WO2023111648A1 - Method of separation of bioactive elements and compounds of saffron plant by temperature shock and high pressure - Google Patents

Abstract

Saffron has unique nutritional and medicinal properties due to the presence of effective compounds, namely crocin, picrocrocin and safranal. Therefore, the extraction and purification of these compounds is of great importance for use in the food and pharmaceutical industries. Methods of soaking, steam distillation, ultrasound, supercritical fluid, solid phase extraction and crystallization to extract saffron compounds have been investigated. Consumption of high volume of solvent, loss of volatile compounds, low efficiency, long extraction time, degradation of unsaturated compounds and retention of toxic solvent are the disadvantages of existing extraction methods. For this reason, a combination of several extraction methods is usually used to purify saffron compounds. The method is as follows: supercritical fluid (CO2) is used and by means of a cycle of cooling, short-term soaking and heating with the help of radio frequency waves and applying pulse pressure on the solution, saffron is extracted to high purity.

Description

Description

1- Title of invention: Method of separation of bioactive elements and compounds of saffron plant by temperature shock and high pressure

2- Technical and practical field: Chemical and biological

3- Background and Summary of Invention:

Project Summary: Saffron has unique nutritional and medicinal properties due to the presence of effective compounds, namely crocin, picrocrocin and safranal. Therefore, the extraction and purification of these compounds is of great importance for use in the food and pharmaceutical industries. Methods of soaking, steam distillation, ultrasound, supercritical fluid, solid phase extraction and crystallization to extract saffron compounds have been investigated. Consumption of high volume of solvent, loss of volatile compounds, low efficiency, long extraction time, degradation of unsaturated compounds and retention of toxic solvent are the disadvantages of existing extraction methods. For this reason, a combination of several extraction methods is usually used to purify saffron compounds. The method is as follows: supercritical fluid (CO2) is used and by means of a cycle of cooling, short-term soaking and heating with the help of radio frequency waves and applying pulse pressure on the solution, saffron is extracted to high purity.

Background:

• W02000057981 - PROCESS FOR OBTAINING AN EXTRACT OF SAFFRON:

In this patent, the stigma of the saffron plant is ground to a size of less than 4 mm. After that, a mixture (in a ratio of 1: 1) ionized water / 96% alcohol is provided in a ratio of 16 liters per 1 kg of ground stigma. Stir the mixture and heat for about two hours at a temperature of approximately sixty degrees Celsius. The mixture is filtered and an enriched solution is obtained, mixed again and mixed again with ionized water / 96% alcohol (1: 1) for about two hours at 60 ° C. The mixture is mixed again with the mentioned percentage of water and alcohol and mixed for about one hour at a temperature of sixty degrees Celsius.

Impurities obtained in the previous steps are filtered through 5 micron tubes. The solution is obtained under vacuum at a relatively low temperature, preferably between 35 and 40 ° C, in order to obtain a liquid extract with solids between 2 and 3. Percentages are concentrated. This liquid extract obtained in this way has a composition of 8 kg of extract per kg of saffron stigma consumed. Then, to obtain a solid extract, dehydration by lyophilization, freezing of the liquid extract is performed at -50 ° C, and then sublimation under vacuum at 0 ° C, to remove the existing water, then at the desired temperature between 35 and 38 Celsius is poured and heated to remove residual moisture. Rest the solid product for 48 hours. The resulting product is a solid extract, especially powdered extract, with a ratio of approximately 50% of usable ingredients. That is, from one kilogram of saffron stigma, approximately half a kilogram of solid or powdered extract is obtained. In this method, the purpose is to extract the solid and liquid solution of saffron for use in the food industry, where the process time is about 53 hours and the purity is 50%, and in which the soaking and crystallization method is used, while in our method The method of supercritical fluid, application of pulse pressure and heating with radio waves has been used and the duration of the process is a maximum of three hours and the purity of the solution obtained is up to 93%.

• IN2869/DEL/2013 - A NOVEL METHOD OF PREPARTION OF SAFFRON EXTRACT AND COMPOSITION THEREOF: In this patent, first the total amount of saffron is mixed with twice the distilled water in a ratio of 1: 2 and kept at 10 ° C for 24 hours. The resulting mixture is kept at 40 ° C for 48 hours. After 48 hours of re-storage, 8: 1 liter of propylene glycol is mixed with the previous mixture and the resulting mixture is kept at 20 ° C for 24 hours. Finally, the mixture is filtered with a vacuum pump and then the liquid saffron extract is obtained. As it is clear, only the soaking method is used and the process time is 96 hours (4 days), while in our method, the supercritical fluid method, applying pulse pressure and heating with radio waves are used, and The process takes a maximum of three hours and the purity of the solution is up to 93%.

