CN110590543A - A method for extracting hydroxycitrate from garcinia cambogia fruit - Google Patents

Abstract

The present invention relates to the field of plant extraction, in particular to a method for extracting hydroxycitrate from Garcinia cambogia fruit, comprising the following steps: S1. Pulverizing the Garcinia cambogia fruit; S2. In the Garcinia cambogia fruit pulverized in S1, according to The ratio of material to liquid is 1:13~15, add water, boil at 110~120°C for 10~20min, filter and retain the filtrate after cooling; S3. Concentrate the filtrate in step S2 by 4~6 times, add 1 to 2 times of 90% to 95% ethanol; S4. step S3 is filtered, and 90% to 95% ethanol is added to the filtrate to adjust the alcohol content to 58 to 62; S5. to add alkaline solution in step S4, Adjust the pH of the solution to 11.5-12.5; S6. Filter, wash, evaporate, and dry step S5. This method keeps hydroxycitric acid in an alkaline atmosphere instead of directly forming a neutral salt with an alkaline substance. Since the carboxyl group cannot form a carboxyl salt, alkaline conditions are not conducive to the esterification reaction, effectively reducing the formation of lactones, and improving the purity and biological activity of the hydroxycitrate.

Description

A method for extracting hydroxycitrate from garcinia cambogia fruit

technical field

The invention relates to the field of plant extraction, in particular to a method for extracting hydroxycitrate from garcinia cambogia fruit.

Background technique

Garcinia cambogia ( Garcinia cambogia L.) is a kind of Garcinia cambogia tree distributed in south subtropical regions. The peel of its fruit is rich in hydroxycitric acid, and the fruit has a sour taste. It is often used as food seasoning and traditional medicine in South Asian countries. Hydroxycitric acid is a strong competitive inhibitor of ATP citrate lyase, which can inhibit fat synthesis and promote glycogenogenesis, increase the utilization rate of pyruvic acid and lactic acid, and suppress appetite. Therefore, hydroxycitric acid is often used as an active ingredient in weight loss functional foods. Recent studies have shown that hydroxycitric acid can also inhibit the formation of calcium oxalate crystals in the kidney.

Hydroxycitric acid is an organic acid containing 2 hydroxyl groups and 3 free carboxyl groups. It is easy to condense into lactone during the separation and extraction process, which reduces the biological activity of the product. In industrial preparation, there are three main methods for extracting hydroxycitric acid from Garcinia cambogia: water extraction, organic solvent extraction and ion exchange chromatography extraction. Garcinia cambogia extracts in domestic and foreign markets are mainly prepared by the above-mentioned methods, and the main component of the product is hydroxycitric acid lactone instead of free hydroxycitric acid.

Contents of the invention

The purpose of the present invention is to overcome the problem that lactone is easily formed in the extracted hydroxycitrate in the prior art, and provides a method for the hydroxycitrate of Garcinia cambogia fruit.

The purpose of the present invention is achieved through the following technical solutions:

A method for extracting hydroxycitrate from garcinia cambogia fruit, comprising the following steps:

S1. pulverizing Garcinia cambogia fruit;

S2. In the garcinia cambogia fruit crushed in S1, add water according to the solid-liquid ratio of 1:13~15, and boil at 110~120°C for 10~20min, filter and retain the filtrate after cooling;

S3. Concentrating the filtrate in step S2 by 4 to 6 times, adding 90% to 95% ethanol of 1 to 2 times the volume of the concentrated solution;

S4. Step S3 is filtered, and 90% to 95% ethanol is added to the filtrate to adjust the alcohol content to 58 to 62;

S5. Add alkali solution in step S4, adjust the pH of the solution to 11.5 ~ 12.5;

S6. Filtering, washing, evaporating and drying step S5.

The above method for extracting citrate from Garcinia cambogia fruit is to extract hydroxycitric acid from Garcinia cambogia fruit, and the final product is in the form of salt due to the alkali solution added during the extraction process. Adding water in step S2 according to the ratio of solid to liquid is 1:13~15 means adding 13~15mL of water per gram of Garcinia cambogia fruit according to the mass volume ratio. The boiling temperature in step S2 is 110-120° C., which is realized by means of pressurization according to different temperature requirements. Above-mentioned by adding 90%～95% ethanol to regulate the alcohol degree of solution, be to measure with 40～70 degree alcohol meter. In addition, in step S6, the final oily precipitate was formed. After the supernatant was discarded, the precipitate was washed three times with 95% ethanol, and then washed twice with absolute ethanol, then placed at room temperature overnight, and then placed at 60°C under reduced pressure. Evaporate under pressure and dry.

