A water-soluble turmeric powder and its preparation method

Abstract

This invention provides a water-soluble turmeric powder and its preparation method. The raw materials used are turmeric powder, β-cyclodextrin, γ-cyclodextrin, cassia seed extract, and edible salt. 95wt% edible alcohol and purified water are used as processing aids. The mass ratio of the raw materials and auxiliary materials is: turmeric powder: β-cyclodextrin: γ-cyclodextrin: cassia seed extract: edible salt = (1-6): (20-30): (45-55): (1-5): (0.05-0.15). This invention uses turmeric as the raw material and common food ingredients as auxiliary materials, employing physical processing technology. The resulting product has good water solubility, high bioavailability, and good safety, and can be widely used as a raw material for food, oral medicines, external medicines, cosmetics, disinfectants, medical devices, etc.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine, specifically to a water-soluble turmeric powder and its preparation method. Background Technology

[0002] Turmeric is the dried rhizome of the ginger family plant Curcuma longa L. It is harvested in winter when the stems and leaves wither, washed, boiled or steamed until thoroughly cooked, sun-dried, and the fibrous roots are removed.

[0003] Curcumin is a polyphenolic substance found in turmeric, a major component of the traditional Chinese medicine turmeric, containing approximately 3% to 6%. It is a rare diketone pigment found in the plant kingdom, belonging to the diketone class of compounds. Clinically, it is widely used for promoting blood circulation, regulating qi, and relieving pain. It is one of the natural food colorings permitted for use both domestically and internationally. Recent studies have revealed its physiological and pharmacological effects, including antioxidant, antitumor, Alzheimer's disease prevention, anti-inflammatory, immunomodulatory, anti-atherosclerotic, hypoglycemic, and hypolipidemic effects, with the advantage of minimal side effects and definite efficacy. Curcumin is a natural compound with free radical scavenging capabilities. It has been reported that curcumin can directly scavenge free radicals (ROS and RNS) both in vitro and in vivo. Compared with other polyphenolic compounds (such as resveratrol), curcumin exhibits better ROS scavenging effects. Curcumin is associated with transcription factor (Nrf2) and can increase intracellular reduced glutathione levels. The antioxidant activity of curcumin is related to its inhibition of lipid peroxidation and maintenance of the activity of various antioxidant enzymes, such as superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH).

[0004] Lipid peroxidation refers to a series of free radical reactions in unsaturated fatty acids within biological membranes induced by oxygen free radicals, and is one of the harmful changes induced by oxygen free radicals. Curcumin inhibits lipid peroxidation by scavenging reactive free radicals involved in the peroxidation reaction. The preventive and therapeutic effects of curcumin on some diseases are also believed to be related to its antioxidant efficacy.

[0005] Numerous studies have demonstrated the immense anti-cancer potential of curcumin. Curcumin can inhibit the proliferation of various cancer cells, and the National Cancer Institute (NCI) has listed it as a third-generation chemopreventive drug for cancer. Further research has revealed that one of its key anti-tumor mechanisms is the induction of tumor cell apoptosis. Studies of drug-resistant tumor cells have shown that curcumin participates in reversing tumor cell drug resistance by inducing apoptosis. Curcumin regulates cell proliferation, DNA damage, and apoptosis by modulating the caspase protein family, the Bcl-2 family of apoptosis-inhibiting proteins, and the important tumor suppressor pathway p53. It also regulates various factors, including the nuclear transcription factor NF-κB, to induce apoptosis and inhibit proliferation in tumor cells. In recent years, with a deeper understanding of its physiological and pharmacological activities, the efficacy of curcumin in disease prevention and control has attracted attention, particularly the discovery of its therapeutic effects on cancer, HIV, and COVID-19. This has not only enhanced the unique efficacy of curcumin in disease prevention and treatment but also greatly promoted the development of traditional Chinese medicine. In terms of external application, curcumin can scavenge various forms of free radicals, regulate the activity of peroxidases, inhibit the activity of enzymes that produce reactive oxygen species, and can easily transfer electrons or donate hydrogen atoms to scavenge reactive free radicals involved in peroxidation reactions, activate cell protection signals, and reverse oxidative damage to epidermal cells. Curcumin can also optimize the wound repair process by reducing inflammatory responses, allowing damaged skin to more easily enter the later healing stages, such as proliferation and remodeling.

[0006] The bottleneck in the use of turmeric: Curcumin in turmeric has very low water solubility (11 ng / mL, 25℃), only a few parts per billion, resulting in poor absorption and low bioavailability in the body, which greatly limits its application in food, pharmaceuticals, and other related fields. Therefore, improving the water solubility and bioavailability of curcumin in turmeric is the key to developing its potential application value.

