that selectivity is good Point, but sensitivity is low.Enzyme Electrode generally only detects a kind of carbohydrate, and specificity is good, but detection efficiency is not high.From Sub- chromatography has many advantages, such as easy, quick, sensitive, accurate, is the ideal method of a kind of novel multicomponent while detection.

Currently, although the method that people detect sugar a variety of in fruit juice, sugar alcohol and alcohols simultaneously possesses some special knowledge, but in mesh The group only detected to certain a kind of component (such as simple carbohydrate or alcohols) or simultaneously that can be retrieved in preceding data bank Dosis refracta is less (only the sugar to 10 kinds or so and sugar alcohol are analyzed simultaneously), is not able to satisfy growing detection demand and therefore grinds Study carefully and establish good, high sensitivity a variety of sugar of a kind of strong applicability, selectivity, the method that sugar alcohols and alcohols detect simultaneously, has Important realistic meaning and application value.

Summary of the invention

The purpose of the present invention is in view of the above shortcomings of the prior art, using integrated pulsed amperometric-chromatography of ions, optimize Pre-treatment detection method establishes disposable while detecting in fruit juice 22 kinds by the research of the influence factor separated to component The method of carbohydrate, sugar alcohol, alcohols, this method is quick, simplicity, high sensitivity, stability are good, realizes quickly analysis and green is divided Analysis solves the technical issues of sugar, sugar alcohol and alcohols Simultaneous Analysis for Multicomponent.

In order to achieve the above objectives, the invention discloses 22 kinds of sugar, sugar alcohol and alcohols in a kind of fruit juice simultaneously detect method, Its detecting step is as follows；

Step 1: the configuration of mixed standard solution:

By 2,3- butanediol, propylene glycol, methanol, glycerine, antierythrite, xylitol, rhamnose, arabite, sorbierite, Trehalose, galactitol, mannitol, 2-deoxy-D-glucose, arabinose, melibiose, glucose, galactolipin, fructose, core Sugar, sucrose, gossypose, maltose standard solution configure hybrid standard stock solution as needed；

Step 2: sample to be tested pre-treatment:

It takes 0.100 g of fruit juice measuring samples in 100 mL beakers, about 80 mL of ultrapure water is added, mix and stand 10 min, use 0.10 mol/L sodium hydroxide solution adjusts dilution pH value to 6.5~8.6, is fully transferred in 100 mL volumetric flasks, Scale is settled to using ultrapure water, shakes up, stand 15 min, takes 10.0 mL solution, first crosses 0.22 μm of filter membrane, after in advance The IC-RP10 column of activation discards preceding 3 mL, collects back segment clear liquid, to be detected.

Step 3: ion chromatography condition and multistage gradient elute condition

Analytical column: CarboPac MA1 (4 × 250 mm)；

Guard column: CarboPac MA1 (4 × 50 mm)；

Working electrode: Gold；

Reference electrode: AgCl electrode；

Mobile phase: A:1000 mmol/L NaOH solution, B: ultrapure water；

Flow velocity: 0.40 mL/min；

Column temperature: 30 DEG C；

Sample volume: 10 μ L；

Multistage gradient elutes condition:

Multistage gradient rinse procedures are as follows: when 0.0-80.0 min, leacheate NaOH concentration is 500.0 mmol/L；80.0-85.0 Min, leacheate NaOH concentration are 500.0-600.0 mmol/L；85.0-105.0 min, leacheate NaOH concentration is 600.0 mmol/L；105.0-110.0 min, leacheate NaOH concentration is 600.0-500.0 mmol/L；110.0-120.0 min, leaching Washing lotion NaOH concentration is 500.0 mmol/L；See Table 1 for details.

