CN109596736A - The detection method of yellow class coloring agent in a kind of prepared slices of Chinese crude drugs - Google Patents
The detection method of yellow class coloring agent in a kind of prepared slices of Chinese crude drugs Download PDFInfo
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- 238000001514 detection method Methods 0.000 title claims abstract description 40
- 239000003814 drug Substances 0.000 title claims abstract description 36
- 229940079593 drug Drugs 0.000 title claims abstract description 26
- 239000003086 colorant Substances 0.000 title claims abstract description 25
- 239000002872 contrast media Substances 0.000 claims abstract description 66
- 239000000243 solution Substances 0.000 claims abstract description 34
- 238000003556 assay Methods 0.000 claims abstract description 19
- 239000012085 test solution Substances 0.000 claims abstract description 18
- 244000248349 Citrus limon Species 0.000 claims description 38
- 235000005979 Citrus limon Nutrition 0.000 claims description 38
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 34
- 238000000862 absorption spectrum Methods 0.000 claims description 30
- 230000014759 maintenance of location Effects 0.000 claims description 30
- 239000007788 liquid Substances 0.000 claims description 26
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 18
- 235000019441 ethanol Nutrition 0.000 claims description 18
- 238000000034 method Methods 0.000 claims description 16
- 230000003595 spectral effect Effects 0.000 claims description 14
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 12
- 238000010828 elution Methods 0.000 claims description 12
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 11
- 239000000706 filtrate Substances 0.000 claims description 9
- 239000000843 powder Substances 0.000 claims description 9
- 239000013558 reference substance Substances 0.000 claims description 9
- 238000004587 chromatography analysis Methods 0.000 claims description 7
- USFZMSVCRYTOJT-UHFFFAOYSA-N Ammonium acetate Chemical compound N.CC(O)=O USFZMSVCRYTOJT-UHFFFAOYSA-N 0.000 claims description 6
- 239000005695 Ammonium acetate Substances 0.000 claims description 6
- 229960000583 acetic acid Drugs 0.000 claims description 6
- 229940043376 ammonium acetate Drugs 0.000 claims description 6
- 235000019257 ammonium acetate Nutrition 0.000 claims description 6
- 239000012362 glacial acetic acid Substances 0.000 claims description 6
- 238000005259 measurement Methods 0.000 claims description 6
- YTJSFYQNRXLOIC-UHFFFAOYSA-N octadecylsilane Chemical compound CCCCCCCCCCCCCCCCCC[SiH3] YTJSFYQNRXLOIC-UHFFFAOYSA-N 0.000 claims description 6
- 239000000047 product Substances 0.000 claims description 6
- 239000000945 filler Substances 0.000 claims description 5
- 239000000377 silicon dioxide Substances 0.000 claims description 5
- 239000003153 chemical reaction reagent Substances 0.000 claims description 4
- 238000001914 filtration Methods 0.000 claims description 4
- 238000010521 absorption reaction Methods 0.000 claims description 3
- 238000000605 extraction Methods 0.000 claims description 3
- 238000002360 preparation method Methods 0.000 claims description 3
- 238000001228 spectrum Methods 0.000 claims description 3
- 238000010998 test method Methods 0.000 abstract description 2
- 239000012071 phase Substances 0.000 description 40
- 238000007689 inspection Methods 0.000 description 7
- 239000003795 chemical substances by application Substances 0.000 description 5
- 239000007791 liquid phase Substances 0.000 description 5
- 239000000203 mixture Substances 0.000 description 3
- 235000013305 food Nutrition 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000002203 pretreatment Methods 0.000 description 2
- 239000013589 supplement Substances 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 238000010200 validation analysis Methods 0.000 description 1
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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
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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
- G01N30/62—Detectors specially adapted therefor
- G01N30/74—Optical detectors
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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
- G01N30/86—Signal analysis
- G01N30/8624—Detection of slopes or peaks; baseline correction
- G01N30/8631—Peaks
-
- 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
- G01N30/86—Signal analysis
- G01N30/8675—Evaluation, i.e. decoding of the signal into analytical information
- G01N30/8679—Target compound analysis, i.e. whereby a limited number of peaks is analysed
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- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
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- Immunology (AREA)
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- Library & Information Science (AREA)
- Engineering & Computer Science (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
The invention discloses a kind of detection methods of yellow class coloring agent in prepared slices of Chinese crude drugs, comprising the following steps: the step of preparing contrast agents solution prepares the step of the step of the step of test solution, assay, result judgement.The detection method is the universal test method of yellow class multicomponent coloring agent, to solve Chinese medicine detection method problem that is different, and keeping detection efficiency slow in different cultivars.
