CN102507533A - Capillary electrophoresis electrochemiluminescence detection method for arecoline in traditional Chinese medicine betelnut extract - Google Patents
Capillary electrophoresis electrochemiluminescence detection method for arecoline in traditional Chinese medicine betelnut extract Download PDFInfo
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- CN102507533A CN102507533A CN201110301068XA CN201110301068A CN102507533A CN 102507533 A CN102507533 A CN 102507533A CN 201110301068X A CN201110301068X A CN 201110301068XA CN 201110301068 A CN201110301068 A CN 201110301068A CN 102507533 A CN102507533 A CN 102507533A
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Abstract
The invention provides a capillary electrophoresis electrochemiluminescence detection method for arecoline in a traditional Chinese medicine betelnut extract. The method applies capillary electrophoresis ruthenium pyridine electrochemiluminescence analysis technology to the detection of arecoline in the traditional Chinese medicine betelnut extract for the first time, and has the advantages that the operation is simple, the sensitivity is high, the detection limit of arecoline is 5*10<-9> mol/L, the separation function is strong, the pollution is avoided, and the field amplified injection is realized and the separation selectivity of arecoline and coexisting media in the betelnut extract is improved by using ionic liquid BMImBF4 in a buffer. Electrochemiluminescence reagent ruthenium pyridine is reversible in reaction and high in photon yield, thus the reagent consumption is greatly reduced and the detection cost is lowered. The analysis method is green, environmentally friendly and low in sample consumption. The analysis can be finished by adopting a capillary electrophoresis electrochemiluminescence comprehensive analyzer and chemical reagents which are common in laboratory and requiring nonoliter-scale injection volume.
Description
Technical field
The invention belongs to the capillary electrophoresis electrochemical light-emitting analysis technical field, be specifically related to the detection method that capillary electrophoresis electrochemical light-emitting detects arecaline in the Chinese medicine betelnut extract.
Background technology
Betel nut is one of China's famous and precious " four Da Nan medicines ", cures mainly malnutrition due to parasitic infestation, dyspepsia, the stagnation of the circulation of vital energy, dysentery, drives illnesss such as ascarid, glaucoma.Excess is taken and is prone to poison, and can stimulate digestion produces vomiting, increases glandular secretion, and influences central nervous system and urinary system.Arecaline is an important biological composition in the betel nut, and the compartment analysis of arecaline is an important indicator of estimating Chinese medicine betel nut quality in the betelnut extract.Because active component content is low in the betel nut, therefore the sensitive arecaline detection technique of development all is one of research focus of Chinese medicine analysis field all the time.
At present, the modal analytical approach of arecaline is high efficiency liquid phase chromatographic analysis method [1], mass-spectrometric technique [2], and high performance liquid chromatography mass spectrometric hyphenated technique [3,4].(list of references [1] Cox S, Piatkov I, Vickers ER, Ma G.j.Chromatogr.A 2004; 1032:93-95. [2] Feng CH, Lu CY.Anal.Chim.Acta 2009; 649:230-235. [3] Pellegrini M, Marchei E, Rossi S, Vagnarelli F, Durgbansh A, Garcia-Algar S, Vall O, Pichini S.Rapid Commun.Mass Spectrom.2007; 21:2693-2703. [4] Marchei E, Durgbanshi A, Rossi S, Garcia-Algar S, Pichini S.Rapid Commun.Mass Spectrom.2005; 19:3416-3418.)
The high performance liquid chromatography mass spectrometric hyphenated technique is an active ingredient of Chinese herbs analysis analysis means commonly used.Yet because high performance liquid chromatography adopts organic composition to be measured that is separated that flows, it is one of subject matter of existing of this technology that environment is polluted, and people are making great efforts exploration environmental protection, efficient, sensitive active ingredient of Chinese herbs compartment analysis new technology always for this reason.
