[specific embodiment]
Embodiment 1:
Come further to set forth health food preparation of soft capsule method of the present invention by the following examples:
A) take by weighing each raw material propolis propolis, Fructus Momordicae charantiae extract, Folium Ginkgo extract, chromium picolinate, olive oil, standby;
B) Fructus Momordicae charantiae extract, chromium picolinate are pulverized 120 mesh sieves, used the coubling dilution mixing, made mixture A;
C) Folium Ginkgo extract, the propolis of described weight proportion were pulverized 120 mesh sieves, mix homogeneously makes mixture B;
D) combined mixture A and mixture B make mixture C;
E) olive oil of described weight proportion is put rustless steel ingredients pot internal heating to 50 ℃-70 ℃, added the mixture C mix homogeneously,, take out bubble, just be prepared into health food of the present invention after the cooling through colloid mill 2-3 time.
F) weight proportion that needs by production takes by weighing pharmagel 750 weight portions, medicinal glycerin 450 weight portions, pure water 300 weight portions, in the inputization glue jar, and water-bath, heating, dissolving is stirred, and is incubated standby behind the evacuation in the glue bucket;
G) in 100,000 grades of dust proof workshops of constant temperature and humidity, the health food of step c) gained and the soft capsule shell material of step e) gained are dropped into soft capsule pellet press, pressing cost invention health food soft capsule;
H) the dry back of the soft capsule that step f) made typing is with the 95% edible ethanol flush away surface oil stain that meets the GB/T39412-1994 standard, and with the sterilization of 75% medical ethanol, 28 ℃ of blowing dryings of aerator are bottled and promptly finished the preparation of health food of the present invention.
Embodiment 2
Health food soft capsule blood lipid regulation effect test report of the present invention
1, material and method
1.1 sample: provide by Beijing Yipintang Pharmaceutical Technology Co., Ltd., be the sepia capsule, directly use the sepia content in the capsule to be mixed with the suspension of testing desired concn, human body recommended amounts 2.0g/ days during test with Semen Maydis oil.
1.2 enzymatic assays TC, TG, HDL-C test kit: German HaoMai Co., Ltd and Great Wall, Baoding clinical reagent company produce.
1.3 laboratory animal: select 48 of the cleaning level male SD rats (credit number is SCXK (Ji) 2003-1-003) that Hebei province's Experimental Animal Center breeds for use, body weight 180-220g.
1.4 high lipid food: normal feedstuff 79%, cholesterol 1%, Adeps Sus domestica 10%, fresh-laid egg yellow liquor (normal saline dilution in 1: 1) 10%.
1.5 dosage is selected: the soft capsule human body recommended amounts of health food system of the present invention is 2.0g/ day/60kg body weight.The dose,equivalent of rat is equivalent to 5 times of human body recommended amounts, be every day edible 0.17g/kg body weight be low dosage, respectively establish a dosage group by 3 times and 6 times of low dosage again, be 0.50g/ day/kg body weight (middle dosage) and 1.00g/ the day/kg body weight (high dose), establish the Semen Maydis oil contrast simultaneously, give the Semen Maydis oil of 1.0ml/100g.bw.
1.6 experimental technique: after observing 5 days with normal feedstuff feed rat, get tail blood on an empty stomach, enzymatic assays serum total cholesterol (TC), triglyceride (TG), HDL-C (HDL-C), according to the TC level animal is divided into 4 groups at random: high fat matched group, three are tried thing group (0.17,0.50,1.00g/kg.bw).Tried thing and be mixed with suspension by desired concn with Semen Maydis oil.Begin from formal test, each treated animal is used high lipid food instead, tried the thing group and irritated the soft capsule that stomach gives the health food system of the present invention of various dose, high fat matched group filling stomach gives the Semen Maydis oil with volume, irritate the stomach amount and press 1.0ml/100g.bw calculating, once a day, after continuous 45 days, get the every blood lipids index of tail hematometry on an empty stomach.
