CN112129721A - Ultraviolet visible light rapid detection method for artemisinin - Google Patents
Ultraviolet visible light rapid detection method for artemisinin Download PDFInfo
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- BLUAFEHZUWYNDE-NNWCWBAJSA-N artemisinin Chemical compound C([C@](OO1)(C)O2)C[C@H]3[C@H](C)CC[C@@H]4[C@@]31[C@@H]2OC(=O)[C@@H]4C BLUAFEHZUWYNDE-NNWCWBAJSA-N 0.000 title claims abstract description 72
- 229960004191 artemisinin Drugs 0.000 title claims abstract description 71
- 229930101531 artemisinin Natural products 0.000 title claims abstract description 71
- 238000001514 detection method Methods 0.000 title claims abstract description 15
- 238000000034 method Methods 0.000 claims abstract description 35
- 238000002360 preparation method Methods 0.000 claims abstract description 15
- 238000010438 heat treatment Methods 0.000 claims abstract description 13
- 239000012490 blank solution Substances 0.000 claims abstract description 12
- 239000012088 reference solution Substances 0.000 claims abstract description 8
- 238000002137 ultrasound extraction Methods 0.000 claims abstract description 8
- 239000003513 alkali Substances 0.000 claims abstract description 7
- 238000005070 sampling Methods 0.000 claims abstract description 4
- 239000000243 solution Substances 0.000 claims description 69
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 42
- 239000000523 sample Substances 0.000 claims description 36
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 33
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 26
- 240000000011 Artemisia annua Species 0.000 claims description 19
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 18
- 235000001405 Artemisia annua Nutrition 0.000 claims description 15
- 235000019441 ethanol Nutrition 0.000 claims description 13
- 239000003208 petroleum Substances 0.000 claims description 13
- 239000000843 powder Substances 0.000 claims description 13
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 13
- 239000012488 sample solution Substances 0.000 claims description 12
- 238000001914 filtration Methods 0.000 claims description 11
- 239000013558 reference substance Substances 0.000 claims description 10
- 238000002835 absorbance Methods 0.000 claims description 9
- 238000005303 weighing Methods 0.000 claims description 9
- 238000001816 cooling Methods 0.000 claims description 8
- 239000000706 filtrate Substances 0.000 claims description 8
- 238000007865 diluting Methods 0.000 claims description 7
- 238000002798 spectrophotometry method Methods 0.000 claims description 7
- 238000006243 chemical reaction Methods 0.000 claims description 6
- 238000010790 dilution Methods 0.000 claims description 5
- 239000012895 dilution Substances 0.000 claims description 5
- 238000000605 extraction Methods 0.000 claims description 5
- 239000012535 impurity Substances 0.000 claims description 5
- 238000005259 measurement Methods 0.000 claims description 5
- 239000012528 membrane Substances 0.000 claims description 5
- 238000009210 therapy by ultrasound Methods 0.000 claims description 4
- 238000001035 drying Methods 0.000 claims description 3
- 238000007873 sieving Methods 0.000 claims description 3
- 238000001291 vacuum drying Methods 0.000 claims description 3
- 239000002994 raw material Substances 0.000 abstract description 13
- 241000049464 Artemisia apiacea Species 0.000 abstract description 11
- 235000011570 Artemisia caruifolia var apiacea Nutrition 0.000 abstract description 11
- 238000012216 screening Methods 0.000 abstract description 6
- 238000009395 breeding Methods 0.000 abstract description 5
- 230000001488 breeding effect Effects 0.000 abstract description 5
- 238000012544 monitoring process Methods 0.000 abstract description 5
- 230000002349 favourable effect Effects 0.000 abstract description 4
- 238000012360 testing method Methods 0.000 description 18
- 238000011084 recovery Methods 0.000 description 6
- 238000004128 high performance liquid chromatography Methods 0.000 description 4
- 235000015700 Artemisia abrotanum Nutrition 0.000 description 3
- 244000249062 Artemisia abrotanum Species 0.000 description 3
- 238000010521 absorption reaction Methods 0.000 description 3
- 241000196324 Embryophyta Species 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 239000012085 test solution Substances 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 1
- 239000002585 base Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000002425 crystallisation Methods 0.000 description 1
- 230000008025 crystallization Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000010812 external standard method Methods 0.000 description 1
- 238000009313 farming Methods 0.000 description 1
