CN111398453A - Method for simultaneously detecting content of effective components in angelica sinensis - Google Patents

Method for simultaneously detecting content of effective components in angelica sinensis Download PDF

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CN111398453A
CN111398453A CN202010202553.0A CN202010202553A CN111398453A CN 111398453 A CN111398453 A CN 111398453A CN 202010202553 A CN202010202553 A CN 202010202553A CN 111398453 A CN111398453 A CN 111398453A
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angelica
senkyunolide
content
effective components
solution
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熊毅
邓雪琪
李俊
饶高雄
陈超
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Yunnan Haochen Pharmaceutical Co ltd
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    • G01N30/02—Column chromatography
    • 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
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Abstract

The invention relates to a method for simultaneously detecting the content of effective components in Chinese angelica, wherein the effective components comprise organic acids, phthalides, amino acids and coumarins; the method comprises the following steps: preparing a test solution, preparing a reference solution and detecting a sample. The method provided by the invention can be used for simultaneously detecting the index components of organic acids, phthalides, amino acids and coumarins in the angelica for the first time, so that the quality of the angelica medicinal material can be more comprehensively evaluated and controlled. The detection indexes selected by the invention are all the components with higher content and pharmacological activity in the angelica sinensis medicinal material. Can provide basis for the safety and effectiveness of the clinical application of the angelica sinensis medicinal material. The detection time and cost are saved, and the method is accurate, simple and feasible. The method has the advantages of good stability, good repeatability, high recovery rate and strong specificity.

Description

Method for simultaneously detecting content of effective components in angelica sinensis
Technical Field
The invention relates to a method for simultaneously detecting the content of effective components, in particular to a method for simultaneously detecting the content of the effective components in Chinese angelica, and belongs to the field of quality detection of Chinese medicinal materials.
Background
Angelica sinensis is a dry root of Angelica angelicae sinensis (Oliv.) Diels, which is a plant of the family Umbelliferae, and is mainly produced in Gansu, and is also planted in Yunnan, Sichuan, Hubei provinces, and in recent years, with the increase of demand of Angelica sinensis market, Angelica sinensis in Gansu, which is a major component of the Angelica sinensis market due to the excellent quality of Angelica sinensis in the area of property of the way, has gradually become an important part of the Angelica sinensis market, and the planting area has been continuously expanded. At present, except regaining, the Chinese angelica is mostly sold in the market. The angelica is used as a medicinal and edible medicinal material, has long medicinal history and wide clinical application, and has the effects of tonifying qi, stopping bleeding, promoting blood circulation, nourishing blood and harmonizing blood.
The chemical components of Chinese angelica mainly comprise organic acids, phthalides, coumarins and the like, and also comprise some amino acid components. Wherein ferulic acid is the main active ingredient of radix Angelicae sinensis organic acid, and has antiinflammatory, senile dementia resisting, blood replenishing and blood circulation promoting effects; chlorogenic acid has wide pharmacological activities of protecting liver, benefiting gallbladder, resisting tumor, resisting virus, etc. The phenolphthalein compounds comprise ligustilide, senkyunolide A, senkyunolide I, butenyl phthalide, etc., which are main components of radix Angelicae sinensis volatile oil, wherein the content of ligustilide is the highest, and the compounds have various pharmacological effects of resisting tumor, resisting oxidation, inhibiting angiogenesis, diminishing inflammation, relieving pain, etc. The coumarin compounds include osthole, psoralen, isoimperatorin, bergapten, imperatorin, etc., and have various pharmacological activities of resisting tumor, hypertension, myocardial ischemia, inflammation, and pain. Tryptophan is an essential amino acid for the human body, and cannot be synthesized by the human body.
The existing standard takes single component ferulic acid as a detection index, is relatively single, cannot comprehensively reflect the quality of the angelica medicinal material, and is not beneficial to the quality control of the angelica medicinal material and the use safety and stability of the angelica.
Therefore, on the basis of the existing research, the establishment of a rapid, comprehensive and highly targeted quality detection method for the angelica sinensis medicinal material has important significance.