[0007] The currently used improvement methods are: (1) Curcumin liposomes, micelles, nanoemulsions, etc. The product is an emulsion liquid, and the relative bioavailability of curcumin is improved. The disadvantage is that a large amount of surfactant is used, and the hemolytic side effect of the surfactant makes it unsuitable for long-term use in food and drugs. (2) Curcumin amorphous solid dispersions, the product is a water-insoluble dispersion, which improves the relative bioavailability of curcumin. However, the synthesized polymer dispersions have an irritating effect on the gastrointestinal tract and are subject to restrictions by food and drug regulations, thus limiting their practical application. (3) Chemical modification of curcumin and chemical formation of complexes are used to improve the solubility and relative bioavailability of curcumin. The disadvantages are that the preparation process is complex and the cost is high, and there are residual impurities and organic solvents, which limits its widespread use in food and drugs. Summary of the Invention

[0008] To increase the solubility and bioavailability of curcumin in turmeric and fully utilize its efficacy, this invention provides a water-soluble turmeric powder and its preparation method. Using turmeric as raw material and common food ingredients as excipients, the powder is produced using physical processing technology. The resulting product has good water solubility, high bioavailability, and good safety, and can be widely used as a raw material for food, oral medicines, external medicines, cosmetics, disinfection products, medical devices, etc.

[0009] A water-soluble turmeric powder and its preparation method are disclosed. The turmeric used is the national food safety standard for food additives - turmeric (GB 1886.60), which is a powder that passes through an 80-mesh sieve and is a fat-soluble component. β-cyclodextrin and γ-cyclodextrin have unique cage-like structures that can encapsulate curcumin molecules to form inclusion complexes. At this time, the drug molecules are contained in the cavities of β-cyclodextrin and γ-cyclodextrin molecules, resulting in high dispersibility. At the same time, due to the hydrophilicity of the external polyhydroxyl groups of β-cyclodextrin and γ-cyclodextrin, the inclusion complexes have good wettability, thereby achieving the solubilization effect on poorly soluble curcumin.

[0010] Because β-cyclodextrin and γ-cyclodextrin have different cavity sizes, they can encapsulate groups of different sizes in curcumin molecules separately, resulting in a synergistic effect. Cassia seed gum in cassia seed extract acts as a stabilizer, contributing to product stability. Edible salt contains various polar ions such as iron, calcium, zinc, potassium, sodium, iodine, and chloride, which enhance solubility through ionization.

[0011] A water-soluble turmeric powder and its preparation method are disclosed. The product has good solubility, with 1g dissolving in 100mL of water at a water bath temperature (98-100℃). It has excellent bioavailability to the human body, about 120 times that of ordinary turmeric.

[0012] To achieve the above objectives, the present invention adopts the following technical solution:

[0013] A water-soluble turmeric powder and its preparation method are disclosed. The raw materials used are turmeric powder, β-cyclodextrin, γ-cyclodextrin, cassia seed extract, and edible salt as food-grade excipients, and 95wt% edible alcohol and purified water as processing aids. The mass ratio of the raw materials and excipients used is: turmeric powder (1-6): β-cyclodextrin (20-30): γ-cyclodextrin (45-55): cassia seed extract (1-5): edible salt (0.05-0.15).

[0014] Preferred, the mass ratio of the raw materials used, namely turmeric powder, β-cyclodextrin, γ-cyclodextrin, cassia seed extract, and edible salt, is: turmeric powder (2-5): β-cyclodextrin (22-28): γ-cyclodextrin (48-53): cassia seed extract (2-4): edible salt (0.08-0.12).

[0015] Preferred, the mass ratio of the raw materials used, turmeric powder, β-cyclodextrin, γ-cyclodextrin, cassia seed extract, and edible salt, is: turmeric powder (3): β-cyclodextrin (25): γ-cyclodextrin (50): cassia seed extract (3.5): edible salt (0.10).

[0016] The turmeric mentioned is: turmeric powder that has passed through an 80-mesh sieve, as per the national food safety standard for food additives (GB 1886.60).

[0017] The β-cyclodextrin mentioned is: National Food Safety Standard for Food Additive β-cyclodextrin (GB 1886.180); the γ-cyclodextrin mentioned is: National Food Safety Standard for Food Additive γ-cyclodextrin (GB 1886.353); and the edible salt mentioned is: National Food Safety Standard for Edible Salt (GB 2721).

[0018] The cassia seed extract mentioned is: Cassia seed extract (T / CCCMHPIE 1.84) (China Chamber of Commerce for Pharmaceuticals & Health Products Import & Export Standard).

[0019] Preferably, the cassia seed extract is purchased from Xi'an Zhongkeda Biotechnology Co., Ltd., with a specification of 10:1 (%). The purified water is bottled drinking water (GB 17323).

[0020] The edible alcohol mentioned is: edible alcohol (GB 31640)

[0021] The preparation method of water-soluble turmeric powder includes the following steps:

[0022] (1) Under light-protected conditions, take the amount of turmeric powder in the formula, add 6-15 times the weight of 95wt% edible ethanol, stir for 10 minutes to form a uniform solution for later use.

[0023] (2) Add purified water to the reactor, and add the prescribed amounts of β-cyclodextrin, γ-cyclodextrin, cassia seed extract and edible salt while stirring. Prepare a 30wt%-50wt% solution of edible salt.

[0024] (3) Set the stirring speed to 150r / min-250r / min, and slowly add the turmeric alcohol solution from the feed port while stirring. The addition should be completed in 20 minutes.

[0025] (4) Stir continuously for 1.5-2.5 hours.

