CN109580823A - The detection method of red colour system coloring agent in a kind of medicinal material - Google Patents
The detection method of red colour system coloring agent in a kind of medicinal material Download PDFInfo
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- CN109580823A CN109580823A CN201811572496.4A CN201811572496A CN109580823A CN 109580823 A CN109580823 A CN 109580823A CN 201811572496 A CN201811572496 A CN 201811572496A CN 109580823 A CN109580823 A CN 109580823A
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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
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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
- G01N30/8675—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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Abstract
The invention discloses a kind of detection methods of red colour system coloring agent in medicinal material, the following steps are included: 1) famille rose, amaranth, erythrosine standard reagent is taken to add the ethyl alcohol of 60-80% that the first standard solution is made, acid red 73, New Fuchsine, 808 scarlet, tonyred Ι, SudanⅣ standard reagent is taken to add ethyl alcohol that the second standard solution is made；2) each 1mL of the first, second standard solution of step 1) is taken, dilutes, obtains compound standard solution；3) the compound standard solution injection liquid chromatograph for taking step 2) to obtain, measures in the Detection wavelength of 500-520nm；4) it takes sample to be tested to be analyzed, doubtful peak is checked using ultraviolet-visible Optical Chromatography.This method can in fast qualitative detection product whether contain red colour system coloring agent, each signal peak sharply without hangover, it is noiseless, reduce the requirement to equipment, improve detector efficiency, have more optimistic market prospects.
Description
Technical field
The present invention relates to a kind of a kind of detection methods of red colour system coloring agent in analysis method more particularly to medicinal material.
Background technique
Chinese medicine addition coloring agent is a kind of illegal addition behavior.The Chinese medicine red serial coloring agent of detection mainly has:
Famille rose, amaranth, erythrosine, acid red 73, New Fuchsine, 808 scarlet, tonyred Ι, SudanⅣ.For this purpose, national medicine prison is total
Office issued multiple Chinese medicines dyeing detection method, but these methods be mostly be directed to single medicinal material material detected it is (as follows
Table 1), when facing several kinds of Chinese medicinal materials, because of the diversity of method, lead to the complexity of pre-treatment, He Shangji analytic process, detection effect
Rate is slow, therefore gropes a kind of detection method of red staining agent in Chinese medicine, finds general detection method,
1 red staining agent of table and detection method
Above-mentioned detection method is mostly single coloring agent detection, and detection efficiency is lower, for this reason, it may be necessary to Chinese medicine red colour system
Column coloring agent multicomponent is groped with the method that the time is detected, and establishing one kind, to be able to carry out the red serial coloring agent of Chinese medicine more
The universal test method that component is detected with the time.
Summary of the invention
For overcome the deficiencies in the prior art, the purpose of the present invention is to provide in a kind of high-throughput, efficient medicinal material
The detection method of red colour system coloring agent.
The purpose of the present invention adopts the following technical scheme that realization:
The detection method of red colour system coloring agent in a kind of medicinal material, comprising the following steps:
1) it takes famille rose, amaranth, erythrosine standard reagent to add the ethyl alcohol of 60-80% that the first standard solution is made, takes acid
Property is red 73, New Fuchsine, 808 scarlet, tonyred Ι, SudanⅣ standard reagent add ethyl alcohol that the second standard solution is made；
2) the first standard solution and each 1mL of the second standard solution of step 1) are taken, dilutes, obtains compound standard solution；
3) the compound standard solution injection liquid chromatograph for taking step 2) to obtain, measures in the Detection wavelength of 500-520nm；
The parameter of the liquid chromatograph are as follows:
Chromatographic column: C18Reverse-phase chromatographic column；
Mobile phase: A acetonitrile；
B 0.02mol/L ammonium acetate solution；
Elution program: t0-15min, mobile phase A is gradually increased to 10-20vt% by 5vt%；
t15-40min, the ratio increase 60vt% of mobile phase A；
t40-41min, the ratio increase 10vt% of mobile phase A；
t41-50min, the ratio of mobile phase A increases to 95vt%；
t>50min, constant mobile phase A is 95vt%；
4) sample to be tested is taken, the ethyl alcohol with step 1) with concentration is added, ultrasonic extraction takes supernatant, and filtering obtains to be measured molten
Liquid；
5) solution sample introduction to be measured is eluted using chromatographic condition identical with step 3), to doubtful peak using it is ultraviolet-can
See that absorption spectrum is checked.
