Extraction method of basil seed total polyphenol and basil seed total polyphenol extract

Abstract

The invention provides a basil seed total polyphenol extraction method and a basil seed total polyphenol extract.The extraction method comprises the following steps that a basil seed raw material is subjected to degreasing treatment through a degreasing solvent, and a degreased raw material is prepared; carrying out ultrasonic extraction on the degreased raw material by adopting an ethanol water solution to obtain basil seed total polyphenol; wherein the degreasing solvent comprises petroleum ether; the volume ratio of ethanol in the ethanol aqueous solution is 40-80%; based on g / mL, the material-liquid ratio of the degreased raw material to the ethanol water solution is 1: (20-70); the ultrasonic extraction time is 20 to 70 minutes. According to the extraction method, the extraction efficiency of the basil seed total polyphenol can be improved, and damage to polyphenol activity is reduced.

Description

Technical Field

[0001] This application relates to the field of traditional Chinese medicine extraction technology, and in particular to a method for extracting total polyphenols from basil seeds and a total polyphenol extract from basil seeds. Background Technology

[0002] Basil seeds are derived from the fruit of the Lamiaceae plant *Ocimum basilicum* L. As a commonly used medicinal material in Uyghur medicine, basil seeds have a medicinal history spanning several centuries, with their medicinal value first recorded in ancient Uyghur medical texts such as *The White Palace*, *The Compendium of Medicine*, and *The Garden of Medicinal Herbs*. In modern standards, this medicinal material is also included in the *Standards for Uyghur Medicinal Materials* (Volume 1, 1993 edition) and the *Uyghur Medicine Section of the Ministry of Health's Drug Standards*.

[0003] In terms of medicinal properties, basil seeds are classified as having a primary dry-heat nature, possessing the effects of stopping diarrhea, clearing blockages, stopping bleeding, refreshing the body and mind, and dispersing qi. Clinically, this herb can be used to treat various heart conditions, palpitations, hemorrhoidal bleeding, chronic diarrhea, and can also be used to relieve headaches from colds, chest tightness, amenorrhea or irregular menstruation in women, and gastrointestinal discomfort such as bloating in hot weather. Modern pharmacological research has confirmed that basil seed oil possesses pharmacological activities such as anti-tumor, hypoglycemic, hypotensive, prolonging coagulation time, and prolonging sleep time. Modern pharmacological research has also determined that basil seeds exhibit excellent performance in anti-inflammatory, anti-diabetic, antioxidant, antibacterial, and anticancer applications. The chemical composition of basil seeds is complex and diverse, mainly consisting of volatile oils, polyphenols and their glycosides, coumarins, and also containing polysaccharides, fatty acids, triterpenoids, saponins, alkaloids, and other components.

[0004] Basil seeds contain polyphenols, which are the active ingredients responsible for their antioxidant and anti-inflammatory effects. These are compounds with strong physiological activity, possessing pharmacological effects such as antioxidant activity, anti-inflammatory activity, regulation of glucose and lipid metabolism, cardiovascular protection, anti-tumor activity, and neuroprotection. In recent years, their application in healthcare and food has become increasingly widespread.

[0005] Currently, the extraction process of total polyphenols from basil seeds suffers from problems such as low extraction efficiency and impaired antioxidant activity of polyphenols. Summary of the Invention

[0006] Therefore, it is necessary to provide a method for extracting total polyphenols from basil seeds and a total polyphenol extract from basil seeds to improve the extraction efficiency of total polyphenols from basil seeds and reduce the damage to the antioxidant activity of polyphenols.

[0007] The first aspect of this application provides a method for extracting total polyphenols from basil seeds, comprising the following steps:

[0008] Basil seeds were degreased using a degreasing solvent to prepare a degreased raw material.

[0009] The defatted raw material was subjected to ultrasonic extraction using an ethanol-water solution to obtain the total polyphenols from basil seeds.

[0010] The defatting solvent includes petroleum ether; the volume percentage of ethanol in the aqueous ethanol solution is 40%-80%; the ratio of the defatting raw material to the aqueous ethanol solution is 1:(20-70) in g / mL; and the ultrasonic extraction time is 20-70 min.

[0011] In some embodiments, the petroleum ether comprises petroleum ether with a boiling range of 60°C-90°C.

[0012] In some embodiments, the ratio of the basil seed raw material to the defatting solvent is 1:(10-30), expressed in g / mL.

[0013] In some embodiments, the degreasing process includes one or more of the following: soaking degreasing, ultrasonic degreasing, Soxhlet extraction degreasing, and vibration degreasing.

