CN103190500B - Herbal tea for clearing away heat and wetting throat and preparation method of herbal tea - Google Patents

Abstract

The invention relates to herbal tea for clearing away heat and wetting throat and a preparation method of the herbal tea. The herbal tea comprises activated components which comprise the following ingredients: houttuynia cordata, chrysanthemum, mint, emblic leafflower fruit and the seed of boat-fruit sterculia. The herbal tea for clearing away heat and wetting throat has the effects of clearing away heat and toxic materials, promoting the secretion of saliva, wetting throat, relieving restlessness and bringing down the heat. The effect is better than that of the prior art.

Description

Cold tea that a kind of heat-clearing wets one's whistle and preparation method thereof

Technical field

The present invention relates to cold tea, be specifically related to cold tea that a kind of heat-clearing wets one's whistle and preparation method thereof.

Background technology

Cold tea, refers to that the Chinese herbal medicine cold and cool property of medicine and that can clear up in human body heat is decocted to water to be cooked beverage and drink, to eliminate the summer heat in human body in summer, or the illness such as the dry throat pain causing for the treatment of winter.Except clearing heat and detoxicating, cold tea also can dry promote the production of body fluid, relieve inflammation or internal heat, improving eyesight, dissipating bind, detumescence etc., also can control hot eyes headache, dizziness and tinnitus, furuncle swelling toxin and hypertension, can work as cold drink completely summer and drink.

Cordate houttuynia is the dry herb of saururaceae plant houttuynia cordata, has heat-clearing, removing toxic substances, diuresis, detumescence effect, and the antipyretic beverage of exploitation take cordate houttuynia as major ingredient will produce significant Social benefit and economic benefit.

Cold tea Patents about cordate houttuynia has:

Chinese patent application 200910044662.8 discloses a kind of cordate houttuynia cold tea, and it is take cordate houttuynia as major ingredient, and prepared slices of Chinese crude drugs peppermint, Radix Glycyrrhizae, chrysanthemum are auxiliary material, adds sucrose, Sucralose, citric acid, Tea Polyphenols formulated.

Chinese patent application 200910103303.5 discloses a kind of herbal tea, and it is to be that raw material is prepared from by yellow chrysanthemum flower, honeysuckle, the tuber of dwarf lilyturf, cordate houttuynia, dried peppermint leaf, sweet wormwood, Radix Glycyrrhizae, golden cypress.

Chinese patent application 200810170826.7 discloses a kind of medicine-food two-purpose flu medicinal herb tea, and it is to be that raw material is prepared from by Momordica grosvenori, honeysuckle, the sterculia seed, cordate houttuynia, purple perilla, Buddha's hand, mulberry leaf.

At present, cold tea kind take cordate houttuynia as major ingredient is not abundant, especially at the how wet weather of partial heat in summer, and easily gastrointestinal disorder, add somebody and have a liking for pungent, the highly seasoned food of food, can occur to some extent getting angry unavoidably, aphthae, abscess of throat, the symptom such as be off one's feed.

Summary of the invention

The object of this invention is to provide the cold tea that a kind of heat-clearing wets one's whistle.

Another object of the present invention is to provide the preparation method of the cold tea that a kind of heat-clearing wets one's whistle.

The cold tea that a kind of heat-clearing provided by the invention wets one's whistle, contains active component, and its active component contains following composition: cordate houttuynia, chrysanthemum, peppermint, emblic and the sterculia seed.

Concrete, the active component of described cold tea contains the composition of following weight portion: cordate houttuynia 1-12 part, chrysanthemum 0.2-3 part, peppermint 0.2-2 part, emblic 0.2-2 part and sterculia seed 0.2-2 part.

Preferably, the composition that the active component of described cold tea contains following weight portion: cordate houttuynia 3-10 part, chrysanthemum 0.5-2.5 part, peppermint 0.5-1.5 part, emblic 0.5-1.5 part and sterculia seed 0.5-1.5 part.

