CN110879262A - HPLC (high Performance liquid chromatography) determination method for high hazy lignin content in Cumin Beeherman medicinal material - Google Patents
HPLC (high Performance liquid chromatography) determination method for high hazy lignin content in Cumin Beeherman medicinal material Download PDFInfo
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Abstract
The invention belongs to the technical field of medicinal material measurement, and discloses an HPLC (high performance liquid chromatography) determination method for high hazy lignin content in a Cuminum cyminum medicinal material, which comprises the steps of taking Cuminum cyminum medicinal material powder, precisely weighing, placing the powder in a conical flask with a plug, precisely adding methanol, weighing, carrying out ultrasonic or reflux treatment, cooling, weighing again, complementing the weight loss by methanol, shaking uniformly, filtering, taking subsequent filtrate, and filtering by a 0.45-micrometer microporous filter membrane to obtain the finished product. The HPLC determination method for the content of high hazy lignin in the Cumin Bayer medicinal material can retain chromatographic peaks with consistent time, and has good separation degree and good instrument precision; and the components to be detected can be stably presented, the components to be detected are basically stable within 24 hours, and the accuracy of the detection result is good. The method can accurately reflect the component content in the medicinal materials, and more effectively evaluate the quality of the medicinal materials from different sources.
Description
Technical Field
The invention belongs to the technical field of measurement, and particularly relates to an HPLC (high performance liquid chromatography) determination method for high hazy lignin content in a Cuminum celery Beeherman medicinal material.
Background
The plant is a commonly used medicinal material in Xinjiang, is a plant in the genus of Salvia in the family of Labiatae, a plant in the genus of Salvia, a plant which grows for two or more years, has a lignified root part and a height of about 1 meter, and has a stem which is quadrangular, upright, fine hair or green purple and is multi-branched; the leaves are opposite, and have long stems, elliptic or oblong leaves. The round umbrella inflorescence has 5-6 small flowers per round, flower amphipathy, small nut ovoid, non-brown and smooth. The flowering period is 7-9 months, and the seed maturation period is 8-10 months; the Cumin Bayhelman is used as a medicine by root, and has the effects of generating heat by drying, tonifying heart and strengthening yang, enriching body and essence, refreshing heart and nourishing mind, eliminating dampness and securing essence, eliminating jaundice, warming uterus and generating brilliance and the like. Can be used for treating damp-cold or mucus diseases, such as palpitation, sexual impotence, marasmus, oligospermia, vexation, premature ejaculation, spermatorrhea, jaundice, cold womb, and dim complexion. In the prior art, relevant researches or reports about high hazy lignin which is a main component of the Cuminum celery Beehermann medicinal material are found, and the method also belongs to the industrial blank. In the preparation process of the medicine, the determination of the high hazy lignin content in the Cumin Beehermann medicinal material is a very key step.
At present, a method for accurately measuring the content of hazy lignin in Cumin Byhermann medicinal materials is lacked.
The significance of solving the technical problems is as follows: the high hazy lignin is separated from the Cuminum cuminum medicinal material for the first time, has high content, and is an important way for controlling the quality of the medicinal material.
Disclosure of Invention
Aiming at the problems in the prior art, the invention provides an HPLC (high performance liquid chromatography) determination method for high hazy lignin content in a Cuminum celery medical material.
The invention is realized in this way, an HPLC determination method of high hazy lignin content in Cumin Beehermann medicinal material, comprising the following steps:
taking a sample of Cumin Beeherman, and weighing;
step two, adding methanol and weighing;
step three, reflux extraction and cooling;
weighing again, complementing the weight loss with methanol, shaking up and filtering; filtering the filtrate with microporous membrane;
and fifthly, repeating the sample feeding for many times under the optimized chromatographic condition, determining the peak area of the high haze lignin peak, taking the average value of the peak areas of 3 times, and calculating the amount of the high haze lignin according to a standard curve.
Further, in the first step, 1.0g of a Cuminum cyminum sample is weighed and placed in a round-bottom flask.
Further, in the second step, the concentration of the methanol is 80%, and the addition amount is 50 ml.
Further, in the third step, the reflux extraction time is 30 minutes.
Further, in step four, it was weighed again and the weight loss was made up with 80% methanol.
Further, in the fourth step, the diameter of the microporous filter membrane is 0.45 μm.
