CN111388611A - Method for extracting total flavonoids from lemongrass - Google Patents
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Abstract
The invention provides a method for extracting total flavonoids by utilizing lemongrass, which comprises the following steps: extracting total flavonoids from Cymbopogon citratus by ultrasonic method with ethanol as extractive solution; the feed-liquid ratio of the lemongrass to the ethanol is 1: 20-50; the concentration of the ethanol is 50% -70%; the extraction temperature is 40-80 ℃; the extraction time is 0.5-2.5 h. The invention utilizes the ultrasonic method to extract the total flavonoids in the lemongrass, not only shortens the extraction time, but also utilizes the acoustic cavitation of the ultrasonic wave on the solvent and the sample to smash the sample, so that the sample is fully contacted with the solvent, and the mass transfer rate is improved, thereby improving the extraction rate of the total flavonoids in the lemongrass.
Description
Technical Field
The invention relates to the technical field of materials, in particular to a method for extracting total flavonoids by utilizing lemongrass.
Background
Cymbopogon citratus (DC.) stapf is perennial herb of Cymbopogon of Gramineae, and is native to south India and Sri Lanka. The citronella essential oil is widely used as a natural aromatizing agent in the aspects of cosmetics, perfumes, bath foam and other daily chemical products, has antibacterial and antioxidant functions, and is added into tobacco to increase aroma, improve cigarette quality and reduce irritation. In south-east asia, it is also an important food seasoning. Citronella is also a common mosquito repellent, is a main active ingredient in the American natural herbal mosquito repellent, can be developed into a botanical pesticide, and has the functions of dispelling wind, dredging collaterals, warming middle-jiao, relieving pain and stopping diarrhea when being used as a medicine.
A total of 9 plants of the genus yunnan citronella have been reported, namely, rutabaga (cymbopogon. distans), cymbopogon culiginosus (citrus c. goeringii), corymbosa (cymbopogon martini c. martini), coleus contortus (c. torulis), lemongrass (c. citrus), cymbopogon nardus (c. nardus), lemongrass (c. flexuosus), citronella javanica (c. winterianus), and citronella viridis (c. caesius). Lemon grass (Cymbopogon citratus) and citronella (Cymbopogon flexuosus) are mostly used for producing the Yunnan citronella oil.
The citronella essential oil has the highest content of Citral (Citral), which contains two components which are stereoisomers of each other, α -Citral (Geranial Geranial) and β -Citral (Neral), and secondly, myrcene, linalool, geraniol, citronellol, dipentene, limonene and the like, the citronella is a lemon type sweet, pungent and warm nature, has the effects of dispelling wind and dredging collaterals, relieving pain, restoring consciousness and relieving diarrhea, and the citronella has different varieties, aromatic oil of the citronella can contain different main components, such as Citral, citronella mainly contains citronellal, citronella alcohol, geraniol and the like, the citronella oil is a lemon type, and the main chemical components are β -myrcene, Neral, citronella essential oil is a citronella oil which is a botanical antioxidant, a citronella essential oil which is a citronella oil, a citronella oil which is a chemical ingredient, a citronella oil which is a flavonoid, a plant, a flavonoid, a plant essence, a plant, a.
The conventional method for extracting the total flavonoids in the lemongrass comprises the steps of firstly, accurately weighing 20g of a lemongrass powder sample, performing reflux extraction for 2 hours by using 60% ethanol water solution, performing reflux extraction for 1 hour for the second time, wherein the liquid-material ratio is 10: 1m L/g, performing reduced pressure distillation on an extract liquid, concentrating the extract liquid into an extract, and performing constant volume to 500m L by using 80% methanol water solution for later use.
Disclosure of Invention
The invention aims to solve the defects in the prior art and provides a method for extracting total flavonoids by utilizing lemongrass, which has short extraction time and high extraction rate.
A method for extracting total flavonoids from Cymbopogon citratus comprises: extracting total flavonoids from Cymbopogon citratus by ultrasonic method with ethanol as extractive solution;
the feed-liquid ratio of the lemongrass to the ethanol is 1: 20-50; the concentration of the ethanol is 50% -70%; the extraction temperature is 40-80 ℃; the extraction time is 0.5-2.5 h.
Further, in the method for extracting total flavonoids by using lemongrass as described above, the concentration of the ethanol is 55%.
Further, according to the method for extracting the total flavonoids by utilizing the lemongrass, the feed-liquid ratio of the lemongrass to the ethanol is 1: 35.
Further, the method for extracting the total flavonoids by utilizing the lemongrass as described above has the extraction temperature of 65 ℃.
Further, the method for extracting the total flavonoids by utilizing the lemongrass as described above has the extraction time of 1.5 h.