[0014] In some embodiments, the soaking temperature for degreasing is 22℃-25℃, and the time is 12h-16h.

[0015] In some embodiments, the ultrasonic power of ultrasonic degreasing is 200W-400W, the ultrasonic frequency is 30KHz-50KHz, and the time is 10min-30min.

[0016] In some embodiments, the Soxhlet extraction defatting time is 3-5 hours, and the water bath temperature is 60℃-90℃.

[0017] In some embodiments, the oscillation frequency of the degreasing process is 120 rpm to 160 rpm, and the time is 12 h to 16 h.

[0018] In some embodiments, the basil seed raw material is in powder form.

[0019] In some embodiments, the volume percentage of ethanol in the aqueous ethanol solution is 40%-80%.

[0020] In some embodiments, the ratio of the defatted raw material to the aqueous ethanol solution is 1:(30-70), expressed in g / mL.

[0021] In some embodiments, the ultrasonic frequency for ultrasonic extraction is 30kHz-50kHz.

[0022] In some embodiments, the ultrasonic power for ultrasonic extraction is 200W-400W.

[0023] The second aspect of this application provides a total polyphenol extract of basil seeds, which is obtained by the extraction method of total polyphenols of basil seeds according to the first aspect of this application.

[0024] The above-mentioned method for extracting total polyphenols from basil seeds first involves defatting the basil seed raw material with petroleum ether to eliminate oil interference, and then performing ultrasonic extraction on the basil seed raw material. This method can efficiently extract total polyphenols while maximizing the preservation of its high antioxidant properties, laying the foundation for the pharmaceutical application of basil seeds. Detailed Implementation

[0025] To facilitate understanding of this application, a more complete description of the application will be provided below with reference to relevant embodiments. Preferred embodiments of the application are given below. However, the application can be implemented in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided to provide a thorough and complete understanding of the disclosure of this application.

[0026] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the application.

[0027] As used herein, the terms "and / or," "or / and," and "and / or" encompass any one of two or more of the related listed items, as well as any and all combinations of the related listed items. These arbitrary and all combinations include any two related listed items, any more related listed items, or a combination of all related listed items. It should be noted that when at least three items are connected using at least two conjunctions selected from "and / or," "or / and," and "and / or," it should be understood that, in this application, the technical solution undoubtedly includes solutions connected by "logical AND," and also undoubtedly includes solutions connected by "logical OR."

[0028] In this application, the technical features described in an open-ended manner include both closed technical solutions consisting of the listed features and open technical solutions that include the listed features.

[0029] In this application, numerical ranges are referred to as continuous unless otherwise specified, and include the minimum and maximum values ​​of the range, as well as every value between the minimum and maximum values. Furthermore, when the range refers to integers, it includes every integer between the minimum and maximum values ​​of the range. Additionally, when multiple ranges are provided to describe a feature or characteristic, the ranges may be merged. In other words, unless otherwise specified, all ranges disclosed herein should be understood to include any and all subranges to which they are incorporated.

[0030] This document only specifically discloses some numerical ranges. However, any lower limit can be combined with any upper limit to form an unspecified range; and any lower limit can be combined with other lower limits to form an unspecified range, just as any upper limit can be combined with any other upper limit to form an unspecified range. Furthermore, each individually disclosed point or single value can itself serve as a lower or upper limit and be combined with any other point or single value or with other lower or upper limits to form an unspecified range.

[0031] Unless otherwise specified, the temperature parameters in this application may be either constant temperature processing or processing within a certain temperature range. The constant temperature processing allows temperature fluctuations within the precision range controlled by the instrument, such as ±5°C, ±4°C, ±3°C, ±2°C, or ±1°C.

[0032] In this document, the term "suitable" as used in phrases such as "suitable combination," "suitable method," and "any suitable method" refers to the ability to implement the technical solution of this application, solve the technical problem of this application, and achieve the expected technical effect of this application.

[0033] In this application, terms such as "further," "even further," and "particularly" are used to describe purposes and indicate differences in content, but should not be construed as limiting the scope of protection of this application.

[0034] In this application, "optionally," "optionally," and "optional" mean that something is optional, that is, it means that it is selected from either "with" or "without." If there are multiple "optional" entries in a technical solution, unless otherwise specified, and there are no contradictions or mutual constraints, each "optional" entry shall be independent.

[0035] In the description of the application, "multiple" means at least two, such as two, three, etc., unless otherwise explicitly specified.

[0036] Unless otherwise specified, all embodiments and optional embodiments of this application can be combined to form new technical solutions. Unless otherwise specified, all technical features and optional technical features of this application can be combined to form new technical solutions.