Disclosure of Invention
In order to comprehensively control the quality of the angelica sinensis medicinal material, the invention aims to solve the technical problems that the angelica sinensis medicinal material in the prior art has single detection index and is not beneficial to the quality control of the angelica sinensis medicinal material, and further provides a quick, comprehensive and strong-pertinence detection method of the angelica sinensis medicinal material. The technical scheme of the invention is as follows:
a method for simultaneously detecting the content of effective components in angelica sinensis comprises the steps of detecting the content of the effective components, wherein the effective components comprise organic acids, phthalides, amino acids and coumarins; the method comprises the following steps:
step (1), preparation of a test solution:
precisely weighing the powder, precisely adding methanol, sealing, weighing, ultrasonically treating or refluxing, cooling, weighing again, adding methanol to make up for lost weight, shaking, standing, collecting supernatant, filtering with 0.4 μm microporous membrane, and collecting filtrate to obtain sample solution;
step (2), preparation of a reference solution:
precisely weighing a proper amount of a reference substance of the component to be detected, and adding 50-100% methanol to prepare a reference substance solution; step (3), sample detection:
the chromatographic conditions were as follows:
the chromatographic column takes carbon octadecyl bonded silica gel as a filler, the flow rate is 0.5-2 m L/min, the column temperature is 20-40 ℃, the detection wavelength is one or two wavelengths of 260-316 nm, the sample injection amount is 5-20 mu l, the mobile phase is a mixed solvent consisting of methanol or acetonitrile and a formic acid aqueous solution or a phosphoric acid aqueous solution, the concentration of the formic acid or the phosphoric acid aqueous solution containing formic acid or phosphoric acid is 0.01-0.5 percent, and gradient elution is carried out according to the following table;
Figure BDA0002419881410000021
respectively sucking 5-20 mu L of the mixed reference solution and the test solution, injecting into a liquid chromatograph, and measuring.
Further, the effective components are Tryptophan (Tryptophan), Chlorogenic Acid (Chlorogenic Acid), Ferulic Acid (Ferulic Acid), senkyunolide I (Senkyunolide I), senkyunolide H (Senkyunolide H), Imperatorin (impactorin), Z-ligustilide (Z-L igutilide) and butenyl phthalide (Butyleephthahalide) in the angelica sinensis.
Further, in the step (1), sieving the powder with a third sieve, taking 0.2-5.0 g of the powder, precisely weighing, placing the powder in a conical flask with a plug, precisely adding 20-50 m L of 50-100% methanol, sealing the plug, weighing, ultrasonically treating or refluxing for 30-60 minutes, cooling, weighing again, supplementing the lost weight with 70-100% methanol, shaking up, standing, taking supernatant, filtering, and taking a subsequent filtrate to obtain the product.
Furthermore, in the step (2), every 1m L reference solution contains 4.22-84.40 μ g of tryptophan, 6.02-120.40 μ g of chlorogenic acid, 4.60-92.00 μ g of ferulic acid, 1.27-25.30 μ g of senkyunolide I, 0.44-8.72 μ g of senkyunolide H, 16.08-321.60 μ g of imperatorin, 44.00-880.00 μ g of Z-ligustilide and 4.11-82.26 μ g of butenyl phthalide.
Further, in the step (2), every 1m L reference solution contains tryptophan 50 μ g, chlorogenic acid 70 μ g, ferulic acid 55 μ g, senkyunolide I16 μ g, senkyunolide H5 μ g, imperatorin 190 μ g, Z-ligustilide 500 μ g, and butenyl phthalide 50 μ g.
Furthermore, in the step (3), the chromatographic conditions comprise a flow rate of 1.0m L/min, a column temperature of 30 ℃, a detection wavelength of 280nm, a sample amount of 10 μ l, mobile phases of A-acetonitrile and B-0.085% formic acid or phosphoric acid aqueous solution, and gradient elution is carried out according to the following table.