Further preferably, the active component of described cold tea contains the composition of following weight portion: cordate houttuynia 4-8 part, chrysanthemum 1-2 part, peppermint 0.8-1.2 part, emblic 0.8-1.2 part and sterculia seed 0.8-1.2 part.

In above-mentioned cold tea, also contain auxiliary material, concrete consumption is 80-250 part, and preferably, consumption is 100-200 part, more preferably 120-180 part.

In above-mentioned cold tea:

Described weight portion can be the known unit of weights of field of medicaments such as μ g, mg, g, kg, can be also its multiple, as 1/10,1/100,10 times, 100 times etc.

Described auxiliary material refers to and in cold tea preparation process, for flavoring, adds required supplies.

Described auxiliary material is sweetener, is selected from one or more in white granulated sugar, fructose, stevioside, glycyrrhizin, xylitol, D-sorbite, maltitol, Aspartame.

The present invention also provides a kind of method of preparing cold tea, and the method comprises the following steps:

According to proportioning, take each composition, active component water is decocted 2-3 time, add the water that active component gross weight 8-20 doubly measures at every turn, each decocting time is 20-120 minute, cold heavy, centrifugal after merging filtrate, filter, add sweetener, add water, high-temperature instantaneous sterilization, packing, obtains.

Preferably, the preparation method of cold tea provided by the invention, the method comprises the following steps:

According to proportioning, take each composition, active component is mixed, water decocts 2-3 time, the water that each active component gross weight 8-20 doubly measures, each decocting time is 20-120 minute, and decoction temperature is 70-80 ℃, filters, merging filtrate, cold heavy 8-16 hour in 4 ℃ of environment, with the centrifugal 20-40min of speed of 3000-5000r, filter, add sweetener, stir, add water, 120-140 ℃ of sterilization 1-20 second, filling after liquid is cooled to 0-95 ℃, be inverted coolingly, packing, obtains.

The present invention also provide cold tea clearing heat and detoxicating in preparation treatment, promote the production of body fluid wet one's whistle, the application in the health products of relieving restlessness annealing.

Cold tea provided by the invention can pack in pop can or plastic bottle, and concentration is 2-10mg crude drug/mL, is preferably 3-6mg crude drug/mL.

The cold tea that heat-clearing provided by the invention wets one's whistle has the following advantages:

1, in the cold tea that heat-clearing provided by the invention wets one's whistle:

Cordate houttuynia is fresh herb or the dry aerial parts of saururaceae plant houttuynia cordata.Taste is pungent, cold nature.Have clearing heat and detoxicating, the carbuncle that disappears apocenosis, the effect of inducing diuresis for treating strangurtia.Be used for the treatment of lung carbuncle pyemesis, phlegm heat panting is coughed, hot dysentery, and heat is drenched, carbuncle sore tumefacting virus.

Peppermint is the dry aerial parts of labiate peppermint.Taste is pungent, cool in nature.There is dispelling wind and heat from the body, the clear sharp head, relieve sore throat, promoting eruption, the effect of soothing the liver promoting the circulation of qi.Be used for the treatment of anemopyretic cold, wind-warm syndrome is from the beginning of, headache, hot eyes, and larynx numbness, aphtha, rubella, measles, the chest side of body is swollen vexed.

Chrysanthemum is the dry capitulum of feverfew chrysanthemum.Taste is sweet, bitter, cold nature.There is loose wind heat-clearing, flat liver improving eyesight, clearing heat and detoxicating effect.Be used for the treatment of anemopyretic cold, have a headache dizzy, red eye, swell pain, eyes is dim-sighted, sore pyogenic infections.

Emblic is the dry mature fruit of euphorbia plant emblic.Taste is sweet, sour, puckery, cool in nature.There is clearing heat and cooling blood, digestion-promoting spleen-invigorating, the effect of the cough-relieving of promoting the production of body fluid.Be used for the treatment of blood-head blood stasis, indigestion, abdominal distension, cough, laryngalgia, dry.

