CN105248718B - A kind of health beverages and the preparation method and application thereof with liver protecting - Google Patents
A kind of health beverages and the preparation method and application thereof with liver protecting Download PDFInfo
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Abstract
The invention belongs to food processing technology fields, and in particular to a kind of health beverages and the preparation method and application thereof with liver protecting.The health drink is health protection tea, is made of the raw material of following weight parts：5~10 parts of green tea, kamuning spend 1~5 part, 1~5 part of feather cockscomb inflorescence.Preparation method is：Kamuning flower is taken to add water to cook 3 times, 1 hour every time, collecting decoction filtered, and filtrate is concentrated into the thick paste that suitable density is 1.25；Feather cockscomb inflorescence is taken to be ground into coarse powder, it is dry with thick paste mixing, green tea, mixing is added, sieving is fitted into tea bag filter paper bag, is sealed.
Description
Technical field
The invention belongs to food processing technology fields, are related to a kind of health drink, it is specially a kind of have to hepatopathy centainly control
The health protection tea for the treatment of effect.
Background technology
Fatty liver, refer to due to various reasons caused by the excessive lesion of fat accumulation in liver cell.Fatty liver disease is just
The health for seriously threatening compatriots becomes the second largest hepatopathy for being only second to virus hepatitis, has been acknowledged as concealment hepatic sclerosis
Common cause.Fatty liver is a kind of common clinical picture rather than a kind of independent disease.Its clinical manifestation less serious case is asymptomatic,
The severe one state of an illness is violent.In general, fatty liver category invertibity disease, early diagnosing and treating in time can often restore normal.
Nonalcoholic fatty liver refers to lesion main body in lobuli hepatis, mainly with the fatty lesion of liver cell and fat
It is stored as pathological character, and one group clinical pathology syndrome of the patient without excessive drinking history.Now with Chinese people all living creatures
The flat continuous improvement of running water, dietary structure are changing, therefore the incidence presentation of nonalcoholic fatty liver improves year by year
Trend.Diet modification is clinically mainly taken for the treatment of nonalcoholic fatty liver, adheres to that motion exercise is main side
Method, but the effect of clinical treatment is general.
Up to the present, Western medicine there is no the active drug of prevention fatty liver and nonalcoholic fatty liver, be adjusted for a long time with Chinese medicine
The treatment of rationality is best, but the Chinese medicine treatment time for the treatment of fatty liver and nonalcoholic fatty liver is long at present, and curative effect is general
All over poor.
In recent years, inventor is achieved using health tea of the present invention for treating fatty liver and nonalcoholic fatty liver
Preferable curative effect.
In the present invention：
Kamuning flower is Rutaceae Murraya plant kamuningMurraya exotica L. [M. Paniculata (L.) Jack. var. exotica (L.) Huang]And murraya paniculataJackM. paniculata (L.) Jack. [Chalcas paniculata L.]Flower.
Feather cockscomb inflorescence is the inflorescence of Amaranthaceae feather cockscomb platymiscium feather cockscomb Celosia argentea L..
Invention content
The object of the present invention is to provide a kind of health drink with liver protecting effect, which is health care
Tea.
It is a further object of the present invention to provide the preparation methods of the health protection tea.
The purpose of the present invention is what is be accomplished by the following way：
A kind of health drink with liver protecting effect, the health drink are health protection tea, are by following weight parts
Raw material be made：5~10 parts of green tea, kamuning spend 1~5 part, 1~5 part of feather cockscomb inflorescence.
The health drink is made of the raw material of following weight parts：10 parts of green tea, kamuning spend 5 parts, 5 parts of feather cockscomb inflorescence.
The health drink can also be made of the raw material of following weight parts：10 parts of green tea, kamuning spends 2 parts, feather cockscomb
Spend 3 parts.
Above-mentioned health drink is made of following methods：Kamuning flower, feather cockscomb inflorescence are taken, is dried, crushes, with green tea mixing, adopts
Loose tea, compressed tea is made with conventional method.
Above-mentioned health drink can also be made of following methods：Kamuning flower, feather cockscomb inflorescence are taken, is dried, is crushed, it is mixed with green tea
It is even, tea bag is made using conventional method.
Above-mentioned health drink can also be made of following methods：It takes kamuning flower to add water to cook 3 times, 1 hour every time, merges
Decocting liquid, filtration, filtrate are concentrated into the thick paste that suitable density is 1.25；Feather cockscomb inflorescence is taken to be ground into coarse powder, it is dry with thick paste mixing,
Green tea, mixing is added, sieving is fitted into tea bag filter paper bag, is sealed, every bag of weight 2g.
