CN112557532A - Detection method of composition containing theanine - Google Patents
Detection method of composition containing theanine Download PDFInfo
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Abstract
The invention relates to a method for detecting a composition containing theanine, which comprises the steps of sample treatment, standard curve preparation and chromatographic conditions, wherein a sample solution is injected into a high performance liquid chromatograph, the sample solution is qualitative through retention time and ultraviolet-visible absorption spectrum, the sample solution is quantitative through peak area, and the corresponding concentration of the theanine can be obtained from the standard curve through the peak area of a chromatographic peak.
Description
Technical Field
The invention relates to a detection method of a composition containing theanine, belonging to the field of analysis methods.
Background
Theanine is a special amino acid in tea, and has multiple physiological functions and pharmacological activities, such as lowering blood pressure, inhibiting tumor, relaxing nerve, protecting brain injury, reducing blood lipid, reducing weight, relieving fatigue, protecting nerve cell, improving menstrual period syndrome, and improving learning memory. Theanine can inhibit secretion of 5-hydroxytryptamine with inhibitory effect on central nervous system, and promote secretion of Dopamine (DA) with exciting effect; after a human body absorbs theanine elements, the proportion of the theanine in the blood can be increased, so that a nervous system can be stimulated to generate exciting feeling, certain resistance can be generated to fatigue, and the fatigue resistance of a user is effectively improved; theanine can reduce the damage of liver caused by exercise, and its intrinsic mechanism is to increase the oxygen content in blood of the exercise fracture, thereby relieving fatigue. Meanwhile, theanine can also quickly and effectively reduce the content of substances generated by movement in the body, such as blood serum and urea nitrogen.
The fructus Lycii is dry mature fruit of Solanaceae plant, is traditional Chinese medicine, and has effects of invigorating kidney, invigorating qi, moistening lung, and promoting sperm production. The lycium barbarum polysaccharide is a soluble polysaccharide extracted from lycium barbarum, is a main active substance of lycium barbarum, can increase the content of hepatic glycogen and myoglycogen of a mouse, improve the motor capacity of the mouse, improve the SOD activity of the mouse, reduce the content of blood urea nitrogen and blood lactic acid, and relieve fatigue symptoms. The rhizoma polygonati contains various components, wherein the rhizoma polygonati polysaccharide is a characteristic component of plants in the genus polygonatum and is also one of main bioactive components. The polygonatum polysaccharide can obviously increase the reserve of muscle glycogen and liver glycogen of a mouse, improve the total activity of blood lactate dehydrogenase before and after exercise, reduce the increase of blood urea nitrogen of the mouse after strenuous exercise, accelerate the removal rate of the blood urea nitrogen after exercise, and have very obvious effects on improving the adaptability of load exercise, resisting fatigue and accelerating the elimination of fatigue. However, the traditional Chinese medicine of rhizoma polygonati and medlar contains a plurality of amino acids, and the amino acids have serious adverse effects on the analysis and identification of theanine. Therefore, the development of a product has important significance in resisting fatigue, supplementing energy and relieving malnutrition.
For the development of compositions containing theanine, there is a need to develop an assay to exclude the serious adverse effects of other ingredients on the analytical identification of theanine, to monitor the synergistic effects of the ingredients, to decrease blood urea nitrogen and blood oxygen acid levels, to increase liver glycogen and muscle glycogen levels, and to alleviate fatigue symptoms in the product composition.
Disclosure of Invention
The invention discloses a detection method of a composition containing theanine, which relates to the following steps:
sample treatment:
injecting the sample solution into a high performance liquid chromatograph, and determining the quality by retention time and ultraviolet-visible absorption spectrum, and determining the quantity by peak area, wherein the corresponding theanine concentration can be obtained from a standard curve by the peak area of a chromatographic peak.