- 201000004792 malaria Diseases 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000010992 reflux Methods 0.000 description 1
- 239000004065 semiconductor Substances 0.000 description 1
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- 238000010998 test method Methods 0.000 description 1
- 238000002834 transmittance Methods 0.000 description 1
- 238000000870 ultraviolet spectroscopy Methods 0.000 description 1
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
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- G01N1/00—Sampling; Preparing specimens for investigation
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- G01N1/34—Purifying; Cleaning
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- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/40—Concentrating samples
- G01N1/4022—Concentrating samples by thermal techniques; Phase changes
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
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- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/33—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using ultraviolet light
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
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- G01N2001/4027—Concentrating samples by thermal techniques; Phase changes evaporation leaving a concentrated sample
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Abstract
The application relates to the technical field of artemisinin content detection, in particular to an artemisinin visible light rapid detection method, which comprises the following steps: s1: ultrasonic extraction; s2: concentrating under reduced pressure; s3: dissolving and fixing the volume; s4: sampling; s5: adding alkali; s6: heat treatment; s7: preparation of a reference solution: s8: preparing a blank solution; s9: and (5) determining the artemisinin content. Compared with the traditional weight method, the method for rapidly detecting the artemisinin by visible light only needs to consume a very small amount of raw material samples, can detect the content of the artemisinin in the raw materials more simply, conveniently, rapidly and accurately, and is favorable for screening out high-content artemisinin to carry out artemisia apiacea seed breeding and quality monitoring when manufacturers purchase artemisia apiacea raw materials on site.
Description
[ technical field ] A method for producing a semiconductor device
The application relates to the technical field of artemisinin content detection, in particular to an artemisinin ultraviolet visible light rapid detection method.
[ background of the invention ]
The artemisinin series of medicines are the most safe and effective medicines for treating malaria at present, because the use amount of the southernwood is huge in the world and the market supply is not in demand, some illegal vendors use the poor southernwood or the fake southernwood to be sold as the certified products on an opportunity, and great loss is caused to artemisinin extraction manufacturers. Secondly, in the aspect of cultivating high-content sweet wormwood seeds, every plant of thousands of sweet wormwood plants in a base is screened one by one every year, the sweet wormwood plants are screened when the sweet wormwood plants bloom and grow vigorously in farming season, and in addition, the original plant of sweet wormwood plants is not damaged during screening, and the detection accuracy is guaranteed.
Therefore, there is a need to develop a simple, fast and accurate analysis method for quality monitoring of field purchase of Artemisia annua raw materials and screening high-content Artemisia annua for breeding. The conventional detection of herba Artemisiae Annuae is mainly weight method (reflux extraction of organic solution such as petroleum ether, crystallization and weighing), and high performance liquid chromatography. However, the gravimetric method requires a large amount of samples, is tedious, time-consuming, volatile and pollutes the environment, and the HPLC chromatography pretreatment is troublesome and expensive, so that the method is not suitable for on-site rapid detection.
[ summary of the invention ]
The application aims to provide the method for quickly detecting the artemisinin through ultraviolet and visible light, only a small amount of raw material samples are consumed, the content of the artemisinin in the raw materials can be detected more simply, conveniently, quickly and accurately, and the method is favorable for screening high-content artemisia apiacea to carry out seed breeding and quality monitoring when manufacturers purchase the artemisia apiacea raw materials on site.