Preferably, in the step S1, the Garcinia cambogia fruit is crushed into 30-50 mesh particles.

Preferably, the ratio of solid to liquid in the step S2 is 1:14.

Preferably, in the step S2, boiling at 115° C. for 15 minutes is adopted.

Preferably, the concentration factor of the filtrate in step S3 is 5 times.

Preferably, the alcohol content in step S4 is 60.

Preferably, the pH of the solution is adjusted to 12 in the step S5.

Preferably, the alkali solution in the step S5 is KOH or NaOH solution. KOH solution is preferably used, and the mass percentage of KOH is 30%~50%.

Compared with the prior art, the present invention has the following technical effects:

The method for extracting hydroxycitrate from garcinia cambogia fruit provided by the present invention adopts alkaline solution, especially KOH solution, to make the extract to be extracted under suitable alkaline conditions. Under this alkaline condition, the carboxyl group in hydroxycitric acid is in the Neutral salts are formed directly under an alkaline atmosphere rather than directly with alkaline substances. Since the carboxyl group cannot form a carboxyl salt, it is not conducive to the progress of the esterification reaction, effectively reduces the formation of lactones, and improves the purity and biological activity of the hydroxycitrate.

Description of drawings

The high-performance liquid chromatogram of Fig. 1 hydroxycitric acid standard;

The high performance liquid chromatogram of Fig. 2 citric acid;

The high performance liquid chromatogram of the product that Fig. 3 embodiment 1 makes;

The high performance liquid chromatogram of the Garcinia cambogia extract reference substance of Fig. 4 NowFoods company;

Figure 5 is a color reaction diagram of ammonium metavanadate; and

Figure 6 Thin layer chromatogram.

Explanation of reference signs:

1-hydroxycitric acid standard substance, 2-citric acid, 3-the product prepared in Example 1, 4-NowFoods garcinia cambogia extract reference substance.

Detailed ways

In order to make the purpose, technical solution and advantages of the present invention clearer, the technical solution of the present invention will be described in detail below in conjunction with specific examples and comparative examples. Apparently, the described embodiments are only some of the embodiments of the present invention, but not all of them. Based on the embodiments of the present invention, all other implementations obtained by persons of ordinary skill in the art without making creative efforts fall within the protection scope of the present invention.

Unless otherwise specified, the equipment used in the examples, comparative examples and experimental examples are conventional experimental equipment, and the materials and reagents used are all commercially available.

Example 1

S1. Take Garcinia cambogia fruit powder that is crushed to 40 meshes;

S2. In step S1, add water according to the ratio of material-to-liquid ratio of 1:14, then cook under high pressure at 115°C for 15 minutes, after cooling, filter and squeeze out the filtrate with 5 layers of medical gauze, and centrifuge the filtrate at 4000×g for 15 minutes;

S3. Concentrate the supernatant by evaporation under reduced pressure at 55°C, the concentration factor is 5 times, and then add 95% ethanol to precipitate pectin in the concentrated solution at a ratio of 1:1 (v/v);

S4. Filter the mixture obtained in step S3 with gauze and centrifuge at 4000×g for 15 min to remove pectin. Add 95% ethanol to the supernatant and adjust it at room temperature with a 40-70 degree alcohol meter (Hebei Wuqiang Huapeng Instrument Factory) The alcohol content of the solution is up to 60;

S5. Then add 40% KOH to adjust the pH to 12;

S6. Discard the supernatant after the oily precipitate formed in step S5, wash the precipitate with 95% ethanol three times, and then wash the precipitate with absolute ethanol twice, then place it at room temperature overnight, then evaporate it under reduced pressure at 60°C, and dry it.

Example 2

S1. Take Garcinia cambogia fruit powder that is crushed to 30 meshes;

S2. In step S1, add water according to the ratio of material-to-liquid ratio of 1:15, then cook under high pressure at 110°C for 20 min, after cooling, filter and extrude the filtrate with 5 layers of medical gauze, and centrifuge the filtrate at 4000×g for 15 min;

S3. Concentrate the supernatant by evaporation under reduced pressure at 55°C, the concentration factor is 6 times, and then add 90% ethanol to precipitate pectin in the concentrated solution at 1:2 (v/v);

S4. Filter the mixture obtained in step S3 with gauze and centrifuge at 4000×g for 15 min to remove pectin. Add 95% ethanol to the supernatant and adjust it at room temperature with a 40-70 degree alcohol meter (Hebei Wuqiang Huapeng Instrument Factory) The alcohol content of the solution is up to 58;

S5. Then add 40% KOH to adjust the pH to 11.5;

S6. Discard the supernatant after the oily precipitate formed in step S5, wash the precipitate with 95% ethanol three times, and then wash the precipitate with absolute ethanol twice, then place it at room temperature overnight, then evaporate it under reduced pressure at 60°C, and dry it.