[0037] Unless otherwise specified, all steps of this application may be performed sequentially or randomly, but sequentially is preferred.

[0038] The high oil content of basil seeds easily forms an oil film on the surface, hindering the penetration of extraction solvents and resulting in low polyphenol extraction rates. Furthermore, traditional thermal extraction methods readily oxidize and degrade heat-sensitive polyphenols, weakening their antioxidant activity.

[0039] Based on the above problems, this application first uses petroleum ether to defatt the basil seed raw material to eliminate oil interference, and then uses a non-thermal extraction method of ultrasonic extraction with ethanol aqueous solution. The volume ratio of ethanol in the ethanol aqueous solution, the material-liquid ratio of ultrasonic extraction, and the ultrasonic extraction time are all controlled within appropriate ranges to achieve efficient extraction of total polyphenols from basil seeds while preserving their high antioxidant properties to the greatest extent.

[0040] One or more embodiments of this application provide a method for extracting total polyphenols from basil seeds, comprising the following steps: defatting basil seed raw material with a defatting solvent to prepare defatted raw material; ultrasonically extracting the defatted raw material with an ethanol aqueous solution to obtain total polyphenols from basil seeds; wherein the defatting solvent includes petroleum ether; the volume percentage of ethanol in the ethanol aqueous solution is 40%-80%; the material-to-liquid ratio of the defatted raw material to the ethanol aqueous solution is 1:(20-70) in g / mL; and the ultrasonic extraction time is 20 min-70 min.

[0041] As an example, the volume percentage of ethanol in the aqueous ethanol solution can be, but is not limited to, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, or any range between two of the above volume percentages. This is beneficial for improving the extraction efficiency of total polyphenols from basil seeds and for maximizing the preservation of the high antioxidant properties of total polyphenols from basil seeds.

[0042] The volume percentage of ethanol in the ethanol-water solution is 40%-80%, expressed in g / mL. The material-to-liquid ratio of the defatted raw material to the ethanol-water solution can be, but is not limited to, 1:20, 1:25, 1:30, 1:35, 1:40, 1:45, 1:50, 1:55, 1:60, 1:65, 1:70, or any two of the above ratios. Using an ethanol-water solution for ultrasonic extraction within the above-mentioned ratios is beneficial for improving the extraction efficiency of total polyphenols from basil seeds and for maximizing the preservation of the high antioxidant properties of these polyphenols.

[0043] The ultrasonic extraction time is 20-70 minutes; for example, it can be, but is not limited to, 20, 30, 40, 50, 60, or 70 minutes, or any range between two of the above times. This is beneficial for improving the extraction efficiency of total polyphenols from basil seeds and for maximizing the preservation of the high antioxidant properties of the total polyphenols from basil seeds. Understandably, the extraction method for total polyphenols from basil seeds in this application first uses petroleum ether to defatt the basil seed raw material to eliminate oil interference, and then performs ultrasonic extraction on the basil seed raw material. This method can efficiently extract total polyphenols while maximizing their high antioxidant properties, laying the foundation for the pharmaceutical application of basil seeds.

[0044] In some embodiments, the petroleum ether includes petroleum ether with a boiling range of 60°C-90°C. Therefore, petroleum ether with a boiling range within this range has a strong ability to dissolve fat-soluble impurities, efficiently removing oils, waxes, and other components from basil seeds, reducing interference from subsequent separation of target active ingredients such as polyphenols and flavonoids. Furthermore, the moderate boiling point range of petroleum ether, below the thermal decomposition temperature of plant polyphenols and flavonoids, prevents damage to the structure and activity of target components during the degreasing process. Simultaneously, petroleum ether has good volatility; after degreasing, solvent residues can be completely removed by low-temperature rotary evaporation or natural evaporation, avoiding interference with subsequent component analysis and activity detection.

[0045] As one possible implementation method, the ratio of basil seed raw material to degreasing solvent, expressed in g / mL, is 1:(10-30); for example, it can be, but is not limited to, 1:10, 1:12, 1:14, 1:16, 1:18, 1:20, 1:22, 1:24, 1:26, 1:28, 1:30, or any range between two of the above ratios. When the ratio of basil seed raw material to degreasing solvent is within the above range, it avoids incomplete degreasing due to insufficient solvent usage, reduces waste caused by excessive solvent and subsequent energy consumption for recovery, and lowers experimental costs.

[0046] In some embodiments, the degreasing process employs one or more of the following methods: soaking degreasing, ultrasonic degreasing, Soxhlet extraction degreasing, and vibration degreasing.

[0047] In some embodiments, the soaking and degreasing temperature is 22°C-25°C; for example, it can be, but is not limited to, 22°C, 23°C, 24°C, 25°C or any two of the above temperatures.