The application is realized by the following technical scheme:
an artemisinin ultraviolet-visible light rapid detection method comprises the following steps:
s1: ultrasonic extraction: taking 0.5-2 g of dry leaf powder, putting the dry leaf powder into ultrasonic equipment, adding petroleum ether, wherein the dosage of the petroleum ether in milliliters is 30-40 times of the mass of the dry leaf powder, completely and ultrasonically extracting for three times, wherein the extraction time is 30min, 20min and 20min in sequence, filtering by quantitative filter paper, and combining to obtain a first filtrate;
s2: and (3) concentrating under reduced pressure: taking the first filtrate, and concentrating under reduced pressure in an evaporator under the condition of-0.3 Pa to separate out artemisinin paste;
s3: dissolving and fixing volume: adding an alcohol preparation to dissolve the artemisinin paste, and transferring to a 25mL, 50mL, 100mL or 150mL volumetric flask;
s4: sampling: taking 0.5-2 mL of the solution obtained in the step S3 into a 25-100 mL measuring flask as a sample;
s5: adding alkali: putting the sample into 0.2% sodium hydroxide solution according to the proportion of 1:4, shaking up, and filtering in a 10mL measuring flask by using a 0.22 mu m microporous filter membrane;
s6: and (3) heat treatment: placing the sample after alkali filtration in a constant-temperature water bath box at 50 +/-1 ℃ for heating reaction for 25-40 min, taking out the sample, and cooling the sample to room temperature to obtain a sample solution;
s7: preparation of a reference solution:
1) precisely weighing 20-25 mg of artemisinin reference substance subjected to vacuum drying at 80 ℃ and constant weight, placing the artemisinin reference substance into a 25mL measuring flask, and adding absolute ethyl alcohol;
2) performing ultrasonic treatment for 5min to dissolve the artemisinin reference substance in the step 1), cooling to room temperature, adding absolute ethanol to dilute to scale, and shaking;
3) precisely measuring 1-2 mL of the solution obtained in the step 2), putting the solution into a 25mL measuring flask, adding absolute ethyl alcohol to dilute the solution to a scale, and shaking up;
4) precisely measuring 1-2 mL of the solution obtained in the step 3) and placing the solution in a 10mL measuring flask;
5) diluting the solution obtained in the step 4) to scale with 0.2% sodium hydroxide solution, shaking up, and placing in a 50 +/-1 ℃ constant-temperature water bath box for heating reaction for 20-40 min to serve as a reference solution;
s8: preparing a blank solution;
s9: and (3) determination: correcting the baseline with blank solution, scanning with visible light ultraviolet spectrophotometry in the wavelength range of 190-400nm to obtain artemisinin content, and substituting into the formula:
M＝(Asample (A)×WTo pair×nSample (A))/(ATo pair×WSample (A)×nTo pair)
Wherein M is the content of artemisinin, ASample (A)Absorbance of the sample solution, ATo pairAbsorbance of control solution, WTo pairMeasured amount of artemisinin as a control solution, WSample (A)Measurement of artemisinin for sample solution, nSample (A)Being sample solutionsDilution multiple, nTo pairIs the dilution multiple of the control solution.
The method for rapidly detecting artemisinin by ultraviolet and visible light comprises the following steps of: precisely measuring 1-5 mL of ethanol or methanol solution, placing the solution in a 25mL measuring flask, diluting the solution to a scale with 0.2% sodium hydroxide solution, shaking the solution uniformly, and placing the solution in a 50 +/-1 ℃ constant-temperature water bath box for heating and reacting for 20-40 min to obtain a blank solution.
The ultraviolet-visible light rapid detection method for artemisinin takes the absorbance at the wavelength of 292nm as a calculated value when measuring the content of artemisinin by a spectrophotometry.
In the method for quickly detecting artemisinin by ultraviolet and visible light, the petroleum ether is No. 6 gasoline in step S1.
In the above method for rapidly detecting artemisinin by ultraviolet and visible light, the size of the screen in step S1 is 20 meshes.
In the method for rapidly detecting artemisinin by ultraviolet and visible light, the power of the ultrasonic equipment in the step S1 is 40w/cm 2.
In the method for rapidly detecting artemisinin by ultraviolet and visible light, the alcohol preparation in step S3 is methanol or ethanol.
The method for rapidly detecting artemisinin by ultraviolet and visible light comprises the following steps of: taking fresh sweet wormwood leaves, drying the fresh sweet wormwood leaves in the sun until the water content is less than or equal to 12%, removing impurities, crushing and sieving to obtain dry leaf powder.
Compared with the prior art, the method has the following advantages:
compared with the traditional gravimetric method and high performance liquid chromatography, the method only needs to consume a very small amount of raw material samples, can detect the content of the artemisinin in the raw materials more simply, conveniently, quickly and accurately, and is favorable for screening out high-content artemisia apiacea to carry out seed breeding and quality monitoring when manufacturers purchase the artemisia apiacea raw materials on site.
[ description of the drawings ]
In order to more clearly illustrate the technical solutions in the embodiments of the present application, the drawings needed to be used in the description of the embodiments are briefly introduced below, and it is obvious that the drawings in the following description are only some embodiments of the present application, and it is obvious for those skilled in the art to obtain other drawings based on these drawings without creative efforts.
FIG. 1 is a graph of the content of artemisinin by scanning with visible light ultraviolet spectrophotometry as described in the examples of the present application.
FIG. 2 is a line graph of the linearity test of artemisinin contained in Artemisia annua as described in the examples of the present application.
[ detailed description ] embodiments
In order to make the technical problems, technical solutions and advantageous effects solved by the present application more clear and obvious, the present application is further described in 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 present application and are not intended to limit the present application.