Figure BDA0002419881410000031
The invention is further illustrated by the following experimental studies.
(1) And (4) selecting a chromatographic column, wherein detection indexes selected by the HP L C detection method established by the invention comprise components with large polarity, medium polarity and small polarity, and are all fat-soluble compounds, so that a reversed-phase C18 chromatographic column is selected.
(2) Selection of detection wavelength: performing full-wavelength scanning on 8 components such as tryptophan, wherein the tryptophan is 227nm and 280 nm; chlorogenic acid is at 327nm and 296 nm; ferulic acid at 320nm and 280 nm; senkyunolide I is at 212nm and 280 nm; senkyunolide H is at 214nm and 280 nm; the imperatorin has maximum absorption at 248nm and 301nm, the Z-ligustilide has maximum absorption at 280nm and 320nm, and the butenyl phthalide has maximum absorption at 226nm and 260nm, the maximum absorption wavelengths of the components are combined, and simultaneously the components with lower content are taken into consideration, and finally one or two wavelengths of 260-316 nm are selected as the detection wavelength.
(3) Selection of mobile phase system: the mobile phase systems of the acetonitrile (A) -phosphoric acid (B) solution and the acetonitrile-formic acid (B) solution are taken as factors for investigation, and the results show that different mobile phase systems are used, the influence on the peak shape and the separation degree of each effective component is different, the separation degree, the tailing factor and the theoretical plate number of each component in the two mobile phase systems are better, the separation degree, the tailing factor and the theoretical plate number of each component in the mobile phase system of the acetonitrile (A) -phosphoric acid (B) solution are better, and the mobile phase systems can be adopted.
(4) Selecting a treatment mode of the test sample: as the selected detection indexes comprise components with large polarity, medium polarity and small polarity, and also comprise thermal instability components, the chemical properties of the components are comprehensively considered, three solvents of 50% methanol, 70% methanol and 100% methanol, and two modes of ultrasonic extraction and reflux extraction are preliminarily selected for investigation. As a result, 50-100% methanol is used as the solvent to completely extract the detected components, so that 50-100% methanol is used as the extraction solvent.
(5) Chromatographic conditions and system applicability test: octadecylsilane chemically bonded silica is used as a filling agent; acetonitrile is taken as a mobile phase A, 0.085 percent phosphoric acid aqueous solution is taken as a mobile phase B, and gradient elution is carried out according to the specification in the following table; the detection wavelength is 280 nm; the column temperature was 30 ℃.
Figure BDA0002419881410000041
(6) A special experiment: under the same test conditions, a mixed reference substance solution and an angelica test substance solution are used as a reference, and a 70% methanol solution is used as a negative reference. The measurement was carried out by referring to the method described in the step (5). Results the peaks of the components of the mixed control solution appeared in the test solution containing the crude drug of angelica, but not in the negative control solution, indicating that the experimental method has specificity, the results of the mixed control solution are shown in fig. 1, the results of the angelica test solution are shown in fig. 2, and the results of the negative control solution are shown in fig. 3.
(7) The linear relation is that a proper amount of each reference substance is precisely weighed and placed in a brown measuring flask, methanol is added for dissolution, and solution containing 84.40 mu g of tryptophan, 120.40 mu g of chlorogenic acid, 92.00 mu g of ferulic acid, 25.30 mu g of senkyunolide I, 8.72 mu g of senkyunolide H, 321.60 mu g of imperatorin, 880.00 mu g of Z-ligustilide and 82.26 mu g of butenyl phthalide per 1m L is prepared to be used as stock solution of the reference substance.