[0026] (5) Pump the liquid into a vacuum concentrator for concentration, control the concentration temperature at 65±3℃, maintain the vacuum at 0.01-0.06Mpa, concentrate the liquid to a solid content of 65wt%-70wt%, and store it for later use.

[0027] (6) The concentrated material is dried by hot air circulation at atmospheric pressure at a temperature of 84±3℃ until it is dried into a blocky dry material with a moisture content of ≤5.0wt%.

[0028] (7) The dried block was crushed and sieved through a 60-mesh sieve using a pulverizer to obtain the final water-soluble turmeric powder. Detailed Implementation

[0029] To make the content of this invention easier to understand, the technical solution of this invention will be further described below with reference to specific embodiments, but this invention is not limited thereto.

[0030] Example 1

[0031] Step 1, Weighing: Weigh 3 parts turmeric powder, 25 parts β-cyclodextrin, 50 parts γ-cyclodextrin, 3.5 parts cassia seed extract, and 0.10 parts edible salt according to the mass ratio.

[0032] Step 2: Under light-protected conditions, take the prescribed amount of turmeric powder, add 10 times its weight of 95wt% edible ethanol, and stir for 10 minutes to form a homogeneous solution for later use.

[0033] Step 3: Add purified water to the reaction vessel, and while stirring, add the prescribed amounts of β-cyclodextrin, γ-cyclodextrin, cassia seed extract, and edible salt, and prepare a 42wt% solution of edible salt.

[0034] Step 4: Set the stirring speed to 190 r / min, and slowly add the turmeric alcohol solution from the feed port while stirring. The addition should be completed in 20 minutes.

[0035] Step 5: Continue stirring for 2.0 hours.

[0036] Step 6: Pump the liquid into a vacuum concentrator for concentration, control the concentration temperature at 65±3℃, maintain the vacuum at 0.02-0.05Mpa, concentrate the liquid to a solid content of 68wt%, and refrigerate for later use.

[0037] Step 7: Place the concentrated material in a stainless steel pan for hot air circulation drying at a temperature of 84±3℃, resulting in a moisture content of 4.7wt%.

[0038] Step 8: Crush and sieve the dried lumps through a 60-mesh sieve using a pulverizer to obtain the final water-soluble turmeric powder.

[0039] Step 9: Submit for testing

[0040] Take samples for testing in a timely manner. Fill out a testing form for the products to be tested, specifying the product name, batch number, specifications, quantity, production date, and other information.

[0041] Example 2

[0042] Step 1, Weighing: Weigh 2 parts turmeric powder, 28 parts β-cyclodextrin, 53 parts γ-cyclodextrin, 2 parts cassia seed extract, and 0.12 parts edible salt according to the mass ratio.

[0043] Step 2: Under light-protected conditions, take the prescribed amount of turmeric powder, add 15 times its weight of 95wt% edible ethanol, and stir for 10 minutes to form a homogeneous solution for later use.

[0044] Step 3: Add purified water to the reaction vessel, and while stirring, add the prescribed amounts of β-cyclodextrin, γ-cyclodextrin, cassia seed extract, and edible salt, and prepare a 50wt% solution of edible salt.

[0045] Step 4: Set the stirring speed to 250 r / min, and slowly add the turmeric alcohol solution from the feed port while stirring. The addition should be completed in 20 minutes.

[0046] Step 5: Continue stirring for 1.5 hours.

[0047] Step 6: Pump the liquid into a vacuum concentrator for concentration, control the concentration temperature at 65±3℃, maintain the vacuum at 0.02-0.06Mpa, concentrate the liquid to a solid content of 65wt%, and refrigerate for later use.

[0048] Step 7: Place the concentrated material in a stainless steel pan for hot air circulation drying at a temperature of 84±3℃, resulting in a moisture content of 4.3wt%.

[0049] Step 8: Crush and sieve the dried lumps through a 60-mesh sieve using a pulverizer to obtain the final water-soluble turmeric powder.

[0050] Step 9: Submit for testing

[0051] Take samples for testing in a timely manner. Fill out a testing form for the products to be tested, specifying the product name, batch number, specifications, quantity, production date, and other information.

[0052] Example 3

[0053] Step 1, Weighing: Weigh out 5 parts turmeric powder, 22 parts β-cyclodextrin, 48 parts γ-cyclodextrin, 4 parts cassia seed extract, and 0.08 parts edible salt according to the following mass ratios.

[0054] Step 2: Under light-protected conditions, take the prescribed amount of turmeric powder, add 8 times its weight of 95wt% edible ethanol, and stir for 10 minutes to form a uniform solution for later use.

[0055] Step 3: Add purified water to the reaction vessel, and while stirring, add the prescribed amounts of β-cyclodextrin, γ-cyclodextrin, cassia seed extract, and edible salt, and prepare a 35wt% solution of edible salt.

[0056] Step 4: Set the stirring speed to 150 r / min, and slowly add the turmeric alcohol solution from the feed port while stirring. The addition should be completed in 20 minutes.

[0057] Step 5: Continue stirring for 2.5 hours.