• US20040126467 - Method for the extraction of saffron pigments and flavor concentrate:

In this patent, saffron is mixed with a food solvent or a combination of more than one solvent, and after soaking and centrifugation, it is cooled and extracted in such a way that, as mentioned above, saffron (10 g) with butanol (250 ml), water (100 ml) and methanol (50 ml) were mixed and soaked at 5 ° C to 5 ° C for 10 h and centrifuged. Liquid materials that were reduced under pressure at a temperature in the range of -30 ° C were lyophilized. The raw material (6.92 g) had a pigment concentration of 16.6%. If in our method, the method of supercritical fluid, pulse pressure and heating with radio waves is used and the duration of the process is a maximum of three hours and the purity of the solution obtained is up to 93%.

• EP1106223 - PROCESS FOR OBTAINING AN EXTRACT OF SAFFRON:

In this patent, saffron is first ground to a size of less than 4 mm. Then, we add a mixture (in a ratio of 1: 1) of ionized water / 96 degree alcohol at a ratio of 16 liters per kilogram of ground saffron stigma. Stir the mixture and heat for 2 hours. The enriched mixture is filtered, and stirred again with ionized water / alcohol mixture at 96 degrees (1: 1) for 2 hours. The filtrate is extracted and the concentrated solution of the previous step is extracted and the stigmas are mixed again with ionized water / alcohol at 96 ° (1: 1) for one hour. The previously obtained heterogeneous solution is filtered through 5 pm tubes and the impurities are removed. Finally, the concentrated extract obtained is concentrated in vacuum at a low temperature of about 40 ° C until the liquid extracts with the material Solid between 2 to 3% is obtained. The obtained product is solid extracts, especially powdered extracts, which is approximately 50% in proportion to the usable materials, ie 0.5 kg of solid or powdered extract is obtained from one kilogram of saffron stigma. Lyophilization, freezing of liquid extract at -50 ° C and then vacuum sublimation at 0 ° C, removal of available water, evacuation and heating at less than 38 ° C to remove moisture, solid product obtained for 48 Rest time is given. In which the method of soaking and centrifugation is used and the duration of the extraction process is about 5 hours. In our method, the method of supercritical fluid, pulse pressure and heating with radio waves is used and the duration of the process is a maximum of three hours and the purity of the solution obtained is up to 93%.

• W02004056201 - METHOD FOR THE PREPARATION OF SAFFRON PIGMENT AND FLAVOR CONCENTRATE:

In this patent, saffron (10 g), diethyl ether (200 ml) and methanol (400 ml) were mixed and soaked at 20 ° C for 14 hours with the addition of distilled water (200 ml) and centrifuged to a small amount. Material is obtained. It evaporates at a temperature in the range of 10 inches under reduced pressure. The raw materials obtained were transferred through the Sephadex column to saffron pigments and flavor concentrates (5.98 g; pigment concentration 17.6%). In our method, the method of supercritical fluid, pulse pressure and heating with radio waves is used and the duration of the process is a maximum of three hours and the purity of the solution obtained is up to 93%.

4- Description: Saffron is the dried part of Crocus Sativus plant that can be used as additives and natural food colors, traditional medicines and in food industries such as sausage production, confectionery, textile and dairy industries. 75,000 saffron blossoms are needed to produce one kilogram of saffron spice, and due to the difficulty of harvesting saffron, saffron is one of the most expensive plants in the world. In terms of total saffron production, Iran is the largest producer of saffron in the world, followed by Spain, India, Italy, Greece and Morocco. The saffron plant has different parts that only the stigma of commercial saffron is. This part of the plant Saffron (stigma) has a variety of chemical compounds, including: carbohydrates, minerals, mucilage, vitamins (especially riboflavin and thiamine) pigments including crocin, anthocyanin, carotene, lycopene, zeaxanthin, a fragrant essential oil called turpentine and saffron Donor (picrocrocin).

There are more than 150 active chemical compounds in this golden spice, the most important of which are responsible for the taste, taste, smell and more importantly the effect of saffron are:

Carotenoids: A group of chemical compounds in plants that are responsible for the color of their leaves and petals. They are also responsible for absorbing light in plants.

Crocetin and crocin: Crocetin and crocin are carotenoids that produce the yellow and red color of saffron.

Other important carotenoids in saffron include lycopene, zeaxanthin, and beta-carotene. Lycopene, zeaxanthin, and beta-carotene.

Essential oils (Volatile oils): Oil obtained from plants, which is also called essential oil or fragrance.

• Picrocrocin: The main responsible for the taste of saffron

Safranal: The main responsible for the aroma and aroma of saffron

Saffron color: The main factor in creating color strength in saffron is a compound called crocin with the chemical formula C44H64O24. Crocin is one of the few limited carotenoids found in nature that dissolves easily in water. This solubility is one of the reasons for its widespread use as a colorant in food and medicine compared to other carotenoids. In addition to crocin, saffron contains crocin aglycone freely and small amounts of anthocyanin pigment. It also contains fat-soluble pigments including lycopene, alpha-carotene, beta-carotene and zeaxanthin. The color strength of saffron is one of the main parameters determining the quality of saffron, which is evaluated by spectrophotometer by measuring the amount of color compounds in it at a wavelength of 443 nm.