The invention discloses a detection method of a theanine-containing composition, which comprises the following steps:
sample treatment:
accurately weighing 0.5g of the product containing theacric acid, adding 60mL of water into a conical flask, heating for 30min on a constant-temperature water bath kettle at 80 ℃, cooling, filtering, transferring to a 100mL volumetric flask, adding water to a constant volume to a scale, and uniformly mixing; filtering with 0.45 μm filter membrane;
preparation of a standard curve:
accurately weighing 20.00mg of a theanine reference substance, dissolving the theanine reference substance in water, transferring the theanine reference substance into a 20mL volumetric flask, and metering the volume to a scale with the water to obtain a theanine standard stock solution with the concentration of 1 mg/mL;
accurately sucking out 0.0mL, 0.1mL, 0.2mL, 0.5mL, 1.0mL, 1.5mL and 2.0mL of theanine standard stock solution respectively, and diluting to 10mL with water to obtain standard theanine use solutions with concentrations of 0.00mg/mL, 0.01mg/mL, 0.02mg/mL, 0.05mg/mL, 0.10mg/mL, 0.15 and 0.20mg/mL respectively;
chromatographic conditions
A chromatographic column: c18 column, 5 μm,4.6 × 250mm or equivalent column;
detection wavelength: 203 nm;
mobile phase: aqueous trifluoroacetic acid (ph 2.7);
flow rate: 1.0 mL/min;
sample introduction amount: 10.0 μ L;
column temperature: 35 ℃;
sample measurement
Injecting the sample solution into a high performance liquid chromatograph, and determining the quality by retention time and ultraviolet-visible absorption spectrum, and determining the quantity by peak area, wherein the corresponding theanine concentration can be obtained from a standard curve by the peak area of a chromatographic peak.
The composition containing theanine for relieving physical fatigue, which is prepared by the detection method, is characterized by comprising the following raw materials in parts by weight: 20-40 parts of theanine, 20-40 parts of medlar extract, 30-50 parts of sealwort extract, 6-10 parts of beeswax and 140 parts of vegetable oil.
The composition containing theanine for relieving physical fatigue, which is prepared by the detection method, is characterized by comprising the following raw materials in parts by weight: 30 parts of theanine, 30 parts of a Chinese wolfberry extract, 40 parts of a sealwort extract, 7 parts of beeswax and 130 parts of vegetable oil; wherein the composition is in the form of a soft capsule.
The invention discloses a composition containing theanine for relieving physical fatigue, which contains a plurality of raw materials as functional components, wherein the components have obvious synergistic effect on the aspect of physical fatigue relieving function.
The invention discloses a composition containing theanine for relieving physical fatigue, which comprises theanine, a medlar extract and a sealwort extract.
The invention discloses a composition containing theanine for relieving physical fatigue, which comprises the following components in part by weight:
component 1: 40g of theanine dry powder, 40g of medlar extract and 30g of sealwort extract.
And (2) component: 40g of theanine dry powder, 20g of medlar extract and 60g of sealwort extract.
And (3) component: 40g of theanine dry powder, 80g of medlar extract and 15g of sealwort extract.
The invention discloses a composition containing theanine for relieving physical fatigue, which is characterized by being prepared from the following raw materials in parts by weight: 20-40 parts of theanine, 20-40 parts of medlar extract, 30-50 parts of sealwort extract, 6-10 parts of beeswax and 140 parts of vegetable oil.
Preferably, the composition containing theanine for relieving physical fatigue is characterized by being prepared from the following raw materials in parts by weight: 30 parts of theanine, 30 parts of a Chinese wolfberry extract, 40 parts of a sealwort extract, 7 parts of beeswax and 130 parts of vegetable oil.
Wherein the composition is in the form of a soft capsule.
Wherein the vegetable oil is added in the form of soybean oil, corn oil, safflower seed oil.
Wherein, the composition also comprises pharmaceutically acceptable auxiliary materials, including gelatin, glycerol and purified water.
Wherein the composition is prepared by the following method: preparing content material liquid, preparing capsule skin material liquid, pelleting, shaping, wiping pills, drying and selecting pills.
The invention also provides an extraction method of theanine in the composition, which comprises the following steps:
(1) leaching: crushing tea leaves, putting the crushed tea leaves into an extractor, adding a certain volume of water, and heating and refluxing for extraction;
(2) and (3) filtering: filtering the product obtained in the step (1) by a 60-mesh screen to obtain a primary filtrate, and then carrying out secondary filtration on the primary filtrate, wherein the aperture is 0.45 um;
(3) adsorption: adsorbing the filtrate obtained in the step (2) by a chromatographic column, eluting, and collecting effluent liquid;
(4) concentration: concentrating the effluent by using a reverse osmosis membrane with the molecular weight cutoff of 80, and then performing secondary concentration by using an evaporator;
(5) and (3) drying: and (4) carrying out spray drying on the concentrated solution to obtain the theanine dry powder.