Description
Technical field
The present invention relates to a kind of a kind of inspections of yellow class coloring agent in traditional Chinese medicine quality detection technique more particularly to prepared slices of Chinese crude drugs
Survey method.
Background technique
In recent years, the case where prepared slices of Chinese crude drugs dye is very serious.National Yao Jian general bureau has issued the dyeing of plurality of Chinese medicine materical crude slice
Detection method, this method are mostly to be directed to single medicinal material medicine materical crude slice to be detected, as shown in table 1.
" state food pharmaceuticals administration general bureau's drug inspection supplement inspection that table 1 is detected about Yellow series coloring agent
Proved recipe method and inspection project approval part ",
When facing plurality of Chinese medicine materical crude slice while needing detection data, because of the diversity of method, lead to pre-treatment, He Shangji
The complexity of analytic process, detection efficiency is slow, and versatility is not strong.
Summary of the invention
For overcome the deficiencies in the prior art, the purpose of the present invention is to provide yellow class coloring agents in a kind of prepared slices of Chinese crude drugs
Detection method.The detection method is the universal test method of yellow class multicomponent coloring agent, to solve Chinese medicine in different cultivars
Material detection method problem that is different, and keeping detection efficiency slow.
The purpose of the present invention adopts the following technical scheme that realization: the detection side of yellow class coloring agent in a kind of prepared slices of Chinese crude drugs
Method, comprising the following steps:
The step of preparing contrast agents solution: accurately weighed lemon yellow, sunset yellow, bright orange contrast agents are taken respectively, add second
Mixed reference substance solution is made in alcohol, shake up to get；
The step of preparing test solution: prepared slices of Chinese crude drugs sample powder to be measured is taken, is set in conical flask, ethyl alcohol is added, shaking mentions
Take, filter, take subsequent filtrate to get；
The step of assay: it is accurate respectively to draw contrast agents solution and each 10 μ l of test solution, inject liquid phase color
Spectrometer, measurement record liquid chromatogram；
The step of result judgement: in sample chromatogram, there is not allowed that identical with contrast agents chromatography main peak retention time
Chromatographic peak；If there is the identical chromatographic peak of retention time, using diode array detector compare test sample with accordingly compare
Uv-visible absorption spectra of the chromatographic peak of reagent in 260-500nm wave-length coverage, the absorption of test sample and corresponding contrast agents
Spectrum is identical, then can determine that and contain corresponding coloring agent in test sample.
Further, in the step of preparing contrast agents solution, the preparation method of the contrast agents solution is as follows: point
Accurately weighed lemon yellow, sunset yellow, bright orange contrast agents are not taken, and adding volumetric concentration is 50% ethyl alcohol, and every 1ml is made and contains
The mixed reference substance solution of 0.2mg, shake up to get.
Further, in the step of preparing test solution, the extraction process of the test solution is as follows: taking to be measured
Prepared slices of Chinese crude drugs sample powder 2g, sets in conical flask, and the ethyl alcohol 25ml that volumetric concentration is 50% is added, and shaking is extracted 5min, filtered,
Take subsequent filtrate to get.
Further, in the assay the step of, the liquid chromatograph is to fill out with octadecylsilane chemically bonded silica
Agent is filled, using acetonitrile as mobile phase A, using the 0.05mol/L ammonium acetate solution for the glacial acetic acid for being 0.5% containing volumetric concentration as mobile phase
B, the sum of mobile phase A and Mobile phase B volume are 100%, carry out gradient elution.