Summary of the invention
In order to solve the problem that prior art exists, the present invention provides the capillary electrophoresis electrochemical light-emitting detection method of arecaline in a kind of Chinese medicine betelnut extract.Capillary Electrophoresis is a kind of high performance liquid chromatogram differential from technology, compares with high performance liquid chromatography, and the sample feeding amount is merely receives upgrading (the high performance liquid chromatography sample size is a micro updating).Capillary Electrophoresis is that driving force is implemented to separate with the electric osmose, and separation electrophoresis solution has been avoided the use of organic solvent; The fine inner diameter kapillary separates can apply higher separation voltage, therefore can realize comprising positive and negative electrical ion and the multicomponent high efficiency separation of neutral compound.
Capillary Electrophoresis can with the purpose of many detection means couplings to realize that sample separation is analyzed.The pyridine ruthenium electrochemical is luminous to be a kind of detection technique of sensitivity, and high, the no bias light of photon productive rate disturbs, and in the reaction of aqueous phase generation electrochemiluminescence, has favorable compatibility with composition to be measured.Capillary electrophoresis separation and Ru (bpy)
3 2+The electrochemiluminescence coupling technique is a kind of novel analytical technology that has application potential that has both high efficiency separation and Sensitive Detection.This isolation technics more is applicable to the compartment analysis of the trace composition to be measured that comprises active ingredient of Chinese herbs.Advantages such as this method has efficiently, sensitive, quick are a kind of new methods of analyzing active ingredient of Chinese herbs.
The present invention provides the capillary electrophoresis electrochemical light-emitting detection method of arecaline in a kind of Chinese medicine betelnut extract, and step and condition are following:
1.1 the preparation of article to be checked
1.1.1 the preparation of arecaline standard solution
Be dissolved in the arecaline standard items in the secondary water, making concentration is 5.0 * 10
-3The arecaline standard solution of mol/L;
1.1.2 the preparation of betelnut extract related solution
1.1.2.1 the preparation of betelnut extract solution
Get 0.3g Chinese medicine Areca Seed, with 2.0mL secondary water ultrasonic extraction 30min in ultrasonic pond, extracting operation repeats 3 times, and combining extraction liquid with the membrane filtration of 0.45 μ m, obtains betelnut extract solution;
1.1.2.2 the preparation of betelnut extract dilute solution
The betelnut extract solution that obtains to step 1.1.2.1 adds secondary water, is settled to 250mL, obtains arecaline extract dilute solution;
1.1.2.3 the preparation of betelnut extract dilution mark-on solution
In the betelnut extract solution that step 1.1.2.1 obtains, adding the concentration that is obtained by step 1.1.1 respectively is 5.0 * 10
-3The arecaline standard solution of mol/L is settled to 250mL with the dilution of secondary water, obtains betelnut extract dilution mark-on solution;
1.2 detect step and condition
1.2.1 apparatus
58cm length, internal diameter 50 μ m not coating melt silicon capillary; Diameter 500 μ m platinum dish working electrodes; Capillary electrophoresis electrochemical light-emitting synthesis analyzer (the auspicious analytical instrument advanced in years in Xi'an Ltd produces);
1.2.2 reagent
The standard items of arecaline, (Ru (bpy)
3Cl
2.6H
2O), NaH
2PO
4, Na
2HPO
4, NaOH and secondary water; It is pure that agents useful for same is analysis; The solution that below prepares is using before all through 0.45 μ m membrane filtration;
1.2.3 the preparation of solution
1.2.3.1 the preparation of background electrolyte
(1) preparation of PBS
At first compound concentration is the NaH of 200.0mmol/L
2PO
4With concentration be the Na of 200.0mmol/L
2HPO
4, again will be with the NaH of concentration
2PO
4And Na
2HPO
4Mix being made into the PBS that concentration is 100.0mmol/L, adjustment PBS pH value is 7.50;
(2) Ru (bpy)
3 2+The preparation of solution
With secondary water preparation 10.0mmol/L Ru (bpy)
3 2+Solution;