Has the advantages that:
1. the time for extracting the total flavonoids in the lemongrass by using the ultrasonic method is longer than that of the heating extraction method, but the extraction rate is higher than that of the heating extraction method.
2. The invention utilizes the ultrasonic method to extract the total flavonoids in the lemongrass, not only shortens the extraction time, but also utilizes the acoustic cavitation of the ultrasonic wave on the solvent and the sample to smash the sample, so that the sample is fully contacted with the solvent, and the mass transfer rate is improved, thereby improving the extraction rate of the total flavonoids in the lemongrass.
Drawings
FIG. 1 is a graph of the effect of different ethanol concentrations (50%, 60%, 70%, 80% and 90%) on the lemongrass flavone extraction rate;
FIG. 2 shows the effect of different feed-liquid ratios (1: 10, 1:20, 1: 30, 1: 40, 1: 50) on the extraction rate of citronella flavone;
FIG. 3 shows the effect of different extraction times (0.5h, 1h, 1.5h, 2h, 2.5h) on the extraction rate of lemongrass flavones;
FIG. 4 shows the effect of different extraction temperatures (40 deg.C, 50 deg.C, 60 deg.C, 70 deg.C, 80 deg.C) on the extraction rate of lemongrass flavones;
FIG. 5 shows the lemongrass flavone extract and VCHas effect on hydroxyl radical clearance rate.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the technical solutions of the present invention are described clearly and completely below, and it is obvious that the described embodiments are some, not all embodiments of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Experimental example:
1. and (4) preparing a rutin standard solution.
Weighing 0.0146g of dried rutin, heating and dissolving with 80% (volume fraction) ethanol solution, and metering to 50m L brown volumetric flask to obtain 0.292 mg/L rutin control solution.
1.1 selection of measurement wavelength.
Accurately measuring 2.0m L rutin standard solution into a 10m L volumetric flask, diluting the solution to 5m L by using 80% ethanol water solution, adding 5% sodium nitrite solution 0.3m L, shaking up, standing for 6min, adding 10% aluminum nitrate solution 0.3m L, uniformly mixing, standing for 6min, adding 4% sodium hydroxide solution 1.5m L, adding 80% ethanol water solution, fixing the volume to 10m L, shaking up, standing for 15min, taking another group of reagent blank as a reference, carrying out ultraviolet spectrum scanning in a wavelength range of 200-650 nm, finding that a maximum absorption peak appears at 500nm, and determining the wavelength as the optimal wavelength.
1.2 Standard Curve plotting.
Accurately measuring rutin standard solutions 0.0, 0.5, 1.0, 2.0, 3.0, 4.0 and 5.0m L to 10m L volumetric flasks in sequence, adding 80% ethanol water solution to 5m L, shaking up, respectively adding 0.3m L5% sodium nitrite solution, shaking up, standing for 6min, adding 10% aluminum nitrate solution 0.3m L, standing for 6min after uniform mixing, adding 4% sodium hydroxide solution 1.5m L, adding 80% ethanol water solution to reach 10m L, taking another group of reagent blank as a reference, measuring absorbance at the wavelength of 500nm, and drawing a standard working curve by taking rutin content (g) as a horizontal coordinate and absorbance as a vertical coordinate to obtain a linear regression equation, wherein Y is 17.102X-0.0082, and the correlation coefficient r is 0.9996.
2. Extraction of total flavonoids
2.1 Single factor test. Extracting total flavone from herba Cymbopogonis Citrari with ethanol by ultrasonic method. Crushing by a high-speed crusher, sieving by a 60-mesh sieve, accurately weighing a certain amount of lemongrass, and researching the influence of ethanol concentration, material-liquid ratio, extraction time and extraction temperature on the extraction rate of total flavonoids in the lemongrass under a certain condition; on the basis of preliminary experiments and reference to other relevant documents, the following experiments were designed:
(1) under the conditions that the ratio of the fixed material to the liquid is 1:20, the extraction temperature is 70 ℃, and the extraction time is 1.5h, the influence of different ethanol concentrations (50%, 60%, 70%, 80% and 90%) on the extraction rate of the lemongrass flavone is discussed.
(2) Under the conditions that the concentration of fixed ethanol is 70%, the extraction temperature is 70 ℃ and the extraction time is 1.5h, the influence of different material-liquid ratios (1: 10, 1:20, 1: 30, 1: 40 and 1: 50) on the extraction rate of the lemongrass flavone is discussed.
(3) Under the conditions of a fixed feed-liquid ratio of 1:20, an extraction temperature of 70 ℃ and an ethanol concentration of 70%, the influence of different extraction times (0.5h, 1h, 1.5h, 2h and 2.5h) on the extraction rate of the lemongrass flavone is discussed.
