CN100480684C - Portable injection chemiluminescence analysis method of isoflavone compound - Google Patents
Portable injection chemiluminescence analysis method of isoflavone compound Download PDFInfo
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- CN100480684C CN100480684C CNB2006100784781A CN200610078478A CN100480684C CN 100480684 C CN100480684 C CN 100480684C CN B2006100784781 A CNB2006100784781 A CN B2006100784781A CN 200610078478 A CN200610078478 A CN 200610078478A CN 100480684 C CN100480684 C CN 100480684C
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Abstract
The invention discloses a method for quantitatively analyzing isoflavones by fluidic injection chemical lighting technique, comprising: selectively oxidizing the isoflavones with K3Fe(CN)6 under alkali condition, and applying chemical lighting characteristic of the product to quantitatively analyze the isoflavones; and the method can quantitatively analyze isoflavones, including flavones and structurally similar compounds, especially compounds with phenolic group structure able to convert into quinone-type structure. And the method can be applied to determination of isoflavone content in food or drug; quality control analysis in the producing and circulating courses of the related products containing isoflavones; and detection of isoflavones metabolism level in an organism, playing a role of monitoring metabolism.
Description
Technical field
The present invention relates to chemical analysis technology, concrete, the present invention relates to the method that the applied chemistry luminescence technology is analyzed isoflavonoid, this method comprises isoflavonoid K
3Fe (CN)
6Selective oxidation is used the chemiluminescent properties of products therefrom and the qualitative and/or quantitative test that the flow injection technology is carried out isoflavonoid.
This method can be used for the isoflavonoid quantitative measurement.Therefore, can be used for isoflavonoid Determination on content in food or the medicine; Be used for containing the Related product production run of osajin and product Quality Control Analysis in the process of circulation; Be used for isoflavones level in the biosome is detected, play the effect of monitoring metabolism; Also can be used to measure the biologically active of isoflavonoid.
Background technology
Isoflavones is a kind of phytoestrogen, can promote the secretion of hormone and growth factor, has the activity of improving menopausal women syndrome, prevention of osteoporosis.The glucosides of isoflavones also has medical value, but tool medical value is not and the sugared aglycon that combines.Isoflavonoid is present in various legumes, particularly in the middle of soybean.Isoflavones in the soybean is divided into the glucoside unit (Aglycon) of free type and glucosides (Glycosides) two classes of mating type, aglycon accounts for 2%-3% of total amount, comprises daidzein (Daidzein), genistein (Genistein), Glycitein (Glycitein).Glucosides accounts for 97%-98% of total amount, mainly contains: daidzin (Daidzin), genistin (Genistin) and soya bean xanthosine (Glycitin) (referring to the gloomy good fortune man of family etc., soybean イ ソ Off ラ ボ Application [M], good fortune study etc.).
The metabolism of isoflavones is associated with hormone in biosome, so change and can play supervisory function bit to the metabolism of people, animal and biosomes such as microorganism and plant according to isoflavone content.To be applied to health is example, because the isoflavone content level is relevant with cancer, inoxidizability and women's climacteric syndrome in the known human body, therefore measuring isoflavone content can understand the health degree that is associated with human body.
Because above-mentioned reason, isoflavonoid have obtained widely using in many aspects, for example in the production and the application of food or medicine industry, and what therefore occur is height and the low content plant culture and the breeding etc. of target with the isoflavones.
Along with the widespread use of isoflavones in fields such as food, medicine and animal husbandry, in producing and using, must adopt the method for scientific and effective fast detecting isoflavonoid to comprise the fast detecting of antioxidation activity.