2, result
2.1 the soft capsule of health food system of the present invention is to the influence (seeing Table 1) of rat body weight
The soft capsule of table 1 health food system of the present invention is to the influence of rat body weight
Table 1 as seen, in entire test, each dosage treated animal body weight there is no unusually.
2.2 the soft capsule of health food system of the present invention is to the influence of rat fat (see Table 2, table 3)
The soft capsule of table 2 health food system of the present invention is to the influence of Serum TC, TG, HDL-C
Compare * P<0.05 * * P<0.01 with oily matched group
By table 2 as seen, give the soft capsule 45 days of the health food system of the present invention of the above-mentioned various dose of rat oral gavage, middle and high dosage group can make Serum TC significantly reduce (P<0.05, P<0.01), each dosage group all can reduce rat blood serum TG (P<0.01), HDL-C high dose group and matched group significantly raise (P<0.05).
The soft capsule of table 3 health food system of the present invention is to the influence of rat fat level
By table 3 as seen, give the soft capsule 45 days of the health food system of the present invention of the above-mentioned various dose of rat, the basic, normal, high dosage group serum TC percentage rate that on average descends is respectively 0.0,9.0 and 11.0; Basic, normal, high dosage group serum TG decline percentage rate is respectively 23.6,28.3 and 37.7.This shows that the soft capsule of health food system of the present invention can obviously reduce the Serum TC level.
3 brief summaries
With 0.17,0.50, the soft capsule of the health food system of the present invention of 1.00g/kg.bw dosage irritates stomach and gave rat 45 days, can obviously reduce Serum TC, TG level, illustrates that this product has certain auxiliary lipid-lowering function.
Embodiment 3
Health food soft capsule auxiliary hyperglycemic effect test report of the present invention
1, material and method
1.1 sample: the soft capsule that health food system of the present invention is provided by Beijing Yipintang Pharmaceutical Technology Co., Ltd., outward appearance is the sepia capsule, directly use the sepia content in the capsule to make the required suspension of test with Semen Maydis oil during test, the human body recommended amounts is 2.0g/ day/60kg body weight.
1.2 laboratory animal: select for use cleaning level Kunming mouse card that Hebei province's Experimental Animal Center provides number to be SCXK (Ji) 2003-1-003.Male, body weight 24-26g.
1.3 dosage is selected: the soft capsule human body recommended amounts of health food system of the present invention is 2.0g/ day/60kg.bw, the dose,equivalent of mice is equivalent to 10 times of human body recommended amounts, be that to take in 0.33g/kg.bw every day be low dosage, respectively establish a dosage group by 2 times and 3 times of low dosage again, be 0.67g/kg.bw (middle dosage) and 1.00g/kg.bw (high dose), matched group is given Semen Maydis oil, and every day, equal-volume was irritated stomach once, tried thing after 30 days, measured every index.
1.4 instrument and reagent:
Alloxan: SIGMA company produces
The blood sugar detection instrument: Britain Bao Ling Man produces
1.5 experimental technique:
1.5.1 influence to the normal mouse fasting glucose
By the mouse blood sugar value grouping of fasting 3h, select 1 to be matched group at random, 1 is high dose group, every group of 10 animals.Tried thing continuously 30 days, fasting was surveyed fasting blood sugar after 3 hours.
1.5.2 influence to alloxan diabetes model mice fasting glucose
Animal fasting 24h is after the tail vein gives 43mg/kg.bw alloxan, and the 7th day fasting 4h surveys blood glucose value, and the mice of choosing blood glucose value 10-25mmol/L is the hyperglycemia model mice.It is divided into four groups at random, 1 model control group wherein, 3 dosage groups, every group of equal 12 mices (matched group is given Semen Maydis oil).Each dosage group was tried thing 30 days continuously, and fasting was surveyed fasting blood sugar after 3.5 hours.