Further, the condition of gradient elution is as follows: when 0-20min, the volume of mobile phase A is 2 → 55%, mobile phase
The volume of B is 98 → 45%；When 20-25min, the volume of mobile phase A is 55%, and the volume of Mobile phase B is 45%.
Further, in the assay the step of, the detector of the liquid chromatograph is diode array detector,
Detection wavelength 420nm.
Further, in the assay the step of, the column temperature of the liquid chromatograph is 20 DEG C, flow velocity 0.7ml/
min。
Further, in the assay the step of, number of theoretical plate is calculated by lemon yellow spectral peak should be not less than 5000, reason
5000 should be not less than by calculating by plate number by sunset yellow spectral peak, and number of theoretical plate is calculated by bright orange chromatographic peak should be not less than 5000.
Further, in the result judgement the step of, specific decision process is as follows: if sample chromatogram occurs and lemon yellow
The identical chromatographic peak of contrast agents chromatographic retention, then using diode array detector compare in test sample with lemon yellow pair
According to the identical chromatographic peak of reagent chromatographic retention absorption spectrum and lemon yellow contrast agents in 260-500nm wave-length coverage
Uv-visible absorption spectra can determine that if absorption spectrum is consistent and contain lemon yellow in test sample；If sample chromatogram goes out
Now chromatographic peak identical with sunset yellow contrast agents chromatographic retention is then compared in test sample using diode array detector
The absorption spectrum of chromatographic peak identical with sunset yellow contrast agents chromatographic retention is with sunset yellow contrast agents in 260-
The uv-visible absorption spectra of 500nm wave-length coverage can determine that if absorption spectrum is consistent and contain sunset yellow in test sample；If
There is chromatographic peak identical with bright orange contrast agents chromatographic retention in sample chromatogram, then uses diode array detector ratio
Compared with chromatographic peak identical with bright orange contrast agents chromatographic retention in test sample absorption spectrum and bright orange contrast agents
The uv-visible absorption spectra of 260-500nm wave-length coverage can determine that in test sample if absorption spectrum is consistent containing bright orange.
Compared with prior art, the beneficial effects of the present invention are:
" state food pharmaceuticals administration general bureau's drug inspection supplements the method for inspection and examines project approval part " is for Huang
The detection method official written reply of color system coloring agent has 6, and plurality of Chinese medicine materical crude slice is then needed, and individually carries out pre-treatment and upper machine point
Analysis, this method are general Yellow series multicomponent coloring agent detection method, can detect plurality of Chinese medicine materical crude slice simultaneously, to improve inspection
Survey efficiency.Detection limit, durability etc. meet the requirement of methodology validation simultaneously.
Detailed description of the invention
Fig. 1 is lemon yellow of the present invention, sunset yellow, the bright orange chromatogram for mixing contrast agents；
Fig. 2 is the sample chromatogram of 1 Cortex Phellodendri of the embodiment of the present invention, 180506591 sample；
Fig. 3 is the sample chromatogram of 2 Cortex Phellodendri of the embodiment of the present invention, 180700231 sample；
Fig. 4 is the sample chromatogram of 3 Cortex Phellodendri of the embodiment of the present invention, 180805631 sample.
It wherein, 1 is lemon yellow in Fig. 1,2 be sunset yellow, and 3 be bright orange.
Specific embodiment
In the present invention, if not refering in particular to, all parts, percentage are unit of weight, used equipment and raw material etc.
It is commercially available or commonly used in the art.Method in following embodiments is unless otherwise instructed the normal of this field
Rule method.
The detection method of yellow class coloring agent in a kind of prepared slices of Chinese crude drugs, comprising the following steps:
The step of preparing contrast agents solution: accurately weighed lemon yellow, sunset yellow, bright orange contrast agents are taken respectively, add second
Mixed reference substance solution is made in alcohol, shake up to get；
The step of preparing test solution: prepared slices of Chinese crude drugs sample powder to be measured is taken, is set in conical flask, ethyl alcohol is added, shaking mentions
Take, filter, take subsequent filtrate to get；
The step of assay: it is accurate respectively to draw contrast agents solution and each 10 μ l of test solution, inject liquid phase color
Spectrometer, measurement record liquid chromatogram；
The step of result judgement: in sample chromatogram, there is not allowed that identical with contrast agents chromatography main peak retention time
Chromatographic peak；If there is the identical chromatographic peak of retention time, using diode array detector compare test sample with accordingly compare
Uv-visible absorption spectra of the chromatographic peak of reagent in 260-500nm wave-length coverage, the absorption of test sample and corresponding contrast agents
Spectrum is identical, then can determine that and contain corresponding coloring agent in test sample.
