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Supercritical CO2 extraction of maca (Lepidium meyenii)

Maca

Maca is a one-year or two-year-old herbaceous plant in the cruciferous family. It enjoys the reputation of “Peruvian Ginseng” and “Peru National Treasure”. It is native to the Andes Mountains above 3500 m above sea level in Peru. It grows normally in a unique environment with large temperature difference between day and night and long-term freezing.

For thousands of years, as one of the important food sources for local indigenous people, it has been regarded by the local people as a gift from the Andean gods due to its rich nutritional value and medicinal effects, especially its functions in enhancing energy and fertility. Precious gift.

Health effects

The health benefits of Maka are inseparable from the nutrients such as protein, amino acids, reducing sugars, and minerals (zinc, iron, calcium).

Supercritical CO2 extraction of maca
Supercritical CO2 extraction of maca

However, most researchers believe that macaamide, benzyl glucosinolates and their decomposition products benzyl isothiocyanate are their main active ingredients and characteristic substances.

Due to its rich nutritional value and medicinal effects, the research and development of maca is becoming more and more popular, but so far the research on maca is not thorough enough, and the related maca products circulating in the market are relatively primitive and of good quality. Jagged.

Research on functional factors

Therefore, to carry out research on the functional factors of Maka, in order to provide a certain scientific basis for the development of high-quality and high-end Maka health food, which has great application value and far-reaching social significance.

In this paper, we studied different extraction methods, including supercritical carbon dioxide extraction, reflux extraction, and steam distillation, to obtain maca fat-soluble parts. The yield of the extract, the types of active ingredients and their relative amounts were used as indicators for comparative analysis. The pros and cons of each extraction method lay a certain foundation for the further in-depth study of Maka and the development of functional factors.

Maca different methods of extraction

SFE-CO2 extraction method

Extraction process: CO2 gas cylinder → extraction kettle → extraction kettle I → extraction kettle II → extraction kettle III → CO2 circulately weigh 500 g of Maca powder (passed through a 40-mesh sieve), and load it into a 5 L extraction kettle. Ⅰ. Separating kettle II and separating kettle III are heated separately, and the chiller is started for refrigeration.

Among them, the SFE-CO2 extraction condition parameters are set as follows: the pressure of the extraction vessel is 32 MPa, the temperature is 45 ℃, the pressure of the separation vessel I is 8 MPa, the temperature is 45 ℃, the pressure of the separation vessel II and the separation vessel Ⅲ are both 6 MPa, and the temperature is both 40 ℃;

When the temperature of the extraction kettle, separation kettle and cooler storage tank all meet the experimental requirements, the CO2 from the CO2 gas cylinder is condensed by the cooler storage tank, and then pumped into the extraction kettle and three separation kettles through the high-pressure pump;

When the pressure of the extraction kettle and each separation kettle reaches the experimental setting value, add the entrainer according to the feeding amount (g): 95% ethanol entrainer volume (ml) of 1:2, and start the cycle timing, total circulation Extract for 2 h;

Discharge, weigh, and calculate the yield from Separation Reactor I, Separation Reactor II, and Separation Reactor III at intervals of 30 minutes; prepare samples for GC/MS analysis and wait for GC/MS analysis and testing;

Traditional extraction method

Petroleum ether reflux extraction method

Weigh 100.0 g of Maca powder (passed through a 40-mesh sieve), put it in a 1,000 ml round-bottomed flask, add 6 times the amount of petroleum ether (30-60 ℃), reflux and extract in a water bath at 39.9 ℃ for 4 h, then depressurize Suction filtration. After recovering the solvent from the filtrate under the conditions of 39.9 ℃ and -0.1 MPa, weigh, calculate the yield, and prepare samples for GC/MS analysis and testing; the extracted drug residues are evaporated to dry petroleum ether and placed separately. Further processing

80% ethanol reflux extraction method

Take the medicine residue extracted with petroleum ether (30-60°C), add 6 times the amount of 80% ethanol, reflux for 3 hours in a water bath at 50°C, and filter under reduced pressure. The filtrate is concentrated to nothing under the conditions of 39.9°C and -0.1 MPa. Alcohol, add 45 ml, 40 ml, 30 ml of ethyl acetate (AR grade) for extraction, combine the ethyl acetate layers, and concentrate to near dryness under the conditions of 39.9 ℃ and -0.1 MPa, weigh, calculate the yield, and Prepare samples for GC/MS analysis and testing;

Steam distillation

Steam distillation method-specific gravity less than 1 part

150.0 g of Maca powder (passed through a 100-mesh sieve), add 1050 ml (ie 7 times) of water, and extract according to the standard method of volatile oil extraction [4] for 7.5 hours;