Preferably, the volume ratio of the tea leaves to the water in the step (1) is 1: 10-25.
Preferably, the leaching temperature in the step (1) is 50-70 ℃.
Preferably, the packing of the chromatographic column used in the step (3) is macroporous adsorption resin.
Preferably, the temperature in the first concentration in the step (4) is normal temperature, and the temperature in the second concentration is 70-90 ℃.
Preferably, the air inlet temperature of the spray drying in the step (5) is 160-.
Test example 1: method for analyzing theanine
1. Sample treatment:
accurately weighing 0.5g of the soft capsule prepared by the formula and the process in the embodiment 1 in a conical flask, adding 60mL of water, heating for 30min in a constant-temperature water bath kettle at 80 ℃, cooling, filtering, transferring to a 100mL volumetric flask, adding water to a constant volume to a scale, and uniformly mixing. Filtering with 0.45 μm filter membrane, and keeping.
2. Preparation of a standard curve:
accurately weighing 20.00mg of theanine reference substance, dissolving with water, transferring into a 20mL volumetric flask, and adding water to constant volume to scale to obtain the standard theanine stock solution with concentration of 1 mg/mL.
Accurately sucking out 0.0mL, 0.1mL, 0.2mL, 0.5mL, 1.0mL, 1.5mL and 2.0mL of theanine standard stock solution respectively, and diluting with water to a constant volume of 10mL to obtain theanine standard use solutions with concentrations of 0.00mg/mL, 0.01mg/mL, 0.02mg/mL, 0.05mg/mL, 0.10mg/mL, 0.15 and 0.20mg/mL respectively.
3. Chromatographic conditions
A chromatographic column: c18 column, 5 μm, 4.6X 250mm or equivalent column.
Detection wavelength: 203 nm.
Mobile phase: aqueous trifluoroacetic acid (pH 2.7).
Flow rate: 1.0 mL/min.
Sample introduction amount: 10.0. mu.L.
Column temperature: 35 ℃ is carried out.
4. Sample measurement
Injecting the sample solution into a high performance liquid chromatograph, and determining the quality by retention time and ultraviolet-visible absorption spectrum, and determining the quantity by peak area, wherein the corresponding theanine concentration can be obtained from a standard curve by the peak area of a chromatographic peak.
Test example 2 Process optimization procedure of theanine analysis method
1. The theanine content in the examples was tested by reference to GB/T23193 "high performance liquid chromatography for determination of theanine in tea".
1.1, sample treatment:
accurately weighing 0.5g of the soft capsule prepared by the formula and the process in the example 1 in a conical flask, adding 100mL of boiling distilled water, placing the mixture on a constant-temperature water bath kettle at 100 ℃ for leaching for 30min, cooling, filtering, transferring the mixture to a 100mL volumetric flask, adding water to a constant volume to scale, and uniformly mixing. Filtering with 0.45 μm filter membrane, and reserving.
1.2, preparing a standard curve:
accurately weighing 20.00mg of theanine reference substance, dissolving with water, transferring into a 20mL volumetric flask, and adding water to constant volume to scale to obtain the standard theanine stock solution with concentration of 1 mg/mL.
Accurately sucking out 0.0mL, 0.1mL, 0.2mL, 0.5mL, 1.0mL, 1.5mL and 2.0mL of theanine standard stock solution respectively, and diluting with water to a constant volume of 10mL to obtain theanine standard use solutions with concentrations of 0.00mg/mL, 0.01mg/mL, 0.02mg/mL, 0.05mg/mL, 0.10mg/mL, 0.15 and 0.20mg/mL respectively.