As further preferred scheme, in the step of preparing contrast agents solution, the preparation of the contrast agents solution
Method is as follows: taking accurately weighed lemon yellow, sunset yellow, bright orange contrast agents respectively, adding volumetric concentration is 50% ethyl alcohol, is made
Every 1ml respectively containing the mixed reference substance solution of 0.2mg, shake up to get.
As further preferred scheme, in the step of preparing test solution, the extraction process of the test solution
It is as follows: to take prepared slices of Chinese crude drugs sample powder 2g to be measured, set in conical flask, the ethyl alcohol 25ml that volumetric concentration is 50% is added, shaking mentions
Take 5min, filter, take subsequent filtrate to get.
As further preferred scheme, the assay the step of in, the liquid chromatograph is with octadecylsilane key
Conjunction silica gel is filler, molten with the 0.05mol/L ammonium acetate for the glacial acetic acid for being 0.5% containing volumetric concentration using acetonitrile as mobile phase A
Liquid is Mobile phase B, and the sum of mobile phase A and Mobile phase B volume are 100%, carries out gradient elution.
As further preferred scheme, the condition of gradient elution is as follows: when 0-20min, the volume of mobile phase A is 2 →
55%, the volume of Mobile phase B is 98 → 45%；When 20-25min, the volume of mobile phase A is 55%, and the volume of Mobile phase B is
45%.
As further preferred scheme, the assay the step of in, the detector of the liquid chromatograph is diode
Array detector, Detection wavelength 420nm.
As further preferred scheme, the assay the step of in, the column temperature of the liquid chromatograph is 20 DEG C, flow velocity
For 0.7ml/min.
As further preferred scheme, the assay the step of in, number of theoretical plate should not by the calculating of lemon yellow spectral peak
Lower than 5000, number of theoretical plate is calculated by sunset yellow spectral peak should be not less than 5000, and number of theoretical plate should not by the calculating of bright orange chromatographic peak
Lower than 5000.
As further preferred scheme, the result judgement the step of in, specific decision process is as follows: if sample chromatogram goes out
Now chromatographic peak identical with lemon yellow contrast agents chromatographic retention is then compared in test sample using diode array detector
The absorption spectrum of chromatographic peak identical with lemon yellow contrast agents chromatographic retention is with lemon yellow contrast agents in 260-
The uv-visible absorption spectra of 500nm wave-length coverage can determine that if absorption spectrum is consistent and contain lemon yellow in test sample；If
There is chromatographic peak identical with sunset yellow contrast agents chromatographic retention in sample chromatogram, then uses diode array detector
The absorption spectrum for comparing chromatographic peak identical with sunset yellow contrast agents chromatographic retention in test sample compares examination with sunset yellow
The uv-visible absorption spectra in 260-500nm wave-length coverage of agent can determine that in test sample and contain if absorption spectrum is consistent
There is sunset yellow；If chromatographic peak identical with bright orange contrast agents chromatographic retention occurs in sample chromatogram, diode is used
Array detector compare in test sample the absorption spectrum of chromatographic peak identical with bright orange contrast agents chromatographic retention with it is bright orange
The uv-visible absorption spectra in 260-500nm wave-length coverage of contrast agents can determine that if absorption spectrum is consistent for examination
Containing bright orange in product.
In the following, being described further in conjunction with attached drawing and specific embodiment to the present invention, it should be noted that not
Under the premise of conflicting, new implementation can be formed between various embodiments described below or between each technical characteristic in any combination
Example.
