CN106701322B

CN106701322B - Extraction method of fermented noni seed essential oil

Info

Publication number: CN106701322B
Application number: CN201710063422.7A
Authority: CN
Inventors: 张玉苍; 徐树英
Original assignee: Hainan University
Current assignee: Hainan University
Priority date: 2017-02-03
Filing date: 2017-02-03
Publication date: 2021-09-10
Anticipated expiration: 2037-02-03
Also published as: CN106701322A

Abstract

The invention discloses an extraction method of fermented noni seed essential oil, which comprises the following steps: (1) crushing, drying, crushing and sieving the fermented noni seeds; (2) mixing and soaking the pretreated fermented noni seed raw material with an ethanol solution, performing suction filtration to obtain a supernatant, and centrifuging by a high-speed centrifuge to obtain the supernatant without impurities. (3) Removing solvent from supernatant by rotary evaporator, and observing no dripping in condensing tube of rotary evaporator to obtain extract; (3) standing the extract at 16 deg.C for 24h to obtain layered oily substance, namely fermented noni seed essential oil. The method adopts the waste in the ferment production process of the noni fruits to ferment the noni fruit seeds to extract the essential oil, has rich raw material sources, belongs to resource recycling, adopts ethanol for normal-temperature extraction, retains the volatile components to the maximum extent, has high extraction rate and little solvent residue, is safe, and can be used as a skin care product additive.

Description

Extraction method of fermented noni seed essential oil

Technical Field

The invention belongs to the technical field of plant extract preparation methods, and particularly relates to an extract from fermented noni seeds

A method for preparing essential oil.

Background

Noni is Morinda of Rubiaceae, Noni (Noni) is a perennial evergreen broad-leaved shrub or small tree growing in tropical regions, 3-10 m high, with a large number of oval leaves (5-17 cm long and 10-40 cm wide). The method is characterized by planting in parts of Hainan province and Taiwan province in China. Its root, bark, stem, leaf and fruit have been used as traditional folk medicine for treating various diseases, such as diabetes, hypertension and cancer. Fresh noni fruit contains 80% moisture, the major dry matter being soluble solids, dietary fiber and protein. In mature fruit, the determined volatile compounds are organic acids, alcohols, esters, ketones and lactones, and the noni fruit also contains active substances such as carotene, vitamin C, amino acids and Xeronine. The noni fruits belong to evergreen tropical fruit trees, and mature fruits can be harvested in one year. The pulp of the corolla bluish white noni (3-10 cm long and 3-6 cm wide in oval shape) has a fleshy and thick appearance and has a relief sense. Generally, the picked fruit is white. The surface of the noni fruit is covered with seeds containing reddish-brown buds. Mature fruits develop a strong stream of butyric acid putrefactive odor. After the fruit ripens, it is egg-shaped, juicy, slightly dark yellow or white, and gelatinous, with many reddish-brown hard triangular pits containing four seeds in each pit. Initially the unripe fruit was green, turned yellow after ripening, softened to translucent upon over ripening and given off an unpleasant odor of butyric acid.

In recent years, noni fruits become a health food industry at home and abroad and have the effects of protecting blood vessels, resisting oxidation, protecting liver, resisting allergy, bacteria, viruses, mold, cancers, enhancing immunity and the like. In 2004, Chinese scholars discovered Chinese noni germplasm resources in the Xisha Yongxing island. In 2005, the Hainan Nonide Industrial park development company started the artificial cultivation of Nonide and the development of a series of products, and at present, a 3000-mu-acre Saxanonie demonstration planting base was built in Hainan, and the self-developed Saxanonie series products including the Saxanonie enzyme, the Saxanonie tea, the Saxanonie soap and the like have been fully introduced into the market. The noni fruits generate a large amount of waste in the process of preparing fermented beverages and unfermented beverages, at present, most of noni products are supplied to the market by fermented liquid of noni fruits, but the fermented fruit residues and seeds of the noni fruits are not fully utilized. At present, few researches on extracting essential oil from fermented noni seeds are carried out, and no relevant patent and literature reports exist.

Disclosure of Invention

The technical scheme adopted for solving the technical problems is as follows: provides a method for extracting fermented noni seed essential oil

A preparation method. The specific technical scheme is as follows:

(1) pretreatment: drying the fermented noni seeds, crushing, sieving and reserving for use;

(2) soaking: weighing pretreated fermented noni seed powder, mixing the powder with a 95% ethanol solution at a solid-to-liquid ratio of 1: 8-12, and soaking at normal temperature for 4-10 h;

(3) and (3) suction filtration: filtering the soaking solution through a Buchner funnel to obtain a supernatant;

(4) high-speed centrifugation: centrifuging the supernatant by using a high-speed centrifuge to obtain supernatant without impurities;

(5) rotary evaporation: evaporating the solvent from the supernatant by rotary evaporator to obtain extract;

(6) high-speed centrifugation: centrifuging the extract with the solvent removed again at a high speed, wherein the rotating speed is 5000-10000 r/min, and the time is 3-10 min to obtain an oily substance;

(7) and (3) refrigerating: and (3) placing the obtained oily substance into a refrigerator for standing and refrigerating at the temperature of 0-8 ℃ for 1-10 hours to obtain a layer of fermented noni seed essential oil, wherein the upper layer is light brown, and the lower layer is dark brown.