(3) preparation of background electrolyte
With the PBS of step (1) preparation and the Ru (bpy) of step (2) preparation
3 2+Solution preparation background electrolyte; Phosphatic concentration is 50.0mmol/L in the background electrolyte, Ru (bpy)
3 2+Concentration be 5.0mmol/L;
1.2.3.2 the preparation of runtime buffer solution
(1) preparation of PBS
Compound concentration is the NaH of 200.0mmol/L
2PO
4With concentration be the Na of 200.0mmol/L
2HPO
4, will use the dilution of secondary water to be made into the PBS of concentration again with the two mixing of concentration as 50.0mmol/L, adjustment PBS pH value is 7.50, obtains PBS;
(2) BMImBF
4The preparation of solution
With BMImBF
4Use the dilution of secondary water to be made into the solution of concentration as 50.0mmol/L;
(3) preparation of runtime buffer solution
The BMImBF that PBS that step (1) is obtained and step (2) obtain
4Solution mixes, and obtains runtime buffer solution with the dilution of secondary water again; Phosphate concn is 20mmol/L in the runtime buffer solution, BMImBF
4Concentration is 10.0mmol/L;
1.2.4 detection step
1.2.4.1 the processing of separation capillary
For activation kapillary inside surface, guarantee the reappearance of analysis result, need separation capillary is done following processing:
New capillary column uses the activation of 100mmol/L NaOH to spend the night, and before experiment every day, capillary column with secondary water flushing 10min, is used the runtime buffer solution equilibria 10min that is obtained by step 1.2.3.2 more then with 100mmol/L NaOH flushing 10min;
1.2.4.2 the detection of arecaline in the Chinese medicine betelnut extract
The betelnut extract dilution mark-on solution that obtains among betelnut extract dilute solution that obtains among the step 1.1.2.2 and the step 1.1.2.3 is detected by following condition:
Detect current potential 1.20V; Electrophoretic separation high pressure 16kV; Add the background electrolyte of step 1.2.3.1 preparation in the detection cell; Electrochemical reaction adopts three-electrode system, and three-electrode system comprises that diameter 500 μ m platinum dish working electrodes, platinum filament are to the utmost point and Ag/AgCl contrast electrode; Long capillary tube 58cm, internal diameter 50 μ m; The runtime buffer solution that electrophoresis uses step 1.2.3.2 to obtain; Adopt styletable electrochemiluminescence detecting pattern; The photomultiplier bias voltage is set in 800V; Obtain the electrophoresis pattern that arecaline detects in the Chinese medicine betelnut extract.
Beneficial effect: the present invention provides the capillary electrophoresis electrochemical light-emitting detection method of arecaline in a kind of Chinese medicine betelnut extract; First Capillary Electrophoresis pyridine ruthenium electrochemical luminesceence analysis technology is applied to the detection of arecaline in the Chinese medicine betelnut extract, adopts ionic liquid BMImBF
4Not only realize electrical field magnified injection, and helped the high efficiency separation and the Sensitive Detection of the complicated betelnut extract of medium.
(1) simple to operate, highly sensitive.
The pyridine ruthenium electrochemical is luminous to be a kind of detection technique of sensitivity, and arecaline has very strong sensitization to the pyridine ruthenium, need not derive to testing sample, and the Chinese medicine betelnut extract can directly be used for the quantitative test of arecaline, and is simple and quick.The present invention adds ionic liquid BMImBF on this basis in the runtime buffer system
4, utilize ion liquid high conductivity, realize the enrichment method electrical field magnified injection, improved detection sensitivity, method of the present invention is limited to 5 * 10 to the detection of arecaline
-9Mol/L.
(2) separation function is powerful, and is pollution-free.
Runtime buffer effects of ion liquid B MImBF
4Use, not only caused electrical field magnified injection, also improved in arecaline and the betelnut extract separation selectivity of coexistence medium, can realize the high efficiency separation of arecaline.Contain the application of phosphate and ionic liquid running buffer, make this analysis method satisfy green, environmental protection requirement.
(3) amount of samples is few, and it is low to detect cost.