Saffron flavor: The main compound that creates a bitter taste in saffron is a glycoside called picrocrocin with a chemical formula. (C16H26O6) This substance is bitter and crystalline and produces hydrolysis of acid, glucose and aldehyde called safranal (C10H14O2).

Aromatic compounds: Saffron has a fragrant smell. The smell of saffron is the result of the aroma of some volatile oils and special essential oils that are present in this substance. Safranal is the main aromatic substance of saffron and constitutes about 60% of the volatile compounds of saffron, which is non-volatile in fresh saffron as picrocrocin, but decomposes due to heat and over time, safranal volatile aldehyde is released. Safranal is a volatile oily liquid. It has a bright yellow stain during water vapor and dissolves well in ethanol, methanol and petroleum ether.

The nutritional value of saffron

One tablespoon of saffron has the following nutritional value:

Carbohydrates: 1.37 grams - Fat: 0.12 grams - Protein: 0.24 grams - Vitamin C: 1.7 mg - Vitamin: B9 0.002 micrograms - Vitamin: B60.02 mg - Vitamin: B30 , 03 mg - Vitamin: B20.01 mg - Iron: 0.23 mg - Magnesium: 6 mg - Manganese: 0.6 mg - Copper: 0.01 mg - Phosphorus: 5 mg - Potassium: 36 mg - Kaimferol: 4.3 mg

Scientific properties of saffron

Saffron increases the levels of proteins needed for body growth and blood pressure, and also helps with brain activity such as brain-derived neurogenic factor (BDNF) and vascular endothelial growth factor (VEGF). This spice has amazing antioxidant properties that kill free radicals that may damage proteins and DNA inside cells. This miracle spice also restricts the growth of amyloid beta proteins that lead to Alzheimer's. In addition, it inhibits the growth of enzymes that disrupt neurotransmission, such as acetylcholinesterase, and lower acetylcholine levels in the body. The properties of saffron are not limited to these and you should know that this golden spice can lower cortisol levels and increase estrogen levels; This helps the bones of the body to be healthy. Many of the healing properties of saffron are due to the presence of crocin molecule in the composition of this spice. Digesting crocin is not an easy task for the body, which is why the human digestive system converts it to crocin.

In general, the healing properties of saffron are as follows:

1- Saffron contains significant types of plant compounds that act as antioxidants. Molecules that protect the body's cells against free radicals and oxidative stress. Notable antioxidants in saffron include crocin, crocetin, safranal, and camper. The antioxidant properties of saffron prevent premature aging and protect the skin from sunlight, which can lead to the growth of cancer cells, which is why saffron is used in sunscreens and lotions.

2- Increasing the body's immunity

One of the most important properties of saffron is to increase the body's immunity. This plant has a lot of vitamin C (ascorbic acid) which is very good for health. The presence of vitamin C or ascorbic acid in saffron increases the production of white blood cells in the blood, which strengthens the immune system. However, it does not lead to an increase in red blood cells, so problems such as blood concentration can not be the effect of excessive consumption of saffron, is.

3- Increasing the body's energy

The carotenoids in saffron increase energy storage and strength in the body muscles. Also, the power of reaction and coordination between the mind and the eyes with these compounds increases. With increasing blood pressure and oxygen reaching the muscles, saffron can be a good choice for athletes.

4 - Reduce the symptoms and difficulties of premenstrual

One of the oldest uses of saffron in reducing premenstrual symptoms and problems such as: imbalance in mood, muscle cramps, bloating and pimples. - Improving heart function

Due to the important role of the heart in the human body, heart health and its protection are very important. One way to protect your heart health and help it function better is to eat foods that have relatively high amounts of potassium and saffron contains a lot of potassium. Potassium dilates the arteries of the body, which prevents high blood pressure and pressure on the arteries of the body, thus preventing risks such as stroke and heart attack. Saffron reduces the risk of heart attack and vascular problems by increasing the pumping capacity of the heart, thanks to the presence of kaimferol. This is probably due to the burning of cholesterol during the conversion of crocin to crocetin. Saffron prevents the increase of cholesterol, triglycerides and blood lipids. Also, in patients with heart attacks, saffron reduces blood clotting in the heart and prevents damage to the heart muscle. The potassium in saffron is very useful for controlling the heart rate and protects the heart by regulating the heart rate. Studies show that the antioxidant properties of saffron reduce blood cholesterol and prevent clogging of blood vessels and arteries.

6- Improving sexual function

7- Wound healing

Saffron ointment is able to increase the number of surface protective molecules of the skin and its anti-inflammatory properties help to heal and repair wounds on the skin.

8- Helping bone health

Saffron helps keep bones healthy by increasing estrogen levels in the body.