Precisely sucking mixed reference substance stock solutions of 0.5-L, 1.0-L, 2.0-L, 4.0-L, 6.0-L, 8.0-L and 10-L respectively, putting the mixed reference substance stock solutions into a 10-L volumetric flask, adding methanol to a constant volume to scale, and shaking uniformly to obtain the mixed reference substance solution, precisely sucking the mixed reference substance solution of 10-L respectively, injecting the mixed reference substance solution into a liquid chromatograph, and measuring according to the method in the step (5), wherein the results show that tryptophan, chlorogenic acid, ferulic acid, senkyunolide I, senkyunolide H, imperatorin, Z-ligustilide and butenyl phthalide respectively have linear relationship of 4.22-84.40 mu g/m L, 6.02-120.40 mu g/m 2, 4.60-92.00 mu g/m L, 1.27-25.30 mu g/m L, 0.44-638.72 mu g/m 92, 16.08-356.00 mu g/m 6 g/m, and 880.00-L-3626 mu g/m respectively.
TABLE 1 results of linear relationship examination
Figure BDA0002419881410000051
(8) Precision investigation: and (4) precisely measuring 10 mu l of mixed reference substance solution under a special investigation item, injecting the mixed reference substance solution into a liquid chromatograph according to the method in the step (5), and continuously injecting the sample for 6 times for determination. The results show that the RSD% of each component is between 0.43 and 2.04. Indicating that the precision of the instrument is good, and the results are shown in table 2.
TABLE 2 precision test results table
Figure BDA0002419881410000052
Figure BDA0002419881410000061
(9) And (3) repeatability inspection: 6 parts of a parallel sample solution was prepared, 10. mu.l of the sample solution was precisely aspirated, and the sample solution was injected into a liquid chromatograph and measured by the method described in (5). The results show that: the RSD percent of each component is 1.18-1.99, and the repeatability is good. The results are shown in Table 3.
TABLE 3 repeatability test results Table
Figure BDA0002419881410000062
(10) And (3) stability investigation: preparing test solution, precisely sucking 10 μ l of the solution at 0h, 2h, 4h, 6h, 8h, 10h and 12h, respectively, and injecting into liquid chromatograph. The result of the determination according to the method in (5) shows that the solution of the test sample is stable within 12 hours at room temperature, and the RSD percent is between 0.98 and 2.08. The results are shown in Table 4.
Table 4 stability test results table
Figure BDA0002419881410000063
Figure BDA0002419881410000071
(11) Sample addition recovery rate test comprises precisely weighing 6 parts of angelica sinensis medicinal material powder (sieved by a sieve III) (tryptophan content 0.0832%, chlorogenic acid content 0.0410%, ferulic acid content 0.1286%, senkyunolide I content 0.0354%, senkyunolide H content 0.0054%, imperatorin content 0.1339%, Z-ligustilide content 1.7182%, butenyl phthalide content 0.0481%), each part is about 0.5g, precisely weighing, adding a mixed reference stock solution 2m L (1m L contains tryptophan 208.4 μ g, chlorogenic acid 101.2 μ g, ferulic acid 323.2 μ g, senkyunolide I88.0 μ g, senkyunolide H13.08 μ g, imperatorin 336.0 μ g, Z-387 5 μ g, butenyl phthalide 122.4 μ g), placing into a 250m L conical bottle, adding 70% methanol 18m L, weighing, sealing, heating for 30min, weighing 70% of ligustilide, weighing.
TABLE 5 sample-application recovery results Table
Figure BDA0002419881410000072
Figure BDA0002419881410000081
Figure BDA0002419881410000091
(12) Durability test: preparing 3 parts of test solution, precisely sucking 10 mu l of test solution, and respectively measuring the volume ratio of the sample solution under different variation factors: injecting samples respectively at the temperature of equipment, a chromatographic column and the column, recording peak areas of 8 components such as tryptophan, calculating the content, and inspecting the durability of chromatographic conditions. Results the RSD% values for the contents of each component were less than 3% under each condition, indicating that the chromatographic conditions were durable. The results are shown in Table 6.
TABLE 6 durability test results table
Figure BDA0002419881410000092
Figure BDA0002419881410000101
Compared with the prior art, the invention has the following beneficial effects:
(1) the method for determining the HP L C content of the angelica sinensis medicinal material, which is disclosed by the invention, can be used for simultaneously detecting the index components of organic acids, phthalides, amino acids and coumarins in the angelica sinensis for the first time and realizing more comprehensive evaluation and control on the quality of the angelica sinensis medicinal material.