Further, in step 1), using 70% ethyl alcohol.
Further, in step 1), concentration of each standard reagent in the first standard solution is 0.08-0.12mg/mL.
Further, in step 1), concentration of each standard reagent in the second standard solution is 0.08-0.12mg/mL.
Further, in step 2), concentration of each standard reagent in compound standard solution is 0.02-0.03mg/mL.
Further, in step 3), solution is by 0.22 μm of filter membrane filtering before sample introduction.
Further, in step 3), the material of filter membrane is polytetrafluoroethylene (PTFE).
Further, in step 3), mobile phase A is gradually increased to 15vt% by 2vt%.
Further, in step 3), Detection wavelength 510nm.
Further, the uv-visible absorption spectra review in step 5), in 260nm-700nm wave-length coverage.
Compared with prior art, the beneficial effects of the present invention are:
Method provided by the invention rapidly can carry out qualitative detection to the coloring agent in medicine, each red colour system coloring agent
Retention time interval is long, and each signal peak is multiple by carrying out uv-visible absorption spectra to doubtful peak sharply without hangover, noiseless
Inspection, reduces the requirement to equipment, improves detector efficiency, has more optimistic market prospects.
Detailed description of the invention
Fig. 1 is the HPLC figure of the standard solution of embodiment 1；
Fig. 2 is the HPLC figure for the madder medicinal material that the lot number of embodiment 2 is 20171126；
Fig. 3 is the HPLC figure for the madder medicinal material that the lot number of embodiment 2 is 20180124；
Fig. 4 is the HPLC figure for the madder medicinal material that the lot number of embodiment 2 is 20180323.
Specific embodiment
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 method of red colour system coloring agent in a kind of medicinal material, comprising the following steps:
1) it takes famille rose, amaranth, erythrosine standard reagent to add the ethyl alcohol of 60-80% that the first standard solution is made, takes acid
Property is red 73, New Fuchsine, 808 scarlet, tonyred Ι, SudanⅣ standard reagent add ethyl alcohol that the second standard solution is made；
The step for sulfonic carmine and amaranth and has phenol hydroxyl by the way of step-wise dissolution, by active group
The erythrosine of base dissolves together, and other red staining agent are dissolved together, can effectively improve dissolved efficiency；
2) the first standard solution and each 1mL of the second standard solution of step 1) are taken, dilutes, obtains compound standard solution；
3) it is filtered using 0.22 μm of Teflon filtration film, the compound standard solution injection liquid phase color for taking step 2) to obtain
Spectrometer is measured in the Detection wavelength of 500-520nm；
The parameter of the liquid chromatograph are as follows:
Chromatographic column: C18Reverse-phase chromatographic column；
Mobile phase: A acetonitrile；
B 0.02mol/L ammonium acetate solution；
Elution program: t0-15Min, mobile phase A are gradually increased to 10-20vt% by 5vt%；
t15-40The ratio of min, mobile phase A increase 60vt%；
t40-41The ratio of min, mobile phase A increase 10vt%；
t41-50The ratio of min, mobile phase A increase to 95vt%；
t>50Min, constant mobile phase A is 95vt%；
Under the elution program, the elution efficiency of each coloring agent is high, and being overlapped or trailing do not occur in the appearance of each coloring agent；
4) sample to be tested is taken, the ethyl alcohol with step 1) with concentration is added, ultrasonic extraction takes supernatant, and filtering obtains to be measured molten
Liquid；
5) it is filtered using 0.22 μm of Teflon filtration film, solution sample introduction to be measured, using chromatography identical with step 3)
Condition is eluted, and is checked using uv-visible absorption spectra doubtful peak.
The result judgement method of step 5) are as follows: in the HPLC chromatogram of sample to be tested, there is not allowed that with contrast agents chromatography master
The identical chromatographic peak of peak retention time.If there is the identical chromatographic peak of retention time, compared using diode array detector
For corresponding chromatographic peak in the uv-visible absorption spectra of 260-700nm wave-length coverage, absorption spectrum should not be identical.
The detection method is by gradient elution so as to carrying out qualitative inspection to the red staining agent in medicinal material in 70min
It surveys, as shown in Figure 1, appearance time of the standard reagent on HPLC is followed successively by amaranth, famille rose, erythrosine, acid red 73, new
Magenta, Sudan red 1,808 scarlet and Sudan IVs.