(4) Under the conditions of fixed extraction time of 1.5h, material-liquid ratio of 1:20 and ethanol concentration of 70%, the influence of different extraction temperatures (40 deg.C, 50 deg.C, 60 deg.C, 70 deg.C, 80 deg.C) on the extraction rate of lemongrass flavone is discussed.
The flavone extraction rate calculation formula is as follows:
R(％)＝(C×N×V)/(m×1000)×100
wherein, C is the concentration of the sample solution; a-absorbance; n-dilution multiple; v is the volume of the extracting solution; m-mass of lemongrass.
Under the conditions of fixed extraction temperature of 70 ℃, material-liquid ratio of 1:20 and extraction time of 1.5h, as can be seen from figure 1, when the ethanol concentration is 50% -60%, the extraction rate of the flavone in the lemongrass is gradually increased, when the ethanol concentration reaches 60%, the extraction rate of the flavone in the lemongrass is the highest (4.480%), and when the ethanol concentration exceeds 60%, the extraction rate of the flavone in the lemongrass is gradually reduced, so that 60% is selected as the optimal ethanol concentration.
Under the conditions of fixed ethanol concentration of 70%, extraction temperature of 70 ℃ and extraction time of 1.5h, as can be seen from fig. 2, the extraction rate of the citronella grass flavone tends to increase and decrease with the increase of the feed-liquid ratio, and when the feed-liquid ratio is 1: 30, the extraction rate of the citronella grass flavone is the highest (3.897%), so that the optimum feed-liquid ratio is selected to be 1: 30.
Under the conditions of fixed feed-liquid ratio of 1:20, extraction temperature of 70 ℃ and ethanol concentration of 70%, as shown in fig. 3, when the ultrasonic extraction time is within 1.5h, the extraction rate of the citronella grass flavone is obviously increased along with the extension of the ultrasonic time, and when the ultrasonic extraction time is within 1.5h, the extraction rate is the highest (5.018%), and when the ultrasonic extraction time exceeds 1.5h, the extraction rate is gradually reduced, and mainly long-time ultrasonic extraction can destroy the total flavone in the citronella grass and influence the extraction rate of the total flavone in the citronella grass, so 1.5h is selected as the optimal ultrasonic extraction time.
Under the conditions of fixed extraction time of 1.5h, material-liquid ratio of 1:20 and ethanol concentration of 70%, as can be seen from fig. 4, when the ultrasonic extraction temperature is from 40 ℃ to 60 ℃, the extraction rate of the total flavonoids in the lemongrass increases with the increase of the temperature, and reaches a maximum value (4.404%) at 60 ℃, and when the ultrasonic extraction temperature exceeds 60 ℃, the extraction rate of the total flavonoids in the lemongrass is obviously reduced, probably because the temperature increases to cause solvent volatilization loss, so 60 ℃ is selected as the optimal extraction temperature.
2.2 orthogonal test from the results of the single-factor test, it can be known that the extraction rate of the lemongrass flavone is greatly influenced by 4 factors of the concentration of an extracting agent (ethanol), the ratio of material to liquid, the extraction temperature and the extraction time, so that the test designs 4 factors 3 level L9 (3) by taking the extraction rate of the total flavone as an investigation index on the basis of the single-factor test4) Orthogonal test to investigate citronellaThe optimum extraction conditions for extracting the total flavonoids from the grass are shown in Table 1.
TABLE 1 orthogonal factor horizontal settings
The orthogonal experimental design is shown in table 2 below:
TABLE 2 Quadrature experimental results of lemongrass flavone extraction process
Optimizing the extraction process conditions of the total flavonoids of the lemongrass by an orthogonal test to obtain the optimal extraction process combination of the lemongrass A3B3C2D1The optimal extraction process conditions of the total flavonoids of the lemongrass are as follows: the ethanol concentration is 55%, the feed-liquid ratio is 1:35, the extraction temperature is 65 ℃, the extraction time is 1.5h, and the obtained relation influencing the flavone extraction factors is as follows: temperature of extraction>Ratio of material to liquid>Concentration of ethanol>And (4) extracting time. To verify A3B3C2D1Whether the optimal condition is met or not is judged, 3 verification tests are carried out, and the result is shown in A3B3C2D1The yield of the lemongrass flavone under the condition is 5.221 percent, which is higher than that of any group in the orthogonal test, and the result of the orthogonal test is accurate.
2.3 comparison of extraction rates between different extraction methods
As can be seen from table 3, the optimal extraction process conditions for the total flavonoids in lemongrass are as follows: the ethanol concentration is 55%, the feed-liquid ratio is 1:35, the extraction temperature is 65 ℃, and the extraction time is 1.5h, and the existing method is utilized: the total flavone in the lemongrass is extracted by a heating reflux method and an ethanol hot-dipping extraction method, and the extraction rate is lower than that of an ultrasonic method.