Further, in step five, the preferred chromatographic conditions specifically include:
octadecylsilane chemically bonded silica is used as a filling agent; taking 0.1% formic acid solution as a mobile phase A and acetonitrile as a mobile phase B, and carrying out gradient elution according to the specification in the following table; the detection wavelength is 289 nm; column temperature: 30 ℃; flow rate: 1 ml/min.
Further, step five further includes: and repeating the sample injection for 3 times under the optimized chromatographic condition, measuring the peak area of the high haze lignin peak by 10 mu l each time, taking the average value of the areas of the 3 peaks, and calculating the amount of the high haze lignin according to a standard curve.
The invention also aims to provide an HPLC (high performance liquid chromatography) measuring device for the high hazy lignin content in the Cuminum celery medical material, which is used for implementing the HPLC measuring method for the high hazy lignin content in the Cuminum celery medical material.
In summary, the advantages and positive effects of the invention are:
the high hazy lignin provided by the invention is obtained and proposed from the medicinal material for the first time. The HPLC method for measuring the content of hazy lignin in the Cumin Bayer medicinal material can retain chromatographic peaks with consistent time, and has good separation degree and instrument precision; and the components to be detected can be stably presented, the components to be detected are basically stable within 24 hours, and the accuracy of the detection result is good. The method can accurately reflect the component content in the medicinal materials, and more effectively evaluate the quality of the medicinal materials from different sources.
Drawings
Fig. 1 is a flow chart of an HPLC method for determining the content of hazy lignin in a cumin bayer process raman medicinal material according to an embodiment of the present invention.
Fig. 2 is a schematic diagram of a relationship between high haze lignin and peak area according to an embodiment of the present invention.
Fig. 3 is a graphical representation of the variation of high haze lignin content with solvent concentration provided by an embodiment of the present invention.
Fig. 4 is a graph illustrating the variation of high haze lignin content with extraction time according to an embodiment of the present invention.
Fig. 5 is a graphical representation of the high haze lignin content as a function of solvent usage provided by an embodiment of the present invention.
Fig. 6 is an HPLC chromatogram of a solution of a cuminum baizmann test sample and a high haze lignin control sample provided in an embodiment of the present invention.
In the figure: 1. a sample of cumin bailmen; 2. high haze lignin control.
FIG. 7 is an HPLC chromatogram 1 of root and rhizome of 15 Cuminum cyminum plants provided by the example of the present invention.
FIG. 8 is an HPLC chromatogram 2 of the root and rhizome of 15 Cuminum cyminum plants provided by the example of the invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following 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 present invention will be described in detail with reference to the accompanying drawings.
The HPLC determination method for the high hazy lignin content in the Cumin Bayer medicinal material provided by the embodiment of the invention comprises the following steps:
weighing Cuminum cyminum medicinal powder, precisely weighing, placing in a conical flask with a plug, precisely adding methanol, weighing, ultrasonically treating or refluxing, cooling, weighing again, supplementing the weight loss with methanol, shaking, filtering, collecting the filtrate, and filtering with 0.45 μm microporous membrane.
As shown in fig. 1, the HPLC method for determining the content of hazy lignin in a cumin bayer raman medicinal material provided in the embodiment of the present invention specifically includes the following steps:
s101, taking about 1.0g of a cumin Bayer sample, precisely weighing, and placing in a round-bottom flask.
S102, 50ml of 80% methanol was added to the round-bottom flask, and the weight was measured.
S103, refluxing and extracting for 30 minutes, and cooling.
S104, weighing again, complementing weight loss with 80% methanol, shaking up, and filtering; the filtrate was then filtered through a 0.45 μm microfiltration membrane.
S105, using octadecylsilane chemically bonded silica as a filler; taking 0.1% formic acid solution as a mobile phase A and acetonitrile as a mobile phase B, and carrying out gradient elution according to the specification in the following table; the detection wavelength is 289 nm; column temperature: 30 ℃; flow rate: 1 ml/min; and repeating the sample injection for 3 times, measuring the peak area of the high-haze lignin peak by 10 mu l each time, taking the average value of the areas of the 3 peaks, and calculating the amount of the high-haze lignin according to a standard curve.
The present invention will be further described with reference to specific examples and experiments.
Example 1:
1. preparation of control solutions
The high hazy lignin control 15mg was weighed accurately, dissolved in methanol and diluted to make a solution containing 0.3mg of control per 1 ml.
2. Preliminary simulation of test solution
Taking about 1.0g of sample powder, precisely weighing, placing in a conical flask with a plug, precisely adding 50ml of methanol, weighing, ultrasonically treating for 30min, cooling, weighing again, supplementing the weight loss with methanol, shaking, filtering, taking the subsequent filtrate, and filtering with 0.45 μm microporous membrane.
