CN106324150A - Method for determining eight sweetening agents in Baijiu by high-performance liquid chromatography-low-temperature evaporative light scattering detector method - Google Patents

Method for determining eight sweetening agents in Baijiu by high-performance liquid chromatography-low-temperature evaporative light scattering detector method Download PDF

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CN106324150A
CN106324150A CN201611052984.3A CN201611052984A CN106324150A CN 106324150 A CN106324150 A CN 106324150A CN 201611052984 A CN201611052984 A CN 201611052984A CN 106324150 A CN106324150 A CN 106324150A
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CN106324150B (en
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刘刚
雷激
张广峰
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Sichuan Provincial Institute Of Food And Drug Administration
Xihua University
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Xihua University
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/04—Preparation or injection of sample to be analysed
    • G01N30/06—Preparation
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/62—Detectors specially adapted therefor
    • G01N30/74—Optical detectors

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Abstract

The invention relates to the technical field of food processing, in particular to a method for determining eight sweetening agents in Baijiu by a high-performance liquid chromatography-low-temperature evaporative light scattering detector method. The method includes the steps of (1), standard solution preparation, (2) sample pretreatment, (3) sample detection, (4) calibration curve establishment and (5) resulting. The method for determining eight sweetening agents in the Baijiu by the high-performance liquid chromatography-low-temperature evaporative light scattering detector method has the advantages of mild detection conditions, simplicity, rapidity, good separation effects and capability of playing a positive role in improving laboratory detection efficiency and reducing detection cost.