Embodiment 1:
A kind of production of the standard diagram of red staining agent, comprising the following steps:
1) respectively take that famille rose, amaranth, that erythrosine standard reagent 1mg adds the ethyl alcohol of 10mL 70% that the first standard is made is molten
Liquid respectively takes acid red 73, New Fuchsine, 808 scarlet, tonyred Ι, SudanⅣ standard reagent 1mg to add the ethyl alcohol system of 10mL 70%
At the second standard solution；
2) the first standard solution and each 1mL of the second standard solution for taking step 1), are diluted to 5mL with 70% ethyl alcohol, obtain
Compound standard solution；
3) it is filtered using 0.22 μm of Teflon filtration film, the compound standard solution injection liquid phase color for taking step 2) to obtain
Spectrometer is measured in the Detection wavelength of 510nm；
The parameter of the liquid chromatograph are as follows:
Chromatographic column: C18Reverse-phase chromatographic column；
Mobile phase: A acetonitrile；
B 0.02mol/L ammonium acetate solution；
Elution program: t0-15min, mobile phase A is gradually increased to 15vt% by 5vt%；
t15-40min, the ratio of mobile phase A increases to 75vt% by 15%；
t40-41min, the ratio of mobile phase A increases to 85vt% by 75%；
t41-50min, the ratio of mobile phase A increases to 95vt% by 85%；
t>50min, constant mobile phase A is 95vt%；
4) ownership of each signal peak is determined.
The HPLC figure of embodiment 1 is shown in Fig. 1, and signal peak 1 is amaranth, retention time 11.6min；Signal peak 2 is
Famille rose, retention time 16.7min；Signal peak 3 is erythrosine, retention time 27.2min；Signal peak 4 is acid red 73,
Retention time is 27.6min；Signal peak 5 is New Fuchsine, retention time 33.4min；Signal peak 6 is Sudan red 1, retention time
For 47.1min；Signal peak 7 is 808 scarlet, retention time 51.3min；Signal peak 8 is Sudan IV, and retention time is
60.5min。
Embodiment 2:
The detection method of red colour system coloring agent in a kind of madder medicinal material, comprising the following steps:
1) 1g sample to be tested is taken, adds 15mL50vt% ethyl alcohol, ultrasonic extraction 10 minutes, centrifuging and taking supernatant, through 0.45 μm
Filter membrane filtering, obtains solution to be measured；
2) through 0.22 μm of Teflon filtration film aspiration step 1) 10 μ L of standard solution, inject liquid chromatograph,
Detection wavelength is to be eluted in 510nm；
Chromatographic column: C18Reverse-phase chromatographic column；
Mobile phase: A acetonitrile；
B 0.02mol/L ammonium acetate solution；
Elution program: t0-15min, mobile phase A is gradually increased to 15vt% by 5vt%；
t15-40min, the ratio increase 60vt% of mobile phase A；
t40-41min, the ratio increase 10vt% of mobile phase A；
t41-50min, the ratio of mobile phase A increases to 95vt%；
t>50min, constant mobile phase A is 95vt%；
In the madder medicinal material that lot number is 20171126, the map of HPLC in retention time and embodiment 1 as shown in Fig. 2, mark
Signal peak is not found within the scope of the retention time ± 0.2min of quasi- reagent, so, in the madder medicinal material that lot number is 20171126 not
Contain red colour system coloring agent.
In the madder medicinal material that lot number is 20180124, the map of HPLC in retention time and embodiment 1 as shown in figure 3, mark
Signal peak is not found within the scope of the retention time ± 0.2min of quasi- reagent, so, in the madder medicinal material that lot number is 20180124 not
Contain red colour system coloring agent.
In the madder medicinal material that lot number is 20180323, the map of HPLC in retention time and embodiment 1 as shown in figure 4, mark
Signal peak is not found within the scope of the retention time ± 0.2min of quasi- reagent, so, in the madder medicinal material that lot number is 20180323 not
Contain red colour system coloring agent.