3. Selection of measurement wavelength
A test solution with a certain concentration and a high-haze lignin reference solution are respectively taken and analyzed and measured on a high performance liquid chromatograph, and the result shows that the two solutions have larger absorption near the wavelength 289nm, so the wavelength 289nm is selected as the measurement wavelength of the experiment.
4. Chromatographic conditions
Octadecylsilane chemically bonded silica is used as a filling agent; taking 0.1% formic acid solution as a mobile phase A and acetonitrile as a mobile phase B, and carrying out gradient elution according to the specification in the following table; the detection wavelength is 289 nm; column temperature: 30 ℃; flow rate: 1 ml/min; sample introduction amount: 10 μ l.
HPLC gradient elution conditions
5. Preparation of Standard Curve for control
Precisely measuring 1ml, 2ml, 3ml, 4ml, 5ml, 6ml, 7ml and 8ml of the reference substance solution, respectively placing in a 10ml volumetric flask, metering to the scale with methanol, and shaking up. Under the above chromatographic conditions, the solution of each concentration was repeatedly injected 3 times, 10. mu.l each time, and the peak area was determined, and the average of the 3 peak areas was taken. Respectively taking the content of the reference substance solution as a horizontal coordinate and the peak area as a vertical coordinate, drawing a standard curve, and calculating a regression equation: Y34.128X-1.2597 (R)20.9997). The result shows that the high haze lignin has a better linear relation with the peak area in the range of 0.3-2.4 mug.
6. Optimization of extraction conditions
6.1 selection of extraction solvent
Precisely weighing about 1.0g of Cumin Beeherman sample powder, 2 parts in total, respectively placing the powder into conical flasks with plugs, precisely adding 25ml of methanol and absolute ethyl alcohol, weighing and recording, ultrasonically extracting for 30min, cooling, weighing again, complementing the weight loss with corresponding solvents, shaking up, filtering, taking subsequent filtrate, and filtering with a 0.45 mu m microporous filter membrane. And (3) repeatedly injecting sample into each sample solution under the chromatographic condition for 3 times, measuring the peak area of the high-haze lignin peak by 10 mu l each time, and taking the average value of the peak areas of 3 times. The extraction efficiencies of these 2 solvents were compared, calculated from the standard curve, and the results showed that the extraction efficiency of methanol as the extraction solvent was slightly higher.
Influence of different extraction solvents on measured amount of high hazy lignin extraction
6.2 selection of extraction method
Precisely weighing about 1.0g of Cumin Beeherman sample powder, 2 parts in total, respectively placing the Cumin Beeherman sample powder into a round-bottom flask, respectively adding 25ml of methanol, weighing and recording, respectively extracting for 30min by an ultrasonic extraction method and a reflux extraction method, cooling, then complementing weight loss by methanol, shaking up, filtering, and taking subsequent filtrate; filtering with 0.45 μm microporous membrane, repeatedly injecting sample into each sample solution for 3 times under the above chromatographic conditions, measuring the peak area of high haze lignin peak, and averaging the peak areas of 3 times. The extraction efficiency of the 2 modes is compared according to the calculation of a standard curve, and the result shows that the extraction efficiency of the reflux extraction method is higher.
Influence of extraction mode on extraction amount of high hazy lignin
6.3 selection of the concentration of extraction solvent
Precisely weighing about 1.0g of Cumin Beeherman sample powder, totally 4 parts, respectively placing the Cumin Beeherman sample powder into a round-bottom flask, respectively adding 25ml of 100% methanol, 90% methanol, 80% methanol and 70% methanol, weighing and recording, performing reflux extraction for 30min at the temperature of about 90 ℃, cooling, complementing weight loss by using a solvent with corresponding concentration, shaking up, filtering, taking subsequent filtrate, and filtering by using a 0.45 mu m microporous membrane. And (3) repeatedly injecting samples into each sample under the chromatographic condition for 3 times, measuring the peak area of the high hazy lignin peak by 10 mu l each time, taking the average value of the areas of the 3 peaks, and calculating the content of the high hazy lignin extracted according to a standard curve. The results showed that the extraction yield was highest with 80% methanol as the extraction solvent.