Description

Eight kinds of sweeting agents in high performance liquid chromatography low-temperature evaporation light scattering determining Chinese liquor Method
Technical field
The present invention relates to food processing technology field, specifically a kind of high performance liquid chromatography low-temperature evaporation light scattering method Measure the method for eight kinds of sweeting agents in Chinese liquor.
Background technology
White wine brewing process is one of non-material cultural heritage of China's preciousness, and the Chinese liquor of high-quality has sweet ice-cold, the alcohol of wine body The feature such as thick, soft, this with excellent White wine brewing process during the higher alcohol that produces of natural fermentation, the material such as polyhydric alcohol have Close relationship.Good wine goes out the traditional fermentation technique regarded for oneself, but the today developed rapidly in current food industry, some wine brewing Workshop or businessman, use edible ethanol or second-rate wine base to hook furnishing and sample wine, in order to cover the bad mouthfeels such as wine body is bitter, puckery Defect, increase sweet sense, in these wine bodies, add artificial synthesis edulcorant without authorization, adulterate.This has not only injured Chinese liquor The vital interests of consumer, and the image of Chinese traditional liquor brewage process is caused bad impact.
Artificial synthesis edulcorant must not expressly provide add, at present to Chinese liquor for country for the fermented wine such as Chinese liquor The detection method of middle sweeting agent mainly have liquid chromatography-mass spectrometry, liquid chromatograph diode array, liquid chromatograph parallax method, from Sub-chromatography, high performance capillary electrophoresis etc..Wherein liquid chromatography-mass spectrometry sensitivity is best, but testing cost is higher, other method Pessimistic and detect in kind owing to using the character of the defect of detector or compound itself to determine its detection sensitivity The most comprehensive.
Summary of the invention
The problem that it is an object of the invention to exist for above-mentioned technology, it is provided that a kind of high performance liquid chromatography-low-temperature evaporation light The method of eight kinds of sweeting agents in scattering method Chinese liquor.The method testing conditions is gentle, has simple, quick, good separating effect Feature, to improve test in laboratory efficiency, reduce testing cost serve positive effect.
For achieving the above object, the technical solution used in the present invention is: a kind of high performance liquid chromatography low-temperature evaporation light scattering Method measures the method for eight kinds of sweeting agents in Chinese liquor, comprises the steps:
(1) standard solution preparation: weigh each 25mg of each standard substance respectively in 25mL volumetric flask, dilute with ultra-pure water and determine Hold to scale, be configured to the standard reserving solution of 1000 μ g/mL, accurate eight kinds of each 0.02mL of sweeting agent standard reserving solution of absorption, 0.05mL, 0.10mL, 0.20mL, 0.50mL, 1.00mL, in 10mL capacity product, are configured to 2.00 μ g/mL, 5.00 μ g/ respectively ML, 10.00 μ g/mL, 20.00 μ g/mL, 50.00 μ g/mL, the hybrid standard working solution of 100.00 μ g/mL, standby；
(2) sample pre-treatments: accurately weigh Wine Sample 5g in 10mL color comparison tube, sample is put into the water of constant temperature 45 DEG C Bath Nitrogen evaporator volatilizes wine body, takes out the sample volatilized and stand to room temperature, with acetic acid water, residue is settled to 1mL, mistake 0.45 μm aqueous phase filter membrane, to be measured；
(3) sample detection: chromatographic column: C18Post is 4.6mm × 250mm, 5 μm；Low-temperature evaporation scatter detector condition: evaporation Pipe temperature 40 DEG C, atomization air pressure 350kPa, Gain value 7, assisted detector photodiode array detection condition: detection wavelength 208nm； Column temperature 30 DEG C；Flow velocity 1.0mL/min；Sampling volume 40 μ l；Flowing is acetonitrile (A)/20mmol/L ammonium acetate solution (B) mutually； Set sampling volume and carry out eluting as 40 μ L；
(4) foundation of calibration trace: use response signal to take the logarithm with standard substance concentration simultaneously and set up calibration curve, Model Y=Xaeb, wherein X is target substance concentration logarithm, and Y is the logarithm of corresponding target substance response peak area, and a, b are logarithm Correlation coefficient；
(5) result: use the calibration curve integration sample chromatogram figure established, result of calculation:
M = C × V m × 1000
Wherein: C is tested component IC, μ g/mL；V is for concentrating constant volume, mL；M is sample volume, g；M is Tested constituent content, g/kg in sample.
Further, in step (2), acetic acid water pH is 4.5.
Further, in step (3), the separated elution program of flowing is:
Further, eight kinds of described sweeting agents are acesulfame potassium, saccharin sodium, cyclamate, sucralose, aspartame, A Li Sweet, neotame and stevioside.
The method have the benefit that: technical solution of the present invention uses low-temperature evaporation scattering detection method to measure in Chinese liquor The eight kinds of sweeting agents contained, the method testing conditions is gentle, has simple, quick, the feature of good separating effect, to improving experiment Room detection efficiency, reduction testing cost serve positive effect.
Accompanying drawing explanation
In order to be illustrated more clearly that the embodiment of the present invention or technical scheme of the prior art, below will be to embodiment or existing In having technology to describe, the required accompanying drawing used is briefly described, it should be apparent that, the accompanying drawing in describing below is only this Some embodiments of invention, for those of ordinary skill in the art, on the premise of not paying creative work, also may be used To obtain other accompanying drawing according to these accompanying drawings.
The evaporative light chromatogram of eight kinds of sweeting agents of Fig. 1 (be respectively from left to right acesulfame potassium, saccharin sodium, cyclamate, sucralose, Aspartame, alitame, neotame, stevioside, concentration 50 μ g/mL)；
Fig. 2 sample mark-on (5 μ g/mL) and positive contrast.
Detailed description of the invention
Below in conjunction with the accompanying drawing in the embodiment of the present invention, the technical scheme in the embodiment of the present invention is carried out clear, complete Describe, it is clear that described embodiment is only a part of embodiment of the present invention rather than whole embodiments wholely.Based on Embodiment in the present invention, it is every other that those of ordinary skill in the art are obtained under not making creative work premise Embodiment, broadly falls into the scope of protection of the invention.
Embodiment
(1) standard solution preparation: weigh each 25mg of each standard substance respectively in 25mL volumetric flask, dilute with ultra-pure water and determine Hold to scale, be configured to the standard reserving solution of 1000 μ g/mL, accurate eight kinds of each 0.02mL of sweeting agent standard reserving solution of absorption, 0.05mL, 0.10mL, 0.20mL, 0.50mL, 1.00mL, in 10mL capacity product, are configured to 2.00 μ g/mL, 5.00 μ g/ respectively ML, 10.00 μ g/mL, 20.00 μ g/mL, 50.00 μ g/mL, the hybrid standard working solution of 100.00 μ g/mL, standby；
(2) sample pre-treatments: accurately weigh Wine Sample 5g in 10mL color comparison tube, sample is put into the water of constant temperature 45 DEG C Bath Nitrogen evaporator volatilizes wine body, takes out the sample volatilized and stand to room temperature, with acetic acid water, residue is settled to 1mL, mistake 0.45 μm aqueous phase filter membrane, to be measured；
(3) preparation of the phase that flows:
The preparation of mobile phase A: mobile phase A is acetonitrile, as flowing phase front ultrasound wave aerofluxus 5min, stand for standby use；
The preparation of Mobile phase B: Mobile phase B is acidifying 20mmol/L ammonium acetate (acetic acid, pH4.5), through 4.5 μm aqueous phases Membrane filtration, standby.
(4) unlatching of detecting instrument and debugging
Open chromatograph of liquid degasser, pump, column oven, low-temperature evaporation light scattering detector and main controller the most respectively and connect Connect detached dowel；
2. liquid chromatographic system aerofluxus；
Start A pump (connection mobile phase A) and B pump (connection Mobile phase B) the most simultaneously, gradually by system flow phase flow velocity by 0mL/min is promoted to 1.2mL/min (forbid risen to by flow velocity suddenly the highest, prevent instantaneous pressure excessive, destroy pillar)；
4. by column temperature in column oven in whole inspection process constant at 30 DEG C, keep constant；
When liquid is closely expired in U-tube in light scattering detector the most to be evaporated, open nitrogen cylinder, slowly atomizing pressure is regulated To 350kPa (2.8L/min)；
6. regulating relative detector response (Gain value) is 7；
7. regulation evaporation tube temperature is 40 DEG C；
Balance system about 15min the most in the above state.
(5) detection of sample
Sampling volume 40 μ l, the separated elution program of flowing clicks step to be carried out:
The evaporative light chromatogram of eight kinds of sweeting agents is as shown in Figure 1.
(6) foundation of calibration trace: use response signal to take the logarithm with standard substance concentration simultaneously and set up calibration curve, Model Y=Xaeb, wherein X is target substance concentration logarithm, and Y is the logarithm of corresponding target substance response peak area, and a, b are logarithm Correlation coefficient；
(7) result: using the calibration curve integration sample chromatogram figure established, result of calculation, result is as shown in table 1,
M = C × V m × 1000
Wherein: C is tested component IC, μ g/mL；V is for concentrating constant volume, mL；M is sample volume, g；M is Tested constituent content, g/kg in sample.
Table 1 testing result
* based on sample weighting amount 5g, constant volume 1mL, sampling volume 40 μ L.
Those skilled in the art of the present technique are appreciated that unless otherwise defined, and all terms used herein (include technology art Language and scientific terminology) have with the those of ordinary skill in art of the present invention be commonly understood by identical meaning.Also should Being understood by, those terms defined in such as general dictionary should be understood that the meaning having with the context of prior art The meaning that justice is consistent, and unless defined as here, will not explain by idealization or the most formal implication.
It should be noted last that: above example is only in order to illustrative not limiting technical scheme, although ginseng According to above-described embodiment, the present invention is described in detail, it will be apparent to an ordinarily skilled person in the art that: still can be to this Invention is modified or equivalent, and any modification or partial replacement without departing from the spirit and scope of the present invention, it is equal Should contain in the middle of scope of the presently claimed invention.