At present, measuring the method for isoflavones normally utilizes the ring texture of isoflavones molecule to have the characteristic of uv absorption, carry out qualitative and quantitative analysis under following three kinds of wavelength: the high performance liquid chromatography under ultraviolet spectrophotometry under the 260nm and 254nm UV-detector is (referring to Wu road silver and Luo Honglin, " extraction of talking isoflavones with separate ", grain processing, 2004,2:45-46); Spectrophotometric method general flavone mensuration under 360nm (referring to Zhao Xiao peak and Wu Rongshu, " health care of isoflavones and DEVELOPMENT PROSPECT ", oil engineering, 2004,8:39-41).These existing analytical approachs exist defective.Because be not to measure hydroxyl,, therefore be difficult to grasp its bioactive variation so can not recognize exactly whether hydroxyl is oxidized.
In order to overcome the defective that prior art exists, the inventor is through experiment repeatedly, and finding to use portable injection chemiluminescence analytical technology of the present invention can the bioactive content of rapid and precise quantitative measurement isoflavonoid.
Portable injection chemiluminescence analysis method is because it is highly sensitive, and the range of linearity is wide, and instrument and equipment is simple, favored by people.Up to the present do not see the report of chemical luminescent detecting isoflavones as yet.Inventor's discovery, in alkaline medium, K
3Fe (CN)
6Can the direct oxidation isoflavones, with the phenol activity hydroxy of isoflavones (4 '-OH) be oxidized to the quinoid structure, this oxidation product can absorb the chemical energy that this reaction emits and be excited, produce chemiluminescence, its values of chemiluminescence is measured and can be carried out (biologically active) quantitative test to described compound.This method not only has enough sensitivity, and good selectivity is arranged.
Summary of the invention
The invention provides a kind of new method of analyzing isoflavonoid, concrete, the invention provides and a kind ofly use the flow injection technology and method that chemiluminescence carries out qualitative and/or quantitative test to isoflavonoid, this method comprises uses K with described isoflavonoid under alkali condition
3Fe (CN)
6Carry out selective oxidation, and the chemiluminescent properties of using resultant product is carried out the qualitative and/or quantitative test of isoflavonoid.
On the other hand, isoflavonoid is carried out quantitative test, or be used to measure the biologically active of isoflavonoid according to the method.
The present invention analyzes in the method for isoflavonoid, and described isoflavonoid for example can be following formula (I) compound:
Wherein, R
1And R
2Be hydrogen, hydroxyl or methoxyl independently of one another.
Preferably, R in described formula (I) compound
1And R
2When being hydrogen, this compound is daidzein (Daidzein);
R in described formula (I) compound
1Be hydroxyl, R
2During for hydrogen, this compound is genistein (Genistein);
R in described formula (I) compound
1Be hydrogen, R
2When being methoxyl, this compound is Glycitein (Glycitein).
In analytical approach of the present invention, described alkaline medium is a sodium hydrate aqueous solution, and the concentration of described sodium hydroxide solution is 0.1-1.0mol/L, preferably approximately is 0.5mol/L concentration.
In order to obtain maximum values of chemiluminescence, the concentration of the K3Fe of oxygenant described in the said method (CN) 6 is generally 3.0-8.0 * 10
-4Mol/L preferably approximately is 5.0 * 10
-4Mol/L concentration.
Analyze in the method for isoflavonoid the K of wherein said isoflavonoid in the present invention
3Fe (CN)
6The chemiluminescence of oxidation product can be carried out with measuring chemiluminescent detection system, and the K of described isoflavonoid
3Fe (CN)
6The maximum emission wavelength of oxidation product is between 350-600nm.Along with the compound difference that is detected, its maximum emission wavelength can change to some extent.
In order to implement analytical approach of the present invention, the specialized equipment of analyzing must be arranged, this equipment comprises having sampling system, can measure chemiluminescent detection system, and to the sampling in the The whole analytical process, oxidation reaction and sample introduction, collection of experiment data and the software systems of carrying out the robotization processing.
Described sampling system can adopt the sampling system of instrumental analysis routine, and preferred flow sample injection system particularly has multichannel flow injection sampling system.
In described sampling system, finish the oxidation reaction process in the analytical approach of the present invention, and the suitable time with sample, the oxidation product that promptly carries out after the oxidation reaction injects detection system; Described detection system is after receiving test sample, and the light signal that can fast this sample be produced is transformed into electric signal, further again amplifying signal, and the basis signal size reaches the purpose of quantitative test.