[0048] As one possible implementation, the soaking and degreasing time is 12h-16h; for example, it can be, but is not limited to, 12h, 13h, 14h, 15h, 16h or any range between the above two times.

[0049] It should be noted that the temperature and time for soaking and degreasing can be combined in any suitable way, and both can be selected from any soaking and degreasing temperature and time described in this article.

[0050] In some embodiments, the ultrasonic power for ultrasonic degreasing is 200W-400W; for example, it can be, but is not limited to, 200W, 220W, 240W, 260W, 280W, 300W, 320W, 340W, 360W, 380W, 400W, or any range between two of the above ultrasonic powers.

[0051] As one possible implementation method, the ultrasonic frequency for ultrasonic degreasing is 30KHz-50KHz; for example, it can be, but is not limited to, 30KHz, 32KHz, 34KHz, 36KHz, 38KHz, 40KHz, 42KHz, 44KHz, 46KHz, 48KHz, 50KHz, or any range between two of the above ultrasonic frequencies.

[0052] In some embodiments, the ultrasonic degreasing time is 10 min to 30 min; for example, it can be, but is not limited to, 10 min, 12 min, 14 min, 16 min, 18 min, 20 min, 22 min, 24 min, 26 min, 28 min, 30 min, or any range between two of the above times.

[0053] It should be noted that the ultrasonic power, ultrasonic frequency, and time of ultrasonic degreasing can be combined in any suitable way, and the three can be selected from any ultrasonic power, ultrasonic frequency, and time described in this article.

[0054] In some implementations, the Soxhlet extraction defatting time is 3-5 hours; for example, it can be, but is not limited to, 3 hours, 4 hours, 5 hours or any range between the two times mentioned above.

[0055] As one possible implementation, the water bath temperature used for Soxhlet extraction defatting is 60℃-90℃; for example, it can be, but is not limited to, 60℃, 65℃, 70℃, 75℃, 80℃, 85℃, 90℃ or any range between two of the above temperatures.

[0056] In some embodiments, the oscillation frequency of the oscillation degreasing is 120 rpm to 160 rpm; for example, it can be, but is not limited to, 120 rpm, 130 rpm, 140 rpm, 150 rpm, 160 rpm or any range between two of the above oscillation frequencies.

[0057] As one possible implementation, the degreasing time is 12h-16h; for example, it can be, but is not limited to, 12h, 13h, 14h, 15h, 16h or any range between the above two times.

[0058] It should be noted that the oscillation frequency and time of the oscillation degreasing can be combined in any suitable way, and both can be selected from any oscillation frequency and time of the oscillation degreasing described in this article.

[0059] In some implementations, the basil seed raw material is in powder form. This is beneficial for further improving the degreasing effect.

[0060] In some optional embodiments, the volume percentage of ethanol in the ethanol-water solution is 40%-80%, and the ratio of the defatting raw material to the ethanol-water solution is 1:(30-70) in g / mL.

[0061] In some implementations, the ultrasonic frequency extracted by ultrasound is 30kHz-50kHz; for example, it can be, but is not limited to, 30KHz, 32KHz, 34KHz, 36KHz, 38KHz, 40KHz, 42KHz, 44KHz, 46KHz, 48KHz, 50KHz or any range between two of the above ultrasonic frequencies.

[0062] As one possible implementation, the ultrasonic power for ultrasonic extraction is 200W-400W; for example, it can be, but is not limited to, 200W, 220W, 240W, 260W, 280W, 300W, 320W, 340W, 360W, 380W, 400W, or any range between two of the above ultrasonic powers.

[0063] One or more embodiments of this application provide a total polyphenol extract of basil seeds, which is obtained by the above-described extraction method for total polyphenols of basil seeds.

[0064] The technical solutions of this application will be described in detail below with reference to specific embodiments. It should be understood that these embodiments are only for illustrating this application and are not intended to limit the scope of this application. For experimental methods in the following embodiments where specific conditions are not specified, please refer to the guidelines given in this application first, or follow experimental manuals or conventional conditions in the field, or follow the conditions recommended by the manufacturer, or refer to experimental methods known in the field.

[0065] In the specific embodiments described below, the measurement parameters involving raw material components may have slight deviations within the weighing accuracy range unless otherwise specified. Temperature and time parameters are subject to acceptable deviations due to instrument testing accuracy or operational precision.