The capillary electrophoresis electrochemical light-emitting synthesis analyzer is the basic operation platform that active ingredient of Chinese herbs is analyzed, and more auxiliary laboratory common chemical reagent can be accomplished analysis task.The capillary electrophoresis electrochemical light-emitting system only need receive the upgrading sample size can satisfy the separation detection demand.The reaction of electrochemiluminescence reagent pyridine ruthenium is reversible, the photon productive rate high, has significantly reduced the consumption of reagent, reduces and detects cost.
Description of drawings
Fig. 1 is the electrophoresis spectrogram that betelnut extract is detected.Wherein a is a betelnut extract dilute solution electrophoresis spectrogram, and b is the betelnut extract dilution mark-on solution electrophoresis spectrogram that adds the arecaline standard items.1 peak is the electrophoresis peak of arecaline.Fig. 1
Embodiment
Following examples adopt the capillary electrophoresis electrochemical light-emitting synthesis analyzer.Detect current potential 1.20V; Electrophoretic separation high pressure 16kV; Add the background electrolyte of step 1.2.3.1 preparation in the detection cell; Electrochemical reaction adopts three-electrode system, and three-electrode system comprises that diameter 500 μ m platinum dish working electrodes, platinum filament are to the utmost point and Ag/AgCl contrast electrode; Long capillary tube 58cm, internal diameter 50 μ m; The runtime buffer solution that electrophoresis uses step 1.2.3.2 to obtain; Adopt styletable electrochemiluminescence detecting pattern; The photomultiplier bias voltage is set in 800V;
The present invention provides the capillary electrophoresis electrochemical light-emitting detection method of arecaline in a kind of Chinese medicine betelnut extract, and step and condition are following:
1.1 the preparation of article to be checked
1.1.1 the preparation of arecaline standard solution
Be dissolved in the arecaline standard items in the secondary water, making concentration is 5.0 * 10
-3The arecaline standard solution of mol/L;
1.1.2 the preparation of betelnut extract related solution
1.1.2.1 the preparation of betelnut extract solution
Get 0.3g Chinese medicine Areca Seed, with 2.0mL secondary water ultrasonic extraction 30min in ultrasonic pond, extracting operation repeats 3 times, and combining extraction liquid with the membrane filtration of 0.45 μ m, obtains betelnut extract solution;
1.1.2.2 the preparation of betelnut extract dilute solution
The betelnut extract solution that obtains to step 1.1.2.1 adds secondary water, is settled to 250mL, obtains arecaline extract dilute solution;
1.1.2.3 the preparation of betelnut extract dilution mark-on solution
In the betelnut extract solution that step 1.1.2.1 obtains, adding the concentration that is obtained by step 1.1.1 respectively is 5.0 * 10
-3The arecaline standard solution of mol/L is settled to 250mL with the dilution of secondary water, obtains betelnut extract dilution mark-on solution;
1.2 detect step and condition
1.2.1 apparatus
58cm length, internal diameter 50 μ m not coating melt silicon capillary; Diameter 500 μ m platinum dish working electrodes; Capillary electrophoresis electrochemical light-emitting synthesis analyzer (the auspicious analytical instrument advanced in years in Xi'an Ltd produces);
1.2.2 reagent
The standard items of arecaline, (Ru (bpy)
3Cl
2.6H
2O), NaH
2PO
4, Na
2HPO
4, NaOH and secondary water; It is pure that agents useful for same is analysis; The solution that below prepares is using before all through 0.45 μ m membrane filtration;
1.2.3 the preparation of solution
1.2.3.1 the preparation of background electrolyte
(1) preparation of PBS
At first compound concentration is the NaH of 200.0mmol/L
2PO
4With concentration be the Na of 200.0mmol/L
2HPO
4, again will be with the NaH of concentration
2PO
4And Na
2HPO
4Mix being made into the PBS that concentration is 100.0mmol/L, adjustment PBS pH value is 7.50;
(2) Ru (bpy)
3 2+The preparation of solution
With secondary water preparation 10.0mmol/L Ru (bpy)
3 2+Solution;