Claims (4)

1. the method for eight kinds of sweeting agents in high performance liquid chromatography low-temperature evaporation light scattering determining Chinese liquor, it is characterised in that include Following steps:
(1) standard solution preparation: weigh each 25mg of each standard substance respectively in 25mL volumetric flask, dilute with ultra-pure water and be settled to Scale, is configured to the standard reserving solution of 1000 μ g/mL, accurately draw eight kinds of each 0.02mL of sweeting agent standard reserving solution, 0.05mL, 0.10mL, 0.20mL, 0.50mL, 1.00mL in 10mL capacity product, be configured to respectively 2.00 μ g/mL, 5.00 μ g/mL, 10.00 μ g/mL, 20.00 μ g/mL, 50.00 μ g/mL, the hybrid standard working solution of 100.00 μ g/mL, standby；
(2) sample pre-treatments: accurately weigh Wine Sample 5g in 10mL color comparison tube, sample is put into the water-bath nitrogen of constant temperature 45 DEG C Blow and instrument volatilizes wine body, take out the sample volatilized and stand to room temperature, with acetic acid water, residue being settled to 1mL, cross 0.45 μm Aqueous phase filter membrane, to be measured；
(3) sample detection: chromatographic column: C18Post is 4.6mm × 250mm, 5 μm；Low-temperature evaporation scatter detector condition: evaporation tube temperature Spend 40 DEG C, atomization air pressure 350kPa, Gain value 7, assisted detector photodiode array detection condition: detection wavelength 208nm；Column temperature 30℃；Flow velocity 1.0mL/min；Sampling volume 40 μ l；Flowing is acetonitrile (A)/20mmol/L ammonium acetate solution (B) mutually；Set Sampling volume is that 40 μ L carry out eluting；
(4) foundation of calibration trace: use response signal to take the logarithm with standard substance concentration simultaneously and set up calibration curve, model Y=Xaeb, wherein X is target substance concentration logarithm, and Y is the logarithm of corresponding target substance response peak area, and a, b are that logarithm is correlated with Coefficient；
(5) result: use the calibration curve integration sample chromatogram figure established, result of calculation:
M = C × V m × 1000
Wherein: C is tested component IC, μ g/mL；V is for concentrating constant volume, mL；M is sample volume, g；M is sample In tested constituent content, g/kg.
The side of eight kinds of sweeting agents in high performance liquid chromatography low-temperature evaporation light scattering determining Chinese liquor the most according to claim 1 Method, it is characterised in that in step (2), acetic acid water pH is 4.5.
The side of eight kinds of sweeting agents in high performance liquid chromatography low-temperature evaporation light scattering determining Chinese liquor the most according to claim 1 Method, it is characterised in that in step (3), the separated elution program of flowing is:
The side of eight kinds of sweeting agents in high performance liquid chromatography low-temperature evaporation light scattering determining Chinese liquor the most according to claim 1 Method, it is characterised in that eight kinds of described sweeting agents be acesulfame potassium, saccharin sodium, cyclamate, sucralose, aspartame, alitame, Neotame and stevioside.
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Cited By (8)