6.4 selection of extraction time
Precisely weighing about 1.0g of Cumin Beeherman sample powder, totally 4 parts, respectively placing the Cumin Beeherman sample powder into a round bottom flask, respectively adding 25ml of 80% methanol solution, weighing and recording, respectively performing reflux extraction for 30min, 60min, 90min and 120min, cooling, weighing again, complementing weight loss with 80% methanol, shaking up, filtering, taking subsequent filtrate, and filtering with a 0.45 mu m microporous membrane. And (3) repeatedly injecting samples for each sample under the chromatographic conditions for 10 mu l each time, measuring the peak area of the high-haze lignin peak, and taking the average value of the peak areas of 3 times. The extraction amount of high haze lignin was calculated according to the standard curve. The result shows that the extraction efficiency gradually decreases with the increase of the extraction time, so the optimal extraction time is 30 min.
6.5 selection of the amount of extraction solvent
Precisely weighing about 1.0g of Cumin Beeherman sample powder, 5 parts in total, respectively placing the Cumin Beeherman sample powder into a round-bottom flask, respectively adding 20 ml, 30 ml, 40 ml, 50ml and 60ml of 80% methanol, weighing and recording, carrying out reflux extraction for 30min, cooling, complementing the weight loss with 80% methanol, shaking up, filtering, taking subsequent filtrate, and filtering with a 0.45 mu m microporous membrane. And (3) repeatedly injecting samples for each sample under the chromatographic conditions for 10 mu l each time, measuring the peak area of the high-haze lignin peak, and taking the average value of the peak areas of 3 times. The results show that the extraction yield is highest when the feed-liquid ratio is 1: 50.
7. Preparation method and content determination method of test solution
Taking about 1.0g of sample powder, precisely weighing, placing in a round bottom flask, precisely adding 50ml of 80% methanol, weighing, reflux extracting for 30min, cooling, weighing again, complementing weight loss with 80% methanol, shaking, filtering, taking the subsequent filtrate, and filtering with 0.45 μm microporous membrane. And (3) repeatedly injecting samples into each sample under the chromatographic condition for 3 times, measuring the peak area of the high hazy lignin peak by 10 mu l each time, taking the average value of the areas of the 3 peaks, and calculating the amount of the high hazy lignin according to a standard curve.
8. Test of
8.1 determination of Components to be tested and System suitability test
The sample solution to be tested and the high hazy lignin reference substance solution with a certain concentration are taken and injected under the chromatographic conditions, and the result shows that the sample solution has chromatographic peaks which have the same retention time with the high hazy lignin reference substance, and the chromatographic peaks have symmetrical peak shapes, good separation degree and high column efficiency.
8.2 precision test
Weighing about 1.0g of Cuminum cyminum sample powder, precisely weighing, placing in a round bottom flask, precisely adding 50ml of 80% methanol, weighing, reflux extracting for 30min, cooling, weighing again, supplementing the lost weight with 80% methanol, shaking, filtering, collecting the subsequent filtrate, and filtering with 0.45 μm microporous membrane. Under the chromatographic conditions, the peak area of the high haze lignin is continuously measured for 6 times, 10 mu l of the peak area is measured each time, and the amount of the high haze lignin is calculated according to a standard curve. The RSD value of the measurement result was 0.79%, indicating that the precision of the apparatus was good.
8.3 reproducibility test
Weighing about 1.0g of Cuminum cyminum sample powder, precisely weighing, paralleling 6 parts, placing in a round bottom flask, precisely adding 50ml of 80% methanol, weighing, reflux extracting for 30min, cooling, weighing again, supplementing the lost weight with 80% methanol, shaking, filtering, taking the subsequent filtrate, and filtering with 0.45 μm microporous membrane. And (3) repeatedly injecting samples into each sample under the chromatographic condition for 3 times, measuring the peak area of the high hazy lignin by 10 mu l each time, taking the average value of the peak areas of 3 times, and calculating the amount of the high hazy lignin according to a standard curve. The RSD value of the measurement result was 1.27%, indicating that the reproducibility of the method was good.
8.4 stability test
Weighing about 1.0g of Cuminum cyminum sample powder, precisely weighing, paralleling 6 parts, placing in a round bottom flask, precisely adding 50ml of 80% methanol, weighing, reflux extracting for 30min, cooling, weighing again, supplementing the lost weight with 80% methanol, shaking, filtering, taking the subsequent filtrate, and filtering with 0.45 μm microporous membrane. And (3) repeatedly injecting samples of each sample under the chromatographic condition for 3 times, wherein each time is 10 mu l, measuring the peak area of the high hazy lignin in 0, 2, 4, 6, 8, 10, 12 and 24 hours respectively, taking the average value of the peak areas of 3 times, and calculating the amount of the high hazy lignin according to a standard curve. The RSD value of the measurement result is 1.16%, which shows that the components to be measured in the test solution prepared by the method are basically stable within 24 hours.