[0066] It should be noted that the method used to determine the total polyphenol content of the extracts obtained in the following examples and comparative examples is as follows:

[0067] 1. Preparation of the reference solution: Accurately weigh 0.01020 mg of gallic acid and place it in a 10 mL brown volumetric flask. Add distilled water to the volumetric flask and shake thoroughly to mix completely. After confirming that the gallic acid has completely dissolved, make up to the final volume to obtain a gallic acid solution with a concentration of 0.933 mg / mL.

[0068] 2. Preparation of 17.5% sodium carbonate solution: Accurately weigh 8.750g of anhydrous sodium carbonate powder and place it in a 50mL volumetric flask. Accurately add distilled water and dilute to the 50mL mark. Shake well until completely dissolved to obtain a 17.5% sodium carbonate solution (prepare immediately before use).

[0069] 3. Colorimetric and Determination Method: Accurately pipette 1000 μL of the test solution into a 10 mL colorimetric tube. Using a pipette, sequentially add 1.0 mL of distilled water, 0.5 mL of 50% Folin-Ciocalteu reagent, and 1.5 mL of 17.5% sodium carbonate solution to the reaction vessel. After the reagents mix in the vessel, a chemical reaction will occur. Continue adding distilled water until the liquid level reaches the calibration mark of the volumetric flask, completing the dilution. To ensure a more complete reaction, place the vessel in a light-protected environment at room temperature and allow it to stand for 90 minutes. After the reaction is complete, pipette an appropriate amount of the standard solution or reaction solution into a clean cuvette, ensuring the solution volume meets the cuvette requirements and is free of air bubbles. Measure the absorbance value at a wavelength of 760 nm.

[0070] 4. Standard Curve Determination: Accurately pipette 0, 0.1, 0.2, 0.4, 0.6, 0.8, 1.0, and 1.2 mL of the reference solution from Section 1 into 10 mL amber volumetric flasks. Add distilled water to the flasks and shake thoroughly to mix completely. Accurately pipette 1000 μL of each reference solution into 10 mL light-protected colorimetric tubes. Then, perform color development according to the method described in Section 3 and measure the absorbance values. After the measurements are completed, plot the absorbance data on the ordinate and the gallic acid reference solution concentration (mg / mL) on the abscissa to construct a standard curve.

[0071] 5. Determination of total polyphenol content in basil seed medicinal material: Take 1000 μL of basil seed test solution into a 10 mL light-protected colorimetric tube, determine the absorbance according to item 3, substitute it into the standard curve to obtain the polyphenol concentration, and then calculate the total polyphenol yield of different batches of basil seed medicinal material:

[0072] Total polyphenol content (mg / g) = C × V × N / (M × 1000)

[0073] In the formula: C is the total polyphenol concentration (μg / mL); V is the sampling volume of the extract (mL); N is the dilution factor of the basil extract; and M is the weight of the basil seed material (g).

[0074] Example 1

[0075] Step S1, Degreasing Treatment: Accurately weigh 3.0 g of basil seed powder and place it in a stoppered conical flask. Add analytical grade petroleum ether with a boiling range of 60℃-90℃ at a material-to-liquid ratio of 1:20 (g / mL). After sealing, perform ultrasonic degreasing for 20 minutes. Use a vacuum filtration device to separate the solid and liquid, and collect the filter residue. Place the degreased basil seed filter residue in a vacuum drying oven and dry it at a low temperature below 40℃ to remove residual petroleum ether. Set aside for later use.

[0076] Step S2, Ultrasonic Extraction: The basil residue was ultrasonically extracted with an ethanol-water solution containing 50% ethanol by volume. The material-to-liquid ratio was 1:15 (g / mL), and the ultrasonic time was 1.5 h. The extracts were combined after two extractions and placed in the corresponding extraction device for extraction. The extracted extracts were then refrigerated for later use.

[0077] Example 2

[0078] The extraction methods in Example 2 and Example 1 are similar, except that in step S1 of Example 2, Soxhlet extraction for 4 hours is used instead of ultrasonic degreasing for 20 minutes; all other steps are the same.

[0079] Example 3

[0080] The extraction methods in Example 3 and Example 1 are similar, except that in step S1 of Example 3, 16 hours of oscillating degreasing is used instead of 20 minutes of ultrasonic degreasing; all other steps are the same.

[0081] Example 4

[0082] The extraction methods in Example 4 and Example 1 are similar, except that in step S1 of Example 4, soaking and defatting for 16 hours is used instead of ultrasonic defatting for 20 minutes; all other steps are the same.

[0083] Comparative Example 1

[0084] The extraction methods of Comparative Example 1 and Example 1 are similar, except that: in Comparative Example 1, the same amount of basil seed slices were used instead of basil seed powder, and the basil seed slices were not degreased in step S1, but were directly extracted by ultrasound in step S2; everything else is the same.