Since the volatile oil extractor did not extract the oil, but a slightly creamy transparent liquid (like water, very strange smell), put it separately (about 180 ml), add ethyl acetate for extraction, 45 ml/ A total of 3 times, divide the ethyl acetate layer, concentrate to dryness under 39.9 ℃,-0.1 MPa conditions, weigh, calculate the yield, and prepare samples for GC/MS analysis and testing;

Steam distillation method-specific gravity greater than 1 part

Filter the decoction liquid under 2.2.3.1 while it is hot. After cooling, add ethyl acetate to the filtrate for extraction, 45 ml/time, 3 times in total. Separate the ethyl acetate layer and heat it at 39.9 ℃, -0.1 Concentrate to dryness under MPa conditions, weigh, calculate the yield, and prepare samples for GC/MS analysis and testing;

Experimental results and discussion

Comparison of samples obtained by different extraction methods

The sample yield obtained by steam distillation is the highest, followed by supercritical carbon dioxide extraction.
But in terms of extraction time, the supercritical carbon dioxide method only takes 2 hours, which is obviously better than steam distillation and reflux method;

At the same time, from the analysis of rational use of medicinal materials, the supercritical CO2 extraction method can obtain a large extract yield per unit time (0.94%), and the subsequent processing of the sample is simple, only the recovery of ethanol is required, and the petroleum ether is refluxed. The yield of the extract obtained by the method is only 0.40%, and the petroleum ether reflux method has the problem of environmental pollution and is not suitable for production scale-up. Therefore, the supercritical CO2 extraction method can obtain the fat-soluble parts of maca to a greater extent and reduce the amount of maca. The waste of resources in turn achieves the purpose of rational use of medicinal resources.

GC/MS analysis SFE-CO2 extraction method

Through GC/MS analysis, a total of 153 peaks were separated from the Marka fat-soluble sample obtained by the SFE-CO2 extraction method. Compared with the standard map database, a total of 40 compounds were identified.

Among them, amides include N-benzylformamide (2.38%), long-chain polyunsaturated olefins (alkynes or acids) include 1,E-11,Z-13-Octadecatriene (0.52%), Z,Z,Z -4,6,9-Nonadecatriene (0.25%), arachidonic acid (1.84%), glucosinolates and their degradation products include benzyl isocyanate (0.32%), 4-methoxybenzaldehyde (0.77 %), 3-methoxybenzeneacetonitrile (1.18%);

GC/MS analysis of petroleum ether reflux method

A total of 156 peaks were separated from the Marka fat-soluble sample obtained by the petroleum ether reflux method. Compared with the standard map database, a total of 31 compounds were identified, as shown in Table 4. Among them, amines include benzylamine (0.23%), Cyclohexaneacetamide, N-(phenylmethyl)- (51.91%), and long-chain polyunsaturated olefins (alkynes or acids) include 9,12,15-Octadecatrienal (0.90%) , Glucosinolates and their degradation products include benzyl isothiocyanate (1.57%) and 3-methoxybenzeneacetonitrile (1.10%);

GC/MS analysis 80% ethanol reflux method

A total of 132 peaks were separated from the Marka fat-soluble sample obtained by the 80% ethanol reflux method. Compared with the standard map database, a total of 14 compounds were identified, as shown in Table 5. Among them, long-chain polyunsaturated olefins (alkynes or acids) include A-linolenic acid (8.57%), and glucosinolates and their degradation products include benzyl isocyanate (0.45%) and benzyl thiocyanate (0.54 %);

GC/MS analysis of parts with specific gravity less than 1 obtained by steam distillation

A total of 157 peaks were separated from the sample with a specific gravity less than 1 obtained by steam distillation. Compared with the standard map database, a total of 21 compounds were identified, as shown in Table 6. Among them, glucosinolates and their degradation products include benzyl isonitrile (85.04%), benzyl isothiocyanate (2.64%), and 3-methoxybenzeneacetonitrile (5.38%);

GC/MS analysis of water vapor distillation method, the proportion is greater than 1 part

A total of 142 peaks were separated from the sample with a specific gravity greater than 1 by steam distillation. Compared with the standard map database, a total of 26 compounds were identified, as shown in Table 7. Among them, amides include benzamide (0.66%), N-benzyl formamide (4.38%), glucosinolates and their degradation products include benzyl isocyanate (0.49%), benzyl thiocyanate (1.25%) %), 4-hydroxyphenylacetonitrile (6.31%), 3-methoxybenzeneacetonitrile (10.63%).

In conclusion

In summary, the maca fat-soluble sample obtained by the supercritical CO2 extraction method not only has a high yield (0.94%), but also requires a short extraction time (2 h), and it is known by GC/MS analysis that the supercritical CO2 extraction method The obtained maca fat-soluble sample not only has more kinds of active ingredients, but also has a relatively high content.

Therefore, compared with the traditional extraction method, the supercritical CO2 extraction method is more in line with today’s green, low-carbon, environmentally friendly and sustainable development goals.