The embodiment of the application provides an artemisinin ultraviolet visible light rapid detection method, which comprises the following steps:
s1: ultrasonic extraction: taking fresh sweet wormwood leaves, drying the fresh sweet wormwood leaves in the sun until the water content is 12%, removing impurities, crushing and sieving to obtain dry leaf powder, wherein the specification of the sieve is 20 meshes, taking 0.5-2 g of the dry leaf powder, putting the dry leaf powder into ultrasonic equipment, adding petroleum ether, wherein the petroleum ether is No. 6 gasoline with the boiling point of 60-90 ℃, the power of the ultrasonic equipment is 40w/cm2, the using amount of the petroleum ether in milliliters is 30-40 times of the mass of the dry leaf powder, completely carrying out ultrasonic extraction for three times, the extraction time is 30min, 20min and 20min in sequence, filtering by quantitative filter paper, and combining to obtain first filtrate;
s2: and (3) concentrating under reduced pressure: taking the first filtrate, concentrating under reduced pressure in an evaporator under the condition of-0.3 Pa until artemisinin paste is separated out, generally separating out 100mL of the paste in 5min, wherein the conventional method needs 2 days;
s3: dissolving and fixing volume: adding an alcohol preparation such as methanol or ethanol to dissolve the artemisinin paste, and transferring to a volumetric flask of 25mL, 50mL, 100mL or 150 mL;
s4: sampling: taking 0.5-2 mL of the solution obtained in the step S3 into a 25-100 mL measuring flask as a sample, preferably taking 1mL of the solution obtained in the step S3 into a 25mL measuring flask;
s5: adding alkali: placing a sample in 0.2% sodium hydroxide solution in a ratio of 1:4, shaking uniformly, and filtering in a 10mL measuring flask by using a 0.22-micron microporous filter membrane, wherein the sodium hydroxide solution can play a role in ring removal and impurity removal, and the light transmittance of the solution is improved by filtering and removing impurities, so that preparation is made for next spectrophotometry detection;
s6: and (3) heat treatment: placing the sample after alkali filtration in a constant-temperature water bath box at 50 +/-1 ℃ for heating reaction for 25-40 min, taking out the sample, and cooling to room temperature (the room temperature is 25 ℃) to be used as a sample solution;
s7: preparation of a reference solution:
1) precisely weighing 20-25 mg of artemisinin reference substance subjected to vacuum drying at 80 ℃ and constant weight, placing the artemisinin reference substance into a 25mL measuring flask, and adding absolute ethyl alcohol;
2) performing ultrasonic treatment for 5min to dissolve the artemisinin reference substance in the step 1), cooling to room temperature, adding absolute ethanol to dilute to scale, and shaking;
3) precisely measuring 1-2 mL of the solution obtained in the step 2), putting the solution into a 25mL measuring flask, adding absolute ethyl alcohol to dilute the solution to a scale, and shaking up;
4) precisely measuring 1-2 mL of the solution obtained in the step 3) and placing the solution in a 10mL measuring flask;
5) diluting the solution obtained in the step 4) to scale with 0.2% sodium hydroxide solution, shaking up, and placing in a 50 +/-1 ℃ constant-temperature water bath box for heating reaction for 20-40 min to serve as a reference solution;
s8: preparing a blank solution: precisely measuring 1-5 mL of ethanol or methanol solution, placing the solution in a 25mL measuring flask, diluting the solution to a scale with 0.2% sodium hydroxide solution, shaking the solution uniformly, and placing the solution in a 50 +/-1 ℃ constant-temperature water bath box for heating and reacting for 20-40 min to obtain a blank solution;
s9: and (3) determination: correcting the baseline with blank solution, scanning with visible light ultraviolet spectrophotometry in the wavelength range of 190-400nm to obtain artemisinin content, and substituting into the formula:
M＝(Asample (A)×WTo pair×nSample (A))/(ATo pair×WSample (A)×nTo pair)
Wherein M is the content of artemisinin, ASample (A)Absorbance of the sample solution, ATo pairAbsorbance of control solution, WTo pairThe amount of artemisinin measured for the control solution,Wsample (A)Measurement of artemisinin for sample solution, nSample (A)Is the dilution multiple of the sample solution, nTo pairIs the dilution multiple of the control solution.
Wherein, from the UV absorption chart of figure 1 of the attached drawings of the specification, artemisinin has maximum absorption at 292nm, other has almost no absorption at the wavelength and does not interfere the measurement, so the absorbance at 292nm is taken as the best calculation value, and the method has better specificity.