Health drink of the present invention can be used following methods and carry out quality testing：
Using high effective liquid chromatography for measuring chiratin（sweroside）Content：
（1）Chromatographic condition：Chromatographic column is C18Chromatographic column；With ratio for 40~60：40~60-1% glacial acetic acid solution of methanol
For mobile phase；300~330nm of Detection wavelength, 25~45 DEG C of column temperature；
（2）The preparation of reference substance solution：Precision weighs chiratin reference substance and is placed in volumetric flask, and methanol is added to dissolve and dilute
To scale, shake up；It pipettes above-mentioned solution and sets volumetric flask, add methanol constant volume, as a contrast product stock solution；
（3）The preparation of test solution：Health protection tea of the present invention, precision weighing is taken to set in conical flask with cover, first is added in precision
Alcohol, close plug, weighed weight are ultrasonically treated, let cool, then weighed weight, the weight of less loss is supplied with methanol, is shaken up, miillpore filter
Filtering, takes subsequent filtrate to get test solution；
（4）It measures：It is accurate respectively to measure above-mentioned test solution, each 5~20 μ L of reference substance solution, inject high-efficient liquid phase color
Spectrometer is detected.
Health drink of the present invention preferably uses following methods to carry out quality testing：
Using high effective liquid chromatography for measuring chiratin content：
（1）Chromatographic condition：Chromatographic column is C18Chromatographic column；With ratio for 45：55-1% glacial acetic acid solution of methanol is flowing
Phase；Detection wavelength 322nm, 25 DEG C of column temperature；Number of theoretical plate is calculated by chiratin peak should be not less than 4000；
（2）The preparation of reference substance solution：Precision weighs chiratin reference substance 10.00mg and is placed in 100ml volumetric flasks, adds first
Alcohol dissolves and is diluted to scale, shakes up；It pipettes above-mentioned solution 10ml and sets 25ml volumetric flasks, add methanol constant volume, product are store as a contrast
Standby liquid；
（3）The preparation of test solution：Health protection tea 3g of the present invention, precision weighing is taken to set in conical flask with cover, precision is added
Methanol 20ml, close plug, weighed weight are ultrasonically treated 30min, let cool, then weighed weight, the weight of less loss is supplied with methanol, is shaken
Even, 0.45 μm of filtering with microporous membrane takes subsequent filtrate to get test solution；
（4）It measures：It is accurate respectively to measure above-mentioned test solution, each 10 μ L of reference substance solution, inject high performance liquid chromatography
Instrument is detected.
The health drink is used to prepare the purposes in treatment fatty liver, nonalcoholic fatty liver health drink.
Experiment one：Experimental study of the health protection tea of the present invention to rats with nonalcoholic fatty liver disease preventive and therapeutic effect
Health protection tea of the present invention is that a kind of dosage form is simple, tea bag convenient to take.This experiment is built by High-fat diet
Vertical Making Rat Models of Nonalcoholic studies preventive and therapeutic effect of the health protection tea of the present invention to rats with nonalcoholic fatty liver disease, to face
Bed treatment provides foundation.
1 material and method
1.1 material
1.1.1 experimental animal
Healthy Wistar rats 62, cleaning grade, 200 ± 20g of weight are carried by Gansu university of TCM Experimental Animal Center
For.
1.1.2 experimental drug
Health protection tea of the present invention：Prescription：30 kilograms of green tea, kamuning spend 15 kilograms, 15 kilograms of feather cockscomb inflorescence；Preparation method：Take nine li
Fragrant flower adds water to cook 3 times, and 1 hour every time, collecting decoction filtered, and filtrate is concentrated into the thick paste that suitable density is 1.25；Take feather cockscomb
Pollen is broken into coarse powder, dry with thick paste mixing, and green tea, mixing is added, and tea bag is made using conventional method in sieving, per pouch
2 grams.
Compare health protection tea A：Prescription：30 kilograms of green tea, kamuning spends 30 kilograms；Preparation method：Kamuning flower is taken to add water to cook 3
Secondary, 1 hour every time, collecting decoction, filtration, filtrate was concentrated into the thick paste that suitable density is 1.25, dry, addition green tea, mixing,
Sieving, tea bag is made using conventional method, per 2 grams of pouch.