[0085] Comparative Example 2

[0086] The extraction methods of Comparative Example 2 and Example 1 are similar, except that: in Comparative Example 2, the basil powder was not degreased in step S1, but was directly extracted by ultrasound according to step S2; all other aspects are the same.

[0087] Comparative Example 3

[0088] The extraction methods of Comparative Example 3 and Comparative Example 1 are similar, except that reflux extraction is used instead of ultrasonic extraction in Comparative Example 3. All other methods are the same.

[0089] Comparative Example 4

[0090] The extraction methods of Comparative Example 4 and Comparative Example 2 are similar, except that reflux extraction is used instead of ultrasonic extraction in Comparative Example 4, while all other methods are the same.

[0091] Comparative Example 5

[0092] The extraction methods of Comparative Example 5 and Example 1 are similar, except that in step S1 of Comparative Example 5, reflux extraction is used instead of ultrasonic extraction, while all other steps are the same.

[0093] Comparative Example 6

[0094] The extraction methods of Comparative Example 6 and Example 2 are similar, except that in step S1 of Comparative Example 6, reflux extraction is used instead of ultrasonic extraction, while all other steps are the same.

[0095] Comparative Example 7

[0096] The extraction methods of Comparative Example 7 and Example 3 are similar, except that in step S1 of Comparative Example 7, reflux extraction is used instead of ultrasonic extraction, while the rest are the same.

[0097] Comparative Example 8

[0098] The extraction methods of Comparative Example 8 and Example 4 are similar, except that in step S1 of Comparative Example 8, reflux extraction is used instead of ultrasonic extraction, while all other steps are the same.

[0099] The total polyphenol content of the extracts obtained in the above examples and comparative examples was determined using the above-described method, and the results are shown in Table 1.

[0100] Table 1

[0101]

[0102] As shown in Table 1, defatting the basil seed raw material before ultrasonic extraction can effectively improve the extraction efficiency of total polyphenols.

[0103] The comparison between Comparative Example 1 and Comparative Example 2 shows that, compared with extraction using basil seed slices, extraction using basil seed powder can effectively improve the extraction efficiency of total polyphenols.

[0104] The results of Examples 1-4 and Comparative Examples 5-8, and the results of Comparative Examples 1-2 and Comparative Examples 3-4 show that ultrasonic extraction can effectively improve the extraction efficiency of total polyphenols compared with reflux extraction.

[0105] Example 5

[0106] Step S1, Degreasing: Accurately weigh 2.5g of basil seed powder and place it in a stoppered conical flask. Add analytical grade petroleum ether (boiling range 60℃-90℃) at a material-to-liquid ratio of 1:20 (g / mL). Seal the flask and ultrasonically degrease for 20 minutes. Use a vacuum filtration device to separate the solid and liquid phases and collect the filter residue. Place the degreased basil seed filter residue in a vacuum drying oven and dry it at a low temperature below 40℃ to remove residual petroleum ether. Set aside for later use.

[0107] Step S2, Ultrasonic Extraction: Weigh 2.0g of defatted and dried basil residue tightly, and use an ethanol aqueous solution with a volume ratio of 40% to extract the basil residue by ultrasound. The material-to-liquid ratio is 1:40 (g / mL), and the ultrasound time is 60min. After extraction, place the extract in the corresponding extraction device and refrigerate for later use.

[0108] Example 6

[0109] The extraction methods in Example 6 and Example 5 are similar, except that in step S2 of Example 6, the volume percentage of ethanol in the ethanol-water solution is 50% during ultrasonic extraction; all other aspects are the same.

[0110] Example 7

[0111] The extraction methods in Example 7 and Example 5 are similar, except that in Example 7, the volume percentage of ethanol in the aqueous ethanol solution is 60% during ultrasonic extraction in step S2; all other aspects are the same.

[0112] Example 8

[0113] The extraction methods in Example 8 and Example 5 are similar, except that in Example 8, the volume percentage of ethanol in the aqueous ethanol solution is 70% during ultrasonic extraction in step S2; all other aspects are the same.

[0114] Example 9

[0115] The extraction methods of Example 9 and Example 5 are similar, except that in Example 9, the volume ratio of ethanol in the ethanol aqueous solution is 80% during ultrasonic extraction in step S2; all other aspects are the same.

[0116] Comparative Example 9

[0117] The extraction methods of Comparative Example 9 and Example 5 are similar, except that the volume percentage of ethanol in the ethanol aqueous solution is 30% during ultrasonic extraction in step S2 of Comparative Example 9; all other aspects are the same.