(3) preparation of background electrolyte
With the PBS of step (1) preparation and the Ru (bpy) of step (2) preparation
3 2+Solution preparation background electrolyte; Phosphatic concentration is 50.0mmol/L in the background electrolyte, Ru (bpy)
3 2+Concentration be 5.0mmol/L;
1.2.3.2 the preparation of runtime buffer solution
(1) preparation of PBS
Compound concentration is the NaH of 200.0mmol/L
2PO
4With concentration be the Na of 200.0mmol/L
2HPO
4, will use the dilution of secondary water to be made into the PBS of concentration again with the two mixing of concentration as 50.0mmol/L, adjustment PBS pH value is 7.50, obtains PBS;
(2) BMImBF
4The preparation of solution
With BMImBF
4Use the dilution of secondary water to be made into the solution of concentration as 50.0mmol/L;
(3) preparation of runtime buffer solution
The BMImBF that PBS that step (1) is obtained and step (2) obtain
4Solution mixes, and obtains runtime buffer solution with the dilution of secondary water again; Phosphate concn is 20mmol/L in the runtime buffer solution, BMImBF
4Concentration is 10.0mmol/L;
1.2.4 detection step
1.2.4.1 the processing of separation capillary
For activation kapillary inside surface, guarantee the reappearance of analysis result, need separation capillary is done following processing:
New capillary column uses the activation of 100mmol/L NaOH to spend the night, and before experiment every day, capillary column with secondary water flushing 10min, is used the runtime buffer solution equilibria 10min that is obtained by step 1.2.3.2 more then with 100mmol/L NaOH flushing 10min;
1.2.4.2 the detection of arecaline in the Chinese medicine betelnut extract
The betelnut extract dilution mark-on solution that obtains among betelnut extract dilute solution that obtains among the step 1.1.2.2 and the step 1.1.2.3 is detected by following condition:
Detect current potential 1.20V; Electrophoretic separation high pressure 16kV; Add the background electrolyte of step 1.2.3.1 preparation in the detection cell; Electrochemical reaction adopts three-electrode system, and three-electrode system comprises that diameter 500 μ m platinum dish working electrodes, platinum filament are to the utmost point and Ag/AgCl contrast electrode; Long capillary tube 58cm, internal diameter 50 μ m; The runtime buffer solution that electrophoresis uses step 1.2.3.2 to obtain; Adopt styletable electrochemiluminescence detecting pattern; The photomultiplier bias voltage is set in 800V; Obtain the electrophoresis pattern (see figure 1) that arecaline detects in the Chinese medicine betelnut extract.
Claims (1)
1. the capillary electrophoresis electrochemical light-emitting detection method of arecaline in the Chinese medicine betelnut extract, step and condition are following:
1.1 the preparation of article to be checked:
1.1.1 the preparation of arecaline standard solution: be dissolved in the arecaline standard items in the secondary water, making concentration is 5.0 * 10
-3The arecaline standard solution of mol/L;
1.1.2 the preparation of betelnut extract related solution:
1.1.2.1 the preparation of betelnut extract solution: get 0.3g Chinese medicine Areca Seed, with 2.0mL secondary water ultrasonic extraction 30min in ultrasonic pond, extracting operation repeats 3 times, and combining extraction liquid with the membrane filtration of 0.45 μ m, obtains betelnut extract solution;
1.1.2.2 the preparation of betelnut extract dilute solution: the betelnut extract solution that obtains to step 1.1.2.1 adds secondary water, is settled to 250mL, obtains arecaline extract dilute solution;
1.1.2.3 the preparation of betelnut extract dilution mark-on solution: in the betelnut extract solution that step 1.1.2.1 obtains, adding the concentration that is obtained by step 1.1.1 respectively is 5.0 * 10
-3The arecaline standard solution of mol/L is settled to 250mL with the dilution of secondary water, obtains betelnut extract dilution mark-on solution;
1.2 detect step and condition:
1.2.1 apparatus: 58cm length, internal diameter 50 μ m not coating melt silicon capillary; Diameter 500 μ m platinum dish working electrodes; The capillary electrophoresis electrochemical light-emitting synthesis analyzer;