9- Treatment of asthma

The anti-inflammatory properties of saffron help calm the lungs.

10- Help treat diabetes

There is manganese in saffron. Manganese in saffron significantly reduces blood sugar levels in people with type 2 diabetes. The quercetin in saffron also improves insulin resistance and helps lower blood sugar. On the other hand, saffron can effectively reduce anxiety in diabetics and help insulin function in the body.

11- Helping eyesight

Saffron has one of the key elements in preventing vision loss and blindness. Saffron improves blood flow in the eyes and prevents eye aging. This process occurs due to the presence of crocin in it. Saffron has many effects on genes that regulate the function of visual cells. Researchers have found that saffron has a great effect on improving people with age-related macular degeneration, which is the most common cause of blindness in old age. Accordingly, saffron extract not only protects photoreceptor cells in the eye, but can also prevent and improve diseases such as macular degeneration and retinal pigment inflammation. Saffron is effective in regulating the function of vision cells and protects vision cells against damage. It also prevents the progression of eye diseases and regenerates damaged cells. On the other hand, saffron is effective in improving retinal yellow spot disorders that occur in old age. Consumption of saffron in the diet prevents the progression of genetic eye diseases and therefore its balanced consumption is recommended for the elderly and people with eye diseases.

12- Increase blood circulation

The high concentration of iron in saffron increases red blood cells in the blood, which increases blood circulation, metabolism in the body and better oxygen delivery to various organs of the body.

13- Treatment of insomnia

Saffron has natural sedative properties and it is said that consuming saffron syrup before going to bed helps to improve the quality of sleep and treats insomnia at night.

14- Fighting and preventing cancer

Saffron has high antioxidant properties that help neutralize harmful free radicals. In laboratory studies, saffron and its compounds selectively kill or stop cancer cells from selectively destroying healthy cells in the body.

15- Decreased appetite and weight loss

According to studies, saffron prevents false appetite. Consumption of saffron significantly reduces weight and eliminates fat masses around the abdomen and sides.

16- Treatment of Alzheimer's

Alzheimer's is the most common mental process disorder that usually occurs in people over the age of 65. A number of studies have examined the antioxidant properties of Crocus Sativus on Alzheimer's disease. Crocin, a substance derived from saffron, can be effective in reducing brain cell death. This substance prevents the accumulation of aging plaques. During the early stages of Alzheimer's disease, it develops as a mental disorder, in which saffron extract can reduce the symptoms of this type of disorder and prevent its spread. The results of these studies show the antioxidant and anti- amyloidogenic activities of saffron extract as well as the positive effects of saffron compounds on cognitive functions of the brain; The quercetin in saffron is useful in preventing Parkinson's disease. Saffron can also boost memory, and the crocin and crocetin in it are effective in treating Alzheimer's and dementia, and in some cases can even improve memory in Alzheimer's patients. According to studies, the decrease in serotonin levels and its activity in the brain can be one of the effective factors in Alzheimer's disease, and interestingly, the positive effect of saffron on increasing serotonin activity in the brain has been proven.

17- Treatment of fatty liver

Saffron has anti-inflammatory properties due to its high levels of antioxidants, inhibition of the growth of enzymes that produce toxins harmful to the liver and the ability to modulate the cellular structure of the liver, which can help detoxify and cleanse the liver and increase the risk of fatty liver disease. Reduce liver enlargement.

18- Mild to moderate depression: The antidepressant function of saffron and its function on the brain by increasing norepinephrine, serotonin and dopamine has made it known as an herbal antidepressant. On the other hand, saffron compounds such as safranal and crocin also reduce the symptoms of morphine withdrawa syndrome and according to research, the antidepressant properties of this extract have been proven on human and animal models.