In described sampling system, in order to measure isoflavonoid accurately, must adjust sample introduction speed accurately, its selection principle is to adjust sample introduction pump speed and valve pond distance, makes maximum luminous lucky can the detection fully by photomultiplier.
In this chemiluminescence current system, flow velocity causes maximum luminous before flow cell too slowly; Flow velocity causes luminous after flow cell too soon.To measure isoflavone content is example, produces chemiluminescent kinetic curve according to potassium ferricyanide oxidation isoflavones, reach the maximal value time to be about 4 seconds (s) from the sample introduction to the chemiluminescence, but luminous intensity decays near the background 6 seconds the time.Therefore the selection principle of sample introduction flow velocity is, adjusts sample introduction pump speed and valve pond distance, makes sample sample introduction 4 seconds, promptly reaches maximum and can be detected fully by photomultiplier just when luminous.
The test sample of the inventive method comprises the various products that contain isoflavonoid, and the isoflavones of being produced by soybean etc. for example contains the various health foods of isoflavonoid and medicine etc.; Or biological sample, for example blood sample of people etc.In described sample, may contain multiple other components, for example protein, inorganic salts, carbohydrate, saponin(e etc.The inventor tests the interference of analytical approach of the present invention these materials that may exist, experimental results show that because the sensitivity and the selectivity of the inventive method are very good, therefore, less demanding to the pre-treatment of sample, do not need to carry out complicated purification process.Usually, before analyzing, with the sample monobasic primary alconol kind solvent that needs are analyzed, for example ethanol extracts, and makes alcoholic solution, uses for analyzing.
Concrete, the method that the present invention analyzes isoflavonoid comprises:
(1) includes sampling system one, can measure chemiluminescent detection system and can carry out in the analytical instrument of the software systems that robotization handles described sampling system directly being connected with detection system The whole analytical process and collection of experiment data and processing;
(2) in described sampling system, make the sample that need analyze, contain isoflavonoid under alkali condition with K
3Fe (CN)
6Carry out oxidation reaction;
(3) measure the chemiluminescence of oxidation reaction product when the 350-600nm wavelength of above-mentioned steps with measuring chemiluminescent detection system;
(4) the isoflavonoid content of application standard curve calculation analytic sample, or the biologically active of calculating isoflavonoid.
Wherein, state on the implementation before the analytical approach, at first the sample that needs are analyzed extracts with alcohols solvent, makes alcoholic solution.Wherein said alcohols solvent preferred alcohol.
In the oxidation reaction of above-mentioned steps 2, described alkali condition is a sodium hydroxide solution, and particularly under the condition of sodium hydrate aqueous solution, the concentration of described sodium hydroxide solution is 0.1-1.0mol/L, preferably approximately is 0.5mol/L concentration.
In the oxidation reaction of above-mentioned steps 2, described oxygenant K
3Fe (CN)
6Concentration be generally 3.0-8.0 * 10
-4Mol/L, preferably approximately 5.0 * 10
-4Mol/L.
The method that the present invention analyzes isoflavonoid combines physical flow injection and chemical chemiluminescence, utilize chemical substance in the redox reaction process, to produce the characteristic of fluorescence or phosphorescence, the light signal that is produced is transformed into electric signal, further again amplifying signal, and the basis signal size reaches the purpose of quantitative test.The present invention proposes a kind of Chemiluminescence System of measuring isoflavone content according to this principle, and in conjunction with the flow injection technology, sets up a kind of simple fast measuring isoflavone content and/or the bioactive new method of isoflavones.This method not only has high sensitivity, and good selectivity is arranged.
Use known technology in the instrumental analysis field, can realize the robotization of analytic process and computation process, for example can use sampling commonly used in the existing analytical instrument and sampling system, detection system and experimental data collection and disposal system and make The whole analytical process realize robotization.The MPI-B type multiparameter chemiluminescence analysis test macro produced of Xi'an Rui Mai Electronics Co., Ltd. for example, is annotated sample, experimental data collection and is handled and finish by MPI-B type multiparameter chemiluminescence analysis testing system software under the Windows XP system at the sampling in the The whole analytical process.