The detection of yellow class coloring agent in the Cortex Phellodendri prepared slices of Chinese crude drugs that 1 lot number of embodiment is 180506591
The detection method of yellow class coloring agent in the radix scutellariae prepared slices of Chinese crude drugs, comprising the following steps:
The step of preparing contrast agents solution: accurately weighed lemon yellow, sunset yellow, bright orange contrast agents are taken respectively, add body
Product concentration be 50% ethyl alcohol, every 1ml is made respectively containing the mixed reference substance solution of 0.2mg, shake up to get.
The step of preparing test solution: taking prepared slices of Chinese crude drugs sample powder 2g to be measured, set in conical flask, and volumetric concentration is added
For 50% ethyl alcohol 25ml, 5min is extracted in shaking, filtration, take subsequent filtrate to get.
The step of assay: it is accurate respectively to draw contrast agents solution and each 10 μ l of test solution, inject liquid phase color
Spectrometer, measurement, record liquid chromatogram, as shown in Figure 1, for lemon yellow, sunset yellow, bright orange contrast agents chromatogram；Fig. 2 is
The chromatogram of this test agent；The liquid chromatograph is using octadecylsilane chemically bonded silica as filler, using acetonitrile as mobile phase
A, using the 0.05mol/L ammonium acetate solution for the glacial acetic acid for being 0.5% containing volumetric concentration as Mobile phase B, mobile phase A and Mobile phase B
The sum of volume is 100%, carries out gradient elution；Condition of gradient elution is as follows: when 0-20min, the volume of mobile phase A is 2 →
55%, the volume of Mobile phase B is 98 → 45%；When 20-25min, the volume of mobile phase A is 55%, and the volume of Mobile phase B is
45%.The detector of the liquid chromatograph is diode array detector, Detection wavelength 420nm.The liquid chromatograph
Column temperature is 20 DEG C, flow velocity 0.7ml/min.Number of theoretical plate is calculated by lemon yellow spectral peak should be not less than 5000, and number of theoretical plate is pressed
Sunset yellow chromatographic peak, which calculates, should be not less than 5000, and number of theoretical plate is calculated by bright orange chromatographic peak should be not less than 5000.
The step of result judgement: as shown in Fig. 2, in sample chromatogram, do not occur and three contrast agents chromatography main peaks
The identical chromatographic peak of retention time；I.e. there is not color identical with lemon yellow, sunset yellow, bright orange retention time in sample chromatogram
Spectral peak then can determine that in test sample without containing lemon yellow, sunset yellow, bright orange coloring agent composition.
The detection of yellow class coloring agent in the Cortex Phellodendri prepared slices of Chinese crude drugs that 2 lot number of embodiment is 180700231
The detection method of yellow class coloring agent in the radix scutellariae prepared slices of Chinese crude drugs, comprising the following steps:
The step of preparing contrast agents solution: accurately weighed lemon yellow, sunset yellow, bright orange contrast agents are taken respectively, add body
Product concentration be 50% ethyl alcohol, every 1ml is made respectively containing the mixed reference substance solution of 0.2mg, shake up to get.
The step of preparing test solution: taking prepared slices of Chinese crude drugs sample powder 2g to be measured, set in conical flask, and volumetric concentration is added
For 50% ethyl alcohol 25ml, 5min is extracted in shaking, filtration, take subsequent filtrate to get.
The step of assay: it is accurate respectively to draw contrast agents solution and each 10 μ l of test solution, inject liquid phase color
Spectrometer, measurement, record liquid chromatogram, as shown in Figure 1, for lemon yellow, sunset yellow, bright orange contrast agents chromatogram；Fig. 3 is
The chromatogram of this test agent；The liquid chromatograph is using octadecylsilane chemically bonded silica as filler, using acetonitrile as mobile phase
A, using the 0.05mol/L ammonium acetate solution for the glacial acetic acid for being 0.5% containing volumetric concentration as Mobile phase B, mobile phase A and Mobile phase B
The sum of volume is 100%, carries out gradient elution；Condition of gradient elution is as follows: when 0-20min, the volume of mobile phase A is 2 →
55%, the volume of Mobile phase B is 98 → 45%；When 20-25min, the volume of mobile phase A is 55%, and the volume of Mobile phase B is
45%.The detector of the liquid chromatograph is diode array detector, Detection wavelength 420nm.The liquid chromatograph
Column temperature is 20 DEG C, flow velocity 0.7ml/min.Number of theoretical plate is calculated by lemon yellow spectral peak should be not less than 5000, and number of theoretical plate is pressed
Sunset yellow chromatographic peak, which calculates, should be not less than 5000, and number of theoretical plate is calculated by bright orange chromatographic peak should be not less than 5000.