[0118] The total polyphenol content of the extracts from Examples 5-9 and Comparative Example 9 was determined, and the results are shown in Table 2.

[0119] Table 2

[0120]

[0121] As can be seen from the comparison of the results of Examples 5-9 and Comparative Example 9 in Table 2, when ultrasonic extraction is performed, the volume percentage of ethanol in the ethanol aqueous solution is 40%-80%, which is beneficial to further improve the extraction efficiency of total polyphenols.

[0122] Example 10

[0123] The extraction methods in Example 10 and Example 7 are similar, except that the ultrasonic extraction time in step S2 of Example 10 is 20 min; all other steps are the same.

[0124] Example 11

[0125] The extraction methods in Example 11 and Example 7 are similar, except that the ultrasonic extraction time in step S2 of Example 11 is 30 min; all other steps are the same.

[0126] Example 12

[0127] The extraction methods in Example 12 and Example 7 are similar, except that the ultrasonic extraction time in step S2 of Example 12 is 40 min; all other steps are the same.

[0128] Example 13

[0129] The extraction methods in Example 13 and Example 7 are similar, except that the ultrasonic extraction time in step S2 of Example 13 is 50 min; all other steps are the same.

[0130] Example 14

[0131] The extraction methods in Example 14 and Example 7 are similar, except that the ultrasonic extraction time in step S2 of Example 14 is 70 min; all other steps are the same.

[0132] Example 15

[0133] The extraction methods in Example 15 and Example 7 are similar, except that the ultrasonic extraction time in step S2 of Example 15 is 80 min; all other steps are the same.

[0134] Example 16

[0135] The extraction methods in Example 16 and Example 7 are similar, except that the ultrasonic extraction time in step S2 of Example 16 is 90 min; all other steps are the same.

[0136] Table 3

[0137]

[0138] The results of Examples 7, 10-14 and 15-16 show that when the ultrasonic extraction time exceeds 70 min, the total polyphenol content in the extract does not increase significantly. As the ultrasonic extraction time is extended, the temperature of the extract increases, which may damage the stability of the total polyphenols in the extract. Therefore, the ultrasonic extraction time is set to 20 min-70 min.

[0139] Example 17

[0140] The extraction methods of Example 17 and Example 13 are similar, except that the material-to-liquid ratio in step S2 of Example 17 is 1:20; all other aspects are the same.

[0141] Example 18

[0142] The extraction methods of Example 18 and Example 13 are similar, except that the material-to-liquid ratio in step S2 of Example 18 is 1:30; all other aspects are the same.

[0143] Example 19

[0144] The extraction methods of Example 19 and Example 13 are similar, except that the material-to-liquid ratio in step S2 of Example 19 is 1:50; all other aspects are the same.

[0145] Example 20

[0146] The extraction methods of Example 20 and Example 13 are similar, except that the material-to-liquid ratio in step S2 of Example 20 is 1:60; all other aspects are the same.

[0147] Example 21

[0148] The extraction methods of Example 21 and Example 13 are similar, except that the material-to-liquid ratio of ultrasonic extraction in step S2 of Example 21 is 1:70; all other aspects are the same.

[0149] Comparative Example 10

[0150] The extraction methods of Comparative Example 10 and Example 13 are similar, except that the material-to-liquid ratio of ultrasonic extraction in step S2 of Comparative Example 10 is 1:10; all other aspects are the same.

[0151] Table 4

[0152]

[0153] The results of Examples 13, 17-21 and Comparative Example 10 show that when using ethanol-water solution for ultrasonic extraction, a material-to-liquid ratio of 1:(20-70) (g / mL) is beneficial to further improve the extraction efficiency of total polyphenols.

[0154] The DPPH radical scavenging ability and ABTS radical scavenging ability of the extracts from Examples 5-21 and Comparative Examples 9-10 were determined using the following methods:

[0155] DPPH free radical scavenging capacity determination: Accurately weigh 0.0039 g of DPPH and transfer it to a 100 mL volumetric flask. Dissolve it in anhydrous ethanol solution, dilute to volume, and prepare a 0.1 mmol / L DPPH ethanol solution. Store in the dark. Add 1.5 mL of the 0.1 mmol / L DPPH ethanol solution and 1.5 mL of different sample solutions to a test tube, mix, shake, and allow to react for 30 min at room temperature and in the dark. Measure the absorbance (A1) at 517 nm. Using a blank reagent as a control, the DPPH free radical scavenging rate is calculated using the following formula:

[0156] Clearance rate = (A0 - A1) / A0 × 100%;

[0157] In the formula: A0 is the absorbance of the DPPH ethanol solution measured at a wavelength of 517 nm.