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CN108445118A (en) * 2018-03-20 2018-08-24 广东产品质量监督检验研究院（国家质量技术监督局广州电气安全检验所、广东省试验认证研究院、华安实验室） The rapid extraction and detection method of Sucralose in food
CN108918716A (en) * 2018-07-23 2018-11-30 江苏挺卫实业有限公司 A kind of detection method for neotame content in beverage
CN109490452A (en) * 2018-10-31 2019-03-19 中国农业科学院茶叶研究所 Method that is a kind of while detecting 6 kinds of synthetic sweeteners in tealeaves
CN109856282A (en) * 2019-03-11 2019-06-07 安徽瑞思威尔科技有限公司 The quick screening method of external source hazardous material in a kind of white wine
CN110726791A (en) * 2019-11-12 2020-01-24 河南中烟工业有限责任公司 Pretreatment method for detecting sweetener in electronic cigarette liquid or tobacco essence
CN110763797A (en) * 2019-11-12 2020-02-07 河南中烟工业有限责任公司 Method for measuring sweetening agent for cigarettes by high performance liquid chromatography
CN110763796A (en) * 2019-11-12 2020-02-07 河南中烟工业有限责任公司 Method for measuring sweetener for cigarettes by liquid chromatography-evaporative light scattering detector
CN110794073A (en) * 2019-11-12 2020-02-14 河南中烟工业有限责任公司 Method for measuring sweetener by using chromatography and diode array detector

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Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108445118A (en) * 2018-03-20 2018-08-24 广东产品质量监督检验研究院（国家质量技术监督局广州电气安全检验所、广东省试验认证研究院、华安实验室） The rapid extraction and detection method of Sucralose in food
CN108918716A (en) * 2018-07-23 2018-11-30 江苏挺卫实业有限公司 A kind of detection method for neotame content in beverage
CN109490452A (en) * 2018-10-31 2019-03-19 中国农业科学院茶叶研究所 Method that is a kind of while detecting 6 kinds of synthetic sweeteners in tealeaves
CN109856282A (en) * 2019-03-11 2019-06-07 安徽瑞思威尔科技有限公司 The quick screening method of external source hazardous material in a kind of white wine
CN110726791A (en) * 2019-11-12 2020-01-24 河南中烟工业有限责任公司 Pretreatment method for detecting sweetener in electronic cigarette liquid or tobacco essence
CN110763797A (en) * 2019-11-12 2020-02-07 河南中烟工业有限责任公司 Method for measuring sweetening agent for cigarettes by high performance liquid chromatography
CN110763796A (en) * 2019-11-12 2020-02-07 河南中烟工业有限责任公司 Method for measuring sweetener for cigarettes by liquid chromatography-evaporative light scattering detector
CN110794073A (en) * 2019-11-12 2020-02-14 河南中烟工业有限责任公司 Method for measuring sweetener by using chromatography and diode array detector
CN110763797B (en) * 2019-11-12 2022-04-19 河南中烟工业有限责任公司 Method for measuring sweetening agent for cigarettes by high performance liquid chromatography

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