As detected object, this method can satisfy in the biosome and the quantitative test of relevant isoflavones (biologically active) in all products method of the present invention with isoflavonoid.In addition, principle of work according to this method, the application extension of this method can be arrived the assay determination of Flavonoid substances fully, the for example assay determination of Flavonoid substances, or expand to other and have the similar structures compound, the assay determination of compound that promptly has the phenolic hydroxyl group of oxidable one-tenth quinoid structure, the assay determination of for example close steroids compound with molecular structure.
Before carrying out above-mentioned analysis, the sample that needs should be analyzed extracts with alcohols solvent, and making the wherein said alcohols solvent of alcoholic solution is the monobasic primary alconol.Because analytical approach of the present invention has high sensitivity and high selectivity, so analytic sample does not need to carry out complicated purification step.
Analytical approach of the present invention can be applied to isoflavonoid Determination on content in food or the medicine; The Related product production run and the Quality Control Analysis of product in the process of circulation that are used for the isoflavone-containing compounds; Being used for the isoflavones is the quality monitoring of height and low content plant culture and breeding process of target; And the isoflavone content in the biosome monitored, so that the bioactive variation according to isoflavone content and/or isoflavonoid is played supervisory function bit to the metabolism of people, animal and biosomes such as microorganism and plant, for example, because the isoflavone content level is relevant with cancer, inoxidizability and women's climacteric syndrome in the known human body, therefore measures the biologically active of isoflavone content and/or isoflavonoid and can understand the health degree that is associated with human body.
Description of drawings
Fig. 1 is that potassium ferricyanide oxidation isoflavones produces chemiluminescent kinetic curve.Wherein, the concentration of isoflavones is 5.0 * 10
-6G/mL, the concentration of sodium hydroxide solution is 5mol/L, and the concentration of potassium ferricyanide solution is 5.0 * 10
-4Mol/L.
Fig. 2 is that potassium ferricyanide concentration is to chemiluminescent influence.The result shows, when the concentration of the potassium ferricyanide is 5.0 * 10
-4During the mol/L left and right sides, the luminous intensity maximum.
Fig. 3 is the emission spectrum of isoflavones (a), the potassium ferricyanide (b) and isoflavones+potassium ferricyanide (c).
Embodiment
1, the selection of instrument and equipment
1.1, instrument: MPI-B type multiparameter chemiluminescence analysis test macro, comprising flow injection hyperchannel sampling system, Rui Mai Electronics Co., Ltd. in Xi'an makes.
Sampling in the The whole analytical process, notes sample, experimental data collection and processing are finished by MPI-B type multiparameter chemiluminescence analysis testing system software under the WindowsXP system.
1.2, the UV-1600 ultraviolet-visible pectrophotometer: the Beijing Rayleigh Analytical Instrument Co.,Ltd.
2. material and reagent:
2.1, analytic target: the isoflavones that is equipped with is obtained through refining in this laboratory, is solvent with 0.5mol/LNaOH, makes the solution of variable concentrations;
2.2, reagent:
Alkali: NaOH makes the solution that concentration is 0.5mol/L;
The potassium ferricyanide: with 0.5mol/L NaOH is solvent, makes the potassium ferricyanide solution of variable concentrations;
Isoflavones standard solution (0.5g/L): accurately take by weighing isoflavones (Japanese Wako Pure Chemical Industries, Ltd.) 5.0mg and be dissolved in absolute ethyl alcohol, the water constant volume is in the brown volumetric flask of 10mL, and is standby.
(3) it is pure that all the other reagent are analysis, and water is secondary deionized water.