[0058] Step 6: Pump the liquid into a vacuum concentrator for concentration, control the concentration temperature at 65±3℃, maintain the vacuum at 0.01-0.05Mpa, concentrate the liquid to a solid content of 70wt%, and refrigerate for later use.

[0059] Step 7: Place the concentrated material in a stainless steel pan for hot air circulation drying at a temperature of 84±3℃, resulting in a moisture content of 3.6wt%.

[0060] Step 8: Crush and sieve the dried lumps through a 60-mesh sieve using a pulverizer to obtain the final water-soluble turmeric powder.

[0061] Step 9: Submit for testing

[0062] Take samples for testing in a timely manner. Fill out a testing form for the products to be tested, specifying the product name, batch number, specifications, quantity, production date, and other information.

[0063] 1. Product Technical Requirements

[0064] 1.1. Sensory requirements

[0065] Sensory requirements should conform to the specifications in the table below.

[0066]

[0067] 1.2. Solubility

[0068] 1g of this product can be dissolved in 100mL of water at a water bath temperature (98-100℃).

[0069] 1.3. Identification

[0070] The identification criteria should conform to the provisions of the table below.

[0071]

[0072] Procedure: Weigh 0.1g of the sample and dissolve it in 5mL of sodium hydroxide solution (0.05mol / L). The solution will initially appear rose-red. After adding hydrochloric acid (1mol / L) until the solution becomes acidic, the rose-red color will change to bright yellow. Weigh 10mg of the sample and dissolve it in 5mL of ethanol solution (95%). Sonicate for 20 minutes. The solution will initially appear pure yellow with a slight green fluorescence. Adding a small amount of sulfuric acid will then turn the solution rose-red.

[0073] 1.4. Physicochemical Indicators

[0074] The physicochemical properties should meet the requirements of the table below.

[0075]

[0076] 1.5. Pollutant Limits

[0077] The limits for contaminants shall comply with the provisions of GB 2762; indicators that are stricter than the national food safety standards shall meet the requirements of the table below.

[0078] project index Test methods Lead (as Pb), mg / kg < 1.0 GB5009.12

[0079] 1.6. Microbiological Indicators

[0080] Microbiological indicators should meet the requirements of the table below.

[0081] project index Total bacterial count, CFU / g ≤ 30000 Coliform bacteria, MPN / g ≤ 0.92 Molds and yeasts, CFU / g ≤ 50 Staphylococcus aureus ≤ 0 / 25g Salmonella ≤ 0 / 25g

[0082] 2. Product solubility and other test results

[0083]

[0084]

[0085] 3. Product bioavailability test report

[0086] 3.1. Test Conclusion:

[0087] This study used high-performance liquid chromatography-mass spectrometry (HPLC-MS) to determine the plasma concentrations of curcumin and its derivatives at different time points after oral administration of 150 mg (calculated as total curcumin) of water-soluble turmeric (provided by Fujian Mofan Nutritional Food Co., Ltd.), 150 mg (calculated as total curcumin) of imported water-dispersible turmeric (provided by Xiamen Zhenyu Biotechnology Co., Ltd.), and 150 mg (calculated as total curcumin) of food additive turmeric to 19 healthy male volunteers. Plasma concentration-time curves were plotted, and pharmacokinetic parameters were determined. The AUC value was calculated using the trapezoidal method, and the t-value was calculated from the concentration at the elimination phase using a semi-logarithmic plot. 1 / 2 .

[0088] The concentrations of curcumin and its derivatives in plasma were determined by high performance liquid chromatography-mass spectrometry. Endogenous substances in plasma did not interfere with the sample determination. The linear range of the standard curve was 2.2–1076 ng / ml, the limit of quantitation was 2.2 ng / ml, and the extraction recovery rate was 67.0%–70.8%. The intra- and inter-batch precision and accuracy met the relevant regulations and the requirements for biological sample analysis.

[0089] 3.2. The results of bioavailability (F) of water-soluble turmeric, water-dispersible turmeric and food additive turmeric in 19 subjects, calculated using Winnonlin 5.2.1 software, are shown in Table 1.

[0090] Table 1. Bioavailability of water-soluble turmeric, water-dispersible turmeric, and food additive turmeric among 19 subjects.

[0091]

[0092] The relative bioavailability (F) of curcumin in water-dispersible turmeric compared to food additive turmeric was 1260%; the relative bioavailability (F) of curcumin in water-soluble turmeric compared to food additive turmeric was 12000%.

[0093] 4. Materials and Methods

[0094] 4.1. Samples: Imported water-dispersible turmeric (Xiamen Zhenyu Biotechnology Co., Ltd., batch number: N156051518); Food additive turmeric raw material (purity: ≥95%, Hebei Luchuan Biotechnology Co., Ltd., batch number:

[0095] 190316); Water-soluble turmeric (Fujian Mofan Nutritional Food Co., Ltd., batch number: 20190903), store in a cool, dry place.

