A method of ball milling extraction of anthocyanins from aronia melanocarpa fruit or pomace

Abstract

This invention belongs to the field of compound extraction technology, specifically relating to a ball milling extraction method for anthocyanins from the fruit or pomace of *Sorbus nigra* var. *nigra*. The extraction method is as follows: Washed *Sorbus nigra* fruit (or pomace) is dried and pulverized into coarse powder; the coarse powder and extraction solvent are added to a centrifuge tube, and grinding beads are added simultaneously for crushing and extraction; after extraction, the mixture is filtered and centrifuged, and the supernatant is collected; the supernatant is rotary evaporated to a certain volume and then freeze-dried to obtain *Sorbus nigra* anthocyanin extract; the extract sample is dissolved, filtered through a 0.22 μm filter membrane, and analyzed and quantified using ultra-high performance liquid chromatography-mass spectrometry (UHPLC). This process is simple to operate, environmentally friendly, and achieves a high anthocyanin extraction rate.

Description

Technical Field

[0001] This invention belongs to the field of compound extraction technology, specifically relating to a ball milling extraction method for anthocyanins from the fruit or pomace of *Sorbus nigra* var. *nigra*. Background Technology

[0002] Black-fruited glandular-ribbed shoga (Aronia prunifolia Viking. and Aronia melanocarpa) is a perennial deciduous shrub belonging to the genus Aronia in the family Rosaceae. Native to northeastern North America, it is now widely distributed in Europe and in Liaoning, Hebei, Beijing, Shandong, and Shanxi provinces of China. Its fruits and pomace are rich in anthocyanins, which possess powerful antioxidant properties and also exhibit various health benefits, including anti-cancer effects, cardiovascular disease prevention, anti-hyperglycemia effects, anti-radiation effects, and immune regulation. These components are widely used in the food, pharmaceutical, and cosmetic industries.

[0003] Currently, commonly used anthocyanin extraction methods include flash extraction, ultrasonic extraction, and microwave extraction. However, with rising energy prices and increased focus on reducing carbon dioxide emissions, there is a growing demand for new technologies that use less energy and require less time to extract natural products. Ball milling extraction utilizes the kinetic energy provided by the high-speed rotation of grinding media in a container to break down sample cells, accelerating the release of anthocyanins and thus increasing the extraction rate. Compared to traditional extraction methods, it offers significant advantages in terms of shorter extraction time, higher yield, cost savings, environmental friendliness, and preservation of the structural integrity of the extracted material. Summary of the Invention

[0004] The purpose of this invention is to overcome the shortcomings of existing extraction technologies and provide a ball milling extraction process for anthocyanins from the fruit or pomace of *Sorbus nigra*.

[0005] The technical solution adopted in this invention is:

[0006] A method for ball milling extraction of anthocyanins from the fruit or pomace of *Sorbus nigra* var. *nigra*, comprising the following steps:

[0007] 1) Drying: Wash the black chokeberry fruit or pomace and dry it until the weight remains unchanged;

[0008] 2) Grinding: Grind the dried black chokeberry fruit or pomace into coarse powder for later use;

[0009] 3) Extraction: Place the coarse powder in the extraction solvent, add grinding beads, and pulverize and extract using a ball mill; filter the extracted sample, centrifuge, take the supernatant and rotary evaporate to a thick paste, freeze dry to obtain black chokeberry anthocyanin extract.

[0010] Furthermore, in the ball milling extraction method for anthocyanins from the fruit or pomace of *Sorbus nigra* described above, the drying temperature in step 1) is 50°C.

[0011] Furthermore, in the ball milling extraction method for anthocyanins from the fruit or pomace of *Sorbus nigra* described above, step 3) uses a 50% (v / v) ethanol-water solution as the extraction solvent.

[0012] Furthermore, in the ball milling extraction method for anthocyanins from the fruit or pomace of *Sorbus nigra* described above, in step 3), the ratio of the coarse powder to the extraction solvent is 1 g:10 mL to 1 g:60 mL.

[0013] Furthermore, in the above-mentioned method for ball milling extraction of anthocyanins from the fruit or pomace of *Sorbus nigra*, in step 3), the vibration frequency of the ball mill is 5–60 Hz.