3. experimental technique
3.1, two passages of the active efflux of MPI-B type instrument are respectively with current-carrying (H
2O) and K
3Fe (CN)
6Solution carries out threeway with the flow velocity of 45r/min by corresponding passage and mixes, the input analytic system.After treating that baseline steadily, secondary moving pump flows into the flow velocity of isoflavones solution with 35r/min in the current-carrying by six logical introduction valves (annotating the sample time: 10 seconds), with K
3Fe (CN)
6Solution mixes, and produces chemiluminescence in flow cell, and is quantitative according to the strong and weak external standard method that adopts of chemiluminescence.
3.2, studied of the influence of following each material respectively to the isoflavones luminous intensity: the kind of oxygenant, NaOH concentration, surfactant, flow velocity and interference experiment.
4. result and discussion:
4.1 chemiluminescence kinetic curve
K
3Fe (CN)
6The oxidation isoflavones produces chemiluminescence (referring to Fig. 2), reaches the maximal value time to be about 4s from the sample introduction to the chemiluminescence, and luminous intensity decays near the background during 6s.
4.2 measure the selection of isoflavones condition
4.2.1 the selection of oxygenant
At acid medium (1mol/L H
2SO
4)-KMnO
4And alkaline medium (0.5mol/LNaOH)-K
3Fe (CN)
6Or H
2O
2Carry out the direct oxidation isoflavones as oxygenant respectively and produce chemiluminescent research, studies show that acid medium-KMnO
4And alkaline medium-K
3Fe (CN)
6When making oxygenant, produce chemiluminescence.Test findings shows, the KMnO in the acid medium
4Oxidisability be better than alkaline K
3Fe (CN)
6, acid KMnO
4Not only can the oxidation isoflavones, can also the oxidation polyhydroxy-alcohol, materials such as sugar, vitamin C, influence test findings.And K
3Fe (CN)
6Oxidation isoflavones optionally only in this system.Therefore, method of the present invention is preferably used alkaline medium-K
3Fe (CN)
6Measure isoflavones as oxygenant.
4.2.2 K
3Fe (CN)
6With NaOH concentration to chemiluminescent influence
Experiment showed, K
3Fe (CN)
6With the concentration of NaOH the chemiluminescence intensity of this system there is very big influence.
Under the concentration of having fixed isoflavones and alkaline medium, The effect the K of variable concentrations
3Fe (CN)
6(0～1.0 * 10
-3Mol/L) and NaOH (1.0～1.0 * 10
-5Mol/L) to the influence of this chemiluminescence reaction.The result shows, works as K
3Fe (CN)
6Concentration be 5.0 * 10
-4During the mol/L left and right sides, luminous intensity maximum (as Fig. 3).K
3Fe (CN)
6Only in alkaline medium, could produce chemiluminescence by the oxidation isoflavones.Test shows that the optium concentration of NaOH is about 0.5mol/L.
4.2.3 the influence of sample introduction flow velocity
In this chemiluminescence current system, flow velocity causes maximum luminous before flow cell too slowly; Flow velocity causes luminous after flow cell too soon.The selection principle of sample introduction flow velocity is to reach maximal value to need 4s from the sample introduction to the chemiluminescence, adjusts sample introduction pump speed and valve pond distance, makes maximum luminous lucky can the detection fully by photomultiplier.
Rotations per minute with direct peristaltic pump and secondary peristaltic pump is 0～99, their revolution has been carried out combination set and find that best revolution is respectively 45r/min and 35r/min.
4.2.4 the range of linearity, detectability and sensitivity: under above-mentioned top condition, measured the relation of peak area and isoflavones concentration, the result shows that isoflavones concentration is 1.0 * 10
-2Have good linear relationship in the scope of～0.5g/L, its regression equation is:
A＝193305C(mg/L)+229.97，r
2＝0.9962
To 5.0 * 10
-3G/L isoflavones solution METHOD FOR CONTINUOUS DETERMINATION, each 3 peak values repeat 7 times, and the relative standard deviation is 2.46%.According to the IUPAC suggestion, detecting of this method is limited to 4.6 * 10
-4G/L.