To test the accuracy, repeatability, etc. of the method, the following methodological tests were also performed:
1. accuracy test (recovery test)
The test is carried out by a sample recovery method. Precisely weighing 1-2g of sweet wormwood leaves (sieved by a 20-mesh sieve), precisely weighing (n is 6), placing the sweet wormwood leaves in a triangular flask with a plug, precisely adding about 10mg of artemisinin with known purity (99.5%), adding petroleum ether (60-90 ℃) for ultrasonic extraction for 3 times, 30-40 ml/time, performing ultrasonic extraction for 30-20 min each time, combining filtrates, washing residues with a small amount of petroleum ether, merging the residues into the filtrate, recovering the solvent under reduced pressure, dissolving the residues with an appropriate amount of ethanol, transferring the residues into a 25ml measuring flask, adding ethanol to scale, shaking up, precisely weighing 1ml to 25ml measuring flasks, adding 4ml of absolute ethanol, diluting to scale with 0.2% NaOH solution, shaking up, filtering the solution with a 0.22 mu m microporous filter membrane in a 10ml measuring flask, placing the solution in a constant-temperature water bath at 50 ℃ and 1 ℃ for 30min, taking out, cooling the solution to room temperature, and using the solution as a sweet wormwood leaf sample solution; injecting 1mg/ml artemisinin reference solution by the same method. And calculating the content (%) and recovery (%) of the sample by an external standard method. The test results show that the accuracy of the measurement is good, see table 1.
TABLE 1 determination of the accuracy of the artemisinin content in Artemisia annua leaves
2. Repeatability test
Preparing a test solution: taking about 0.5-1-2g of artemisia apiacea leaves (passing through a 20-mesh sieve) and placing the artemisia apiacea leaves into a triangular flask with a plug, precisely weighing, adding petroleum ether (60-90 ℃) for ultrasonic extraction for 3 times, and preparing an artemisia apiacea leaf test solution according to a recovery test method.
Preparation of control solution, preparation of blank solution and determination method: the same recovery test was conducted.
TABLE 2 repeatability test for determination of artemisinin content in Artemisia annua leaves
3. Linear test
The method comprises the following steps: accurately weighing appropriate amount of artemisinin reference, and dissolving with anhydrous ethanol to obtain 5mg/ml solution. Precisely measuring the reference substance storage solutions 2, 3, 4, 5 and 6ml respectively, placing into a measuring flask containing herba Artemisiae Annuae solution with a certain concentration to 25ml, adding anhydrous ethanol to about 25ml respectively, performing ultrasonic treatment for 5min, cooling to room temperature, adding anhydrous ethanol to scale, and performing other operations in the same recovery test. The data obtained are shown in Table 3:
TABLE 3 Artemisinin Linearity test
The data in table 3 are plotted as shown in fig. 2, and the linear result shows that the sample preparation adopts a filter membrane method, so that the requirement of measuring the content of the artemisinin can be met.
a. Intermediate precision test
The method comprises the following steps: for the same repeatability test, TU-1901 was used as an ultraviolet-visible spectrophotometer.
TABLE 4 first intermediate precision test data
TABLE 5 first intermediate precision test data
4. Determination of the concentration Range
The method is applied in the range of 50-150% (4-12 mu g/ml) of the specified determination concentration (8 mu g/ml) by combining linearity (4-12 mu g/ml), precision (6.4-9.6 mu g/ml) and accuracy test (6.4-9.6 mu g/ml).
In summary, the present application has, but is not limited to, the following beneficial effects:
compared with the traditional gravimetric method and high performance liquid chromatography, the method only needs to consume a very small amount of raw material samples, can detect the content of the artemisinin in the raw materials more simply, conveniently, quickly and accurately, and is favorable for screening out high-content artemisia apiacea to carry out seed breeding and quality monitoring when manufacturers purchase the artemisia apiacea raw materials on site.
The foregoing is illustrative of one or more embodiments provided in connection with the detailed description and is not intended to limit the disclosure to the particular forms disclosed. Similar or identical methods, structures, etc. as used herein, or several technical inferences or substitutions made on the concept of the present application should be considered as the scope of the present application.