In general, the methods of extracting chemical compounds and saffron extract are as follows: - Aqueous solvent method (cold): In this method, methanol and ethanol, aqueous and water are usually used to extract the bioactive compounds of saffron. After removing the fat with diethyl ether, the stigma is used 2 to 9 times using ethanol or 90-70% of methanol is extracted. The highest extraction efficiency of saffron extract is related to 50% methanol, followed by 50% ethanol, 25% ethanol and water. It rests for 24 hours and then passes through a filter of 5-10 micrometers. A noteworthy point in this method is the need to soak powdered saffron in solvent for about 24 hours, but on the other hand, with increasing the extraction time to 24 hours, the color strength decreases. Also, the extraction step of the extract affects its color strength. Put. In this way, if the filtering is done after dilution, the color strength will be reduced. - Dry ice or GC x GC-ToF-MS method: In this method, saffron lipid compounds are extracted using diethyl ether in a dry ice bath. After centrifugation and filtration, the concentrated extract and volatile compounds are separated and recovered using gas chromatography. - Steam distillation method: This method is used for fresh plants that contain essential oils. After collection, the plant is put directly into the distillation machine and instead of soaking the plant, it is poured into special baskets. This is because fresh plants contain a significant amount of water. It passes between the plant and the essential oil, along with water vapor, is collected in the cooling part of the device and then separated from the water. In this method, due to direct steam distillation, some essential compounds are hydrolyzed and some are decomposed by high heat. Therefore, to perform a good distillation, steam must penetrate into plant tissues and cells with high pressure to minimize the breakdown of essential oils. - UAE ultrasonic extraction: In this method, ultrasonic waves with a frequency of 20 to 2000 kHz are used. The main extraction mechanism with ultrasound is related to the phenomenon of cavitation. As a sound wave passes through an elastic medium, it causes the particles to move longitudinally, acting as a piston on the surface of the medium, resulting in a sequence of contraction and expansion phases. The molecules are temporarily detached from their original position and pass through as sound waves that can collide with the surrounding molecules. Then, during the expansion phase, the first group of molecules is attracted back to their original position and their kinetic energy is further pulled back. Thus, expansion zones are created in the environment, and since each medium has a critical molecular distance, when this distance is exceeded, the molecular interactions are broken and holes are created in the fluid. The cavities created in the environment are cavitation bubbles caused by ultrasound that are able to grow during the expansion stages and decrease in size during the contraction cycles. When the size of these bubbles reaches a critical point, they disintegrate during the contraction cycle, releasing large amounts of energy. One of the disadvantages of this method is the destructive and sometimes known effects of ultrasonic wave energy of more than 20 kHz on the active compounds of medicinal plants through the formation of free radicals and as a result of undesirable changes in medicinal molecules. Ultrasound is the process of extracting plant compounds. And facilitates and accelerates mass transfer. - SFE supercritical fluid: A supercritical fluid is a compound that is compressed and heated above its critical temperature and pressure Tc, Pc. The nature and nature of supercritical fluid in terms of thermodynamics is that it is similar to gases (with high diffusion coefficient) and similar to liquids and has a wide range of solubility for compounds. This phase has a solubility of about liquid, while it has the usual transfer properties in gases. The solubility behavior of the fluid is close to that of the liquid phase, while the diffusion into the solid is provided by the transfer properties similar to those of a liquid gas. The solubility of the material in this phase changes with the change of density which is under the control of temperature and pressure, achieved. A supercritical fluid is actually a dense gas, a liquid above the critical temperature and pressure to a certain value. For a supercritical fluid to be, the temperature drop must not exceed 1.2 degrees or 1.3 degrees Celsius. While it is possible to reduce high pressure within technical limits, the important points in the extracritical fluid extraction method are the choice of solvent. From carbon dioxide due to low price, toxicity and low flammability and favorable critical conditions (temperature 304 degrees Kelvin and pressure 7.3 MPa), the ability to eliminate after extraction and high solubility capacity of non-polar molecules to extract compounds Non-polar is used as hydrocarbons. Another advantage of this method is the solubility of supercritical fluid, which under the influence of changed temperature and pressure, provides the possibility of extracting certain compounds, and at the end of the solvent work can be easily recovered. Due to the use of safe solvents in this method, environmental pollution is prevented, as well as increasing the performance and quality of volatile oils and aromatic substances are other advantages of this method.- Microwave (MAE): Microwave waves are electromagnetic waves with a frequency of 0.3 to 300 GHz. These waves penetrate plant tissue and react with polar molecules such as water to generate heat. In fact, polar molecules such as water absorb these waves and cause high internal heat in plant matter, resulting in cell destruction. By absorbing a substance, the microwave electromagnetic energy is converted into heat energy. Microwaves penetrate directly into the plant material and cause internal heat by causing abrasion and friction between the molecules. This friction is caused by the rotation of bipolar ions within the polar solvent and the movement of unresolved ions. This creates a quick and selective heat throughout the plant material without the need to overheat the air. In this process, fresh or dried plant material is immersed in a solvent that transmits microwaves, and then the plantsolvent mixture is poured into an extraction tank that is exposed to microwaves. When a medicinal plant containing essential oils is exposed to microwave waves, these waves are selectively absorbed only by the liquid part of the plant cells (essential oils, water molecules, etc.). In this case, the temperature of the liquids in the plant material rises rapidly, causing the wall of the glands containing the essential oil to disintegrate, allowing the essential oil to leave the plant and enter the solvent. The resulting extract is then refined and converted to pure essential oil by subsequent processing. This technique can be combined with various extraction systems and solvents. The choice of a suitable solvent in this method depends on both the solubility and the dielectric constant of the solvent. Therefore, mixtures of solvents are usually used and water is added to increase the polarity of solvents such as hexane, xylene and toluene. Extraction of compounds Biologically active substances are different from plant materials. The temperature and particle size as well as the moisture content of the plant material are important factors in this method. For example, the higher the moisture content of the material, the more waves it absorbs and the more heat it generates, resulting in increased