(2) The detection indexes selected by the invention are tryptophan, chlorogenic acid, ferulic acid, senkyunolide I, senkyunolide H, imperatorin, Z-ligustilide and butenyl phthalide, which are not only representative chemical components of organic acids, phthalides, coumarins and amino acids in Chinese angelica medicinal materials, but also main pharmacological active components. The method simultaneously detects the content of 8 components such as ferulic acid and the like for the first time through the high performance liquid chromatography, saves the detection time and cost, and is not only accurate, but also simple and feasible. The quality of the angelica sinensis medicinal material is comprehensively evaluated and controlled, so that the effectiveness and safety of clinical medication of the angelica sinensis medicinal material are guaranteed. The method has the advantages of good stability, good repeatability, high recovery rate and strong specificity.
Drawings
FIG. 1 is a liquid chromatogram of a mixed control solution in a specific experiment;
FIG. 2 is a liquid chromatogram of a sample solution of Angelica sinensis in a specific experiment;
FIG. 3 is a liquid chromatogram of a negative control solution in a specificity experiment.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is described in further detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention. The examples do not specify particular techniques or conditions, and are performed according to the techniques or conditions described in the literature in the art or according to the product specifications. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available by purchase.
Example 1
In the method for simultaneously detecting the content of the effective components in the angelica sinensis, the contents of tryptophan, chlorogenic acid, ferulic acid, senkyunolide I, senkyunolide H, imperatorin, Z-ligustilide and butenyl phthalide in the Yunnan Weixi angelica sinensis are measured by HP L C, and the method specifically comprises the following steps:
step (1) preparation of a reference solution:
accurately weighing appropriate amount of tryptophan, chlorogenic acid, ferulic acid, senkyunolide I, senkyunolide H, imperatorin, Z-ligustilide and butenyl phthalide, adding 70% methanol to prepare solution containing tryptophan 50 μ g, chlorogenic acid 70 μ g, ferulic acid 55 μ g, senkyunolide I16 μ g, senkyunolide H5 μ g, imperatorin 190 μ g, Z-ligustilide 500 μ g and butenyl phthalide 50 μ g per 1m L.
Step (2) preparation of a test solution:
taking test samples of different producing areas, taking about 1.0g of powder (passing through a third sieve), precisely weighing, placing in a conical flask with a plug, precisely adding 20m L% of 70% methanol, sealing the plug, weighing, heating, reflux-extracting for 30min, cooling, weighing again, supplementing the lost weight with 70% methanol, shaking, standing, taking supernatant, filtering with 0.4 μm microporous membrane, and taking the subsequent filtrate.
And (3) detecting a sample:
octadecylsilane chemically bonded silica gel as filler, acetonitrile as mobile phase A, 0.085% phosphoric acid aqueous solution as mobile phase B, at flow rate of 1.0m L/min, and gradient eluting according to the specification in the following table, with detection wavelength of 280nm and column temperature of 30 deg.C.
Figure BDA0002419881410000111
Respectively sucking 10 μ L of the mixed reference solution and the sample solution, injecting into liquid chromatograph, and measuring.
The origin and content of the angelica sinensis sample of this example are shown in table 7.
TABLE 7 measurement results of the content of Angelica sinensis samples
Figure BDA0002419881410000112
Figure BDA0002419881410000121
According to the content detection result, the method can be stably used for detecting the content of the angelica sinensis medicinal material.