Compare health protection tea B：Prescription：30 kilograms of green tea, 30 kilograms of feather cockscomb inflorescence；Preparation method：It takes feather cockscomb inflorescence to be ground into coarse powder, is added
Tea bag is made using conventional method in green tea, mixing, sieving, per 2 grams of pouch.
Methionine compound choline piece, Jilin Province Huinan Changlong Biochemistry Medicine Co., Ltd production, authentication code：Traditional Chinese medicines
Quasi- word H22026151.
Propylthiouracil Tablets, Suzhou Dawnrays Pharmaceutical Co., Ltd.'s production, authentication code：Chinese medicines quasi-word H20013331.Courage is solid
Alcohol, Tianjin great Mao chemical reagent factories.Lard is commercially available.
1.1.3 experiment reagent
It is total cholesterol assay kit, triglyceride determination kit, high-density lipoprotein cholesterol assay kit, low
Density lipoprotein-cholesterol assay kit, above four kits are provided by Zhejiang Dong Ou bioengineering Co., Ltd.Third
Histidine amino group converting Enzyme（ALT）And Aspartate amino converting Enzyme（AST）Reagent used above is that analysis is pure.
1.1.4 laboratory apparatus
UV-3000 type UV detectors, Shimadzu Corp.II type supercentrifuges of LXJ-, medical point of Shanghai
Analyse instrument plant.BP310S type electronic balances, German Sai Duolisi.SC-329GA type refrigerators, China's Haier Haier.HH-W21 types
Electric heating constant temperature water temperature case, Shanghai Medical constant-temperature instrument factory.OLYMPUS automatic clinical chemistry analyzers, Japan's production.
1.1.5 high lipid food formula
Basal feed 87.8%, cholesterol 2%, propylthiouracil (PTU) 0.2%, lard 10%.（By animal housing of Chinese radiation research institute
Manufacture）
1.2 experimental method
Take Wistar rats 65（Half male and half female）, 200 ± 20g of weight, with basal feed adaptable fed rat one week.
One weekend, random grouping, result were blank control group 10 for the first time according to rat body weight, gave normal diet nursing；Experimental group 52
Only, high lipid food nursing is given.8 weekends randomly selected experimental group female mice and male each 1 of mouse kills inspection, and liver is taken to make Histopathology
It checks, it was demonstrated that reconstructed model success.50 rats of experimental group are randomly divided into model group, positive controls, comparison health care from 9 weeks
Tea A groups, comparison health protection tea B groups, health protection tea group of the present invention, 5 groups, every group each 10, and be numbered and be administered.Methionine compound
Choline piece positive controls give methionine compound choline piece 0.945g/kg daily, and comparison health protection tea A groups are daily to compare health care
Tea A concentrate 2.5g/kg gastric infusions（Gavage volume is 20mL/kg）, it is dense to compare health protection tea B daily to compare health protection tea B groups
Contracting liquid 2.5g/kg gastric infusions（Gavage volume is 20mL/kg）, health protection tea group of the present invention is daily with health protection tea concentrate of the present invention
2.5g/kg gastric infusion（Gavage volume is 20mL/kg）, blank control group and model group gavage give same volume distilled water.Even
Continuous administration 30 days.It weighs in every 2 weeks 1 time.
1.3 observation index and detection method
By rat in fasting 12 hours after the last administration, urethane（0.8g/kg）Intraperitoneal injection of anesthesia, back of the body position are fixed, edge
Abdomen median line is cut, and inferior caval vein puts to death after taking blood 5ml, wins liver, carry out as follows：1. claiming liver wet weights, calculate
Liver weight in wet base index（=liver wet weights/rat body weight × 100% when killing inspection）.2. detaching serum：3000 turns of minutes centrifuge 15 minutes,
Clarification.3. weighing liver lobus dexter site tissue 100mg, add ethyl alcohol：Acetone（1：1）Extract 2.5ml, is homogenized with homogenizer, 4 DEG C
Centrifugation 3000 turns points, 30 minutes, is extracted supernatant, is surveyed according to liver tissue homogenate TG, liver tissue homogenate's TG kit specifications
It is fixed.4. taking tissue in the middle part of 0.5 lobus dexter liver, is fixed with formalin, send Lanzhou pathology department of institute of traditional Chinese medicine, do HE stained slices,
In the degree of light microscopic observation hepatic steatosis, criterion becomes the percentage progress fat change journey that cell accounts for liver cell according to fat
Degree classification.
1.4 statistical method
Experimental data with± s indicates that the variance analysis of measurement data multiple-group analysis compares use Student- two-by-two
Newman-Keulsfa methods q is examined.Statistics is carried out by SAS8.2 software packages.Using two-sided test, P<0.05 has for its difference
Significant.