The step of result judgement: as shown in figure 3, in sample chromatogram, do not occur and three contrast agents chromatography main peaks
The identical chromatographic peak of retention time；I.e. there is not color identical with lemon yellow, sunset yellow, bright orange retention time in sample chromatogram
Spectral peak then can determine that in test sample without containing lemon yellow, sunset yellow, bright orange coloring agent composition.
The detection of yellow class coloring agent in the Cortex Phellodendri prepared slices of Chinese crude drugs that 3 lot number of embodiment is 180805631
The detection method of yellow class coloring agent in the radix scutellariae prepared slices of Chinese crude drugs, comprising the following steps:
The step of preparing contrast agents solution: accurately weighed lemon yellow, sunset yellow, bright orange contrast agents are taken respectively, add body
Product concentration be 50% ethyl alcohol, every 1ml is made respectively containing the mixed reference substance solution of 0.2mg, shake up to get.
The step of preparing test solution: taking prepared slices of Chinese crude drugs sample powder 2g to be measured, set in conical flask, and volumetric concentration is added
For 50% ethyl alcohol 25ml, 5min is extracted in shaking, filtration, take subsequent filtrate to get.
The step of assay: it is accurate respectively to draw contrast agents solution and each 10 μ l of test solution, inject liquid phase color
Spectrometer, measurement, record liquid chromatogram, as shown in Figure 1, for lemon yellow, sunset yellow, bright orange contrast agents chromatogram；Fig. 4 is
The chromatogram of this test agent；The liquid chromatograph is using octadecylsilane chemically bonded silica as filler, using acetonitrile as mobile phase
A, using the 0.05mol/L ammonium acetate solution for the glacial acetic acid for being 0.5% containing volumetric concentration as Mobile phase B, mobile phase A and Mobile phase B
The sum of volume is 100%, carries out gradient elution；Condition of gradient elution is as follows: when 0-20min, the volume of mobile phase A is 2 →
55%, the volume of Mobile phase B is 98 → 45%；When 20-25min, the volume of mobile phase A is 55%, and the volume of Mobile phase B is
45%.The detector of the liquid chromatograph is diode array detector, Detection wavelength 420nm.The liquid chromatograph
Column temperature is 20 DEG C, flow velocity 0.7ml/min.Number of theoretical plate is calculated by lemon yellow spectral peak should be not less than 5000, and number of theoretical plate is pressed
Sunset yellow chromatographic peak, which calculates, should be not less than 5000, and number of theoretical plate is calculated by bright orange chromatographic peak should be not less than 5000.
The step of result judgement: as shown in figure 4, in sample chromatogram, do not occur and three contrast agents chromatography main peaks
The identical chromatographic peak of retention time；I.e. there is not color identical with lemon yellow, sunset yellow, bright orange retention time in sample chromatogram
Spectral peak then can determine that in test sample without containing lemon yellow, sunset yellow, bright orange coloring agent composition.
The above embodiment is only the preferred embodiment of the present invention, and the scope of protection of the present invention is not limited thereto,
The variation and replacement for any unsubstantiality that those skilled in the art is done on the basis of the present invention belong to institute of the present invention
Claimed range.