Example 2
In the method for simultaneously detecting the content of the effective components in the angelica, the contents of tryptophan, chlorogenic acid, ferulic acid, senkyunolide I, senkyunolide H, imperatorin, Z-ligustilide and butenyl phthalide in the Weixi angelica are measured by HP L C, and the method specifically comprises the following steps:
step (1) preparation of a reference solution:
accurately weighing a proper amount of tryptophan, chlorogenic acid, ferulic acid, senkyunolide I, senkyunolide H, imperatorin, Z-ligustilide and butenyl phthalide reference substances, adding 100% methanol to prepare a solution containing 4.22 mu g of tryptophan, 6.02 mu g of chlorogenic acid, 4.60 mu g of ferulic acid, 1.27 mu g of senkyunolide I, 0.44 mu g of senkyunolide H, 16.8 mu g of imperatorin, 44.0 mu g of Z-ligustilide and 4.11 mu g of butenyl phthalide per 1m of L, and obtaining the test solution in the step (2):
weighing about 0.2g of the powder (sieved by a third sieve), precisely weighing, placing in a conical flask with a plug, precisely adding 50m L of 100% methanol, sealing the plug, weighing, heating, refluxing and extracting for 60 minutes, cooling, weighing again, supplementing the loss by 100% methanol, shaking uniformly, standing, taking supernatant, filtering with a 0.4 mu m microporous filter membrane, and taking subsequent filtrate.
Step (3) sample detection
Octadecylsilane chemically bonded silica gel as filler, acetonitrile as mobile phase A, 0.01% formic acid water solution as mobile phase B, and gradient elution at flow rate of 2.0m L/min, detection wavelength of 260nm, and column temperature of 20 deg.C.
Figure BDA0002419881410000131
Respectively sucking 20 μ L of the mixed reference solution and the sample solution, injecting into liquid chromatograph, and measuring.
The origin and content of the angelica sinensis sample of this example are shown in Table 8.
TABLE 8 measurement results of the content of Angelica sinensis
Figure BDA0002419881410000132
Figure BDA0002419881410000141
According to the content detection result, the method can be stably used for detecting the content of the angelica sinensis medicinal material.
Example 3
In the method for simultaneously detecting the content of the effective components in the angelica, the contents of tryptophan, chlorogenic acid, ferulic acid, senkyunolide I, senkyunolide H, imperatorin, Z-ligustilide and butenyl phthalide in Gansu, Yunnan Lijiang, Reliang and Chinese angelica are measured by HP L C, and the method comprises the following steps:
step (1) preparation of a reference solution:
accurately weighing appropriate amount of tryptophan, chlorogenic acid, ferulic acid, senkyunolide I, senkyunolide H, imperatorin, Z-ligustilide and butenyl phthalide reference substances, adding 50% methanol to prepare solution containing 84.40 μ g of tryptophan, 120.40 μ g of chlorogenic acid, 92.00 μ g of ferulic acid, 25.30 μ g of senkyunolide I, 8.72 μ g of senkyunolide H, 321.60 μ g of imperatorin, 880 μ g of Z-ligustilide and 82.26 μ g of butenyl phthalide per 1m L.
Step (2) preparation of a test solution:
taking test samples of different producing areas, taking about 5.0g of the powder (passing through a third sieve), precisely weighing, placing in a conical flask with a plug, precisely adding 30m L of 50% methanol, sealing the plug, weighing, ultrasonically treating for 40 minutes, cooling, weighing again, supplementing the lost weight with 50% methanol, shaking up, standing, taking the supernatant, filtering with a 0.4 mu m microporous filter membrane, and taking the subsequent filtrate.
And (3) detecting a sample:
octadecylsilane chemically bonded silica gel as filler, acetonitrile as mobile phase A, 0.5m L/min flow rate as mobile phase B, and 0.5% phosphoric acid water solution as mobile phase B, and performing gradient elution according to the specification in the following table, wherein the detection wavelength is 280nm and 316nm, and the column temperature is 40 deg.C.
Figure BDA0002419881410000142
Figure BDA0002419881410000151
(4) The determination method comprises respectively sucking 5 μ L mixed reference solution and sample solution, injecting into liquid chromatograph, and determining.
(5) As a result: the results of the angelica sinensis sample production area and content measurement are shown in table 9.
TABLE 9 measurement results of the content of Angelica sinensis sample
Figure BDA0002419881410000152
According to the content detection result, the method can be stably used for detecting the content of the angelica sinensis medicinal material.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.