1.3 chromatographic conditions
A chromatographic column: c18 column, 5 μm, 4.6X 250mm
Detection wavelength: 210nm
Mobile phase: purified water + acetonitrile
Flow rate: 1.0mL/min
Sample introduction amount: 10.0. mu.L
Column temperature: 35 deg.C
Gradient elution conditions:
1.4, measurement of sample
And after the flow rate and the column temperature are stable, blank operation is carried out, 10uL of theanine standard use solution is accurately sucked and injected into HPLC, 10uL of test solution is injected under the same chromatographic condition, the test solution is quantified by a peak area, and the corresponding theanine concentration can be obtained from a standard curve by the peak area of a chromatographic peak.
1.5 test results
As can be seen from FIG. 1, the target peak and the adjacent peaks before and after the target peak do not reach the baseline separation, and the quantification cannot be accurately performed.
2. Chromatographic condition optimization
2.1, the pH of the purified water was adjusted to 3.0 with trifluoroacetic acid to prepare a mobile phase, the detection wavelength was changed to 203nm, and the sample of 1.1 was subjected to sample measurement, and the results are shown in FIG. 2.
As can be seen from FIG. 2, the target peak and the adjacent peak still do not reach the baseline separation, and can not be accurately quantified, but the separation effect is better than that of FIG. 1.
2.2 purified water was adjusted to pH2.7 with trifluoroacetic acid as fluidity, and the measurement wavelength was 203nm, and a sample of 1.1 was subjected to the measurement of the sample, and the result is shown in FIG. 3.
The pH of the purified water was adjusted to 2.5, 2.6 and 2.8 with trifluoroacetic acid, respectively, and the mobile phase was detected at a wavelength of 203nm and the pH of the mobile phase was screened, as shown in FIGS. 4, 5 and 6.
As can be seen from fig. 4, 5, and 6, the pH of the mobile phase was set to 2.7 because the target peak and the adjacent peak were separated from each other at the baseline at pH2.5, 2.6, and 2.7, and the target peak and the adjacent peak were not separated from each other at pH2.8, and the baseline was floated. The sample content is obviously lower than the theoretical amount through calculation, and the sample treatment method needs to be optimized.
3 sample processing method optimization
Accurately weighing 0.5g of the soft capsule prepared by the formula and the process in the embodiment 1 in a conical flask, adding 60mL of water, heating for 30min in a constant-temperature water bath kettle at 80 ℃, cooling, filtering, transferring to a 100mL volumetric flask, adding water to a constant volume to a scale, and uniformly mixing. Filtering with 0.45 μm filter membrane, and keeping.
Standard Curve preparation is as same as 1.2
A chromatographic column: c18 column, 5 μm, 4.6X 250mm
Detection wavelength: 203nm
Mobile phase: trifluoroacetic acid aqueous solution (pH2.7)
Flow rate: 1.0mL/min
Sample introduction amount: 10.0. mu.L
Column temperature: 35 deg.C
The calculated sample content is consistent with the theoretical amount, which indicates that the sample processing method is more reasonable.
Test example 3 composition of Contents
The invention relates to a composition containing theanine for relieving physical fatigue, wherein a capsule core consists of theanine dry powder, a Chinese wolfberry extract and a sealwort extract, and the components comprise, by weight, 40g of theanine dry powder, 40g of Chinese wolfberry extract, 30g of sealwort extract, 8g of beeswax and 140g of soybean oil. The capsule shell consists of 90g of gelatin, 90g of purified water and 45g of glycerol.
component 2 of the contents: 40g of theanine dry powder, 20g of medlar extract, 60g of sealwort extract, or,
component 3 of the contents: 40g of theanine dry powder, 80g of medlar extract and 15g of sealwort extract.
The remaining steps were carried out in the same manner as in example 1 to prepare a soft capsule
Respectively taking 9 parts of each of a soft capsule 1 (component 1 of the content), a soft capsule 2 (component 2 of the content) and a soft capsule 3 (component 3 of the content), wherein each part of the soft capsule is 0.5g, adding 60mL of artificial gastric juice into a conical flask, heating for 10 min, 30min and 60min on a constant-temperature water bath kettle at 37 ℃, cooling, filtering, transferring to a 100mL volumetric flask, diluting to a constant volume with water to a certain volume, and uniformly mixing. Filtering with 0.45 μm filter membrane, detecting theanine, and heating with soft capsule of component 1 of the content for 30min to obtain the final product with highest theanine content.