2 results
2.1 each group rat ordinary circumstances
For each group animal without death, model group rats weight is in rising trend at the end of experiment, and activity is reduced, and hair is dilute
It dredges.
The comparison of 2.2 each group rat liver weight in wet base indexes, hepatic tissue cholesterol and triglycerides is shown in Table 1-1.
Influences of the table 1-1 to liver wet weights index, hepatic tissue cholesterol and triglycerides（± s, n=10）
Note：Compared with blank group, ##P<0.01；Compared with model group, * * P<0.01；*P<0.05；
The comparison of 2.3 each group rat fats
It the results are shown in Table 1-2.
Influence of the table 1-2 health protection teas of the present invention to blood fat（± s, n=10, mmol/L）
Note：Compared with blank group, ##P<0.01；Compared with model group, * * P<0.01；*P<0.05；
2.4 each group rat apolipoproteins than, liver function, Histopathology quantify score comparison
It the results are shown in Table 1-3.
Table 1-3 to apolipoprotein than, liver function, Histopathology quantify score influence（±s）
Note：Compared with blank group, #P<0.05, ##P<0.01；Compared with model group, * P<0.05, * * P<0.01；
3 conclusions
From table 1, table 2, table 3 it can be seen that compared with model group, health protection tea of the present invention can significantly reduce serum total cholesterol
（TC）, triglycerides（TG）, high-density lipoprotein cholesterol（HDL-C）, low density lipoprotein cholesterol（LDL-C）, hepatic tissue
Total cholesterol in homogenate（TC）, triglycerides in liver tissue homogenate（TG）, alanine aminotransferase（ALT）And asparatate
Aminotransferase（AST）, increasing high density lipoprotein cholesterol（HDLC）, apolipoprotein ratio（=HDL-CT/C）, to liver wet weights
Index is without influence.
Pathological examination is shown：Health protection tea of the present invention can significantly mitigate rat model liver cell fatty liver denaturation degrees, with sun
Property comparison medicine methionine compound choline piece piece is compared, no significant difference（P>0.05）.Show health care tea set of the present invention
There is good lipotropic effect：It adjusts blood fat and improves the degree of intrahepatic fat denaturation.
Experiment two：The quality standard research of health protection tea of the present invention
Using chiratin content in high effective liquid chromatography for measuring health protection tea of the present invention
The preparation of 1 sample solution
The preparation of 1.1 reference substance solutions
Precision weighs chiratin reference substance 10.00mg and is placed in 100ml volumetric flasks, adds methanol to dissolve and is diluted to scale,
It shakes up.It pipettes above-mentioned solution 10ml and sets 25ml volumetric flasks, add methanol constant volume, as a contrast product stock solution（40.80 μ g/ of chiratin
ml）.
The preparation of 1.2 test solutions
Prescription：30 kilograms of green tea, kamuning spend 15 kilograms, 15 kilograms of feather cockscomb inflorescence.
Preparation method：Kamuning flower is taken to add water to cook 3 times, 1 hour every time, collecting decoction filtered, and filtrate is concentrated into suitable density
For 1.25 thick paste；Feather cockscomb inflorescence is taken to be ground into coarse powder, it is dry with thick paste mixing, green tea, mixing, sieving, using conventional side is added
Tea bag is made in method, per 2 grams of pouch.
Health protection tea 3g of the present invention, precision weighing is taken to set in conical flask with cover, methanol 20ml is added in precision, and close plug is weighed heavy
Amount is ultrasonically treated 30min, lets cool, then weighed weight, the weight of less loss is supplied with methanol, is shaken up, 0.45 μm of miillpore filter mistake
Filter, takes subsequent filtrate to get test solution.
The preparation of 1.3 negative controls
It takes remaining ingredient in composition in addition to kamuning is spent that the negative controls spent without kamuning are made, presses
Negative control solution is made in " 1.2 ".Go out peak position in chiratin and do not occur absorption peak, illustrate in health protection tea of the present invention other at
Divide and the assay of chiratin is not interfered with.
2 chromatographic conditions
Chromatographic column is SHIMADZU C18Chromatographic column（5 μm, 4.6mm × 150.0mm, Shimadzu Corporation）；With -1% ice vinegar of methanol
Acid solution（45：55）For mobile phase；Detection wavelength 322nm, 25 DEG C of column temperature.Number of theoretical plate should be not less than by the calculating of chiratin peak
4000；With this condition, chiratin separation is good in health protection tea of the present invention.