cell destruction. The center is for the removal of residual solids during the process and the poor performance for the extraction of non-polar, low-polarity or volatile compounds. - Electric field method (PEF): Electric field method with strong pulses is a process that is performed by applying high voltage pulses between two electrodes (20-80 kV / cm) in food. The use of PEF as a membrane permeability technique has received considerable attention because of its potential to increase extraction or create alternatives to conventional methods in food extraction and processing. High voltage pulse electric field is a technology in food processing such as pasteurization, compression and extraction and so on. - Soxhlet extractor: In Soxhlet extraction or hot continuous extraction method, a very small plant is poured into a porous bag (short metal tube) made of a strong paper filter or cellulose, and then in its place, inside the chamber of the device. Soxle is placed. The extraction solvents at the bottom of the hot flask evaporate into the sample, and after condensation inside the condenser, drip into the liquid. When the Soxhlet tank is completely filled, the chamber is automatically emptied by the side siphon, and the solvent is poured back into the distillation chamber. This cycle may need to be repeated for hours or days. In each cycle, some of the non-volatile materials are dissolved in the solvent. After many cycles, the desired compound is concentrated in a distillation vessel. The advantage of this system is that instead of using a large amount of solvent, the same solvent that has passed through the sample is recycled. After extraction, the solvent is removed using a rotary evaporator and the result is the desired composition. The insoluble portion of the solid remains inside the porous sac, which is usually discarded. - Extraction with CO2: Carbon dioxide gas due to many advantages such as suitable physical properties such as low temperature and critical pressure (PC = 8.72 bar, TC = / 06.31 C), neutral, non-toxic, impervious Flammability and explosiveness, non-corrosive in dry environments, high selectivity, abundance, cheapness and the possibility of preparation with high purity, for work on an industrial scale, has received more attention. Because carbon dioxide is a low molecular weight and solvent nonpolar, it is a good solvent for low molecular weight components with low polarity. According to a general estimate, supercritical carbon dioxide has the ability to dissolve soluble compounds in normal hexane and chloroform, and the solubility of the components in this solvent decreases with increasing molecular mass (or increasing molecule size) and decreasing vapor pressure. Therefore, in the extraction of polar compounds and compounds with high molecular mass, to achieve greater solubility or better selectivity, it is possible to add a small amount of polar solvents such as methanol, ethanol, acetone or water to carbon dioxide in most essential oils. This method is used on the basis of terpene origin. The principles of this extraction method are based on the critical point of CO2 gas. Liquid carbon dioxide is able to dissolve aromatic substances in itself and form two stages in the gaseous state. Expansion, which is accompanied by the change of carbon dioxide from a liquid to a gaseous state, separates the essential oils from the aromatic substances. 0- Soaking: Soaking involves placing plant material (coarse or powder) in a sealed container with solvent at room temperature for a minimum of three days. During this period, the stirring process takes place regularly. This process is intended to soften and break down the cell wall of plants to release soluble phytochemicals. After 3 days, the mixture is pressed or filtered by filtration. Extraction period, extraction filtration stage, type of filter and extraction solvents are influential factors in this method. 1- Emulsion liquid membrane (ELM): This method is one of the best separation methods for the extraction of metal contaminants and molecular species and has high mass transfer rate, high sensitivity, low solvent requirement and low investment. The ELM process includes the membrane phase, the dispersed or internal phase, and the external phase. Both phases are aqueous solutions, while the membrane phase is an organic liquid and somewhat hydrophobic, forming a thin layer with the inner phase. Usually the external phase contains a compound that must be extracted. Usually, the membrane phase can not be mixed with either the external phase or the internal phase, and contains additives, carriers, and surfactants that are such as to increase the selectivity, stability, and permeability of the membrane layer. By using ELM, it is possible to extract more compounds by using more factors. On the other hand, one of the disadvantages of solvent extraction methods is the use of high speed mixers up to 1000 rpm for two-phase mixing, but in ELM low speeds of 300 rpm and above are used. 2- Crystallization: Crystallization is the solidification of atoms or molecules and their transformation into a regular structure called a crystal. Of course, in processes such as freezing and direct deposition of a gas, crystallization is also used. Crystallization of materials is used in chemical laboratories to purify solids. Crystallization of materials has a good efficiency when the amount of impurities is small and less than 5% molar in solid. Also, if the temperature range of the impurity is very different from the material in question, crystallization can be used for purification. The general procedure is that an impure solid is completely dissolved in a small amount of hot and boiling solvent and The resulting solution is allowed to cool slowly. The crystals that will eventually form as a result are of very high purity, and the impurities of the solid remain in the saturated solution. Finally, solid crystals can be separated from impurities by filtration. This method has also been used to extract and purify crocin from saffron. - Accelerated solvent extraction method (ASE): Accelerated solvent extraction from organic solvents or aqueous solvents at high temperature and pressure to obtain complete extraction of analytes from solid and semi-solid samples in short periods of time and Uses small amounts of solvents. Accelerated solvent extraction is used in many applications such as extraction from pesticides and bio / nutrient active compounds. This method uses high temperature and pressure during the extraction process. High pressures greater than 1000 psi allow the solvent to reach higher temperatures at its boiling point, which increases the penetration rate, disrupts the strong effects of the solution and reduces the viscosity of the liquid solvent, allowing better penetration into the solution and subsequent recovery. Water and ethanol are able to extract different classes of compounds such as polyphenols depending on the temperature used in the accelerated solvent extraction. The advantages of this method are high speed, cheap price, high yield and also avoidance of toxic organic residues in the extracted materials. But the disadvantages of this method are the need for large amounts of water and relatively high temperatures during the process. In this method, the extraction chamber is filled with the solid sample and placed in a furnace with controllable temperature. After the solvent is added, the chamber is heated to a constant pressure (adjustable between 0.3 and 20 MPa) up to a maximum temperature of 200 ° C and kept for a period of time to maintain equilibrium. The extract is then transferred to a sample tube. A sample often goes through several extraction cycles.