Test example 4 safety and functional verification
First, safety detection
Acute toxicity test:
1. sample preparation: the soft capsule is prepared according to the formula and the process described in the example 1.
2. Experimental animals: SPF-grade Kunming and Wistar rats.
3. Oral acute toxicity test (MTD) in mice: 20 SPF-grade Kunming mice of 18-22g, half of each sex, are taken and orally administered with 20g/kg & BW for one-time intragastric administration, and the administration is continuously observed for 14 days. The signs of intoxication and death were recorded.
4. Oral acute toxicity test (MTD) in rats: 180-220g SPF-level Kunming mice are selected, and the male and female halves of the mice are subjected to oral intragastric lavage at the dose of 20g/kg & BW once, and are continuously observed for 14 days after administration. The signs of intoxication and death were recorded.
5. As a result: both sexes of large and small mice were gavaged at a dose of 20 g/kg. BW for 14 days. No obvious toxic manifestation was observed during the experiment, and no death occurred during the observation period. The acute oral toxicity (MTD) of the tested substance to the big mouse and the small mouse of both sexes is more than 20g/kg & BW, and belongs to the non-toxic grade according to the acute toxicity grading of food safety toxicology evaluation program and method (2003 edition), and the specific results are detailed.
Acute toxicity test results of sample mice
Acute toxicity test results of sample rats
Feeding test for 30 days:
1. sample preparation: samples prepared according to the formulation and procedure described in example 1. The intended dose was 3.0g/60kg BW daily.
2. Experimental animals: 70-100g of weight and 80 SPD-grade Wistar rats with half of male and female are selected.
3. The test method comprises the following steps: rats were randomly divided into three test groups and one control group, 20 per group, each half male and female. The dose design is as follows: the low, medium and high dose groups were 1.248, 2.502 and 5.004g/kg & BW (corresponding to 25 times, 50 times and 100 times of the intended dose in humans, respectively). The control group was given an equal amount of vegetable oil and observed continuously for 30 days.
4. Observation of indices and results
4.1 general case observations: animals were observed daily for appearance, behavior, signs of toxicity, and mortality. Weighing body weight and food intake weekly, and calculating food utilization rate, total food intake and total weight gain weekly. Results animals in each group grew normally, worked normally, and were free of toxic manifestations and death, and animals in each group had no statistical differences in weekly weight and weight gain, weekly food availability, and total food availability (P > 0.05).
4.2 hematological examination: hemoglobin content (Hgb), Red Blood Cell (RBC) and White Blood Cell (WBC) counts, white blood cell classifications (lymphoid, monocyte, neutrophil, eosinophil, basophil) were determined. All indexes of hematology at the end of experiment are within normal value range, and the hemoglobin, erythrocyte, leucocyte count and leucocyte classification of each group of animals are not statistically different (P is more than 0.05) compared with the control group.
4.3 biochemical index determination: serum alanine Aminotransferase (ALT), aspartate Aminotransferase (AST), urea nitrogen (BUN), Cholesterol (CHO), Triglycerides (TG), blood Glucose (GLU), Total Protein (TP), Albumin (ALB), Creatinine (CRE) were determined. All biochemical indexes of the experimental animals at the end of the experiment are within normal value ranges, and the blood biochemical indexes of all groups of animals have no statistical difference (P is more than 0.05) compared with those of the control group.
4.4 gross observations and pathological tissue examinations: at the end of the experiment, the animals were sacrificed by cervical dislocation and the gross pathological changes of the major organs, the chest and the abdominal cavity were observed. The livers, kidneys, spleens, and testicles of all animals were removed, weighed, and organ coefficients calculated. The livers, kidneys, spleens, testicles (or ovaries), stomachs, and duodenum of the control and high dose animals were removed, fixed with 12% formalin, paraffin-embedded, sectioned, HE-stained, and examined histologically under light. No significant pathological changes were observed in all major organs (heart, liver, spleen, lung, kidney, stomach, intestine, etc.).
The acute toxicity test and the 30-day feeding test result show that the product is nontoxic and can be taken for a long time.