In the technical scheme, the fermentation method for fermenting the noni seeds comprises the following steps: picking white noni fresh fruits, cleaning, putting into a fermentation tank, controlling the anaerobic fermentation temperature to be 20-40 ℃, and fermenting for 100-150 d; the PH value is 6.8-7.2; the fermentation can be completed by utilizing endophyte of noni fruit and the strain CICC10883 acetobacter cacao, and solid matters in the fermentation product are taken for standby. Among them, the identification and biological properties of the bacterium CICC10883 used have been studied in the literature: identification and biological characteristics of 1 strain of Simonoi fruit puree bacterium CICC10883, Kao Cao Yan Hua, No. 8 of 2016 (report on Chinese food Co., Ltd.).

In the technical scheme, the drying temperature is 50-60 ℃, and the sieving is a 20-100-mesh sieve.

In the technical scheme, the centrifugation in the step (4) is to centrifuge the supernatant for 3-10 min at 5000-10000 r/min.

In the above technical solution, the rotary evaporation is completed when no droplet drips in the condensation tube of the rotary evaporator.

The method is simple, the fermented noni seed essential oil has extremely strong inoxidizability, the processing process is safe and environment-friendly, and the fermented noni seed essential oil can be used as a cosmetic additive.

Drawings

FIG. 1: a technical route schematic diagram;

FIG. 2: an oily substance after ethanol extraction; wherein: a is an extract obtained by extracting unfermented noni seeds with 95% ethanol; b is an extract obtained by extracting fermented noni seeds with 95% ethanol; as can be seen from FIG. 2, the extraction rate of the essential oil of the fermented noni seeds after 95% ethanol extraction is higher than that of the unfermented noni seeds.

FIG. 3: and (3) extracting a total ion flow diagram of the fermented noni seed essential oil by using ethanol. The effective components obtained by ethanol extraction and fermentation are few, and most of the organic matters have large molecular weight and long retention time. Shows that the internal chemical components and the content of the noni subjected to fermentation treatment are greatly changed. The esters in the fermented noni seed essential oil are caprylic esters; eugenol is found in the fermented noni seed essential oil, the content of the eugenol in the fermented essential oil is 0.89%, and researches show that the eugenol has the oxidation resistance.

FIG. 4: and (3) carrying out DPPH free radical and ethanol extraction on the effective components at the lower layer of the fermented noni seed essential oil. In FIG. 4A, it can be analyzed that DPPH has 3 pairs of characteristic peaks at 3475-3675. The signal intensity was around 140000. This characteristic peak is consistent with literature reports. The noni effective component has no obvious characteristic peak (the peak at the magnetic field intensity of 3600 is an air peak). Paramagnetic detection is performed on the DPPH solution at the lower layer of the fermented noni seed essential oil fermentation extracted by the organic solvent with the volume fraction of 4 per mill, and the result is shown in fig. 4B. By analyzing the paramagnetic wave spectrum, 3 pairs of characteristic peaks with 140000 signal intensity at 3475-3675 are found to disappear, so that the DPPH free radical can be concluded to be completely eliminated. The noni active ingredient with the volume fraction of 4 per mill has extremely high oxidation resistance.

Detailed Description

The following non-limiting examples are presented to enable one of ordinary skill in the art to more fully understand the present invention and are not intended to limit the scope thereof

The invention is limited in any way. Within the technical scope of the present disclosure, any person skilled in the art can easily understand the technical scope of the present invention

The technical scheme and the inventive concept of the invention are equivalent to replace or change the technical scheme and the inventive concept of the invention.

Example 1

1. The fermentation method of the noni seeds comprises the following steps: picking white noni fresh fruits, cleaning, putting into a fermentation tank, controlling the anaerobic fermentation temperature at 25 ℃, and fermenting for 150 d; the PH value is 6.8-7.2; the fermentation can be completed by utilizing endophyte of noni fruit and the strain CICC10883 acetobacter cacao, and solid matters in the fermentation product are taken for standby.