The sterculia seed is the dry mature seed of the Sterculiaceae plant sterculia seed.Taste is sweet, cold in nature.There are clearing heat and moistening lung, relieving sore-throat to restore voice, the effect relaxing bowel.Be used for the treatment of lung heat celostomia, dry cough without phlegm, pharyngodynia with dryness, thermojunction just closes, headache hot eyes.

Mentioned component share total clearing heat and detoxicating, promote the production of body fluid wet one's whistle, the effect of relieving restlessness annealing.

2,, in cold tea provided by the invention, in peppermint, chrysanthemum and the sterculia seed, the volatile oil material of fragrance can significantly be covered the fishlike smell of cordate houttuynia, makes cold tea have taste fragrance, free from extraneous odour.

The preparation technology of cold tea provided by the invention uses 70-80 ℃ of water extraction to get and 120-140 ℃ of high-temperature short-time sterilization, has extracted fully the active ingredient in medicinal material, and it is refrigerant, sweet and have back a sweet mouthfeel to have retained cold tea.

3, that the cold tea that heat-clearing provided by the invention wets one's whistle has is clearing heat and detoxicating, promote the production of body fluid wet one's whistle, the effect of relieving restlessness annealing, effect is better than prior art.

The specific embodiment

Following examples are used for illustrating the present invention, but are not used for limiting the scope of the invention.

Embodiment 1: the cold tea that heat-clearing wets one's whistle

1, preparation method:

Cordate houttuynia 10kg, the chrysanthemum 2kg, peppermint 2kg, emblic 2kg, the sterculia seed 2kg that get wash clean, mix, and adds the decocting of 8 times of amounts of weight summation to boil 3 times, each decocting time is 20 minutes, and decocting temperature is 70 ℃, filters, merging filtrate, cold sinking 8 hours in 4 ℃ of environment, with the centrifugal 20min of speed of 3000r, filter, add the aqueous solution containing 800kg white granulated sugar, stir, add water to 4500L, 125 ℃ of sterilizations 5 seconds, filling after liquid is cooled to 95 ℃, be inverted coolingly, packing, obtains.

2, specification: 500ml/ plastic bottle, containing crude drug 4.0mg/mL.

Embodiment 2: the cold tea that heat-clearing wets one's whistle

1, preparation method:

Get cordate houttuynia 12kg, chrysanthemum 3kg, peppermint 2kg, emblic 2kg, the sterculia seed 2kg of wash clean, mix, add the decocting of 20 times of amounts of weight summation to boil 2 times, each decocting time is 30 minutes, and decocting temperature is 80 ℃, filters, merging filtrate, cold sinking 10 hours in 4 ℃ of environment, with the centrifugal 30min of speed of 4000r, filters, add the Aspartame aqueous solution containing 250kg, stir, add water to 4800L, 130 ℃ of sterilizations 15 seconds, it is filling after liquid is cooled to 0 ℃, be inverted coolingly, packing, obtains.

2, specification: 300ml/ plastic bottle, containing crude drug 4.38mg/mL.

Embodiment 3: the cold tea that heat-clearing wets one's whistle

1, preparation method:

Get cordate houttuynia 3kg, chrysanthemum 0.5kg, peppermint 0.5kg, emblic 0.5kg, the sterculia seed 0.5kg of wash clean, mix, add the decocting of 10 times of amounts of weight summation to boil 3 times, each decocting time is 50 minutes, and decocting temperature is 80 ℃, filters, merging filtrate, cold sinking 12 hours in 4 ℃ of environment, with the centrifugal 20min of speed of 4000r, filters, add the stevioside sweet solution containing 100kg, stir, add water to 1250L, 140 ℃ of sterilizations 1 second, it is filling after liquid is cooled to 45 ℃, be inverted coolingly, packing, obtains.

2, specification: the bottling of 330ml/ metal can, containing crude drug 4.0mg/mL.