1.2.2 reagent: the standard items of arecaline, (Ru (bpy)
3Cl
2.6H
2O), NaH
2PO
4, Na
2HPO
4, NaOH and secondary water; It is pure that agents useful for same is analysis; The solution that below prepares is using before all through 0.45 μ m membrane filtration;
1.2.3 the preparation of solution:
1.2.3.1 the preparation of background electrolyte: the preparation of (1) PBS: at first compound concentration is the NaH of 200.0mmol/L
2PO
4With concentration be the Na of 200.0mmol/L
2HPO
4, again will be with the NaH of concentration
2PO
4And Na
2HPO
4Mix being made into the PBS that concentration is 100.0mmol/L, adjustment PBS pH value is 7.50; (2) Ru (bpy)
3 2+The preparation of solution: with secondary water preparation 10.0mmol/L Ru (bpy)
3 2+Solution; (3) preparation of background electrolyte: with the PBS of step (1) preparation and the Ru (bpy) of step (2) preparation
3 2+Solution preparation background electrolyte; Phosphatic concentration is 50.0mmol/L in the background electrolyte, Ru (bpy)
3 2+Concentration be 5.0mmol/L;
1.2.3.2 the preparation of runtime buffer solution: the preparation of (1) PBS: compound concentration is the NaH of 200.0mmol/L
2PO
4With concentration be the Na of 200.0mmol/L
2HPO
4, will use the dilution of secondary water to be made into the PBS of concentration again with the two mixing of concentration as 50.0mmol/L, adjustment PBS pH value is 7.50, obtains PBS; (2) BMImBF
4The preparation of solution: with BMImBF
4Use the dilution of secondary water to be made into the solution of concentration as 50.0mmol/L; (3) preparation of runtime buffer solution: the BMImBF that PBS that step (1) is obtained and step (2) obtain
4Solution mixes, and obtains runtime buffer solution with the dilution of secondary water again; Phosphate concn is 20mmol/L in the runtime buffer solution, BMImBF
4Concentration is 10.0mmol/L;
1.2.4 detection step:
1.2.4.1 the processing of separation capillary: new capillary column uses the activation of 100mmol/L NaOH to spend the night; Before experiment every day; Capillary column is with 100mmol/L NaOH flushing 10min; With secondary water flushing 10min, use the runtime buffer solution equilibria 10min that obtains by step 1.2.3.2 then again;
1.2.4.2 the detection of arecaline in the Chinese medicine betelnut extract: the betelnut extract dilution mark-on solution that obtains among betelnut extract dilute solution that obtains among the step 1.1.2.2 and the step 1.1.2.3 is detected by following condition: detect current potential 1.20V; Electrophoretic separation high pressure 16kV; Add the background electrolyte of step 1.2.3.1 preparation in the detection cell; Electrochemical reaction adopts three-electrode system, and three-electrode system comprises that diameter 500 μ m platinum dish working electrodes, platinum filament are to the utmost point and Ag/AgCl contrast electrode; Long capillary tube 58cm, internal diameter 50 μ m; The runtime buffer solution that electrophoresis uses step 1.2.3.2 to obtain; Adopt styletable electrochemiluminescence detecting pattern; The photomultiplier bias voltage is set in 800V; Obtain the electrophoresis pattern that arecaline detects in the Chinese medicine betelnut extract.
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| CN106770217A (en) * | 2017-03-30 | 2017-05-31 | 陕西理工大学 | A kind of capillary electrophoresis electrochemical light-emitting detection method of flavone compound |
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| CN103478719A (en) * | 2013-09-27 | 2014-01-01 | 许启太 | Method for extracting betel nut biotin form fresh betel nuts |
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| CN106770217A (en) * | 2017-03-30 | 2017-05-31 | 陕西理工大学 | A kind of capillary electrophoresis electrochemical light-emitting detection method of flavone compound |
| CN106770217B (en) * | 2017-03-30 | 2019-07-02 | 陕西理工大学 | A kind of capillary electrophoresis electrochemical light-emitting detection method of flavone compound |
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Application publication date: 20120620 |