Benefits of this invention:

1- Low solvent volume (about one liter per step)

2- Short extraction time (maximum 3 hours)

3- Percentage of purity up to 93%

4- Improving the purity of the extract in the extraction process, because the lower the temperature and the shorter the time, the less damage to the composition and quality of the produced extract will be;

5- Low cost and better extraction performance

6. In this method, radio frequency waves are used for heating.

7- The method of applying pressure to the solution is pulsed and depending on the amount of opening of the drain valves, which as a result causes shock and better extraction of the extracts.

8- With a special technique (pressure changes in order to separate the gas from the heavy particles) and its repetition, no residue of saffron particles remains in the chamber and the device.

9- In this technique, the short-term soaking method (about one hour) is used, unlike other methods (which lasts up to 48-24).

10- In this technique, the supercritical fluid method is used and the desired fluid is CO2. The reason for its use is low price, toxicity and low flammability, favorable critical conditions (temperature 304 degrees Kelvin and pressure 7.3 MPa), Ability to remove it after extraction and high solubility capacity of non-polar molecules to extract non-polar compounds such as hydrocarbons. Process instructions:

In our method, a CO2 tank is installed (Figure 1-2) which, by connecting to a motor pump (Figure 2-4), compresses the CO2 gas as much as possible. After this stage, there are two other tanks called EXI and EX2 (Figure 2-9) in which our main ingredient, saffron, is placed along with the solvent, but initially the saffron is powdered up to 50 microns to be well extracted. The solvent we are looking for is made up of watanol water (Figure 2-6-2-7), which is mixed in a ratio of 50-50 and poured into one liter of each tank. At this stage, the temperature of the tanks is measured by radio waves. Increase the pulse frequency (Figure 2-5) to 40 degrees Celsius. At the same time, discharge the compressed gas in the compression tank, which should have a pressure of 50 MPa, through the interface pipes and in the form of pulses into the two tanks EXI and EX2. . This process is repeated several times to cause a shock shock to our saffron solution and increase the pressure of these two tanks by 45 MPa. In these environmental conditions, ie temperature of 40 degrees Celsius and pressure of 45 MPa, a one-hour time pause is performed, until the saffron compounds are completely separated. As mentioned, in addition to the safranal in saffron itself, there are volatile gases in our solvent such as ethanol that evaporate quickly, so the next step is to have two chambers called (SI Figure 2-12) and S2 (Figure 2- 15). It is installed that first we increase its temperature by radio frequency waves (Figure 2-11 and 14- 14) to 80 degrees Celsius and the solution of saffron and gases in tanks EXI and EX2 with a low pressure of 8 to 10 megabytes Pascal is transferred to chamber SI by the drain valve (Figure 2-10) and the connecting pipes (the drain valve opens slightly at 10 degrees) and at this stage, unlike the previous step, it enters the heat shock solution. At the end of this chamber is a drain tank (Figure 2-16) into which our fluid is drained from inside chamber SI. This step is performed to separate terpenes and volatile gases. As a result, the expanded gas inside chamber SI is transferred back to chamber S2 and we pause for 5-6 minutes to separate the gas from the suspended particles in it which are heavier and into the discharge tank at the end of chamber S2 (Figure 2-16). ) To be entered. In the second step, the gas accumulated at the top of the s2 chamber is returned to the CO2 compression tank by a pump (Figure 19-2) and the liquid extracted in the previous step is combined with one and a half liters of solvent in each tank. (The percentage of solvent composition in this step is 70% ethanol and 30% water). Drain with a pressure of 45 MPa as a pulse into two tanks EXI and EX2. After that, a one-hour pause is performed and the connection valve between the two tanks EXI and EX2 and chamber SI is rotated up to 50 degrees until the pressure SI reaches 20 times. At this stage, the temperature of the two chambers SI and S2 is the same as the previous step up to 80 degrees C is increased by radiofrequency waves. After 10 minutes, the CO2 gas expands into the SI tank and the pressure rises to 25 MPa. At this time, the SI outlet valve opens into the drain tank and the picrocrocin (with a molar mass of 330 g / mol) is separated. After complete removal of the liquid, the remaining gas inside SI is transferred to S2 (with a pause of 5-6 minutes), and once again the suspended particles in the gas settle and enter the discharge tank S2 and the remaining gas is returned to the CO2 compression tank. Is. And in the third step to separate the crocin, the liquid obtained from the previous step is combined with one liter of solvent in each tank with a ratio of 70 to 30 ethanol and water and poured into tanks EXI and EX2. The gas is discharged at a pressure of 45 MPa as a pulse into tanks EXI and EX2. The pause time for separation at this stage is 30 minutes. Then open the drain valve of EXI and EX2 to SI until the pressure inside the tank SI reaches 30 MPa. Pause again for 15 minutes, to expand the gas inside chamber SI at 80 ° C and increase the pressure to 35 MPa. At this time, the outlet valve SI is opened and crocin is extracted with a molar mass of 970 g / mol. After the liquid is completely removed, the gas is transferred again from SI to S2 and we wait for a while until the heavy particles suspended in the gas move downwards and we take the crocin out of S2 and the rest of the gas from S2 to the compression tank. We turn. Due to the high concentration of crocin, we repeat this cycle three times in the same way from the beginning until the crocin is dehydrated and the color of saffron powder becomes completely colorless and white and only pure crocin solution is obtained. Finally, chromatography with HPLC device Equipped with UV detector, the amount of compounds and purity percentage of saffron extract is determined.