1.5.3 influence to alloxan diabetes model mice carbohydrate tolerance
Animal fasting 24h is after the tail vein gives 43mg/kg.bw alloxan, and the 7th day fasting 4h surveys blood glucose value, and the mice of choosing blood glucose value 10-25mmol/L is the hyperglycemia model mice.It is divided into four groups at random, 1 model control group wherein, 3 dosage groups, every group of equal 12 mices (matched group is given Semen Maydis oil).Each dosage group was tried thing 30 days continuously, and per os gives glucose 2.0g/kg.bw after last is irritated stomach 15-20min, the blood glucose value of mensuration 0,0.5,2h.1.6 date processing: adopt t check carrying out statistical disposition
2 results
2.1 the soft capsule of health food system of the present invention sees Table 1 to the influence of hyperglycemia model mice body weight
Table 1 is respectively organized the initial body weight of mice, mid-term body weight, body weight in latter stage (g)
By table 1 as seen, per os gives the soft capsule 30 days of the health food system of the present invention of mice various dose, and the body weight of each test group mice is compared there was no significant difference (P>0.05) with matched group.
2.2 the soft capsule of health food system of the present invention is to the influence of normal mouse fasting glucose: see Table 2.
The soft capsule of table 2 health food system of the present invention is to the influence of normal mouse fasting glucose
By table 2 as seen, per os gives the soft capsule 30 days of the health food system of the present invention of normal mouse high dose, the fasting glucose of mice change with administration before there was no significant difference (P>0.05) relatively.
2.3 the soft capsule of health food system of the present invention is to the influence of hyperglycemia mice serum fasting glucose: see Table 3
The soft capsule of table 3 health food system of the present invention is to the influence of hyperglycemia model mice fasting glucose
By table 3 as seen, per os gives the soft capsule 30 days of the health food system of the present invention of mice various dose, and the fasting blood sugar of each dosage group mice reduces than model control group, but difference does not have significance (P>0.05).
2.4 the soft capsule of health food system of the present invention is to the influence of hyperglycemia model mice carbohydrate tolerance: see Table 4.
The soft capsule of table 4 health food system of the present invention is to the influence of hyperglycemia model mice carbohydrate tolerance
With model control group than * P<0.05
By table 4 as seen, per os gives the soft capsule 30 days of the health food system of the present invention of mice various dose, gives each dosage group mice 2.0g/kg.bw glucose, and 0.5h blood glucose value high dose group significantly is lower than model control group (P<0.05).
2.5 the influence of the soft capsule hyperglycemia mice carbohydrate tolerance of health food system of the present invention: see Table 5
The soft capsule of table 5 health food system of the present invention is to the influence of hyperglycemia model mice area under curve
By table 5 as seen, per os gives the soft capsule 30 days of the health food system of the present invention of mice various dose, and from the carbohydrate tolerance experimental result as can be seen, area is compared obvious reduction under the high dose group blood glucose curve with matched group, and difference has significance (P<0.05).
3 brief summaries: per os gave the soft capsule of health food system of the present invention of normal mouse and hyperglycemia mice various dose after 30 days, each dosage group can not reduce the fasting blood sugar of hyperglycemia mice, the carbohydrate tolerance experimental result shows and gives behind the glucose that area obviously reduces under the high dose group blood glucose curve that difference has significance; The normal mouse fasting glucose there is not obvious influence.According to the regulation of " health food check and assessment technique standard " (version in 2003), judge that the soft capsule of health food system of the present invention has the auxiliary hyperglycemic effect.
Embodiment 4
The soft capsule auxiliary hyperglycemic effect on human body test-meal test report of health food system of the present invention
1, material and method
1.1 sample: the soft capsule of health food system of the present invention is provided by Beijing Yipintang Pharmaceutical Technology Co., Ltd., outward appearance sepia soft capsule.Recommended amounts 2.0g/ people/sky.
1.2 study subject: collection after the treatment of diet control or oral antidiabetic drug stable disease, do not need to change types of drugs and dosage, only take adult type ii diabetes people 110 examples of maintenance dose, age is 18--65 year, complication persons such as no severe cardiac, liver, kidney.