3 methodological studies.
3.1 the range of linearity
Precision draws above-mentioned chiratin reference substance solution, add methanol dilution at chiratin concentration to be respectively 0.816,1.632,
3.264, the solution of 4.896,6.528,8.160 μ g/ml, successively 10 μ l of sample introduction.With peak area（Y）It is dense with sample introduction for ordinate
Degree（X）Linear regression is carried out for abscissa, obtains regression equation：
Y=3.638 × 104X -1.068 × 104, correlation coefficient r=0.9996.Show chiratin concentration 0.816~
With chromatographic peak area in good linear relationship within the scope of 8.160 μ g/ml.
3.2 precision test
The 10 μ l METHOD FOR CONTINUOUS DETERMINATIONs of chiratin reference substance solution 5 times of a concentration of 4.890 μ g/ml are taken, RSD 0.15% shows instrument
Device precision is good.
3.3 stability test
It takes same test solution to distinguish 10 μ l of sample introduction in 0,2,4,6,8h, records peak area, the results showed that：Test sample is molten
Liquid is 1.60% being placed at room temperature for stabilization in 8h, chiratin peak area RSD.
3.4 repetitive test
By health protection tea of the present invention, parallel extraction prepares 6 parts of test solutions, and sample introduction measures respectively, RSD 1.23%（n=
6）, show that this method repeatability is good.
3.5 sample recovery rates are tested
Precision weighs each 9 parts of the health protection tea of the present invention of known content, the chiratin reference substance of corresponding amount is added, by test sample
The preparation method of solution is handled and is measured.Sample recovery rate test result is shown in Table 2-1.Chiratin average recovery rate is 97.7%,
RSD is 1.66%, shows that this method is accurate, reliable.
Table 2-1 recovery test results（n=9）
3.6 sample sizes measure.Take 4 batches, health protection tea sample of the present invention, according to test solution prepare method, 0.45 μm
Sample introduction after filtering with microporous membrane, different lot number health protection tea contents of the present invention are shown in Table 2-2.
Table 2-2 sample measurement results（n=3）
Specific implementation mode：
Embodiment 1：
Prescription：10 kilograms of green tea, kamuning spend 5 kilograms, 5 kilograms of feather cockscomb inflorescence.
Preparation method：Above-mentioned kamuning flower, feather cockscomb inflorescence are taken, dries, is crushed to 20 mesh, and green tea mixing, using conventional method system
At tea bag, per 2 grams of pouch.
Embodiment 2：
Prescription：10 kilograms of green tea, kamuning spend 2 kilograms, 3 kilograms of feather cockscomb inflorescence.
Preparation method：Above-mentioned kamuning flower, feather cockscomb inflorescence are taken, is dried, is crushed, with green tea mixing, be made and pressed of conventional method
Tea.
Embodiment 3：
Prescription：5 kilograms of green tea, kamuning spend 4 kilograms, 5 kilograms of feather cockscomb inflorescence.
Preparation method：Above-mentioned kamuning flower, feather cockscomb inflorescence are taken, is dried, is crushed, with green tea mixing, loose tea is made using conventional method.
Embodiment 4：
Prescription：30 kilograms of green tea, kamuning spend 15 kilograms, 15 kilograms of feather cockscomb inflorescence.
Preparation method：Kamuning flower is taken to add water to cook 3 times, 1 hour every time, collecting decoction filtered, and filtrate is concentrated into suitable density
For 1.25 thick paste；Feather cockscomb inflorescence is taken to be ground into coarse powder, it is dry with thick paste mixing, green tea, mixing, sieving, using conventional side is added
Tea bag is made in method, per 2 grams of pouch.
Detection method：Using high effective liquid chromatography for measuring chiratin content：
（1）Chromatographic condition：Chromatographic column is SHIMADZU C18Chromatographic column；With ratio for 45：55-1% glacial acetic acid of methanol is molten
Liquid is mobile phase；Detection wavelength 322nm, 25 DEG C of column temperature；Number of theoretical plate is calculated by chiratin peak should be not less than 4000；
（2）The preparation of reference substance solution：Precision weighs chiratin reference substance 10.00mg and is placed in 100ml volumetric flasks, adds first
Alcohol dissolves and is diluted to scale, shakes up；It pipettes above-mentioned solution 10ml and sets 25ml volumetric flasks, add methanol constant volume, product are store as a contrast
Standby liquid；
（3）The preparation of test solution：Health protection tea 3g of the present invention, precision weighing is taken to set in conical flask with cover, precision is added
Methanol 20ml, close plug, weighed weight are ultrasonically treated 30min, let cool, then weighed weight, the weight of less loss is supplied with methanol, is shaken
Even, 0.45 μm of filtering with microporous membrane takes subsequent filtrate to get test solution；
（4）It measures：It is accurate respectively to measure above-mentioned test solution, each 10 μ L of reference substance solution, inject high performance liquid chromatography
Instrument is detected；
（5）As a result：Chiratin content is 54.91 μ g in every gram of health protection tea of the present invention.