Embodiment 4: the cold tea that heat-clearing wets one's whistle

1, preparation method:

Get cordate houttuynia 10kg, chrysanthemum 2.5kg, peppermint 1.5kg, emblic 1.5kg, the sterculia seed 1.5kg of wash clean, mix, add the decocting of 18 times of amounts of weight summation to boil 2 times, each decocting time is 60 minutes, and decocting temperature is 70 ℃, filters, merging filtrate, cold sinking 10 hours in 4 ℃ of environment, with the centrifugal 40min of speed of 5000r, filters, add the fructose water solution containing 200kg, stir, add water to 4250L, 120 ℃ of sterilizations 20 seconds, it is filling after liquid is cooled to 60 ℃, be inverted coolingly, packing, obtains.

2, specification: 500ml/ plastic bottle, containing crude drug 4mg/mL.

Embodiment 5: the cold tea that heat-clearing wets one's whistle

1, preparation method:

Get cordate houttuynia 4kg, chrysanthemum 1kg, peppermint 0.8kg, emblic 0.8kg, the sterculia seed 0.8kg of wash clean, mix, add the decocting of 12 times of amounts of weight summation to boil 2 times, each decocting time is 90 minutes, and decocting temperature is 75 ℃, filters, merging filtrate, cold sinking 16 hours in 4 ℃ of environment, with the centrifugal 30min of speed of 4000r, filters, add the aqueous solution containing 120kg white granulated sugar, stir, add water to 1800L, 140 ℃ of sterilizations 5 seconds, it is filling after liquid is cooled to 20 ℃, be inverted coolingly, packing, obtains.

2, specification: the bottling of 300ml/ metal can, containing crude drug 4.11mg/mL.

Embodiment 6: the cold tea that heat-clearing wets one's whistle

1, preparation method:

Get cordate houttuynia 8kg, chrysanthemum 2kg, peppermint 1.2kg, emblic 1.2kg, the sterculia seed 1.2kg of wash clean, mix, add the decocting of 15 times of amounts of weight summation to boil 3 times, each decocting time is 120 minutes, and decocting temperature is 75 ℃, filters, merging filtrate, cold sinking 12 hours in 4 ℃ of environment, with the centrifugal 40min of speed of 5000r, filters, add the aqueous solution containing 180kg xylitol, stir, add water to 3000L, 130 ℃ of sterilizations 10 seconds, it is filling after liquid is cooled to 40 ℃, be inverted coolingly, packing, obtains.

2, specification: 1.5L/ plastic bottle, containing crude drug 4.53mg/mL.

Embodiment 7: the cold tea that heat-clearing wets one's whistle

1, preparation method:

Get cordate houttuynia 6kg, chrysanthemum 1.4kg, peppermint 1kg, emblic 1kg, the sterculia seed 1kg of wash clean, mix, add the decocting of 12 times of amounts of weight summation to boil 3 times, each decocting time is 120 minutes, and decocting temperature is 75 ℃, filters, merging filtrate, cold sinking 12 hours in 4 ℃ of environment, with the centrifugal 40min of speed of 4500r, filters, add the aqueous solution containing 156kg maltitol, stir, add water to 2400L, 130 ℃ of sterilizations 12 seconds, it is filling after liquid is cooled to 40 ℃, be inverted coolingly, packing, obtains.

2, specification: 500ml/ plastic bottle, containing crude drug 4.33mg/mL.

Comparative example 1: prepare cold tea with reference to the embodiment 1 in CN200910044662.8

Comparative example 2: prepare cold tea with reference to the embodiment 2 in CN200810170826.7

Experimental example 1: product detects

To embodiment 1-7 and comparative example 1 detect the amount, total acid content of its soluble solid, containing female amount (total female and organic female content), lead tolerance, copper content, zinc content, stanniferous amount, iron-holder, BHC and DDT residual quantity, methamidophos residue amount, potassium sorbate amount, sulfur dioxide residual quantity, salmonella, staphylococcus aureus, Shigella, total plate count, Escherichia coli, yeast and mold number, concrete detection method is:

1, the amount of soluble solid: be GB/T12143-2008 according to standard number, standard name is the amount that the assay method of soluble solid in beverage universaling analysis method is measured soluble solid, and concrete detection method is:

1. reagent and solution:

Ether, ethanol

2. instrument:

Abbe refractometer or other refractometers, tissue mashing machine

3. Specimen Determination:

Before measuring, by specification is proofreaied and correct refractometer, with Abbe refractive power, counts example, other refractometer by specification operations; Separately refractometer two and prism, dips in ether or ethanol is cleaned with absorbent cotton; The glass bar that melts circle with end dips 2-3 of test solutions, drips in refractometer prism facets central authorities (noting not making glass bar touch minute surface); Rapid closing prism, standing 1mh, makes test solution evenly without bubble, and is full of the visual field; Alignment light source, observes objective lens by eyepiece.Regulate alidade, make the visual field be divided into two of light and shades, then rotary fine adjustment spiral, make terminator clear, and make its line of demarcation just on the right-angled intersection point of objective lens.Read percentage or index of refraction in the eyepiece visual field, and record prism temperature.

2, total acid content: be GB/T12456-2008 according to standard number, standard name is that the method in the mensuration of total acid in food is measured total acid content, and concrete detection method is:

1. reagent and solution:

0.1mol/L Standard Volumetric Solutions for Sodium Hydroxide

0.01mol/L Standard Volumetric Solutions for Sodium Hydroxide

0.05mol/L Standard Volumetric Solutions for Sodium Hydroxide

1% phenolphthalein solution

2. instrument:

Tissue mashing machine, water-bath, mortar, condenser pipe

3. the preparation of test solution

By the quick Filter paper filtering of sample, collect filtrate, for measuring.

Take 10g-50g sample, be accurate to 0.001g, be placed in 100ml beaker, the content in beaker is transferred in 250ml volumetric flask with approximately 80 ℃ of water that boiled.Be placed in boiling water bath and boil 30min, take out, be cooled to room temperature, use the water boiling to be settled to 250ml.Use quick Filter paper filtering.Collect filtrate, for measuring.

4. the mensuration of sample

Take 25.000g-50.000g test solution, make it, containing 0.035-0.07g acid, to be placed in 250ml triangular flask.Add 40ml-60ml water and 0.2ml1% phenolphthalein indicator, with 0.1mol/L Standard Volumetric Solutions for Sodium Hydroxide, be titrated to blush 30s and do not fade.Record consumes the numerical value (V of the volume of 0.1mol/L Standard Volumetric Solutions for Sodium Hydroxide ₁).

Blank test: water replaces test solution, record consumes the numerical value (V of the volume of 0.1mol/L Standard Volumetric Solutions for Sodium Hydroxide ₂).

Computing formula:

$X=\frac{c\×(V_1-V_2)\×K\×F}{m}\×1000$

In formula:

X is total acid content (g/kg);

C is the numerical value accurately (mol/L) of Standard Volumetric Solutions for Sodium Hydroxide concentration;

V ₁during for burette test, consume the numerical value (ml) of the volume of Standard Volumetric Solutions for Sodium Hydroxide;

V ₂during for blank test, consume the numerical value (ml) of the volume of Standard Volumetric Solutions for Sodium Hydroxide;

K is sour conversion coefficient: malic acid, 0.067; Acetic acid, 0.060; Tartaric acid, 0.075; Citric acid, 0.064; Citric acid, 0.070(is containing a part crystallization water); Lactic acid, 0.090; Hydrochloric acid, 0.036; Phosphoric acid, 0.049;

F is the extension rate of test solution;

M is the numerical value (g) of the quality of sample.

3, containing female amount: be GB/T5009.11-2003 according to standard number, standard name is that the method in the mensuration of total Arsenic in Food and inorganic arsenic is measured arsenic content.