Example 3

S1. Take Garcinia cambogia fruit powder that is crushed to 50 mesh;

S2. In step S1, add water according to the ratio of material-to-liquid ratio of 1:13, then cook under high pressure at 120°C for 10 minutes, after cooling, filter and squeeze out the filtrate with 5 layers of medical gauze, and centrifuge the filtrate at 4000×g for 15 minutes;

S3. Concentrate the supernatant by evaporation under reduced pressure at 55°C, the concentration factor is 4 times, and then add 90% ethanol to precipitate pectin in the concentrated solution at 1:1 (v/v);

S4. Filter the mixture obtained in step S3 with gauze and centrifuge at 4000×g for 15 min to remove pectin. Add 95% ethanol to the supernatant and adjust it at room temperature with a 40-70 degree alcohol meter (Hebei Wuqiang Huapeng Instrument Factory) The alcohol content of the solution is up to 62;

S5. Then add 40% KOH to adjust the pH to 12.5;

S6. Discard the supernatant after the oily precipitate formed in step S5, wash the precipitate with 95% ethanol three times, and then wash the precipitate with absolute ethanol twice, then place it at room temperature overnight, then evaporate it under reduced pressure at 60°C, and dry it.

Comparative example 1~3

Main steps are identical with embodiment 1. The difference is that the pHs of the solutions in Step S5 of Comparative Example 1 to Comparative Example 3 are 9, 11, and 13 in sequence.

Comparative example 4~6

The main steps are the same as in Example 1. The difference is that the ratio of solid to liquid in step S2 of Comparative Example 4 to Comparative Example 6 is 1:6, 1:10, 1:18 in sequence.

Comparative example 7~9

Main steps are identical with embodiment 1. The difference is that the concentration ratios of the solutions in Step S3 of Comparative Example 7 to Comparative Example 9 are 1, 3, and 7 in sequence.

Comparative example 10~12

Main steps are identical with embodiment 1. The difference is that the alcoholic degrees of the solutions in Step S4 of Comparative Example 10 to Comparative Example 12 are 40, 50, and 70 in sequence.

Example 1

The potassium citrate in Garcinia cambogia fruit was extracted according to the methods of each embodiment and each comparative example, and the yields were calculated respectively. The results are shown in Table 1.

Yield = (mass of potassium hydroxycitrate obtained by extraction/mass of garcinia cambogia fruit) × 100%

Table 1 Yield table

It can be seen from the above table that the ratio of solid to liquid, concentration ratio, alcohol content and pH all have a great influence on the yield of hydroxycitrate. Only when the solid-liquid ratio is 1:13~15, the concentration ratio is 4~6 times, the alcohol content is 58~62, and the pH is 11.5~12.5, the hydroxycitrate can have a good yield.

Experimental example 2

Reference literature (Hui ZF, Xiao H, Hui YH, et al. Determination of hydroxycitric acid in Garcinia cambogia by RP-HPLC [J]. Northwest Pharmaceutical J, 2015,30: 371-372. ) using LC-10Avp plus (Shimadzu ) high-performance liquid chromatography, the chromatographic column is Wondasil C18 (250 mm×4.6 mm, 5 μm), the mobile phase is 0.05 mol/L anhydrous sodium sulfate, the pH is adjusted to 2.3 with sulfuric acid, and the flow rate is 1 mL/min. 50 mg of the sample to be tested was dissolved in 40 mL of 3.0% phosphoric acid solution, ultrasonicated for 30 min, and filtered with a 0.022 μm filter membrane, with a sample volume of 20 μL, and detected at 210 nm by an ultraviolet detector. The above method was used to test the hydroxycitric acid standard (Figure 1), the product prepared in Example 1 of citric acid (Figure 2) (Figure 3), and the NowFoods Garcinia cambogia extract control substance (Figure 4).

It can be seen from the figure that the absorption peak appears at the retention time of 2.90min in Figure 3, which is the same as that of the hydroxycitric acid standard (2.96min) in Figure 1 and the NowFoods Garcinia Cambogia Extract reference substance (2.88min) in Figure 4 the peak position. And the citric acid in Fig. 2 has no absorption peak at 2.90min. Illustrate that the product that embodiment 1 makes is hydroxycitrate. In addition, there is a stronger absorption peak at 5.63min in the Garcinia cambogia extract reference substance of NowFoods Company in Figure 4, which corresponds to the formation of lactone hydroxycitric acid, while Figure 3 has no absorption peak at 5.63min , indicating that the hydroxycitrate obtained by the method of the present invention exists as a pure hydroxycitrate in which no lactone is formed.