Claims (9)
1. the detection method of yellow class coloring agent in a kind of prepared slices of Chinese crude drugs, which comprises the following steps:
The step of preparing contrast agents solution: accurately weighed lemon yellow, sunset yellow, bright orange contrast agents are taken respectively, add ethyl alcohol system
At mixed reference substance solution, shake up to get；
The step of preparing test solution: taking prepared slices of Chinese crude drugs sample powder to be measured, set in conical flask, adds ethyl alcohol, and shaking is extracted, filter
Cross, take subsequent filtrate to get；
The step of assay: it is accurate respectively to draw contrast agents solution and each 10 μ l of test solution, liquid chromatograph is injected,
Measurement records liquid chromatogram；
The step of result judgement: in sample chromatogram, there is not allowed that chromatography identical with contrast agents chromatography main peak retention time
Peak；If there is the identical chromatographic peak of retention time, test sample and corresponding contrast agents are compared using diode array detector
Uv-visible absorption spectra of the chromatographic peak in 260-500nm wave-length coverage, the absorption spectrum of test sample and corresponding contrast agents
It is identical, then it can determine that and contain corresponding coloring agent in test sample.
2. detection method as described in claim 1, which is characterized in that described right in the step of preparing contrast agents solution
It is as follows according to the preparation method of reagent solution: to take accurately weighed lemon yellow, sunset yellow, bright orange contrast agents respectively, add volumetric concentration
For 50% ethyl alcohol, every 1ml is made respectively containing the mixed reference substance solution of 0.2mg, shake up to get.
3. detection method as described in claim 1, which is characterized in that described for examination in the step of preparing test solution
The extraction process of product solution is as follows: taking prepared slices of Chinese crude drugs sample powder 2g to be measured, sets in conical flask, it is 50% that volumetric concentration, which is added,
Ethyl alcohol 25ml, shaking extract 5min, filtration, take subsequent filtrate to get.
4. detection method as described in claim 1, which is characterized in that in the assay the step of, the liquid chromatograph
Using octadecylsilane chemically bonded silica as filler, using acetonitrile as mobile phase A, with the glacial acetic acid for being 0.5% containing volumetric concentration
0.05mol/L ammonium acetate solution is Mobile phase B, and the sum of mobile phase A and Mobile phase B volume are 100%, carries out gradient elution.
5. detection method as claimed in claim 4, which is characterized in that the condition of gradient elution is as follows: when 0-20min, stream
The volume of dynamic phase A is 2 → 55%, and the volume of Mobile phase B is 98 → 45%；When 20-25min, the volume of mobile phase A is 55%,
The volume of Mobile phase B is 45%.
6. detection method as claimed in claim 4, which is characterized in that in the assay the step of, the liquid chromatograph
Detector be diode array detector, Detection wavelength 420nm.
7. detection method as claimed in claim 4, which is characterized in that in the assay the step of, the liquid chromatograph
Column temperature be 20 DEG C, flow velocity 0.7ml/min.
8. detection method as claimed in claim 4, which is characterized in that in the assay the step of, number of theoretical plate presses lemon
Yellow spectral peak, which calculates, should be not less than 5000, and number of theoretical plate is calculated by sunset yellow spectral peak should be not less than 5000, and number of theoretical plate is by bright
Yellow spectral peak, which calculates, should be not less than 5000.
9. detection method as described in claim 1, which is characterized in that in the result judgement the step of, specific decision process is such as
Under: if chromatographic peak identical with lemon yellow contrast agents chromatographic retention occurs in sample chromatogram, use diode array
Detector compares the absorption spectrum and lemon yellow of chromatographic peak identical with lemon yellow contrast agents chromatographic retention in test sample
The uv-visible absorption spectra in 260-500nm wave-length coverage of contrast agents can determine that if absorption spectrum is consistent for examination
Contain lemon yellow in product；If chromatographic peak identical with sunset yellow contrast agents chromatographic retention occurs in sample chromatogram, adopt
Compare the absorption of chromatographic peak identical with sunset yellow contrast agents chromatographic retention in test sample with diode array detector
The uv-visible absorption spectra in 260-500nm wave-length coverage of spectrum and sunset yellow contrast agents, if absorption spectrum is consistent,
It then can determine that and contain sunset yellow in test sample；If there is color identical with bright orange contrast agents chromatographic retention in sample chromatogram
Spectral peak then compares chromatographic peak identical with bright orange contrast agents chromatographic retention in test sample using diode array detector
Absorption spectrum and bright orange contrast agents the uv-visible absorption spectra in 260-500nm wave-length coverage, if absorption spectrum one
It causes, then can determine that in test sample containing bright orange.
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