8.5 sample application recovery test
Taking about 0.5g of Cumin Beeherman sample powder, respectively adding 1.97mg of high hazy lignin control, precisely weighing, paralleling 6 parts, placing in a round bottom flask, precisely adding 25ml of 80% methanol, weighing, reflux extracting for 30min, cooling, weighing again, complementing the lost weight with 80% methanol, shaking up, filtering, taking the subsequent filtrate, and filtering with a 0.45 mu m microporous membrane. And (3) repeatedly injecting samples into each sample under the chromatographic condition for 3 times, measuring the peak area of the high hazy lignin by 10 mu l each time, taking the average value of the peak areas of 3 times, and calculating the amount of the high hazy lignin according to a standard curve. The calculated average recovery was 100.59% and the RSD value was 1.37%, indicating that the method is highly accurate for determining high haze lignin content.
9. Determination of high hazy lignin content in sample
The content of high haze lignin in the dry product was calculated by measuring the dry roots and rhizomes of Cuminum cyminum of Labiatae in 15 parts by the method, as shown in FIG. 7.
Sample source of Cumin Beeherman
| Numbering | Name (R) | Producing area/collecting |
|
| 1 | Kjellman Beeherman | |
|
| 2 | Kjellman Beeherman | Ink jade county |
|
| 3 | Kjellman Beeherman | Uygur hospital in |
|
| 4 | Kjellman Beeherman | |
|
| 5 | Kjellman Beeherman | Uygur medical hospital in |
|
| 6 | Kjellman Beeherman | Turpan |
|
| 7 | Kjellman Beeherman | Aksu |
|
| 8 | Kjellman Beeherman | Uygur Hospital in |
|
| 9 | Kjellman | Kashiuygur hospital | |
| 10 | Kjellman Beeherman | Hospital for |
|
| 11 | Kjellman Beeherman | |
|
| 12 | Kjellman Beeherman | The second |
|
| 13 | Kjellman | Kashi market | |
| 14 | Kjellman Beeherman | Hami |
|
| 15 | Kjellman Beeherman | The He Tian medicinal materials market |
Determination result of high hazy lignin content in 15 medicinal materials
| Sample numbering | Content (mg/g) |
| 1 | 3.38 |
| 2 | 1.94 |
| 3 | 3.86 |
| 4 | 6.81 |
| 5 | 1.96 |
| 6 | 6.81 |
| 7 | 2.87 |
| 8 | 3.86 |
| 9 | 1.84 |
| 10 | 3.98 |
| 11 | 3.00 |
| 12 | 3.47 |
| 13 | 1.05 |
| 14 | 4.23 |
| 15 | 3.36 |
The present invention will be further described with reference to effects.
In the research of measuring the content of high hazy lignin in a sample by HPLC, the peak of the high hazy lignin has larger absorption at the position of wavelength 289nm in a sample solution and a high hazy lignin reference substance by HPLC analysis and detection, so 289nm is selected as the measuring wavelength of the experiment.
In the extraction condition optimization experiment, the extraction solvent is selected, and the results of the measured extraction amount of the high haze lignin by different extraction solvents show that the extraction efficiency of the methanol is slightly higher; in the selection of the extraction mode, the extraction amount of the reflux extraction is obviously higher than that of the ultrasonic extraction, so the reflux extraction is selected in the extraction mode; in the optimization of the concentration of the extraction solvent, four concentration groups of 100 percent, 90 percent, 80 percent and 70 percent are respectively selected, and the result of the extraction amount shows that the extraction efficiency is highest when 80 percent methanol is used as the extraction solvent; in an experiment for researching the extraction time, reflux extraction is respectively carried out for 30min, 60mim, 90min and 120min, the extraction amount of high hazy lignin is gradually reduced along with the increase of time, and the time of components is possibly long and easy to decompose, so that 30min is selected as the extraction time; in the optimization experiment of the feed-liquid ratio, the feed-liquid ratio is respectively selected to be 1: 20,1: 30,1: 40,1: 50,1: 60, refluxing and extracting, wherein the result shows that when the feed-liquid ratio is 1: at 50, the extraction efficiency is highest.