1.2.1 diagnostic criteria: fasting glucose 〉=7.8mmol/L, or 2 hours after the meal blood glucose 〉=11.1mmol/L.
1.2.2 the person's of including in standard: all type ii diabetes patients who meets above-mentioned diagnostic criteria, all can carry out the test-meal test.
1.2.3 eliminator's standard:
1.2.3.1 the age 18--65 year with epigenesist, gestation or nursing women.
1.2.3.2 complication such as severe cardiac, liver, kidney are arranged, or are associated with other serious primary disease, the psychotic.
1.2.3.3 the noncooperationist can't judge the infull person of curative effect or data.
1.3 EXPERIMENTAL DESIGN and grouping: be divided at random and be subjected to examination group and matched group.Each group adopts the own control design, is control design between group between two groups.
1.4 test-meal method: 110 routine experimenters, be divided into two groups at random, be test group (55 example) and placebo group (55 example), two groups of crowds are on former hypoglycemic medicine kind of taking and the equal basis of invariable of dosage, add the soft capsule or the placebo of health food system of the present invention respectively, 4 of every days, take (0.5 gram/grain) at twice.The former dietary habit of examination trencherman is constant.
1.5 instrument and reagent: CYSMEX-SY1800I type blood counting instrument, OLYMPUS-AU600 type fully automatic blood biochemistry analyzer, Changchun Di Rui-H500 full automatic urine analyzer.
2 observation index: each measures once every index when test-meal on-test and end.
2.1 effect observation:
2.1.1 observation of symptoms:
Detailed medical history-taking is understood patient's diet situation and activity.
Observe cardinal symptom: thirsty polydipsia, polyorexia, fatigue and weakness, polyuria etc.Add up integrated value by symptom weight (serious symptom 3 minutes, moderate 2 minutes, light disease 1 minute, nothing 0 minute) before and after test-meal, and improve (improving 1 is divided into effectively) with regard to its cardinal symptom, the improvement rate observes the symptoms.
2.1.2 fasting glucose and 2 hours after the meal blood sugar test.
2.1.3 glucose in urine detects.
2.1.4 blood fat (T-CHOL TC, triglyceride TG) detects.
2.2 safety is observed:
2.2.1 blood routine examination: red, numeration of leukocyte, hemoglobin, platelet are measured.
2.2.2 biochemical indicator is measured: serum albumin ALB, total protein TP, the heart, liver, renal function (glutamate pyruvate transaminase ALT, glutamic oxaloacetic transaminase, GOT AST, uric acid UA).
2.2.3 Abdominal B type ultrasonography, electrocardiogram, the fluoroscopy of chest of X line.
3 effect criterion:
Formulate with reference to " health food check and assessment technique standard " (version in 2003).
3.1 effectively: fasting glucose result judges: test back blood glucose descends 〉=10%; The 2h blood sugar effects is judged after the meal: test back blood glucose descends 〉=10%.
3.2 it is invalid: as not reach These parameters.
4 results
4.1 physical data:
Observe 101 examples altogether, soft capsule group male 21 examples of health food system wherein of the present invention, women's 30 examples, 57.0 ± 7.7 years old mean age, average course of disease 5.2 ± 3.3 years; Placebo group male's 24 examples, women's 26 examples, mean age 58.9 ± 5.3, average course of disease 6.2 ± 3.1 years.(annotate: the 9 example persons of being excluded are the noncooperationist)
Situation comparison as table 1 observation is last (X ± SD)
Table 2 examination trencherman takes the hypoglycemic medicine situation
By table 1,2 as seen, every index no significant difference before two groups of test-meals has comparability.