Claims (6)

1. A method for simultaneously detecting the content of effective components in angelica is characterized in that: the effective components comprise organic acids, phthalides, amino acids and coumarins; the method comprises the following steps:
step (1), preparation of a test solution:
weighing the powder, adding methanol, sealing, weighing, ultrasonic treating or refluxing, cooling, weighing, adding methanol to compensate weight loss, shaking, standing, collecting supernatant, filtering with 0.4 μm microporous membrane, and collecting filtrate;
step (2), preparation of a reference solution:
precisely weighing a proper amount of a reference substance of the component to be detected, and adding 50-100% methanol to prepare a reference substance solution;
step (3), sample detection:
the chromatographic conditions were as follows:
the chromatographic column takes octadecylsilane chemically bonded silica as a filler, the flow rate is 0.5-2 m L/min, the column temperature is 20-40 ℃, the detection wavelength is one or two wavelengths of 260-316 nm, the sample injection amount is 5-20 mu l, the mobile phase is a mixed solvent consisting of methanol or acetonitrile and a formic acid aqueous solution or a phosphoric acid aqueous solution, the concentration of the formic acid or phosphoric acid aqueous solution containing formic acid or phosphoric acid is 0.01-0.5%, and gradient elution is carried out according to the following table;
respectively sucking 5-20 mu L of the mixed reference solution and the test solution, injecting into a liquid chromatograph, and measuring.
2. The method for simultaneously detecting the content of the effective components in the angelica according to claim 1, wherein the method comprises the following steps: the effective components are tryptophan, chlorogenic acid, ferulic acid, senkyunolide I, senkyunolide H, imperatorin, Z-ligustilide and butenyl phthalide in the Chinese angelica medicinal material.
3. The method for simultaneously detecting the content of the effective components in the angelica according to claim 1, wherein the method comprises the following steps:
in the step (1), sieving the powder with a third sieve, taking 0.2-5.0 g of the powder, precisely weighing, placing the powder in a conical flask with a plug, precisely adding 50-100% of methanol 20-50 m L, sealing the plug, weighing, performing ultrasonic or reflux extraction for 30-60 minutes, cooling, weighing again, supplementing the lost weight with 50-100% of methanol, shaking uniformly, standing, taking supernatant, filtering with a 0.4-micron microporous membrane, and taking a subsequent filtrate to obtain the product.
4. The method for simultaneously detecting the content of the effective components in the angelica sinensis as claimed in claim 2, wherein in the step (2), each 1m L reference solution contains 4.22-84.40 μ g of tryptophan, 6.02-120.40 μ g of chlorogenic acid, 4.60-92.00 μ g of ferulic acid, 1.27-25.30 μ g of senkyunolide I, 0.44-8.72 μ g of senkyunolide H, 16.08-321.60 μ g of imperatorin, 44.00-880.00 μ g of Z-ligustanolide and 4.11-82.26 μ g of butenyl phthalide.
5. The method for simultaneously detecting the content of effective components in angelica sinensis as claimed in claim 4, wherein in the step (2), each 1m L reference solution contains solutions of tryptophan 50 μ g, chlorogenic acid 70 μ g, ferulic acid 55 μ g, senkyunolide I16 μ g, senkyunolide H5 μ g, imperatorin 190 μ g, Z-ligustilide 500 μ g, and butenyl phthalide 50 μ g.
6. The method for simultaneously detecting the content of the effective components in angelica sinensis as claimed in claim 4, wherein in the step (3), the chromatographic conditions comprise a flow rate of 1.0m L/min, a column temperature of 30 ℃, a detection wavelength of 280nm, a sample injection amount of 10 μ l, mobile phases of A-acetonitrile and B-0.085% formic acid or phosphoric acid aqueous solution, and gradient elution is carried out according to the following table
Figure 212277DEST_PATH_IMAGE002
。
CN202010202553.0A 2020-03-20 2020-03-20 Method for simultaneously detecting content of effective components in angelica sinensis Pending CN111398453A (en)

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