6- Explain the figures, diagrams, maps and ...

• Figure 1: Extraction process flowchart

1- CO2 tank

2- Extraction tankl

3- Extraction2 tank

4- Separation tankl

5- Separation2 tank

6- Pump

• figure 2

1- Co2 gas tank

2- Liquid CO2 transfer pump

3- Condenser

4- Pressure pump

5- Extraction heater

6- Water tank

7- Ethanol tank

8- Pump

9- Tank EXI and EX2

10- Drain valve

11- Separation heaterl

12- Separatorl (SI) tank

13- Drain valve

14- Separation heater2

15- Separator (S2) tank

16- Evacuation tank

17- Drain valve

18- Filter

19- Compressor Co2 Recycle

Claims

Claim 1. This technique is a process to obtain saffron extract, which includes: grinding the dry stigmas of saffron flowers to increase the extraction efficiency of chemical and bioactive compounds of saffron, mixing the stigmas ground with a mixture of water and ethanol to produce a mixture of stigmas, cooling and heating Mixing and applying pulse pressure to create shock in it, filtering and concentrating the mixture obtained to produce liquid saffron extract.

Claim 2. The process of claim 1, wherein the saffron is pulverized up to 50 microns to be well extracted.

Claim 3. The process of claim 1, wherein the milled stigmas and a solution of water and ethanol are mixed together in several steps at 50-50, 30-70 ratios in EXI and EX2 tanks.

Claim 4: The process of claim 3 wherein the solution of water, ethanol and saffron at each stage for the exit of chemical compounds for one hour in tanks EXI and EX2, rest for about 10 minutes in chamber SI and 5-6 minutes in chamber S2.

Claim 5. The process of claim 4, in which the time interval performed in EXI and EX2 tanks to soak saffron powder in the solvent and its extract is removed and the time interval performed in chambers SI and S2 to separate terpenes and volatile gases as well as heavier suspended particles It is done inside the gas.

Claim 6. The process of claim 1, wherein to apply pressure and generate a cold shock, we discharge CO2 gas at a pressure of 50 MPa into tanks EXI and EX2 containing the solution.

Claim 7. The method of claim 6, in which the cold shock occurs because the tanks EXI and EX2 are initially heated to 40 ° C, as a result of the solution of water, ethanol and saffron, the temperature rises, but after discharging the high-pressure CO2 gas, it decreases. The temperature is momentary and enters the shock solution, which causes a better drainage of saffron extract.

Claim 8. The process of claim 1, which uses radiofrequency waves to heat tanks, apply heat shock, and finally dehumidify, which raises tanks EXI and EX2 to 40 ° C and chambers SI and S2 to 80 ° C.

Claim 9. The process of claim 1, in which a pump is used to return the gas into the CO2 compression tank and repeat the process.

Claim 10. The process of claim 1, in which we repeat the cycle of cooling, heating and applying pressure several times to increase the pressure and completely drain the extract.

Claim 11. The process of claim 1, wherein from the first stage to the final stages of the extraction cycle, the pressure inside the output chambers SI and S2 for further shock and separation of volatile gases varies from (8-10) MPa, 25 MPa and 30 MPa, respectively.

Claim 12. The process of claim 11 wherein the adjustment and pressure changes are controlled by the amount of drain valve opening between the EX2, S2 and SI tanks.