[0096] 4.2. Subjects

[0097] 4.2.1. Inclusion criteria for subjects: (1) Healthy volunteers, male, aged 18 to 40 years; (2) Body mass index (BMI) between 19 and 24 (BMI = weight (Kg) / height (m) squared); (3) No history of heart, liver, lung, kidney, digestive tract, nervous system, mental abnormalities and metabolic abnormalities; (4) No abnormalities in blood pressure, electrocardiogram, respiratory status, liver and kidney function and blood count as determined by physical examination; (5) No other medications taken within 2 weeks prior to the trial; (6) No history of blood donation within 3 months; (7) No test drugs taken within 30 days prior to the trial.

[0098] 4.2.2. Exclusion criteria for subjects: (1) Those under 18 years of age or over 40 years of age; (2) Pregnant or lactating women, or those allergic to this sample; (3) Those with serious diseases of the heart, liver, kidneys and hematopoietic system, or those with mental illness; (4) Those who have taken other drugs in a short period of time, which may affect their judgment of the results; (5)

[0099] Those who do not meet the inclusion criteria, do not consume the test samples as required, or whose incomplete data affects the judgment.

[0100] 4.3. Experimental Design and Grouping: A double-blind crossover experimental design was adopted, taking into account as many major factors affecting the results as possible, such as age, gender, and diet, and conducting a balance test to ensure comparability between groups of subjects.

[0101] 4.4. Dosage and Timing: Nineteen subjects (fasted overnight) successively ingested the three types of turmeric powder, each sample orally administered 150 mg (calculated as total curcumin). A one-week washout period was allowed between each sample intake. After product consumption, blood samples were drawn hourly for 1-8 hours within the first 12 hours, and every 2 hours for 8-12 hours, for analysis (using needle prick sampling) to determine the blood concentration and relative absorption rate of curcumin and its derivatives.

[0102] 4.5. Requirements for Main Instruments, Reagents and Testing Environment

[0103] 4.5.1. Reagents: Curcumin reference standard (batch number: 190112) (purity: 99.9%), Beijing Zhongke Quality Inspection Biotechnology Co., Ltd.; methanol, formic acid, acetonitrile, tert-butyl methyl ether (chromatographic grade); castor oil (analytical grade); anhydrous ethanol (analytical grade); ultrapure water (self-made).

[0104] 4.5.2. Instruments: Agilent high-performance liquid chromatograph; Agilent single quadrupole mass spectrometer; ChemStation Rev. B 03.01 chromatography workstation; electronic balance; analytical balance; centrifuge; micro vortex mixer; nitrogen blowing concentrator; shaker.

[0105] 4.5.3. Electrocardiogram, X-ray fluoroscopy machine, B-ultrasound scanner, biochemical analyzer, blood cell counter, sphygmomanometer, etc.

[0106] 5. Selection and physical examination results

[0107] Twenty-one healthy male volunteers participated in the screening process. After being informed of all possible adverse reactions related to the drug, they signed informed consent forms. Subject 21 (screening number) had an abnormal blood routine test during the screening period and did not meet the inclusion criteria. Ultimately, 20 subjects were enrolled. Subject 13 (screening number) withdrew from the group for personal reasons, so 19 subjects were ultimately enrolled and completed the trial. After the trial, all 19 subjects underwent a comprehensive physical examination. Except for Subject 20 (screening number), whose WBC value was elevated (judged by a physician to be outside the normal range but without clinical significance), all other subjects' physical examination values ​​were normal (including renal function, liver function, blood routine, and urinalysis indicators). The results are shown in Tables 2-1 to 2-4.

[0108] Table 2-1 Subject Screening Results

[0109]

[0110]

[0111] Table 2-2 Subject Information

[0112]

[0113]

[0114] Table 2-3 Results of electrocardiogram, liver and kidney function tests, and routine blood and urine tests of the subjects before the trial.

[0115]

[0116]

[0117] Subject 13 (screening number) withdrew from the group midway due to personal reasons;

[0118] Subject 21 (screening number) was not selected because of abnormal blood routine and did not meet the inclusion criteria.

[0119] Table 2-4 Results of electrocardiograms, liver and kidney function tests, and routine blood and urine tests of the subjects after the trial.

[0120]

[0121]

[0122] *: Values ​​outside the normal range, but deemed clinically insignificant by a doctor.

[0123] 6. Experimental food ingredients

[0124] 6.1. Food additive turmeric: content: 95%, Hebei Lvchuan Biotechnology Co., Ltd., batch number: 190316; 6.2. Water-dispersible turmeric: imported by Xiamen Zhenyu Biotechnology Co., Ltd., batch number: N156051518;

[0125] 6.3. Water-soluble turmeric: Prepared by Fujian Mofan Nutritional Food Co., Ltd., batch number: 20190903.

[0126] 7. Quantity and timing of consumption

[0127] Based on information from foreign marketed products and literature, the daily intake of curcumin is 30 mg. The dosage used in this experiment was 5 times the usual dosage to ensure measurability. The patient had fasted for more than 10 hours before consumption and took the product with 250 ml of warm water on an empty stomach on the morning of the test day. The results were recorded.