[0014] Furthermore, in the above-mentioned method for ball milling extraction of anthocyanins from the fruit or pomace of *Sorbus nigra*, step 3) involves a ball milling and extraction time of 1–30 min.

[0015] Furthermore, in the ball milling extraction method for anthocyanins from the fruit or pomace of *Sorbus nigra* described above, in step 3), the particle size combination of the grinding beads is 3mm+3mm, 3mm+4mm, 4mm+4mm, 3mm+3mm+4mm, or 3mm+4mm+4mm.

[0016] Preferably, in step 3), the ball mill pulverization and extraction time is 22.0 min, the ball mill vibration frequency is 44.0 Hz, and the material-to-liquid ratio of coarse powder to extraction solvent is 1 g:15 mL.

[0017] The beneficial effects of this invention are: compared with existing anthocyanin extraction methods, the method of this invention is simple in process, easy to operate, energy-saving, environmentally friendly and has high extraction efficiency. Attached Figure Description

[0018] Figure 1 This is a response surface optimization diagram of three factors that significantly affect anthocyanin extraction. Detailed Implementation

[0019] Exemplary embodiments will now be described in detail, examples of which are illustrated in the accompanying drawings.

[0020] The embodiments described in the following exemplary embodiments do not represent all embodiments consistent with the disclosure of this invention. Rather, they are merely examples of methods consistent with some aspects of the invention disclosed as detailed in the appended claims. The following embodiments are intended to enable those skilled in the art to better understand the invention, but are not intended to limit the invention.

[0021] Example 1

[0022] Wash the fruit or pomace of *Sorbus nigra* and dry at 50℃ until the weight remains constant. Crush the dried fruit or pomace into coarse powder. Weigh a certain amount of the coarse powder and place it in a 50% (v / v) ethanol aqueous solution. Add grinding beads and extract using a ball mill. The ball milling time is 15 min, the vibration frequency is 20 Hz, and the liquid-to-material ratio is 30 mL / g. Grinding beads with diameters of 3mm+3mm, 3mm+4mm, 4mm+4mm, 3mm+3mm+4mm, and 3mm+4mm+4mm are used in the experiment. Filter the extracted sample, centrifuge, and rotary evaporate the supernatant to a thick paste. Freeze-dry to obtain the anthocyanin extract of *Sorbus nigra*. Dissolve the sample, filter through a 0.22 μm filter membrane, and analyze.

[0023] The extraction rates, calculated by ultra-high performance liquid chromatography-mass spectrometry (UHPLC), were 17.9, 18.4, 20.7, 21.3, and 21.8 mg / g, respectively.

[0024] Example 2

[0025] A certain amount of coarse powder from *Sorbus nigra* was weighed out. A ball mill with 3mm+4mm grinding beads, a vibration frequency of 20 Hz, and a liquid-to-material ratio of 30 mL / g was used for milling at times of 1, 5, 10, 15, 25, and 30 min. The extracted sample was filtered, centrifuged, and the supernatant was rotary evaporated to a thick paste. This paste was then freeze-dried to obtain *Sorbus nigra* anthocyanin extract. The sample was dissolved, filtered through a 0.22 μm filter membrane, and then analyzed.

[0026] The extraction rates, calculated by ultra-high performance liquid chromatography-mass spectrometry (UHPLC), were 18.5, 19.4, 19.7, 18.4, 20.6, and 18.2 mg / g, respectively.

[0027] Example 3

[0028] A certain amount of coarse powder from *Sorbus nigra* was weighed and ball-milled for 15 min using 3mm+4mm grinding beads at a liquid-to-solid ratio of 30 mL / g. The ball mill vibration frequencies were 5 Hz, 10 Hz, 20 Hz, 30 Hz, 40 Hz, and 60 Hz. The extracted sample was filtered, centrifuged, and the supernatant was rotary evaporated to a certain volume and freeze-dried to obtain *Sorbus nigra* anthocyanin extract. The sample was dissolved, filtered through a 0.22 μm filter membrane, and then analyzed.