Second, physical fatigue test
1. Sample preparation: the soft capsule is prepared according to the formula and the process described in the example 1. The intended dose was 3.0g/60kg BW daily.
2. Kunming mouse, weight 18-23 g. Mice were randomized into 4 groups: control group, low, medium, high (0.5 g/kg. BW, 1.0 g/kg. BW, 1.5 g/kg. BW) dose group, 1 mice per group, each group male and female half. After continuous gavage for 30d, experimental observation is started, and animals in a control group are gavaged with distilled water. After the test object is given to the mouse for 30min at the last time, the mouse is placed in a swimming box for swimming, the water depth is about 30cm, the water temperature is 25 +/-1.0 ℃, and the tail root part of the rat is loaded with 5 percent of weight of lead skin. The time from the start of swimming to death of the mouse was recorded as weight-bearing swimming time(s). P <0.05
Effect of samples on mouse weight bearing swimming time (x. + -.s)
The heavy swimming time of the high-dose group mice is obviously longer than that of the control group, which shows that the product has the function of relieving physical fatigue.
Description of the drawings:
FIG. 1: HPLC (high Performance liquid chromatography) with target peak and front and rear adjacent peaks not reaching baseline separation
FIG. 2: HPLC with purified water adjusted to pH 3.0 with trifluoroacetic acid as mobile phase and detection wavelength of 203nm
FIG. 3: HPLC with purified water adjusted to pH2.7 with trifluoroacetic acid as mobile phase and detection wavelength of 203nm
FIG. 4: HPLC at mobile phase pH2.5
FIG. 5: HPLC at mobile phase pH2.6
FIG. 6: HPLC at mobile phase pH2.7
Detailed Description
The following examples further describe embodiments of the present invention in detail. The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention.
Example 1: soft capsule
Raw material dosage:
a composition containing theanine for relieving physical fatigue comprises a capsule core and a capsule core, wherein the capsule core comprises 30 parts by weight of theanine dry powder, 30 parts by weight of Chinese wolfberry extract, 40 parts by weight of rhizoma polygonati extract, 7 parts by weight of beeswax and 130 parts by weight of soybean oil. The capsule shell consists of 90 parts by weight of gelatin, 90 parts by weight of purified water and 45 parts by weight of glycerol.
The preparation method of the soft capsule comprises the following steps:
(1) preparation of content feed liquid
Respectively sieving theanine, the wolfberry extract and the sealwort extract by a sieve of 80 meshes, and weighing the theanine, the wolfberry extract, the sealwort extract, the beeswax and the soybean oil according to the formula; heating soybean oil and beeswax to 70 deg.C, cooling to room temperature after melting beeswax, adding theanine, fructus Lycii extract and rhizoma Polygonati extract, stirring, grinding into slurry with colloid mill, standing, and vacuumizing to obtain content liquid.
(2) Preparation of capsule shell liquid
Adding purified water and glycerol into a gelatin melting tank, mixing uniformly, heating to 60 ℃, adding gelatin, stirring to completely dissolve and mix uniformly, stopping stirring, standing, vacuumizing until no bubbles exist, and preserving heat to obtain a capsule shell material liquid.
(3) Pelleting and shaping
And (3) pelleting the prepared content material liquid and capsule skin material liquid, conveying the pressed soft capsules into a rolling cage by using a conveying belt, and rolling the soft capsules along with the rolling cage for shaping.
(4) Wiping pill
Wiping the shaped soft capsule with 95% edible alcohol.
(5) Drying
Drying the wiped soft capsule in a drying chamber at 20-30 deg.C and relative humidity of 15-40% until the soft capsule has moderate hardness, and stopping drying.
(6) Pill selection
And (4) manually selecting the dried soft capsules, removing unqualified waste pills with unattractive appearance, bubbles and leakage, and packaging qualified soft capsules to obtain finished products.