4.2 blood sugar lowering effect
4.2.1 empty stomach and post-prandial glycemia are relatively
Empty stomach and blood glucose comparison in 2 hours after the meal before and after table 3 test-meal (mmol/L, X ± SD)
*Contrast P<0.01 between the expression group; # represents own control P<0.01
The preceding two groups of fasting glucose of test-meal and 2 hours after the meal blood glucose there was no significant differences (P>0.05), the fasting glucose of the soft capsule group of health food system of the present invention and 2 hours after the meal blood glucose is all than obviously reducing (P<0.01) after the test-meal before the test-meal, and its reduction value and placebo group comparing difference remarkable (P<0.01).
4.2.2 empty stomach glucose in urine result
Test-meal group glucose in urine on an empty stomach on average descended 0.299 ± 1.29 fen, and placebo group on average increases by 0.15 ± 1.52 fen, through the Ridit check, tests preceding two groups of there was no significant differences, and there is significant difference (P<0.05) the test back for two groups.After the glucose in urine test-meal of the soft capsule group of health food system of the present invention than obviously reducing before the test-meal.
4.2.3 carbohydrate tolerance changes
Carbohydrate tolerance variation before and after table 4 test-meal (post-prandial glycemia-fasting glucose mmol/L, X ± SD)
Own control #P<0.01
Change obviously (P<0.05) before and after the carbohydrate tolerance test-meal of the soft capsule group of health food system of the present invention, placebo group changes also not obvious, and two groups of comparing differences are significantly (P>0.05) not.
4.3 effect of lowering blood sugar relatively
Table 5 blood glucose value effect statistics
*Compare P<0.05 between two groups
Table 6 symptom value effect statistics
*Compare P<0.05 between two groups
From the blood glucose value effect, the effective percentage of test-meal group is higher than placebo group, and significant difference (P<0.05) is arranged between the two, aspect each cardinal symptom improves, the soft capsule group of health food system of the present invention all is better than placebo group, and significant difference (P<0.05) is arranged between two groups.The soft capsule group that health food system of the present invention is described has the effect that improves the diabetics clinical symptoms preferably.
4.4 Blood Lipid situation:
Blood Lipid comparison before and after table 8 test-meal (X ± SD)
*With preceding relatively P<0.05 of test
Soft capsule group test back test-meal group TC, the TG of health food system of the present invention reduce than test is preceding, and there is significant difference front and back.And the placebo group variation is not remarkable, and front and back do not have significant difference.
4.5 blood safety index observing
Blood safety index variation comparison before and after table 9 test-meal (X ± SD)
Before and after two groups of test-meals, the every index of blood testing does not all have significant difference in normal range and between two groups.
4.6 Chest X-rays, electrocardiogram, ultrasound diagnosis: all experimenters there is no unusually.
5 brief summaries
5.1 the soft capsule of health food system of the present invention has the effect of tangible reduction type ii diabetes people blood glucose.The result shows, the test-meal group fasting glucose 0.93 ± 1.93mmol/L (descending 8.2%) that on average descends after the test-meal, the post-prandial glycemia 2.54 ± 2.79mmol/L (descending 15.2%) that descends, and before and after these two index test-meals and between group statistical significance is arranged more all.The soft capsule that health food system of the present invention is described has certain auxiliary hyperglycemic effect.
5.2 the soft capsule of health food system of the present invention has a better role to hyperglycemia examination trencherman polyphagia, polydipsia, polyuria, symptom such as weak.
5.3 the soft capsule of health food system of the present invention does not all have obvious influence to hematology's conventional index and blood parameters.
5.4 the soft capsule of health food system of the present invention is not observed allergy and other untoward reaction in the test-meal process.
Embodiment 5
The soft capsule auxiliary antilipemic effect on human body test-meal test report of health food system of the present invention
1, material and method
1.1 sample: the soft capsule of health food system of the present invention is provided by Beijing Yipintang Pharmaceutical Technology Co., Ltd., outward appearance sepia soft capsule.Recommended amounts 2.0g/ people/sky.
1.2 study subject: collection after the treatment of diet control or oral lipid lowerers stable disease, do not need to change types of drugs and dosage, only take adult hyperlipidemia patient 110 examples of maintenance dose, the age is in 18--65 year, complication persons such as no severe cardiac, liver, kidney.