[0128] 8. Biological sample collection

[0129] Subjects fasted for at least 10 hours before consuming the product. On the morning of the test day, the product was consumed on an empty stomach with 250ml of warm water. Five ml of venous blood was collected from the forearm via an indwelling needle before consumption and at 1.0, 2.0, 3.0, 4.0, 5.0, 6.0, 7.0, 8.0, 10.0, and 12.0 hours after consumption. The blood was placed in a heparin-anticoagulated tube, centrifuged at 3000 rpm for 10 minutes, and the plasma was separated and stored at -20°C for later use.

[0130] 9. Determination of curcumin blood concentration

[0131] 9.1. Instruments, reagents, and other materials

[0132] 9.1.1 Instruments

[0133]

[0134]

[0135] 9.1.2 Reagents and Test Chemicals

[0136]

[0137] 9.2. Plasma Sample Analysis Methods

[0138] This study used high-performance liquid chromatography-mass spectrometry (HPLC-MS) to determine the plasma concentrations of curcumin and its derivatives at different time points after consuming three different samples in 19 healthy male volunteers.

[0139] 9.2.1. Preparation of curcumin stock solution and standard series solutions

[0140] Accurately weigh 10.76 mg of curcumin reference standard and place it in a 100 ml volumetric flask. Dissolve and dilute to the mark with methanol to prepare a stock solution of curcumin with a concentration of 107.6 μg / ml. Dilute the curcumin stock solution with methanol-water (1:1) to prepare a series of curcumin standard solutions. The process is shown in Table 3 (all operations are performed under light protection).

[0141] Table 3. Preparation process of curcumin series standard solutions

[0142]

[0143] 9.2.2. Preparation of Curcuma Quality Control Series Standard Solutions

[0144] The curcumin stock solution was diluted with methanol-water (1:1) to prepare curcumin QC series solutions, as shown in Table 4 (all operations were performed under light).

[0145] Table 4. Preparation process of turmeric quality control series standard solutions

[0146]

[0147] 9.2.3. Preparation of internal standard solution

[0148] Accurately weigh 10.51 mg of tinidazole reference standard (equivalent to 10.50 mg of tinidazole), place it in a 50 mL volumetric flask, dissolve and dilute to the mark with methanol to obtain tinidazole internal standard stock solution I with a concentration of 210.0 μg / mL. Accurately measure 0.5 mL of internal standard stock solution I, place it in a 100 mL volumetric flask, and dilute to the mark with methanol to obtain internal standard stock solution II with a concentration of 1.050 μg / mL. Accurately measure 8.0 mL of internal standard stock solution II, place it in a 100 mL volumetric flask, and dilute to the mark with methanol to obtain internal standard solution with a concentration of 84.0 ng / mL.

[0149] Each stock solution, standard series solution, and internal standard solution was prepared in calibrated volumetric flasks of the corresponding volume and stored in a refrigerator at 4°C for later use.

[0150] 9.2.4. Chromatographic and Mass Spectrometry Conditions

[0151] 9.2.4.1. Chromatographic conditions

[0152] Chromatographic conditions, column C 18 XTerra MS column (5 μm, 100 × 2.1 mm, Waters, USA); mobile phase: acetonitrile:water (containing 10 mM ammonium acetate) (80:20, v / v), flow rate: 0.3 mL / min, column temperature: 25 °C. Analysis time: 2.0 min.

[0153] 9.2.4.2. Mass Spectrometry Conditions

[0154] The ion source was an electrospray ionization source (ESI+); the electrospray voltage in positive ion mode was 4500 V; the capillary temperature was 350 °C; the nebulizer temperature was 280 °C; the sheath gas (N2) pressure was 30 psi, the auxiliary gas (N2) pressure was 25 psi, and the collision gas (Ar) pressure was 1.5 mTorr; the peak width was 0.5 Th; and the scan time was 0.3 s. Selected reaction monitoring (SRM) was used for the scan.

[0155] 9.2.4.3. Plasma Sample Processing Methods

[0156] Add 50 μl of the corresponding internal standard tinidazole solution to the plasma, vortex for 10 s to mix, add 0.5 ml of tert-butyl methyl ether, vortex for 120 s, centrifuge at 15000 rpm for 5 min, transfer 400 μl of the organic layer to another clean 1.5 ml centrifuge tube, evaporate to dryness under vacuum at room temperature, add 100 μl of reconstitution solution (acetonitrile:water (containing 10 mM ammonium acetate) (80:20, v / v)) to dissolve, centrifuge at 15000 rpm for 5 min, take 80 μl of the supernatant and transfer it to a sample vial, inject 10 μl for UPLC-MS / MS analysis.

[0157] 9.2.5. Standard Curve and Linear Range

[0158] Accurately measure 1.0 ml of blank plasma and add 100 μl of curcumin series standard solutions to prepare plasma samples with curcumin concentrations of 2.2, 4.3, 10.8, 21.5, 43.0, 107.6, 215.2 ng / ml, and 1076 ng / ml. Perform the same experiments as described in the "Plasma Sample Processing Methods" section. For each concentration, perform dual-sample analysis to prepare a standard curve for curcumin. Plot the analyte concentration (ng / ml) on the x-axis and the peak area ratio of the analyte to the internal standard (AS / Ai) on the y-axis. Perform linear regression using the weighted least squares method (weighting coefficient: 1 / X²) to obtain the standard curve equation. The results show that curcumin exhibits good linearity with the peak area ratio of the analyte and internal standard within the range of 2.2–1076 ng / ml, and the concentration determination results meet the required precision and accuracy. The standard curve results for the three-day method validation are shown in Table 5.