In addition, it should be understood that although this specification is described in terms of embodiments, but not each embodiment is only wrapped
Containing an independent technical solution, this description of the specification is merely for the sake of clarity, and those skilled in the art should
It considers the specification as a whole, the technical solutions in the various embodiments may also be suitably combined, forms those skilled in the art
The other embodiment being appreciated that.
Claims (4)
1. a kind of composition for treating nonalcoholic fatty liver, it is characterised in that the composition is by the raw material system of following weight parts
At：10 parts of green tea, kamuning spend 5 parts, 5 parts of feather cockscomb inflorescence；It is made of following methods：Kamuning flower is taken to add water to cook 1~4 time, often
Secondary 0.5~2 hour, collecting decoction filtered, and filtrate is concentrated into the thick paste that suitable density is 1.10~1.30；Feather cockscomb inflorescence is taken to crush
It is dry with thick paste mixing at coarse powder, green tea, mixing is added, sieving is fitted into tea bag filter paper bag.
2. composition as described in claim 1, it is characterised in that the composition is made of following methods：Take kamuning flower plus water
It decocts 3 times, 1 hour every time, collecting decoction filtered, and filtrate is concentrated into the thick paste that suitable density is 1.25；Feather cockscomb inflorescence is taken to be ground into
Coarse powder, it is dry with thick paste mixing, green tea, mixing is added, sieving is fitted into tea bag filter paper bag.
3. composition as described in claim 1, it is characterised in that the composition is detected using following methods：Using efficient
Liquid chromatography for measuring chiratin content：
（1）Chromatographic condition：Chromatographic column is C18Chromatographic column；With ratio for 40~60：40~60-1% glacial acetic acid solution of methanol is stream
Dynamic phase；300~330nm of Detection wavelength, 25~45 DEG C of column temperature；Number of theoretical plate is calculated by chiratin peak should be not less than 4000；
（2）The preparation of reference substance solution：Precision weighs chiratin reference substance and is placed in volumetric flask, adds methanol to dissolve and is diluted to quarter
Degree, shakes up；It pipettes above-mentioned solution and sets volumetric flask, add methanol constant volume, as a contrast product stock solution；
（3）The preparation of test solution：Composition, precision weighing is taken to set in conical flask with cover, methanol is added in precision, and close plug claims
Determine weight, is ultrasonically treated, lets cool, then weighed weight, the weight of less loss is supplied with methanol, is shaken up, filtering with microporous membrane takes continuous filter
Liquid is to get test solution.
（4）It measures：It is accurate respectively to measure above-mentioned test solution, each 5~20 μ L of reference substance solution, inject high performance liquid chromatography
Instrument is detected.
4. composition as claimed in claim 3, it is characterised in that the composition is detected using following methods：Using efficient
Liquid chromatography for measuring chiratin content：
（1）Chromatographic condition：Chromatographic column is C18Chromatographic column；With ratio for 45：55-1% glacial acetic acid solution of methanol is mobile phase；Inspection
Survey wavelength 322nm, 25 DEG C of column temperature；
（2）The preparation of reference substance solution：Precision weighs chiratin reference substance 10.00mg and is placed in 100ml volumetric flasks, adds methanol molten
Scale is solved and be diluted to, is shaken up；It pipettes above-mentioned solution 10ml and sets 25ml volumetric flasks, add methanol constant volume, as a contrast product stock solution；
（3）The preparation of test solution：Composition 3g, precision weighing is taken to set in conical flask with cover, methanol 20ml is added in precision,
Close plug, weighed weight are ultrasonically treated 30min, let cool, then weighed weight, the weight of less loss is supplied with methanol, is shaken up, 0.45 μm
Filtering with microporous membrane takes subsequent filtrate to get test solution.
（4）It measures：It is accurate respectively to measure above-mentioned test solution, each 10 μ L of reference substance solution, high performance liquid chromatograph is injected, into
Row detection.
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