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 (10)
1. the detection method of red colour system coloring agent in a kind of medicinal material, comprising the following steps:
1) it takes famille rose, amaranth, erythrosine standard reagent to add the ethyl alcohol of 60-80% that the first standard solution is made, takes acid red
73, New Fuchsine, 808 scarlet, tonyred Ι, SudanⅣ standard reagent add ethyl alcohol that the second standard solution is made；
2) the first standard solution and each 1mL of the second standard solution of step 1) are taken, dilutes, obtains compound standard solution；
3) the compound standard solution injection liquid chromatograph for taking step 2) to obtain, measures in the Detection wavelength of 500-520nm；
The parameter of the liquid chromatograph are as follows:
Chromatographic column: C18Reverse-phase chromatographic column；
Mobile phase: A acetonitrile；
B 0.02mol/L ammonium acetate solution；
Elution program: t0-15min, mobile phase A is gradually increased to 10-20vt% by 5vt%；
t15-40min, the ratio increase 60vt% of mobile phase A；
t40-41min, the ratio increase 10vt% of mobile phase A；
t41-50min, the ratio of mobile phase A increases to 95vt%；
t>50min, constant mobile phase A is 95vt%；
4) sample to be tested is taken, the ethyl alcohol with step 1) with concentration is added, ultrasonic extraction takes supernatant, and filtering obtains solution to be measured；
5) solution sample introduction to be measured is eluted using chromatographic condition identical with step 3), is inhaled to doubtful peak using ultraviolet-visible
Receive spectrum review.
2. the detection method of red colour system coloring agent in medicinal material as described in claim 1, in step 1), using 70% ethyl alcohol.
3. the detection method of red colour system coloring agent in medicinal material as described in claim 1, in step 1), each standard reagent is first
Concentration in standard solution is 0.08-0.12mg/mL.
4. the detection method of red colour system coloring agent in medicinal material as described in claim 1, in step 1), each standard reagent is second
Concentration in standard solution is 0.08-0.12mg/mL.
5. the detection method of red colour system coloring agent in medicinal material as described in claim 1, in step 2), each standard reagent is compound
Concentration in standard solution is 0.02-0.03mg/mL.
6. the detection method of red colour system coloring agent in medicinal material as described in claim 1, in step 3), solution passes through before sample introduction
0.22 μm of filter membrane filtering.
7. the detection method of red colour system coloring agent in medicinal material as claimed in claim 6, in step 3), the material of filter membrane is poly-
Tetrafluoroethene.
8. the detection method of red colour system coloring agent in medicinal material as described in claim 1, in step 3), mobile phase A by 2vt% by
It edges up to 15vt%.
9. the detection method of red colour system coloring agent in medicinal material as described in claim 1, in step 3), Detection wavelength 510nm.
10. the detection method of red colour system coloring agent in medicinal material as claimed in claim 9, in step 5), in 260nm-700nm wave
Uv-visible absorption spectra review in long range.
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| CN106990183A (en) * | 2017-05-05 | 2017-07-28 | 蚌埠市疾病预防控制中心 | The method for chromatographic determination of synthetic coloring matter in a kind of food |
| CN107202847A (en) * | 2017-07-21 | 2017-09-26 | 云南中烟工业有限责任公司 | The liquid chromatogram measuring method of eight kinds of colouring agents in a kind of quick-fried pearl wall material of cigarette |
| CN108387657A (en) * | 2018-03-07 | 2018-08-10 | 吉林师范大学 | The detection method of colorant content in Waterelon Frost Lozenges |
-
2018
- 2018-12-21 CN CN201811572496.4A patent/CN109580823A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20130056953A (en) * | 2011-11-23 | 2013-05-31 | (주)아모레퍼시픽 | Simultaneous determination of oxidative hair dye compounds by high performance chromatography |
| CN106990183A (en) * | 2017-05-05 | 2017-07-28 | 蚌埠市疾病预防控制中心 | The method for chromatographic determination of synthetic coloring matter in a kind of food |
| CN107202847A (en) * | 2017-07-21 | 2017-09-26 | 云南中烟工业有限责任公司 | The liquid chromatogram measuring method of eight kinds of colouring agents in a kind of quick-fried pearl wall material of cigarette |
| CN108387657A (en) * | 2018-03-07 | 2018-08-10 | 吉林师范大学 | The detection method of colorant content in Waterelon Frost Lozenges |
Non-Patent Citations (3)
| Title |
|---|
| GERTRUD E.MORLOCK 等: "Rapid Planar Chromatographic Analysis of 25 Water-Soluble Dyes Used as Food Additives", 《MORLOCK & OELLIG: JOURNAL OF AOAC INTERNATIONAL》 * |
| 朱日然 等: "超高效液相色谱-串联质谱法同时测定中药中29种合成着色剂", 《药物分析杂志》 * |
| 苟琰 等: "液质联用法快速筛查中药材及饮片中常见染色掺假物质的方法研究及数据库建立", 《药物分析杂志》 * |
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