TABLE 3 orthogonal factor horizontal settings
3. Hydroxyl radical scavenging experiments
The clearance rate of total flavone and Vc in lemon grass to hydroxyl free radical is measured by salicylic acid method, firstly, the total flavone in lemon grass is extracted by utilizing optimum technological condition, the extract is concentrated by using rotary evaporator, dried by using drying oven under the condition of below 60 deg.C, and matched with Vc whose concentration is 1.0mg/ml by using 55% ethyl alcohol, and compared with clearance effect, the concrete method is that 5 test tubes are respectively added with 10 mmol/L FeSO41.0m L and 5 mmol/L salicylic acid-ethanol 2.0m L, mixing, adding 0.2, 0.4, 0.6, 0.8, 1.0m L1.0 mg/m L flavone solution to make flavone have different concentrations, adding deionized water to 7.0m L, and adding 2.0m L H2O2The reaction was started, incubated at 37 ℃ for 30min and absorbance measured at 510 nm. The clearance rate of flavone to hydroxyl radical with different concentrations is calculated according to the following formula.
E(％)＝[A0-(Am-An)]/A0×100％
Wherein A0 is the light absorption value of blank control solution, Am is the light absorption value after flavone is added, An is no H2O2Absorbance of flavonoids.
As can be seen from FIG. 5, within a certain concentration range, the lemongrass flavone extract and VCHas effect in scavenging hydroxy free radicals, and the Cymbopogon citratus flavone extract has no effect in scavenging hydroxy free radicals as much as VCHowever, the lemongrass flavone extract has a certain scavenging effect on hydroxyl free radicals, and the scavenging effect on the hydroxyl free radicals is continuously enhanced along with the increase of the concentration of the flavone, so that the lemongrass flavone extract has a certain antioxidant activity and provides scientific reference for the medicinal development of the lemongrass.
The invention discusses the optimum process conditions for extracting the total flavonoids through single-factor experiments and orthogonal experiments, and scientifically and reasonably optimizes the extraction conditions of the total flavonoids in the lemongrass. The single-factor test design is a test design method which lists all factors influencing test results, only changes the numerical value of the factor to be examined, and keeps the numerical values of all other factors unchanged. Although the intuition is good, the experiment times are more, the cost is higher, the mutual influence among the factors is not considered in the experiment, the relevance is not considered by default, and systematic errors are easy to generate. The orthogonal test has no strict limitation on the number of the factors, and the design can be utilized when the factors have interaction or not, a preliminary conclusion can be obtained by performing test comparison through an orthogonal table, a specific conclusion can be obtained through variance analysis, and the optimal test condition can be obtained, so that the test times can be greatly reduced through the comprehensive test, and the optimal condition can be found through the test analysis.
In addition, the lemongrass flavone extract has no effect of eliminating hydroxyl free radicals like VCHowever, the lemongrass flavone extract has a certain scavenging effect on hydroxyl free radicals, and the scavenging effect on the hydroxyl free radicals is continuously enhanced along with the increase of the concentration of the flavone, so that the lemongrass flavone extract has a certain antioxidant activity and provides scientific reference for the medicinal development of the lemongrass.
Finally, it should be noted that: the above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions of the embodiments of the present invention.
Claims (5)
1. A method for extracting total flavonoids by utilizing lemongrass is characterized by comprising the following steps: extracting total flavonoids from Cymbopogon citratus by ultrasonic method with ethanol as extractive solution;
the feed-liquid ratio of the lemongrass to the ethanol is 1: 20-50; the concentration of the ethanol is 50% -70%; the extraction temperature is 40-80 ℃; the extraction time is 0.5-2.5 h.
2. The method for extracting total flavonoids from lemongrass according to claim 1, wherein the concentration of ethanol is 55%.
3. The method for extracting total flavonoids from lemongrass according to claim 1, wherein the ratio of the lemongrass to the ethanol is 1: 35.
4. The method for extracting total flavonoids from lemongrass according to claim 1, wherein the extraction temperature is 65 ℃.
5. The method for extracting total flavonoids from lemongrass according to claim 1, wherein the extraction time is 1.5 h.
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| 张雪梅等: "香茅草黄酮提取物对L-NNA诱导的高血压大鼠血压", 《吉林中医药》 * |
| 李青山等: "《药学基本实验技术》", 31 August 2006, 中国医药科技出版社 * |
| 谷瑶等: "柠檬香茅草总黄酮和多糖含量的测定", 《应用化工》 * |
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| CN112972332A (en) * | 2021-03-03 | 2021-06-18 | 李培丰 | Rose whitening essence and preparation method thereof |
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