Experimental example 3

Refer to literature for color reaction (Bainto LC, Dizon EI, Laurena AC, et al. Isolation and quantification of hydroxycitric acid from batuan [Garcinia binucao (Blanco)Choisy] fruit [J]. Int Food Res J, 2018, 25: 706-711 )Methods. Weigh 0.1 g of the sample to be tested and dissolve it in 50 mL of 3.0% phosphoric acid solution. After ultrasonic treatment for 30 min, take 1 mL of the supernatant, add 3 mL of 1% ammonium metavanate, and observe after standing for 30 min.

Ammonium metavanitate was used for color reaction (as shown in Figure 5), and it was found that hydroxycitric acid standard 1, product 3 of Example 1 and Garcinia cambogia extract 4 of NowFoods Company were all reddish brown, while citric acid 2 was yellow .

Experimental example 4

Take 0.1 g of the sample to be tested and dissolve it in 0.9 mL of 3.0% phosphoric acid solution. After ultrasonic treatment for 30 min, filter it with a 0.022 μm filter membrane, and spot the sample on an activated GF254 thin-layer chromatography silica gel plate (2.5×7.5 cm). Use methanol: water = 6:1 (v/v) as the developing solvent. After the chromatography, 5% ammonium metavanitate solution was used for color development. The Rf value is calculated according to the following formula:

Rf = the distance from the origin to the center of the component point ÷ the distance from the origin to the solvent front

As shown in Figure 6, the Rf values of hydroxycitric acid standard 1, citric acid 2, product 3 prepared in Example 1 and NowFoods Garcinia cambogia extract 4 have no significant difference, all between 0.69-0.71, but citric acid The Rf value is slightly lower. It further shows that the extract is hydroxycitrate.

Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention rather than to limit the scope of protection of the present invention. For those of ordinary skill in the art, on the basis of the above description and ideas, they can also make There is no need to and cannot exhaustively list all the implementation manners for other changes or changes in different forms. All modifications, equivalent replacements and improvements made within the spirit and principles of the present invention shall be included within the protection scope of the claims of the present invention.

Claims (8)

1. A method for extracting hydroxycitrate from lucerne fruits is characterized by comprising the following steps:

s1, crushing garcinia cambogia fruits;

s2, adding water into the garcinia cambogia fruits crushed in the step S1 according to the material-liquid ratio of 1:13 ~ 15, boiling for 10 ~ 20min at the temperature of 110 ~ 120 ℃ and 120 ℃, cooling, and filtering to keep filtrate;

s3, concentrating the filtrate obtained in the step S2 by 4-4 ~ 6 times, and adding 90- ~ 95% of ethanol with the volume 1-1 ~ 2 times that of the concentrated solution;

s4, filtering the filtrate obtained in the step S3, and adding 90% to ~ 95% of ethanol into the filtrate to adjust the alcohol degree to 58 ~ 62;

s5, adding an alkali solution in the step S4, and adjusting the pH value of the solution to 11.5 ~ 12.5.5;

s6, filtering, washing, evaporating and drying are carried out on the step S5.

2. The method for extracting hydroxycitrate from Garcinia cambogia fruit as claimed in claim 1, wherein the Garcinia cambogia fruit is crushed into 30 ~ 50 mesh particles in step S1.

3. The method for extracting hydroxycitrate from Garcinia cambogia fruit of claim 1, wherein the ratio of liquid to feed in step S2 is 1: 14.

4. The method for extracting hydroxycitrate from Garcinia cambogia fruit of claim 1, wherein the step S2 is carried out by boiling at 115 ℃ for 15 min.

5. The method for extracting hydroxycitrate from Garcinia cambogia fruit of claim 1, wherein the filtrate is concentrated 5 times in step S3.

6. The method for extracting hydroxycitrate from Garcinia cambogia fruit of claim 1, wherein the alcohol content in step S4 is 60.

7. The method for extracting hydroxycitrate from Garcinia cambogia fruit of claim 1, wherein the pH of the solution is adjusted to 12 in step S5.

8. The method for extracting hydroxycitrate from Garcinia cambogia fruit of claim 1, wherein the alkaline solution in step S5 is KOH or NaOH solution.