2. Drying the fermented noni seeds at 60 ℃, crushing and sieving by a 80-mesh sieve, weighing pretreated fermented noni seed powder, and preparing a 95% ethanol solution by using absolute ethanol and ultrapure water. According to the ratio of the product to the solvent of 1: 10 adding 200mL of 95% ethanol solution into 20g of fermented (unfermented) nutlets, and soaking for 6 h; obtaining supernatant through suction filtration, and obtaining supernatant without impurities by taking the supernatant through a high-speed centrifuge (8000r/min, time 5 min). Evaporating the solvent of the supernatant through a rotary evaporator, and observing that evaporation is finished when no liquid drops drop in a condensation pipe of the rotary evaporator; centrifuging the solvent-removed extract at high speed at 8000r/min for 5 min; and (3) placing the obtained oily substance into a refrigerator for standing and refrigerating at the temperature of 5 ℃ for 2 hours to obtain the fermented noni seed essential oil, wherein the essential oil comprises an upper layer and a lower layer of the oily substance, the upper layer is light brown, the lower layer is dark brown, and the average content of the fermented noni extract is 6.1 mL.

And (3) preparing and detecting the fermented noni seed extract extracted by the organic solvent according to the method to obtain a total ion flow diagram of the sample. See figure 3 and table 1 for the total ion flow. The ethanol extraction fermentation of the noni fruits detected 18 substances in total as the active ingredients, 15 of which were identified, accounted for 83.33% of the total compounds, and the identified compound content accounted for 98.93% of the total extract content. Mainly esters and acids, the esters accounted for 72.09%, the esters mainly accounted for ethyl palmitate (40.50%), di (. alpha. -ethylhexyl) phthalate (18.71%), the acids accounted for 6.97%, the organic acids mainly accounted for octanoic acid (6.63%). Among the active ingredients, squalene (12.95%), a benzene derivative, squalene (1.84%) was found in the identified compound, which has hepatoprotective effect, can promote hepatocyte regeneration and protect hepatocytes, thereby improving liver function, and has antitumor activity, etc. Vitamin E (5.92%). Sterol substance is 1%, which is campesterol (0.45%) stigmasterol (0.55%).

The effective components obtained by 95% ethanol extraction contain organic matters with larger molecular weight, such as isopropyl linoleate, squalene, ethyl palmitate and the like which are obtained in ethanol extraction fermentation and non-fermentation noni. Therefore, according to GCMS data analysis, the main chemical composition obtained by steam distillation is volatile organic ester substances with small molecular weight, and the main chemical composition obtained by 95% ethanol extraction in the method is organic substances with large molecular weight, wherein squalene has biological activities such as anti-tumor and the like.

Example 2

The fermentation method of the noni seeds comprises the following steps: picking white noni fresh fruits, cleaning, putting into a fermentation tank, controlling the anaerobic fermentation temperature at 25 ℃, and fermenting for 150 d; the PH value is 6.8-7.2; the fermentation can be completed by utilizing endophyte of noni fruit and the strain CICC10883 acetobacter cacao, and solid matters in the fermentation product are taken for standby.

2, drying the fermented noni seeds at 50 ℃, crushing and sieving by a 90-mesh sieve, weighing pretreated fermented noni seed powder, and preparing a 95% ethanol solution by using absolute ethanol and ultrapure water. According to the ratio of the product to the solvent of 1: 9 adding 200mL of 95% ethanol solution into 20g of fermented (unfermented) nutlet, and soaking for 8 h; obtaining supernatant through suction filtration, and obtaining supernatant without impurities by taking the supernatant through a high-speed centrifuge (9000r/min, time 10 min). Evaporating the solvent of the supernatant through a rotary evaporator, and observing that evaporation is finished when no liquid drops drop in a condensation pipe of the rotary evaporator; centrifuging the solvent-removed extract at high speed at 8500r/min for 10 min; and (3) placing the obtained oily substance into a refrigerator for standing and refrigerating at 4 ℃ for 3 hours to obtain the fermented noni seed essential oil, wherein the essential oil comprises an upper layer and a lower layer of the oily substance, the upper layer is light brown, the lower layer is dark brown, and the average content of the fermented noni extract is 6.7 mL.

Example 3

1. The fermentation method of the noni seeds comprises the following steps: picking white noni fresh fruits, cleaning, putting into a fermentation tank, controlling the anaerobic fermentation temperature to 25 ℃, and fermenting for 150 d; the PH value is 6.8-7.2; the fermentation can be completed by utilizing endophyte of noni fruit and the strain CICC10883 acetobacter cacao, and solid matters in the fermentation product are taken for standby.