[0029] The extraction rates, calculated by ultra-high performance liquid chromatography-mass spectrometry (UHPLC), were 18.2, 18.9, 18.4, 20.5, 20.7, and 19.8 mg / g, respectively.

[0030] Example 4

[0031] A certain amount of coarse powder from *Sorbus nigra* was weighed out. The grinding beads were selected as 3mm+4mm, the ball mill vibration frequency was 20 Hz, and the ball milling time was 15 min. The liquid-to-solid ratios were 10, 20, 30, 40, 50, and 60 mL / g. After extraction, the sample was filtered, centrifuged, and the supernatant was rotary evaporated to a certain volume and freeze-dried to obtain *Sorbus nigra* anthocyanin extract. The sample was dissolved, filtered through a 0.22 μm filter membrane, and then injected for analysis.

[0032] The extraction rates, calculated by ultra-high performance liquid chromatography-mass spectrometry (UHPLC), were 17.6, 19.6, 18.4, 19.4, 18.1, and 20.3 mg / g, respectively.

[0033] Example 5: Response Surface Optimization

[0034] Based on the results of the single-factor experiments, response surface methodology was used to investigate three factors that significantly affected anthocyanin extraction: ball milling time, ball mill vibration frequency, and liquid-to-solid ratio. The extraction process was optimized using response surface methodology with Design Expert 10 software.

[0035] Mathematical analysis based on the calculated regression model shows that the anthocyanin extraction rate = 21.78 - 0.21A - 0.07B + 0.26C + 0.19AB - 0.01AC + 0.21BC - 2.79A 2 -0.82B 2 -0.76C 2 The optimal extraction parameters for anthocyanins were: extraction time of 22.0 min, ball mill vibration frequency of 44.0 Hz, and liquid-to-solid ratio of 15.0 mL / g. Under these conditions, the theoretical yield was 20.3 mg / g. The optimization results are as follows: Figure 1 As shown.

Claims

1. A method for ball milling extraction of anthocyanins from the fruit or pomace of *Sorbus nigra*, characterized in that, Includes the following steps: 1) Drying: Wash the black chokeberry fruit or pomace and dry it until the weight remains unchanged; 2) Grinding: Grind the dried black chokeberry fruit or pomace into coarse powder for later use; 3) Extraction: Place the coarse powder in the extraction solvent, add grinding beads, and pulverize and extract using a ball mill; filter the extracted sample, centrifuge, take the supernatant and rotary evaporate to a thick paste, freeze dry to obtain black chokeberry anthocyanin extract.

2. The ball milling extraction method for anthocyanins from the fruit or pomace of *Sorbus nigra* as described in claim 1, characterized in that... In step 1), the drying temperature is 50°C.

3. The ball milling extraction method for anthocyanins from the fruit or pomace of *Sorbus nigra* as described in claim 1, characterized in that... In step 3), the extraction solvent is a 50% ethanol aqueous solution by volume.

4. The ball milling extraction method for anthocyanins from the fruit or pomace of *Sorbus nigra* as described in claim 1, characterized in that... In step 3), the ratio of the coarse powder to the extraction solvent is 1 g:10 mL to 1 g:60 mL.

5. The ball milling extraction method for anthocyanins from the fruit or pomace of *Sorbus nigra* as described in claim 1, characterized in that... In step 3), the vibration frequency of the ball mill is 5 to 60 Hz.

6. The ball milling extraction method for anthocyanins from the fruit or pomace of *Sorbus nigra* as described in claim 1, characterized in that, In step 3), the ball mill pulverization and extraction time is 1 to 30 minutes.

7. The ball milling extraction method for anthocyanins from the fruit or pomace of *Sorbus nigra* as described in claim 1, characterized in that... In step 3), the grinding bead particle size combination is 3mm+3mm, 3mm+4mm, 4mm+4mm, 3mm+3mm+4mm or 3mm+4mm+4mm.

8. The ball milling extraction method for anthocyanins from the fruit or pomace of *Sorbus nigra* as described in claim 1, characterized in that... In step 3), the ball mill pulverization and extraction time is 22.0 min, the ball mill vibration frequency is 44.0 Hz, and the material-to-liquid ratio of coarse powder to extraction solvent is 1 g:15 mL.

Images