CN102841172A

CN102841172A - High performance liquid chromatography (HPLC) detection method for azodicarbonamide (ADA)

Info

Publication number: CN102841172A
Application number: CN2012103636448A
Authority: CN
Inventors: 李培; 袁丽红; 丁洪流; 陈英; 张锦; 谢云飞; 姚卫蓉
Original assignee: SUZHOU PRODUCT QUALITY SUPERVISION TESTING INSTITUTE; Jiangnan University
Current assignee: SUZHOU PRODUCT QUALITY SUPERVISION TESTING INSTITUTE; Jiangnan University
Priority date: 2012-09-27
Filing date: 2012-09-27
Publication date: 2012-12-26

Abstract

The invention belongs to the field of detection for a flour modifier, namely azodicarbonamide (ADA). The method comprises the following steps of: immersing and extracting ADA from the flour by using acetone, centrifugating, blowing nitrogen to supernate, concentrating until the supernate is dried, fixing the volume by using a 20-mM ammonium acetate aqueous solution, filtering by a film with the thickness of 0.45 mu m, detecting the ADA in the sample by a high performance liquid chromatography (HPLC), and detecting the sample by selecting a C18 or ODS reversed phase column and a photoelectric diode array detector at the detection wavelength of 246 nm by taking methyl alcohol and the 20-mM ammonium acetate aqueous solution as flowing phases. The ADA detection method is high in detection speed, high in sensitivity and low in detection cost.

Description

The HPLC detection method of a kind of azodicarbonamide (ADA)

Technical field

The invention belongs to the detection range of food additives, is to utilize the high performance liquid chromatography method of detection faces flour modifying agent azodicarbonamide quickly and accurately.

Background technology

Azodicarbonamide (ADA) can improve the physical operations character and the Flour product institutional framework of dough, widespread use abroad as a kind of novel flour-dough improver.Aspect the flour maturation process, the halfcystine in can the oxidation wheat flour, thus flour muscle degree is increased, improve dough gas reservation amount, increase the elasticity and the toughness of baking goods, improve the operability and conditioning property of dough.Because azodicarbonamide belongs to alkaline matter,, may the stomach and the liver of human body be caused damage if excess is used in food.ADA at high temperature decomposes the generation gas foaming, can generate noxious material semicarbazide hydrochloride (SEM) simultaneously.Stipulated among the GB2760-2007 that the maximum use amount of ADA in wheat flour is 45mg/kg.

Though China has stipulated the maximum use amount of azodicarbonamide in flour, also do not see report at present relevant for ADA detection method in the flour, there is not relevant examination criteria yet.

Summary of the invention

The object of the invention be to provide a kind of easy and simple to handle, analyze the high performance liquid chromatography of ADA (HPLC) detection method rapidly.Technical scheme of the present invention is following:

(1) flour sample pre-service

Sample is carried out the immersion extraction of ADA with organic solvent, centrifugal, get the supernatant liquid nitrogen and blow and be concentrated near doing, and with 20mM ammonium acetate solution constant volume, with 0.45 μ m membrane filtration, thus the pre-treatment of completion sample;

(2) HPIC condition

Select C for use ₁₈Or the ODS reverse-phase chromatographic column, adopt photodiode array detector to detect: detecting and receiving wavelength is 246nm; Mobile phase volume ratio: V (CH ₃OH): V (20mM ammonium acetate solution)=15: 85 isocratic elutions; Flow velocity: v=0.8mL/min;

(3) drafting of ADA high performance liquid chromatography examination criteria curve

Accurately take by weighing ADA0.05g, use N, dinethylformamide (DMF) is dissolved in the brown volumetric flask of 50mL surely; Be mixed with the 1000mg/LADA storing solution; Water is diluted to respectively that concentration is respectively 2,5,10,20,30, the standard serial solution of 40mg/L, measures according to the condition of scheme (2), and be horizontal ordinate with ADA concentration; The characteristic peak peak area is an ordinate drawing standard curve, is used for the content of analytic sample ADA;

(4) mensuration of sample

After sample pretreatment in the scheme (1), detect by testing conditions in the scheme (2), and utilize ADA content in the typical curve calculation sample that obtains in the scheme (3).

Advantage of the present invention and beneficial effect are: adopt HPLC method of the present invention to detect ADA in the flour, and use photodiode array detector, can carry out further qualitative to object according to scanning optical spectrum; Convenient and swift; Highly sensitive, detectability is low, helps the fast detecting of ADA; Can be used for extensive sample detection, be suitable for standardization.