[0159] Table 5 Standard Curve (Method Validation)

[0160]

[0161] 9.2.6. Lower limit of quantification

[0162] Accurately measure 1.0 ml of blank plasma and add 100 μl of curcumin standard solution to prepare a plasma sample with a curcumin concentration of 2.2 ng / ml. The same method was used in the "Plasma Sample Processing Method" section. Six samples were analyzed, with three consecutive batches measured. The concentration of each sample was calculated based on the standard curve for that batch. The results show that the limit of quantitation for curcumin in plasma determined by the high-performance liquid chromatography-mass spectrometry method established in this experiment can reach 2.2 ng / ml. See Table 6 for details.

[0163] 9.2.7. Accuracy and Precision

[0164] Accurately measure 1.0 ml of blank plasma and add 100 μl of curcumin standard solution of different concentrations to prepare plasma samples with curcumin concentrations of 4.8, 19.4, and 193.7 ng / ml (six samples for each concentration are analyzed). Perform the same test as under "Plasma Sample Processing Methods," and continuously measure three batches (simultaneously with the standard curve). Calculate the concentration of the plasma samples and determine the accuracy and precision of the method. The results show that the intra-batch and inter-batch precision of the high-performance liquid chromatography-mass spectrometry method established in this experiment meets the experimental requirements. See Table 6 for details.

[0165] Table 6. Lower Limit of Quantification, Accuracy, and Precision

[0166]

[0167]

[0168] 9.2.8. Extraction Recovery Rate

[0169] Accurately measure 1.0 ml of blank plasma and add 100 μl of curcumin standard solution of different concentrations to prepare plasma samples with curcumin concentrations of 4.8, 19.4, and 193.7 ng / ml (six samples for each concentration were analyzed). The same method was used under "Plasma Sample Processing Methods" to obtain the corresponding chromatographic peak areas. Separately, take 800 μl of each corresponding concentration of curcumin standard solution, add 400 μl of tinidazole internal standard solution, and dilute with 400 μl of mobile phase to obtain in vitro samples with the same concentration as the processed plasma samples. Inject 50 μl of each sample for analysis to obtain the corresponding chromatographic peak area (average of six determinations). The extraction recovery rate was calculated based on the peak area ratio of the two processing methods for each concentration. The results show that the extraction recovery rate of curcumin in plasma samples meets the relevant regulations, and the internal standard extraction recovery rate is stable. See Tables 7-1 and 7-2 for details.

[0170] Table 7-1 Curcumin Extraction Recovery Rate

[0171]

[0172] Table 7-2 Recovery rate of internal standard extraction

[0173]

[0174]

[0175] 9.2.9. Matrix effect

[0176] Accurately measure 1.0 ml of blank plasma. Except for the absence of standard series solutions and internal standard, follow the same procedure as described under "Plasma Sample Processing Methods." After drying, add 100 μl of curcumin standard solutions at concentrations of 48.4, 193.7, and 1936.8 ng / ml, 50 μl of internal standard solution, and 50 μl of mobile phase to the obtained residue. Vortex for 1 minute, then transfer the entire sample to a 1.5 ml centrifuge tube and centrifuge for 3 minutes (15000 rpm). Inject 50 μl of the solution for analysis to obtain the corresponding chromatographic peak area (six samples per concentration). Separately, take 800 μl of each of the corresponding concentrations of curcumin standard solutions, add 400 μl of tinidazole internal standard solution, and dilute with 400 μl of mobile phase to obtain in vitro samples with the same concentration as the plasma samples after processing. Inject 50 μl of each sample for analysis to obtain the corresponding chromatographic peak area (average of six measurements). Calculate the matrix effect using the peak area ratio of the two processing methods for each concentration. The results show that the matrix effect of this method is negligible. See Tables 8 and 9 for details.

[0177] Table 8. Matrix Effects of Curcumin

[0178]

[0179] Table 9 Internal Standard Matrix Effect

[0180]

[0181]

[0182] 9.2.10. Accompanying standard curve and quality control during sample determination.

[0183] Plasma samples from subjects were analyzed according to the same method described under "Plasma Sample Processing Methods". To reduce systematic errors, a concurrent standard curve was established for each batch of samples to calculate the drug concentration. The concurrent standard curve was calculated using the weighted least squares method (W = 1 / X). 2 Linear regression was performed, and quality control samples at low, medium, and high concentrations (4.8, 19.4, and 193.7 ng / ml) of curcumin were measured in parallel. The relative efficiency (RE) of the quality control samples was within ±15%, and a maximum of one-third of the quality control samples at different concentrations were allowed to exceed the limit for the entire batch of data to be acceptable. The results showed that the quality control sample results all complied with relevant regulations. See Table 10 for details.

[0184] Table 10. Curcumin accompanying standard curve and quality control samples (QC)

[0185]

[0186] *: Overflow value, not included in statistics.