The method for preparing the theanine dry powder from the tea comprises the following steps:
(1) leaching: crushing tea leaves, putting the crushed tea leaves into an extractor, adding 10 times of water, and heating and refluxing the mixture at 60 ℃ for extraction;
(2) filtering the product obtained in the step (1) by a 60-mesh screen to obtain a primary filtrate, and then carrying out secondary filtration on the primary filtrate, wherein the aperture is 0.45 um;
(3) adsorption: adsorbing the filtrate obtained in the step (2) by a chromatographic column, eluting, and collecting effluent liquid, wherein the filler of the chromatographic column is macroporous adsorption resin;
(4) concentration: performing primary concentration on the effluent by using a reverse osmosis membrane with the molecular weight cutoff of 80 at normal temperature until the solid content is more than or equal to 10%, and performing secondary concentration by using an evaporator at 75 ℃ until the solid content is more than or equal to 40%. Cooling to room temperature;
(5) and (3) drying: and (3) carrying out spray drying on the concentrated solution, wherein the air inlet temperature of the spray drying is 160 ℃, the air outlet temperature is 90 ℃, and the feeding pressure is 7MPa, so as to obtain the theanine dry powder.
Example 2: soft capsule
Raw material dosage:
a composition containing theanine for relieving physical fatigue comprises a capsule core and a capsule core, wherein the capsule core comprises, by weight, 40 parts of theanine dry powder, 40 parts of Chinese wolfberry extract, 30 parts of rhizoma polygonati extract, 8 parts of beeswax and 140 parts of soybean oil. The capsule shell consists of 90 parts by weight of gelatin, 90 parts by weight of purified water and 45 parts by weight of glycerol.
The preparation method of the soft capsule comprises the following steps: same as example 1
The method for preparing the theanine dry powder from the tea comprises the following steps:
(1) leaching: pulverizing folium Camelliae sinensis, placing in an extractor, adding 25 times of water, heating at 50 deg.C, reflux extracting;
(2) filtering the product obtained in the step (1) by a 60-mesh screen to obtain a primary filtrate, and then carrying out secondary filtration on the primary filtrate, wherein the aperture is 0.45 um;
(3) adsorption: adsorbing the filtrate obtained in the step (2) by a chromatographic column, eluting, and collecting effluent liquid, wherein the filler of the chromatographic column is macroporous adsorption resin;
(4) concentration: the effluent is subjected to primary concentration by adopting a reverse osmosis membrane with the molecular weight cutoff of 80 at normal temperature until the solid content is more than or equal to 10 percent, and then is subjected to secondary concentration by using an evaporator at the temperature of 85 ℃ until the solid content is more than or equal to 40 percent. Cooling to room temperature;
(5) and (3) drying: and (3) carrying out spray drying on the concentrated solution, wherein the air inlet temperature of the spray drying is 170 ℃, the air outlet temperature is 100 ℃, and the feeding pressure is 9MPa, so as to obtain the theanine dry powder.
Example 3: soft capsule
Raw material dosage:
a composition containing theanine for relieving physical fatigue comprises a capsule core and a capsule core, wherein the capsule core comprises 20 parts by weight of theanine, 35 parts by weight of wolfberry extract, 35 parts by weight of rhizoma polygonati extract, 9 parts by weight of beeswax and 120 parts by weight of soybean oil. The capsule shell consists of 90 parts by weight of gelatin, 90 parts by weight of purified water and 45 parts by weight of glycerol.
The preparation method comprises the following steps: same as example 1
The method for preparing the theanine dry powder from the tea comprises the following steps:
(1) leaching: pulverizing folium Camelliae sinensis, placing in an extractor, adding 20 times of water, heating at 65 deg.C, reflux extracting;
(2) filtering the product obtained in the step (1) by a 60-mesh screen to obtain a primary filtrate, and then carrying out secondary filtration on the primary filtrate, wherein the aperture is 0.45 um;
(3) adsorption: adsorbing the filtrate obtained in the step (2) by a chromatographic column, eluting, and collecting effluent liquid, wherein the filler of the chromatographic column is macroporous adsorption resin;
(4) concentration: performing primary concentration on the effluent by using a reverse osmosis membrane with the molecular weight cutoff of 80 at normal temperature until the solid content is more than or equal to 10%, and performing secondary concentration by using an evaporator at 80 ℃ until the solid content is more than or equal to 40%. Cooling to room temperature;
(5) and (3) drying: and (3) carrying out spray drying on the concentrated solution, wherein the air inlet temperature of the spray drying is 180 ℃, the air outlet temperature is 100 ℃, and the feeding pressure is 8MPa, so as to obtain the theanine dry powder.