1 ion chromatography of table elutes condition

Step 4: calibration curve is drawn:

The chromatographiccondition and multistage gradient that the mixed standard solution configured in step 1 is provided according to step 3 elute item Part is detected using ion chromatograph, using constituent mass concentration to be measured as abscissa, using component peak area to be measured as ordinate, To draw calibration curve, quantified by external standard method；

Step 5: the detection of sample solution:

Chromatographiccondition and multistage gradient elution condition that gained clear liquid in step 2 is provided according to step 3 are used into ion Chromatograph is detected, and calculated result；

Step 6: result calculates:

By result measured by step 4 mixed standard solution come calculated result measured by aligning step five, to calculate institute Survey each component content in fruit juice.

Preferably, above-mentioned solution uses the ultrapure water that resistivity is 18.2 M Ω cm to prepare.

The utility model has the advantages that

1, sugar, sugar alcohol and alcohols multi-component material in fruit juice are detected simultaneously, are always the technological difficulties of analytical chemistry field, The present invention disposably determines 22 kinds of sugar, sugar alcohol and alcohols in fruit juice simultaneously, solves at present using multiple standards, Duo Zhongyi Device, a variety of pre-treating methods carry out the technical problem of low efficiency when various ingredients measurement, and detection efficiency greatly improved, reduce Testing cost.

2,22 kinds of carbohydrates, sugar alcohol, alcohol component are the very big work of a technical difficulty in separation detection fruit juice simultaneously.For The overall separation degree for improving component, is compared by experimental analysis, and present invention selection is in preceding 80 min using isocratic elution Method, carries out the strong flushing of chromatographic column after 85 min using the sodium hydroxide solution of 600 mmol/L, and removal influences to be measured The chaff interferent of component can increase substantially the accuracy of detection.

3, the present invention investigates analysis by multifactor more matrix, has successively researched and analysed multistage gradient elution condition, initially The experimental factors such as concentration, chromatogram column temperature, pH value are eluted, various ingredients is farthest improved while surveying timing method Accuracy and applicability.

4, the present invention passes through deep study and analysis, it was found that solution ph detects 22 kinds of components in fruit juice important simultaneously It influences, the variation of pH value causes certain constituent contents that fluctuation has occurred.It, can be with by adjusting solution ph to 6.5~8.6 Greatly improve the accuracy and precision of detection.

5, the present invention has studied chromatogram column temperature to the great influence of detection, the minor variations of chromatogram column temperature, same meeting Component is caused to be overlapped.In 30 DEG C of column temperatures, it is able to satisfy the qualitative, quantitative requirement of 22 components just.

6, detection method is easy to operate quickly, detection sensitivity is high, operates, that is, is available on the machine by simple pre-treatment Detection.

7, it does not use or generates in detection method to human health, the harmful solvent of ecological environment, examination Agent, by-product meet the theory of green analysis.

Detailed description of the invention

Fig. 1 is the chromatography of ions figure of 22 kinds of sugar, sugar alcohol and alcohols mixed standard solution.

Fig. 2 is the chromatography of ions figure of mixed standard solution under different initial eluent concentrations.

Fig. 3 is the chromatography of ions figure of mixed standard solution under different column temperatures.

Fig. 4 is the influence that solution ph detects 6 kinds of components.

Fig. 5 is the chromatography of ions figure of 6 kinds of samples of juice.

Corresponding substance: 1. 2,3- butanediols (2,3-Butanediol) is respectively numbered in attached drawing；2. propylene glycol (Propanediol)；3. methanol (Methanol)；4. glycerine (Glycerol)；5. antierythrite (Erythritol)； 6. xylitol (Xylitol)；7. rhamnose (L-rhamnose)；8. arabite (Arabinitol)；9. sorbierite (Sorbitol)；10. trehalose (Trehalose)；11. galactitol (Galactitol)；12. mannitol (Mannitol)；13. 2-deoxy-D-glucose (2-Deoxy-D-glucose)；14. arabinose (Arabinose)；15. Melibiose (Melibiose)；16. glucose (Glucose)；17. galactolipin (Galactose)；18. fructose (Fructose)；19. ribose (Ribose)；20. sucrose (Sucrose)；21. gossypose (Raffinose)；22. maltose (Maltose).

Specific embodiment