Description of drawings

Fig. 1 is the HPLC collection of illustrative plates of ADA standard items; Hypersil ODS2 chromatographic column (5 μ m * 4.6mm * 150mm), the photodiode array UV-detector, absorbing wavelength is 246nm, mobile phase volume compares CH ₃OH: the 20mM ammonium acetate solution is 15: 85 isocratic elutions, flow velocity 0.8mL/min)

Fig. 2 is the typical curve of ADA, and horizontal ordinate is an ADA concentration, and ordinate is an ADA chromatographic peak peak area.

Embodiment

Below in conjunction with embodiment the present invention is explained further details, but the present invention is not limited only to following embodiment.

Embodiment 1

Accurately take by weighing ADA 0.0500g, be dissolved in surely in the brown volumetric flask of 50mL, be mixed with the ADA storing solution of 1000mg/L, place 4 ℃ of refrigerators to preserve with DMF.Get the ADA standard reserving solution of a certain amount of 1000mg/mL; Be diluted to deionized water that concentration is respectively 2,5,10,20,30, the standard serial solution of 40mg/L, utilize HPLC to measure, the detection wavelength is 246nm; With peak area (Y) concentration X (mg/L) is carried out linear regression; Drawing standard curve (Fig. 2), must the typical curve funtcional relationship be: Y=113967X-24180, related coefficient be R ²=0.9996, linearity is good, can be used for the content of ADA in the quantitative sample.

Embodiment 2

Choose and show the flour sample that adds ADA in the list of ingredients, accurately take by weighing flour 1g (being accurate to 0.0001g) in the 50mL centrifuge tube, add 10mL acetone; Vortex is even, and concussion rotation 20min is centrifugal; Getting the supernatant liquid nitrogen blows near dried, with 2mL20mM ammonium acetate solution constant volume.Concentrate is crossed the disposable miillpore filter of 0.45 μ m, and filtrating is injected HPLC and analyzed.

Flour is done three parallel sample, and the result sees table 1.To parallel appearance 1 replication 6 times, analysis result is seen table 2.Know that through analyzing the relative standard deviation of parallel sample (RSD) is 2.44, the relative standard deviation of replication experiment is 2.17%, and repeatability better.

The content of ADA in table 1 flour sample

The parallel appearance of table 21 replication result

Embodiment 3

In the flour of known ADA content, add the ADA powder of inequality; Operate by step in the instance 2, each sample replication 6 times is measured recovery of standard addition and relative standard deviation simultaneously; The result sees table 3; Recovery of standard addition is between 91.50%～98.60%, and relative standard deviation is between 2.42%～4.50%, and method is reliable.

Table 3 sample recovery rate is measured (n=6)

Sample content (ug/g)	Addition (ug/g)	Measured value (ug/g)	The recovery (%)	RSD(％)
					7.50	4.00	11.19	93.17	2.42
7.50	8.00	15.38	98.60	3.97
					7.50	15.00	21.20	91.50	4.50

Above preferred embodiment only is used to explain content of the present invention; In addition; The present invention also has other embodiments; In every case those skilled in the art are because of teachings involved in the present invention, are equal to the technical scheme that replacement or equivalent deformation mode form and all drop in protection scope of the present invention and adopt.

Claims

1. the high-efficiency liquid chromatography method for detecting of an azodicarbonamide is characterized in that: comprise the steps:

(1) sample is carried out the immersion extraction of azodicarbonamide (ADA) with organic solvent, centrifugal, get the supernatant liquid nitrogen and blow and be concentrated near doing, and with 20mM ammonium acetate solution constant volume, with 0.45 μ m membrane filtration, thus the pre-treatment of completion sample;

(2) with ADA in high performance liquid chromatography (HPLC) working sample: select reverse-phase chromatographic column for use, methyl alcohol and 20mM ammonium acetate solution adopt photodiode array detector as moving phase, and being provided with and detecting wavelength is 246nm, and sweep limit is 190-400nm;

(3) with the treated sample of step (1) gained set by step (2) parameter of setting carry out HPLC and detect.

2. the HPLC detection method of ADA according to claim 1 is characterized in that: extract the organic solvent that uses in the said step (1) and be acetone.

3. the HPLC detection method detection method of ADA according to claim 1, it is characterized in that: used reverse-phase chromatographic column can be C in the said step (2) ₁₈Or ODS chromatographic column.

4. the HPLC detection method of ADA according to claim 1 is characterized in that: the volume ratio of moving phase is V (methyl alcohol): V (20mM ammonium acetate solution)=15: 85 in the said step (2).