2. Drying the fermented noni seeds at 55 ℃, crushing and sieving by a 100-mesh sieve, weighing pretreated fermented noni seed powder, and preparing a 95% ethanol solution by using absolute ethanol and ultrapure water. According to the ratio of the product to the solvent of 1: 11 adding 95% ethanol solution into 2 fermented (unfermented) nutlets, and soaking for 4 h; obtaining supernatant through suction filtration, and obtaining supernatant without impurities by taking the supernatant through a high-speed centrifuge (10000r/min, time is 8 min). Evaporating the solvent of the supernatant through a rotary evaporator, and observing that evaporation is finished when no liquid drops drop in a condensation pipe of the rotary evaporator; centrifuging the solvent-removed extract at high speed at 9000r/min for 10 min; and (3) placing the obtained oily substance into a refrigerator for standing and refrigerating at 6 ℃ for 4 hours to obtain the fermented noni seed essential oil, wherein the essential oil comprises an upper layer and a lower layer of the oily substance, the upper layer is light brown, the lower layer is dark brown, and the average content of the fermented noni extract is 5.6 mL.

TABLE 1 effective ingredients and relative contents of the components obtained by ethanol extraction of fermented noni seeds

Example 4

2. Drying the fermented noni seeds at 50 ℃, crushing and sieving by a 60-mesh sieve, weighing pretreated fermented noni seed powder, and preparing a 95% ethanol solution by using absolute ethanol and ultrapure water. According to the ratio of the product to the solvent of 1: 10 adding 95% ethanol solution into the fermented noni seeds, and soaking for 7 hours; and (3) primarily obtaining supernatant through suction filtration, and taking the supernatant to pass through a high-speed centrifuge (8500r/min, time 5min) to obtain supernatant without impurities. Evaporating the solvent of the supernatant through a rotary evaporator, and observing that evaporation is finished when no liquid drops drop in a condensation pipe of the rotary evaporator; centrifuging the solvent-removed extract at high speed at 8000r/min for 8 min; and (3) placing the obtained oily substance into a refrigerator for standing and refrigerating at the temperature of 5 ℃ for 2 hours to obtain the fermented noni seed essential oil, wherein the fermented noni seed essential oil is divided into an upper layer oily substance and a lower layer oily substance, the upper layer oily substance is lighter in color, the lower layer oily substance is darker in color, and the average content of the fermented noni extract is 6.1 mL.

The electron paramagnetic oxidation resistance experiment is carried out on the upper layer essential oil and the lower layer essential oil of the fermented noni seeds, the fact that the ability of removing DPPH free radicals is strong is found, the fact that the fermented noni seed essential oil extracted by 95% ethanol has strong oxidation resistance activity is shown, and meanwhile the oxidation resistance activity of the lower layer essential oil is higher than that of the upper layer essential oil.

Claims

1. The extraction method of the fermented noni seed essential oil is characterized by comprising the following steps: comprises the following steps of the following operation steps,

(1) pretreatment: drying the fermented noni seeds, crushing and sieving;

(2) soaking: taking the pretreated fermented noni seed powder, mixing the powder with a 95% ethanol solution at a solid-to-liquid ratio of 1: 8-12, and soaking at normal temperature for 4-10 hours;

(3) after suction filtration, centrifuging the supernatant and taking the supernatant;

(4) rotary evaporation: evaporating the solvent from the supernatant by rotary evaporator to obtain extract;

(5) high-speed centrifugation: centrifuging the extract at a speed of 5000-10000 r/min for 3-10 min to obtain oily substance;

(6) and (3) refrigerating: putting the obtained oily substance into a temperature of 0-8 ℃, and refrigerating for 1-8 h to obtain the product; obtaining the fermented noni seed essential oil, layering the essential oil, and obtaining an upper oily substance and a lower oily substance, wherein the upper oily substance is light brown, and the lower oily substance is dark brown;

the fermentation method for fermenting the noni seeds comprises the following steps: picking white noni fresh fruits, cleaning, putting into a fermentation tank, controlling the anaerobic fermentation temperature to be 20-40 ℃, and fermenting for 100-150 d; the PH value is 6.8-7.2; fermenting by using endophyte of noni fruit and strain CICC10883 acetobacter cacao, and taking solid matters in a fermentation product for later use;

the drying temperature is 50-60 ℃; the sieving is a 20-100 mesh sieve.

2. The method of extracting fermented noni seed essential oil according to claim 1, wherein: and the suction filtration is to perform suction filtration on the soak solution through a Buchner funnel to obtain a supernatant.

3. The method of extracting fermented noni seed essential oil according to claim 1, wherein: and (3) centrifuging the supernatant in 5000-10000 r/min for 3-10 min.

4. The method of extracting fermented noni seed essential oil according to claim 1, wherein: and (4) finishing the evaporation when no liquid drops drop in a condensation pipe of the rotary evaporator is observed in the rotary evaporation in the step (4).