1.2.1 diagnostic criteria: T-CHOL 〉=5.2mmol/L, or triglyceride 〉=1.65mmol/L.
1.2.2 the person's of including in standard: all hyperlipemic patients that meets above-mentioned diagnostic criteria, all can carry out the test-meal test.
1.2.3 eliminator's standard:
1.2.3.1 the age 18--65 year with epigenesist, gestation or nursing women.
1.2.3.2 complication such as severe cardiac, liver, kidney are arranged, or are associated with other serious primary disease, the psychotic.
1.2.3.3 the noncooperationist can't judge the infull person of curative effect or data.
1.3 EXPERIMENTAL DESIGN and grouping: be divided into test-meal group and placebo group at random.Each group adopts the own control design, is control design between group between two groups.
1.4 test-meal method: 110 routine experimenters, be divided into two groups at random, be test group (55 example) and placebo group (55 example), two groups of crowds are on former types of drugs of taking and the equal basis of invariable of dosage, add the soft capsule or the placebo of health food system of the present invention respectively, 4 of every days, take (0.5 gram/grain) at twice.The former dietary habit of examination trencherman is constant.
1.5 instrument and reagent: CYSMEX-SY1800I type blood counting instrument, OLYMPUS-AU600 type fully automatic blood biochemistry analyzer, Changchun Di Rui-H500 full automatic urine analyzer.
2 observation index: each measures once every index when test-meal on-test and end.
2.1. blood fat (T-CHOL TC, triglyceride TG, high density lipoprotein HDL-C) detects.
2.2 safety is observed:
2.2.1 blood routine examination: red, numeration of leukocyte, hemoglobin, platelet are measured.
2.2.2 biochemical indicator is measured: serum albumin ALB, total protein TP, the heart, liver function (glutamate pyruvate transaminase ALT, glutamic oxaloacetic transaminase, GOT AST, uric acid UA).
2.2.3 Abdominal B type ultrasonography, electrocardiogram, the fluoroscopy of chest of X line.
3 effect criterion:
Formulate with reference to " health food check and assessment technique standard " (version in 2003).
3.1 effectively: TC reduces>10%; TG reduces>15%; HDL-C rising>0.104mmoL/L.
3.2 it is invalid: as not reach effective standard person.
4 results
4.1 physical data:
Observe 103 examples altogether, soft capsule group male 21 examples of health food system wherein of the present invention, women's 30 examples, 56.8 ± 7.8 years old mean age; Placebo group male's 21 examples, women's 31 examples, 56.9 ± 5.4 years old mean age.(annotate: the 7 example persons of being excluded are the noncooperationist)
Situation comparison as table 1 observation is last (X ± SD)
By table 1 as seen, every index no significant difference before two groups of test-meals has comparability.
4.2 fat-reducing effect
Blood Lipid before and after table 2 test-meal (mmol/L, X ± SD)
*Self match and check P<0.05, compare P<0.05 between # two group
The serum TC, the TG that intervene preceding two groups of crowds do not have significant difference, and after intervening 1 month, the TC of the test-meal group 0.72 ± 1.22mmol/L that descends, the TG 0.59 ± 1.05mmol/L that descends and has significant difference before the intervention, with placebo group significant difference is also arranged.
4.3 blood fat reducing function relatively
Table 3 blood fat value effect statistics
*Compare P<0.05 between two groups
The people that the serum TC of test-meal group and TG lowering of concentration reach significant degree has surpassed 50%, with placebo group significant difference is arranged relatively, illustrate that the soft capsule of health food system of the present invention can significantly improve the serum lipid concentrations of people with hyperlipidemia, and do not have significant difference between two groups of HDL-C.
4.4 blood safety index observing
Blood safety index variation comparison before and after table 4 test-meal (X ± SD)
Before and after two groups of test-meals, the every index of blood testing and does not all have significant difference all in normal range before and after between two groups.