1. reagent, test solution:

KI (500g/L)+thiourea solution (50g/L) (1+1)

Sodium hydroxide solution (400g/L) and sodium hydroxide solution (100g/L)

Sulfuric acid (1+1)

Liquor argenti nitratis ophthalmicus (8g/L): take 4.0g silver nitrate in 500ml beaker, add suitable quantity of water dissolve after add 30ml nitric acid, add water to 500ml, store in brown bottle.

Poly-vinyl alcohol solution (4g/L): take 0.4g polyvinyl alcohol (degree of polymerization 1500-1800) in small beaker, add 100ml water, in boiling water bath, heat, be stirred to dissolving, insulation 10min, taking-up lets cool standby.

Absorption liquid: get the each portion of liquor argenti nitratis ophthalmicus (8g/L) poly-vinyl alcohol solution (4g/L), add the ethanol (95%) of two parts of volumes, mix as absorption liquid.During use, now join.

Potassium borohydride sheet: potassium borohydride and sodium chloride are mixed and finely ground by 1:4 mass ratio, after fully mixing, on tablet press machine, make diameter 10mm, the tablet of thick 4mm, every is 0.5g.Avoid compressing tablet when wet weather.

Lead acetate (100g/L) cotton: absorbent cotton is steeped in lead acetate solution (100g/L), squeeze and go redundant solution after several minutes, spread cotton out, after 80 ℃ of oven dry, store in wide-mouth vial.

Citric acid (1.0mol/L)-ammonium citrate (1.0mol/L): take 192g citric acid, 243g ammonium citrate, be diluted to 1000ml after being dissolved in water.

Arsenic standard reserving solution: take through 105 ℃ of dry 1h and be placed in arsenic trioxide 0.1320g that drier is cooled to room temperature in 100ml beaker, add 10ml sodium hydroxide solution (2.5mol/L), after to be dissolved, add 5ml perchloric acid, 5ml sulfuric acid, put and on electric hot plate, be heated to emit white cigarette, after cooling, proceed in 1000ml volumetric flask, and dilute with water is settled to scale.Every milliliter of this solution is containing arsenic (pentavalent) 0.100mg.

Arsenic Standard Applying Solution: draw 1.00ml arsenic standard reserving solution in 100ml volumetric flask, be diluted with water to scale.Every milliliter of this solution is containing arsenic (pentavalent) 1.00 μ g.

Methyl red indicator (2g/L): take 0.1g methyl red and be dissolved in 50ml ethanol (95%).

2. instrument: spectrophotometer, hydrogen arsenide generating device

3. sample pretreating:

Draw 10ml sample in 250ml triangular flask, low-temperature heat adds 2ml perchloric acid, 10ml nitric acid, 2.5ml sulfuric acid (1+1) after removing ethanol or carbon dioxide, after placement a few hours (or spending the night), put on electric hot plate and heat, brown if solution becomes, should add nitric acid makes organic substance decomposing complete, take off and let cool, add 15ml water, then be heated to emit white cigarette, take off, with 20ml moisture, for several times digestive juice is quantitatively proceeded in 100ml arsenic hydride generating bottle.Make reagent blank simultaneously.

The preparation of standard series: in 6 100ml arsenic hydride generating bottles, add successively arsenic Standard Applying Solution 0,0.25,0.5,1.0,2.0,3.0ml, add water to respectively 3ml, then add 2.0ml sulfuric acid (1+1).

4. the mensuration of sample

In sample and standard arsenic hydride generating bottle, add respectively 0.1g ascorbic acid, 2.0ml KI (500g/L) thiourea solution (50g/L), put in boiling water bath, heat 5min(now in bottle a temperature must not exceed 80 ℃), taking-up lets cool, add 1 of methyl red indicator, add about 3.5ml sodium hydroxide solution, with sodium hydroxide solution, be adjusted to solution and be just yellow, add 1.5ml citric acid-ammonium citrate solution, add water to 40ml, add a potassium borohydride tablet, by the conduit that is plugged with lead acetate cotton, be connected with the absorption tube that fills 4.0ml absorption liquid immediately, frequently shake arsenic hydride generating bottle, react and after 5 minutes, add again a potassium borohydride tablet, continue reaction 5min.Take off absorption tube, use 1cm cuvette, at 400nm wavelength, take standard pipe zero pipe, adjust absorbance as zero, measure each pipe absorbance.By each standard series pipe arsenic content to extinction

Degree drawing standard curve or calculating regression equation.