4.2.5 interference experiment: isoflavones derives from soybean, coexistence other components (protein, inorganic salts, carbohydrate, saponin etc.) in leaching process, and they may measure isoflavones to this chemical luminous system certain influence.For this reason, studied 1000 times Na
+, Cl
-, K
+, Br
-, CO
3 2-, SO
4 2-And NO
3 -100 times starch, dextrin; 10 times glucose, fructose, sucrose; Vitamin C doubly is respectively to 5.0 * 10 together
-6The isoflavones of g/mL carries out the interference measurement experiment.The result shows that they are interference measurement not.Have only halfcystine that measured signal is had weak interference (signal to noise ratio (S/N ratio) is less than 1) in the saturated solution of 20 seed amino acids, do not influence test findings; Other amino acid are noiseless.Therefore, when adopting flow injection and chemiluminescence determination isoflavones (biologically active) content, do not need sample is carried out purification process, can be accurately quantitatively.
Though 4.2.6 luminescence mechanism has produced chemical luminescent detecting rutin (referring to Li Baoxin etc., analytical chemistry, 2001,428) with the potassium ferricyanide as the oxygenant direct oxidation.But this paper is not done further investigation for its luminescence mechanism.
In order to disclose the luminescence mechanism of the potassium ferricyanide to the isoflavones oxidation more accurately, the present invention has done following experiment.Analyzed the emission spectrum (Fig. 3) of the potassium ferricyanide, isoflavones titer and the potassium ferricyanide+isoflavones mixed liquor with the LS45 fluorophotometer, the result shows that isoflavones has faint fluorescence, its maximum emission wavelength be 412nm (a) as figure.Figure b is a potassium ferricyanide reflection at peak, is not fluorescence peak, and is identical with first peak among the figure c.After adding the potassium ferricyanide, isoflavones is by potassium ferricyanide oxidation, the fluorescent weakening of isoflavones, and the potassium ferricyanide amount of adding is many more, and the fluorescence of isoflavones is just weak more, and the maximum emission wavelength of its oxidation product is 350～600nm (as figure b).This illustrates that the luminophor of this chemical luminous system is the product of isoflavones oxidation.Oxidation product has absorbed the chemical energy that this reaction emits and has been excited, and this part energy that is dropped to ground state by excited state discharges with the form of photon.
4.2.7 practical measurement
Use chemiluminescence method of the present invention and measure isoflavones in the capsule, and with colourimetry in contrast.
Buy the isoflavones capsule from market, get the powder in 1 capsules at random, the porphyrize mixing spends the night with anhydrous alcohol solution.The centrifugal 10min of 10000rpm next day, it is fixed molten to 100ml with absolute ethyl alcohol to take out supernatant.In the range of linearity, measure triplicate with this chemiluminescence method and 260nm colourimetry respectively.The measurement result of the inventive method is respectively the 5.16mg/ grain; The measurement result of colourimetry is the 5.72mg/ grain, illustrates and compares the accuracy of method of the present invention with known method.
According to above description, identifiable is to compare in the method for prior art, the present invention uses the chemiluminescent properties of isoflavonoid and quantitative analysis method that the flow injection technology is carried out isoflavonoid has higher sensitivity and selectivity, and therefore the advantage of bringing is that analytic sample does not need to carry out complicated purification step.Therefore, analytical approach of the present invention is very practical advanced technology.
Described embodiment of the present invention now in detail, clearly can do a lot of improvement and variation for a person skilled in the art and can not deviate from essence spirit of the present invention.All these changes and improvements think all within the scope of the present invention that its feature is determined by above-mentioned instructions.
Claims (23)
1, the analytical approach of isoflavonoid, this method is included in the alkaline medium, uses K
3Fe (CN)
6Oxidation isoflavonoid, and the chemiluminescence of using resulting oxidation product carry out qualitative and/or quantitative test to isoflavonoid; Wherein said analysis is to include sampling system and can measure in the analytical instrument of chemiluminescent detection system, and measuring emission wavelength is the chemiluminescence of 350-600nm.