[0158] ABTS free radical scavenging capacity determination: Weigh ABTS reagent and prepare a 7.4 mmol / L ABTS stock solution with ultrapure water. Prepare a 2.6 mmol / L potassium persulfate solution with ultrapure water. Mix the ABTS stock solution and potassium persulfate solution in a 1:1 ratio and let stand at room temperature in the dark for 16 h to generate ABTS free radical cations. Before use, dilute with PBS to an absorbance of 0.70 ± 0.02 at 734 nm, which is the ABTS working solution. Take 0.2 mL of sample solution, add 3.8 mL of ABTS working solution, shake well, and react at room temperature in the dark for 6 min. A blank group (0.2 mL anhydrous ethanol + 3.8 mL ABTS working solution) and a control group (0.2 mL sample dilution + 3.9 mL anhydrous ethanol) were also set up. The absorbance values ​​of each group were measured at 734 nm using a UV-Vis spectrophotometer.

[0159] Using a blank reagent as a control, the formula for calculating the ABTS free radical scavenging rate is as follows:

[0160] Clearance rate = (A0 - A1) / A0 × 100%;

[0161] In the formula: A0 is the absorbance measured at 734 nm wavelength for the blank group, and A1 is the absorbance measured at 734 nm wavelength for the sample solution group.

[0162] The results of the DPPH free radical scavenging capacity determination of Examples 5-21 and Comparative Examples 9-10 are shown in Table 5.

[0163] Table 5

[0164]

[0165] The results of the ABTS radical scavenging capacity determination of Examples 5-21 and Comparative Examples 9-10 are shown in Table 6.

[0166] Table 6

[0167]

[0168] As shown in Tables 5 and 6, the total polyphenols extracted from basil seeds using the extraction method of this application exhibit high DPPH and ABTS radical scavenging rates, indicating high antioxidant properties. This suggests that the extraction method of this application causes minimal damage to the antioxidant activity of the total polyphenols.

[0169] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0170] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the protection scope of the present invention. Therefore, the protection scope of this invention patent should be determined by the appended claims.

Claims

1. A method for extracting total polyphenols from basil seeds, characterized in that, Includes the following steps: Basil seeds were degreased using a degreasing solvent to prepare a degreased raw material. The defatted raw material was subjected to ultrasonic extraction using an ethanol-water solution to obtain the total polyphenols from basil seeds. The defatting solvent includes petroleum ether; the volume percentage of ethanol in the aqueous ethanol solution is 40%-80%; the ratio of the defatting raw material to the aqueous ethanol solution is 1:(20-70) in g / mL; and the ultrasonic extraction time is 20-70 min.

2. The method for extracting total polyphenols from basil seeds as described in claim 1, characterized in that, The petroleum ethers include petroleum ethers with a boiling range of 60℃-90℃.

3. The method for extracting total polyphenols from basil seeds as described in claim 1, characterized in that, The ratio of the basil seed raw material to the defatting solvent is 1:(10-30) in g / mL.

4. The method for extracting total polyphenols from basil seeds as described in claim 1, characterized in that, The degreasing process can be carried out by one or more of the following methods: soaking degreasing, ultrasonic degreasing, Soxhlet extraction degreasing, and vibration degreasing.

5. The method for extracting total polyphenols from basil seeds as described in claim 4, characterized in that, The soaking temperature for degreasing is 22℃-25℃, and the time is 12h-16h; and / or, Ultrasonic degreasing uses an ultrasonic power of 200W-400W, an ultrasonic frequency of 30KHz-50KHz, and a duration of 10min-30min; and / or, Soxhlet extraction defatting time is 3-5 hours, using a water bath temperature of 60℃-90℃; and / or, The oscillation frequency for degreasing is 120 rpm-160 rpm, and the time is 12 h-16 h.

6. The method for extracting total polyphenols from basil seeds as described in claim 1, characterized in that, The basil seed raw material is in powder form.

7. The method for extracting total polyphenols from basil seeds as described in claim 1, characterized in that, The ratio of the defatted raw material to the ethanol aqueous solution is 1:(30-70), expressed in g / mL.

8. The method for extracting total polyphenols from basil seeds according to any one of claims 1 to 7, characterized in that, The ultrasonic extraction frequency is 30kHz-50kHz.

9. The method for extracting total polyphenols from basil seeds according to any one of claims 1 to 7, characterized in that, The ultrasonic power for ultrasonic extraction is 200W-400W.

10. A total polyphenol extract of basil seeds, characterized in that, It was obtained by the extraction method of total polyphenols from basil seeds as described in any one of claims 1 to 9.

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