4.5 Chest X-rays, electrocardiogram, ultrasound diagnosis: all experimenters there is no unusually.
5 brief summaries
5.1 the soft capsule of health food system of the present invention makes T-CHOL decline 0.72 ± 1.22mmol/L (rate of descent 10.6%) of hyperlipidemia patient, triglyceride decline 0.59 ± 1.05mmol/L (rate of descent 15.3%) relatively has significant difference with placebo group; And do not have significant difference between high density lipoprotein (HDL-C) and placebo group.Illustrate that health food soft capsule of the present invention has the effect of certain auxiliary antilipemic.
5.2 the soft capsule of health food system of the present invention does not all have obvious influence to hematology's conventional index and blood parameters.
5.3 the soft capsule of health food system of the present invention is not observed allergy and other untoward reaction in the test-meal process.
Embodiment 6
Health food soft capsule of the present invention
The detection method of functional component, content and functional component
One, functional component and content
Project | Index |
Total flavones (in rutin) (mg/g) 〉= | 39.1 |
Total Saponin (in the ginsenoside Re, mg/g) 〉= | 11.5 |
Chromium picolinate (mg/g) 〉= | 0.72 |
Two, the detection method of functional component
(1), the mensuration of total flavones
1 reagent
1.1 Silon
1.2 rutin standard solution: take by weighing the 5.0mg rutin, add dissolve with methanol and be settled to 100mL, promptly get 50ug/ml.
1.3 ethanol methanol analytical pure
2 analytical procedures
2.1 sample is handled:
Take by weighing a certain amount of sample, add the ethanol standardize solution to 25mL, after shaking up, supersound extraction 20min places, and draws supernatant 1.0mL, in evaporating dish, adds the absorption of 1g Silon, flings to ethanol in water-bath, changes chromatographic column then over to.Wash with 20mL benzene earlier, benzene liquid discards, and uses the methanol-eluted fractions flavone then, is settled to 25mL.This liquid is measured absorption value in wavelength 360nm.Be standard substance with the rutin simultaneously, the bioassay standard curve is asked regression equation, calculates general flavone content in the sample.
2.2 rutin standard curve:
Draw the rutin standard solution: 0,1.0,2.0,3.0,4.0,5.0mL in the 10mL color comparison tube, add methanol to scale, shake up, in wavelength 360nm colorimetric.Ask regression equation, calculate general flavone content in the sample.
3 calculating and result represent:
In the formula:
X---content of total flavone in the sample, mg/100g;
A---by standard curve calculate flavone amount in the test solution, ug;
M---sample mass, g;
V1---measure and use volume of sample, mL;
V2---sample standardize solution cumulative volume, mL.
Result of calculation keeps two position effective digitals.
(2), the mensuration (in the ginsenoside Re) of total Saponin
1 reagent
1.1 the Amberlite-XAD-2 macroporous resin, Sigma chemical company, U.S.A..
1.2 n-butyl alcohol, ethanol, perchloric acid, ice ethanol analytical pure.
1.3 neutral alumina chromatography usefulness, the 100-200 order.
1.4 the ginsenoside Re is available from Nat'l Pharmaceutical ﹠ Biological Products Control Institute.
1.5 vanillin solution takes by weighing the 5g vanillin, dissolve with ethanol on the rocks also is settled to 100mL.
1.6 ginsenoside Re's standard solution: accurately take by weighing ginsenoside Re's standard substance 0.020g, with dissolve with methanol and be settled to 10.0mL, promptly every milliliter to contain the ginsenoside Re be 2.0mg.
2 instruments
2.1 tintometer
2.2 chromatographic column
3 experimental procedures
3.1 sample is handled
Take by weighing the sample (it is fixed to contain people's parameter according to sample) about 1.000g, place the 100mL volumetric flask, add a spot of water, ultrasonic 30min, water is settled to 100mL again, shakes up, and places, and draws supernatant 1.0mL and carries out column chromatography.