In the methodology analysis, the determination of the component to be tested and the system applicability test, the precision test, the reproducibility test, the stability test and the sample adding and recovery rate test are respectively carried out. Experiments prove that the sample solution and the high hazy lignin reference solution have chromatographic peaks with consistent retention time and good separation degree; through the investigation of a precision test, the RSD value of the measurement result of the content of the high hazy lignin is 0.79 percent, which indicates that the precision of an instrument is good; in a reproducibility test, six groups of evaluation are respectively carried out, and the result shows that the RSD value of the high haze lignin content measurement is 1.27%, which indicates that the reproducibility of the method is good; through a stability test, the amount of high hazy lignin is measured in 0, 2, 4, 6, 8, 10, 12 and 24 hours respectively, and the RSD value of the measurement result is 1.16%, which indicates that the components to be measured in the test solution prepared by the method are basically stable in 24 hours; in the sample adding and recycling test, the average recycling rate of the high hazy lignin content is 100.59%, and the RSD value of the high hazy lignin content is 1.37%, which shows that the method has good accuracy in measuring the high hazy lignin content.
During the content measurement of the high hazy lignin in the sample, 15 Cuminum birehmann medicinal materials from different sources are respectively crushed, sieved and extracted by refluxing, the peak areas of the high hazy lignin peaks in the 15 different medicinal materials are measured by HPLC, and the amount of the high hazy lignin is calculated according to a standard curve, so that the component content in the 15 medicinal materials can be more accurately reflected, and the quality of the medicinal materials from different sources can be more effectively evaluated.
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 and improvements made within the spirit and principle of the present invention are intended to be included within the scope of the present invention.
Claims (9)
1. An HPLC (high performance liquid chromatography) determination method for the content of hazy lignin in a Cumin Bayer medicinal material is characterized by comprising the following steps of:
taking a sample of Cumin Beeherman, and weighing;
step two, adding methanol and weighing;
step three, reflux extraction and cooling;
weighing again, complementing the weight loss with methanol, shaking up and filtering; filtering the filtrate with microporous membrane;
and fifthly, repeating the sample feeding for many times under the optimized chromatographic condition, determining the peak area of the high haze lignin peak, taking the average value of the peak areas of 3 times, and calculating the amount of the high haze lignin according to a standard curve.
2. The HPLC method for measuring the content of hazy lignin in the Cuminum cuminum Biffman medicinal material as claimed in claim 1, wherein in the step one, 1.0g of a Cuminum cuminum Biffman sample is taken and weighed to be placed in a round bottom flask.
3. The HPLC method for measuring the content of hazy lignin in the Cumin Bayer medicinal material as claimed in claim 1, wherein in the second step, the concentration of the methanol is 80%, and the addition amount is 50 ml.
4. The HPLC method for measuring the hazy lignin content in the Cumin Bayer medicinal material as claimed in claim 1, wherein in the third step, the reflux extraction time is 30 minutes.
5. The HPLC method for measuring the hazy lignin content in the Cumin Bayer process medicine material as claimed in claim 1, wherein in the fourth step, the Cumin Bayer process medicine material is weighed again, and 80% methanol is used for complementing the weight loss.
6. The HPLC method for measuring the content of hazy lignin in the Cumin Bayer medicinal material as claimed in claim 1, wherein in the fourth step, the diameter of the microporous filter membrane is 0.45 μm.
7. The HPLC method for measuring the content of hazy lignin in the Cumin Bayer medicinal material as claimed in claim 1, wherein in the fifth step, the preferred chromatographic conditions specifically include:
octadecylsilane chemically bonded silica is used as a filling agent; performing gradient elution by using a 0.1% formic acid solution as a mobile phase A and acetonitrile as a mobile phase B; the detection wavelength is 289 nm; column temperature: 30 ℃; flow rate: 1 ml/min.
8. The HPLC method for measuring the content of hazy lignin in the Cumin Bayer medicinal material as set forth in claim 1, wherein the fifth step further comprises: and repeating the sample injection for 3 times under the optimized chromatographic condition, measuring the peak area of the high haze lignin peak by 10 mu l each time, taking the average value of the areas of the 3 peaks, and calculating the amount of the high haze lignin according to a standard curve.
9. An HPLC measurement device for measuring the high hazy lignin content in the Cuminum cuminum medicinal material by implementing the HPLC measurement method for measuring the high hazy lignin content in the Cuminum cuminum medicinal material according to claim 1.
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