Example 4: method for preparing theanine dry powder from tea
(1) Leaching: pulverizing folium Camelliae sinensis, placing in an extractor, adding 20 times of water, heating at 65 deg.C, reflux extracting;
(2) filtering the product obtained in the step (1) by a 60-mesh screen to obtain a primary filtrate, and then carrying out secondary filtration on the primary filtrate, wherein the aperture is 0.45 um;
(3) adsorption: adsorbing the filtrate obtained in the step (2) by a chromatographic column, eluting, and collecting effluent liquid, wherein the filler of the chromatographic column is macroporous adsorption resin;
(4) concentration: the effluent is subjected to primary concentration by adopting a reverse osmosis membrane with the molecular weight cutoff of 80 at normal temperature until the solid content is more than or equal to 10 percent, and then is subjected to secondary concentration by using an evaporator at the temperature of 60 ℃ until the solid content is more than or equal to 30 percent. Cooling to room temperature;
(5) and (3) drying: and (3) carrying out spray drying on the concentrated solution, wherein the air inlet temperature of the spray drying is 180 ℃, the air outlet temperature is 100 ℃, and the feeding pressure is 8MPa, so as to obtain the theanine dry powder.
Example 4: method for preparing theanine dry powder from tea
(1) Leaching: pulverizing folium Camelliae sinensis, placing in an extractor, adding 20 times of water, heating at 65 deg.C, reflux extracting;
(2) filtering the product obtained in the step (1) by a 60-mesh screen to obtain a primary filtrate, and then carrying out secondary filtration on the primary filtrate, wherein the aperture is 0.45 um;
(3) adsorption: adsorbing the filtrate obtained in the step (2) by a chromatographic column, eluting, and collecting effluent liquid, wherein the filler of the chromatographic column is macroporous adsorption resin;
(4) concentration: performing primary concentration on the effluent by using a reverse osmosis membrane with the molecular weight cutoff of 80 at normal temperature until the solid content is more than or equal to 10%, and performing secondary concentration by using an evaporator at 80 ℃ until the solid content is more than or equal to 40%. Cooling to room temperature;
(5) and (3) drying: and (3) carrying out spray drying on the concentrated solution, wherein the air inlet temperature of the spray drying is 180 ℃, the air outlet temperature is 100 ℃, and the feeding pressure is 8MPa, so as to obtain the theanine dry powder.
Example 5:
a composition containing theanine for relieving physical fatigue,
component 1: 40g of theanine dry powder, 40g of medlar extract, 30g of sealwort extract, or,
and (2) component: 40g of theanine dry powder, 20g of medlar extract, 60g of sealwort extract, or,
and (3) component: 40g of theanine dry powder, 80g of medlar extract and 15g of sealwort extract.
Claims (3)
1. Detection method of composition containing theanine
Sample treatment:
injecting the sample solution into a high performance liquid chromatograph, and determining the quality by retention time and ultraviolet-visible absorption spectrum, and determining the quantity by peak area, wherein the corresponding theanine concentration can be obtained from a standard curve by the peak area of a chromatographic peak.
2. The composition containing theanine for relieving physical fatigue, which adopts the detection method of claim 1, is characterized by being prepared from the following raw materials in parts by weight: 20-40 parts of theanine, 20-40 parts of medlar extract, 30-50 parts of sealwort extract, 6-10 parts of beeswax and 140 parts of vegetable oil.
3. The composition containing theanine for relieving physical fatigue, which adopts the detection method of claim 1, is characterized by being prepared from the following raw materials in parts by weight: 30 parts of theanine, 30 parts of a Chinese wolfberry extract, 40 parts of a sealwort extract, 7 parts of beeswax and 130 parts of vegetable oil; wherein the composition is in the form of a soft capsule.
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| CN108828122A (en) * | 2018-06-20 | 2018-11-16 | 精晶药业股份有限公司 | A kind of efficient liquid phase detection method of theanine |
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