[0187] 9.3. Methodological Summary

[0188] This study established a high-performance liquid chromatography-mass spectrometry (HPLC-MS / MS) method for determining the concentration of curcumin in plasma at different time points after oral administration of turmeric, and validated the method. Experimental results show that this method has good accuracy and high sensitivity, meets relevant requirements, and can be used for studies on the relative bioavailability of turmeric and curcumin-based products in humans.

[0189] 10. Data Quality Assurance

[0190] During the experiment, on-site technicians monitored the process according to regulations and reported promptly. Data entry was reviewed by at least two people, and Winnonlin 5.2.1 data statistics software was used to ensure data consistency and accuracy.

[0191] 11. Research Results Data

[0192] 11.1. Blood drug concentration data table of turmeric, an edible food additive

[0193] Table 11-1 Blood concentrations (ng / ml) of curcumin at different time points after oral administration of turmeric to 19 subjects.

[0194]

[0195] 11.2. Blood drug concentration data table of water-dispersible turmeric.

[0196] Table 11-2 Blood concentrations of curcumin (ng / ml) at different time points after oral administration of water-dispersible turmeric to 19 subjects.

[0197]

[0198] 11.3. Blood drug concentration data table of edible water-soluble turmeric

[0199] Table 11-3 Blood concentrations (ng / ml) of curcumin at different time points after oral administration of water-soluble turmeric raw material to 19 subjects.

[0200]

[0201]

[0202] 12. Pharmacokinetic parameters

[0203] 12.1. Pharmacokinetic parameters of various types of edible turmeric

[0204] Pharmacokinetic parameters of the test or reference formulations were calculated in 19 subjects using Winnonlin 5.2.1 statistical software. The results are shown in Tables 12-1 to 12-3.

[0205] Table 12-1 Pharmacokinetic parameters of 19 subjects after consuming water-soluble turmeric

[0206]

[0207]

[0208] Table 12-2 Pharmacokinetic parameters of 19 subjects after consuming turmeric

[0209]

[0210]

[0211] Table 12-3 Pharmacokinetic parameters of 19 subjects after consuming water-dispersible turmeric

[0212]

[0213]

[0214] 12.2. Relative bioavailability of water-soluble turmeric compared to turmeric

[0215] Bioavailability F is applied to the AUC of each subject. 0-t and AUC 0-∞ Calculate separately:

[0216] F = (AUC) 0-t ) 水溶性姜黄均值 / (AUC 0-t ) 姜黄均值 ×100%,

[0217] F = (AUC) 0-∞ ) 水溶性姜黄均值 / (AUC 0-∞ ) 姜黄均值 ×100%.

[0218] The results of bioavailability (F) of water-soluble turmeric, water-dispersible turmeric, and turmeric in 19 subjects are shown in Table 13.

[0219] Table 13 Bioavailability of water-soluble turmeric, water-dispersible turmeric, and turmeric in 19 subjects

[0220]

[0221] The above description is only a preferred embodiment of the present invention. All equivalent changes and modifications made within the scope of the claims of the present invention should be included in the scope of the present invention.

Claims

1. A water-soluble turmeric powder, characterized in that, The raw materials used are turmeric powder, β-cyclodextrin, γ-cyclodextrin, cassia seed extract, and edible salt. 95wt% edible alcohol and purified water are used as processing aids. The mass ratio of turmeric powder:β-cyclodextrin:γ-cyclodextrin:cassia seed extract:edible salt used is (2-5):(22-28):(48-53):(2-4):(0.08-0.12).

2. The water-soluble turmeric powder according to claim 1, characterized in that, The raw materials used, namely turmeric powder, β-cyclodextrin, γ-cyclodextrin, cassia seed extract, and edible salt, were in a mass ratio of 3:25:50:3.5:0.

10.

3. The water-soluble turmeric powder according to claim 1, characterized in that, The turmeric powder mentioned is a powder that has passed through an 80-mesh sieve.

4. The method for preparing water-soluble turmeric powder according to any one of claims 1-3, characterized in that, Includes the following steps: (1) Under light-protected conditions, take the amount of turmeric powder in the formula, add 6-15 times the weight of 95wt% edible alcohol and stir for 10 minutes to form a uniform solution for later use. (2) Add purified water to the reactor, and add the formula amount of β-cyclodextrin, γ-cyclodextrin, cassia seed extract and edible salt while stirring. Prepare a 30wt%-50wt% solution of edible salt. (3) Set the stirring speed to 150r / min-250r / min, and slowly add the turmeric alcohol solution from the feed port while stirring, and finish adding it in 20 minutes; (4) Stir continuously for 1.5-2.5 hours; (5) Pump the liquid into a vacuum concentrator for concentration, control the concentration temperature at 65±3℃, maintain the vacuum at 0.01-0.06Mpa, concentrate the liquid to a solid content of 65wt%-70wt%, and store it for later use. (6) The concentrated material is dried by hot air circulation at atmospheric pressure at a temperature of 84±3℃ until it becomes a blocky dry material with a moisture content of ≤5.0wt%. (7) The dried block was crushed and sieved through a 60-mesh sieve using a pulverizer to obtain water-soluble turmeric powder.

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