3.2 column chromatography:
Make the chromatography pipe with the 10mL syringe, interior dress 3cm Amberlite-XAD-2 macroporous resin, on add the 1cm neutral alumina.Earlier wash post with 25mL70% ethanol, discard eluent, reuse 25mL washes post, discard eluent, accurately add the sample solution (seeing 3.1) that 1.0mL has handled well, wash post with 25mL, discard eluent, with 25mL70% ethanol elution ginsenoside, collect eluent in evaporating dish, place 60 ℃ of water-baths to volatilize.Doing colour developing with this uses.
3.3 colour developing:
In the above-mentioned evaporating dish that has volatilized, accurately add 0.2mL5% vanillin ice alcoholic solution, rotate evaporating dish, residue is all dissolved, add 0.8mL perchloric acid again, move into behind the mixing in the 5mL band plug graduated centrifuge tube, heat 10min in 60 ℃ of water-baths, take out, after the ice bath cooling, accurately add ice ethanol 5.0mL, after shaking up, be in standard pipe in the 560nm wavelength with the 1cm colorimetric pool and carry out colorimetric determination.
3.4 standard pipe:
Draw ginsenoside Re's standard solution (2.0mg/mL) 100uL and put in the evaporating dish, be placed on water-bath and volatilize (being lower than 60 ℃), or hot blast drying (it is overheated not make), below operation from " 3.2 column chromatographies .... " rise, identical with sample.Measure absorbance.
4 calculate:
In the formula:
X: total Saponin amount (in the ginsenoside Re) in the sample, mg/100g;
A1: the absorbance of test solution;
A2: the absorbance of titer;
C: standard pipe ginsenoside Re's amount, ug;
V: sample dilution volume, mL;
M: sample mass, g.
Result of calculation keeps two position effective digitals.
(3), the mensuration of chromium picolinate
1 reagent
1.1 the methanol top grade is pure.
1.2 dipotassium hydrogen phosphate, potassium dihydrogen phosphate analytical pure.
1.3 the accurate weighing chromium picolinate of pyridine carboxylic acid chromium standard solution standard substance 0.0100g adds methanol: water=1: 1 also is settled to 100.0mL, if any a small amount of residue, can use the ultrasound wave accelerate dissolution.The every mL of this solution contains the 100ug chromium picolinate.
2 instrument and equipments
2.1 high performance liquid chromatograph: attached UV-detector (UV)
2.2 ultrasonic cleaner
2.3 centrifuge
3 analytical procedures
3.1 sample is handled: get 20 capsules sample mixings, accurately take by weighing a certain amount of sample in the scale test tube, add methanol: water=1: 1 also is settled to 20.0mL, behind the supersound extraction 5min with the centrifugal 3min of 3000rpm/min.Standby behind the 0.45um membrane filtration.
3.2 liquid chromatograph reference conditions
3.2.1 chromatographic column: C18Post 4.6 * 250mm.
3.2.2 column temperature: room temperature
3.2.3 UV-detector: detect wavelength 254nm
3.2.4 mobile phase: 0.125mol/L phosphate buffer: acetonitrile=425: 75
3.2.5 flow velocity: 0.5mL/min
3.2.6 sample size: 10uL
3.2.7 chromatography: measure in 10uL standard solution and the sample solution injecting chromatograph, qualitative with retention time, more quantitative with sample peak height or peak area and standard.
3.3 standard curve prepares, and configuration concentration is 0.0,2.00,5.00,10.0,50.0,100ug/mL chromium picolinate titer, carries out liquid-phase chromatographic analysis under given instrument condition, with peak height or peak area concentration is made standard curve.
3.4 calculate
In the formula: the content of chromium picolinate in the X-sample,
H1-sample peak height or peak area
The C-concentration of standard solution, ug/mL
V-sample constant volume, mL
H2-standard solution peak height or peak area
The M-sample size, g
The foundation of detection method: " health food check and assessment technique standard " (version in 2003)