Claims (8)
1. An artemisinin ultraviolet visible light rapid detection method is characterized by comprising the following steps:
s1: ultrasonic extraction: taking 0.5-2 g of dry leaf powder, putting the dry leaf powder into ultrasonic equipment, adding petroleum ether, wherein the dosage of the petroleum ether in milliliters is 30-40 times of the mass of the dry leaf powder, completely and ultrasonically extracting for three times, wherein the extraction time is 30min, 20min and 20min in sequence, filtering by quantitative filter paper, and combining to obtain a first filtrate;
s2: and (3) concentrating under reduced pressure: taking the first filtrate, and concentrating under reduced pressure in an evaporator under the condition of-0.3 Pa to separate out artemisinin paste;
s3: dissolving and fixing volume: adding an alcohol preparation to dissolve the artemisinin paste, and transferring to a 25mL, 50mL, 100mL or 150mL volumetric flask;
s4: sampling: taking 0.5-2 mL of the solution obtained in the step S3 into a 25-100 mL measuring flask as a sample;
s5: adding alkali: putting the sample into 0.2% sodium hydroxide solution according to the proportion of 1:4, shaking up, and filtering in a 10mL measuring flask by using a 0.22 mu m microporous filter membrane;
s6: and (3) heat treatment: placing the sample after alkali filtration in a constant-temperature water bath box at 50 +/-1 ℃ for heating reaction for 25-40 min, taking out the sample, and cooling the sample to room temperature to obtain a sample solution;
s7: preparation of a reference solution:
1) precisely weighing 20-25 mg of artemisinin reference substance subjected to vacuum drying at 80 ℃ and constant weight, placing the artemisinin reference substance into a 25mL measuring flask, and adding absolute ethyl alcohol;
2) performing ultrasonic treatment for 5min to dissolve the artemisinin reference substance in the step 1), cooling to room temperature, adding absolute ethanol to dilute to scale, and shaking;
3) precisely measuring 1-2 mL of the solution obtained in the step 2), putting the solution into a 25mL measuring flask, adding absolute ethyl alcohol to dilute the solution to a scale, and shaking up;
4) precisely measuring 1-2 mL of the solution obtained in the step 3) and placing the solution in a 10mL measuring flask;
5) diluting the solution obtained in the step 4) to scale with 0.2% sodium hydroxide solution, shaking up, and placing in a 50 +/-1 ℃ constant-temperature water bath box for heating reaction for 20-40 min to serve as a reference solution;
s8: preparing a blank solution;
s9: and (3) determination: correcting the baseline with blank solution, scanning with visible light ultraviolet spectrophotometry in the wavelength range of 190-400nm to obtain artemisinin content, and substituting into the formula:
M＝(Asample (A)×WTo pair×nSample (A))/(ATo pair×WSample (A)×nTo pair)
Wherein M is the content of artemisinin, ASample (A)Absorbance of the sample solution, ATo pairAbsorbance of control solution, WTo pairMeasured amount of artemisinin as a control solution, WSample (A)Measurement of artemisinin for sample solution, nSample (A)Is the dilution multiple of the sample solution, nTo pairIs the dilution multiple of the control solution.
2. The method for rapidly detecting artemisinin by ultraviolet and visible light, as claimed in claim 1, wherein the preparation of blank solution comprises: precisely measuring 1-5 mL of ethanol or methanol solution, placing the solution in a 25mL measuring flask, diluting the solution to a scale with 0.2% sodium hydroxide solution, shaking the solution uniformly, and placing the solution in a 50 +/-1 ℃ constant-temperature water bath box for heating and reacting for 20-40 min to obtain a blank solution.
3. The method for rapidly detecting artemisinin by ultraviolet and visible light as claimed in claim 1, wherein absorbance at 292nm wavelength is taken as a calculated value when measuring artemisinin content by spectrophotometry.
4. The method for rapidly detecting artemisinin by ultraviolet and visible light as claimed in claim 1, wherein the petroleum ether in step S1 is No. 6 gasoline.
5. The method for rapidly detecting artemisinin by ultraviolet and visible light as claimed in claim 1, wherein the size of the screen in step S1 is 20 mesh.
6. The method for rapidly detecting artemisinin by ultraviolet and visible light, as claimed in claim 1, wherein the power of the ultrasonic device in step S1 is 40w/cm2。
7. The method for rapid ultraviolet-visible detection of artemisinin in claim 1, wherein the alcohol preparation in step S3 is methanol or ethanol.
8. The method for rapidly detecting artemisinin by ultraviolet and visible light as claimed in claim 1, wherein the preparation of the dry leaf powder comprises: taking fresh sweet wormwood leaves, drying the fresh sweet wormwood leaves in the sun until the water content is less than or equal to 12%, removing impurities, crushing and sieving to obtain dry leaf powder.
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