2, according to the described analytical approach of claim 1, wherein said isoflavonoid is following formula (I) compound:
Wherein, R
1And R
2Be hydrogen, hydroxyl or methoxyl independently of one another.
3, according to the described analytical approach of claim 2, in wherein said formula (I) compound, R
1And R
2Be hydrogen.
4, according to the described analytical approach of claim 2, in wherein said formula (I) compound, R
1Be hydroxyl, and R
2Be hydrogen.
5, according to the described analytical approach of claim 2, in wherein said formula (I) compound, R
1Be hydrogen, and R
2It is methoxyl.
6, according to the described analytical approach of claim 1, wherein the employed analytical instrument of this method also comprises the software systems of The whole analytical process, collection of experiment data and processing being carried out the robotization processing.
7, according to the described analytical approach of claim 1, wherein said sampling system is the flow injection sampling system.
8, according to the described analytical approach of claim 7, wherein said oxidation reaction is carried out in this sampling system.
9, according to the described analytical approach of claim 8, wherein adjust the sample introduction flow velocity of described sampling system, the oxidation product that makes isoflavonoid detects fully reaching maximum photomultiplier that just can detected system when luminous.
10, according to the described analytical approach of claim 2, wherein at the described K that measures described isoflavonoid in the chemiluminescent detection system that measures
3Fe (CN)
6The chemiluminescence of oxidation product.
11, according to the described analytical approach of claim 1, wherein said K
3Fe (CN)
6Concentration be 3.0-8.0 * 10
-4Mol/L.
12, according to the described analytical approach of claim 11, wherein said K
3Fe (CN)
6Concentration be about 5.0 * 10
-4Mol/L.
13, according to the described analytical approach of claim 1, wherein said alkaline medium is a sodium hydrate aqueous solution, and the concentration of described sodium hydroxide solution is 0.1-1.0mol/L.
14, the analytical approach of stating according to claim 13, the concentration of wherein said sodium hydroxide solution is approximately 0.5mol/L.
15, according to any described analytical approach of claim 1-14, this method comprises:
(1) includes sampling system one, can measure chemiluminescent detection system and can carry out in the analytical instrument of the software systems that robotization handles described sampling system directly being connected with detection system The whole analytical process and collection of experiment data and processing;
(2) in described sampling system, make the sample that need analyze, contain isoflavonoid under alkali condition with K
3Fe (CN)
6Carry out oxidation reaction;
(3) measure the chemiluminescence of oxidation reaction product when the 350-600nm wavelength of above-mentioned steps with measuring chemiluminescent detection system; With
(4) content of isoflavonoid in the application standard curve calculation analytic sample.
16, according to the described analytical approach of claim 15, wherein before analyzing, the sample that needs are analyzed extracts with alcohols solvent, makes alcoholic solution.
17, according to the described analytical approach of claim 16, wherein said alcohols solvent is the monobasic primary alconol.
18, according to the described analytical approach of claim 17, wherein said primary alconol is an ethanol.
19, according to the described analytical approach of claim 15, wherein the described alkali condition of step (2) is meant and has NaOH.
20, according to the application of any described analytical approach of claim 1-19 in isoflavonoid is measured.
21, according to the application of claim 20, wherein said application comprises the detection of isoflavone content in the detection, biosome of the food of quality of production control, isoflavone-containing compounds of isoflavone-containing compounds product or medicine and to the bioactive detection of isoflavonoid.
22, according to the application of claim 21, wherein the isoflavonoid of being measured is following formula (I) compound:
Wherein, R
1And R
2Be hydrogen, hydroxyl or methoxyl independently of one another.
23, according to the application of claim 22, wherein the isoflavonoid of being measured is daidzein, genistein and/or Glycitein.
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| 紫外分光光度法测定大豆异黄酮含量. 王哲等.中国油脂，第30卷第1期. 2005 |
| 紫外分光光度法测定大豆异黄酮含量. 王哲等.中国油脂，第30卷第1期. 2005 * |
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