Computing formula: $X=\frac{A\×1000}{m\×1000}$

In formula:

X is the content (mg/kg or mg/L) of arsenic in sample;

A measures the quality that checks in from calibration curve with digestive juice (μ g);

M is sample mass or volume (g or ml).

4, lead tolerance: be GB/T5009.12-2010 according to standard number, standard name is that the method in the mensuration of food security national standard Pb in food is measured lead tolerance, and concrete detection method is:

1. reagent and solution:

Nitric acid, ammonium persulfate, hydrogen peroxide (30%), perchloric acid, nitric acid (1+1), nitric acid (0.5mol/L), nitric acid (1mo1/L), ammonium dihydrogen phosphate (20g/L);

Mixed acid: nitric acid ten perchloric acid (9+1).Getting 9 parts of nitric acid mixes with 1 part of perchloric acid;

Plumbous standard reserving solution: accurately take 1.000g metallic lead (99.99%), gradation adds a small amount of nitric acid (4.5), heating for dissolving, total amount is no more than 37ml, moves into 1000ml volumetric flask, adds water to scale.Mix.Every milliliter of this solution is containing 1.0mg lead;

Plumbous standard solution: draw plumbous standard reserving solution 1.0ml at every turn in 100ml volumetric flask, add nitric acid (4.6) to scale.So through being repeatedly diluted to every milliliter containing 10.0ng, 20.0ng, 40.0ng, 60.0ng, the standard solution of 80.0ng lead.

2. instrument:

Muffle furnace, balance, dry insulating box, porcelain crucible, adjustable electric furnace, Atomic Absorption Spectrometer, graphite stove and plumbous hollow cathode lamp

3. sample pretreating:

Take sample 1g-5g(and be accurate to 0.001g) in conical flask or high pin beaker, put several beades, add 10ml mixed acid (4.9), add a cover soaked overnight, add a little funnel clears up on electric furnace, if change brownish black, add again mixed acid, until Mao Baiyan, digestive juice is water white transparency or slightly yellow, lets cool, sample digestive juice is washed or is filtered in (depending on the salinity of sample after digestion) 10ml-25ml volumetric flask with dropper, water repeatedly washs conical flask or high pin beaker on a small quantity, and washing lotion is incorporated in volumetric flask and is settled to scale, mixes standby; Make reagent blank simultaneously.

4. the mensuration of sample

Instrument condition: be adjusted to optimum state according to instrument performance separately.Reference conditions are wavelength 283.3nm, slit 0.2nm-1.0nm, lamp current 5mA-7mA, 120 ℃ of baking temperatures, 20s; 450 ℃ of ashing temperatures, continue 15s-20s, atomization temperature: 1700 ℃-2300 ℃, continue 4s-5s, and background correction is deuterium lamp or Zeemen effect.

Specification Curve of Increasing: draw the plumbous standard solution 10.0ng/ml(or the μ g/L that prepare above), 20.0ng/ml(or μ g/L), 40.0ng/ml(or μ g/L), 60.0ng/ml(or μ g/L), 80.0ng/ml(or μ g/L) each 10 μ L, inject graphite furnace, record its light absorption value and try to achieve light absorption value and the one-variable linear regression equation of concentration relationship.

Specimen Determination: draw respectively the each 10 μ L of sample liquid and reagent blank liquid, inject graphite furnace, record its light absorption value, try to achieve lead content in sample liquid in the one-variable linear regression equation of substitution standard series.

The use of matrix modifier: to there being interference sample, injecting appropriate matrix modifier ammonium dihydrogen phosphate (4.8) (be generally 5 μ L or measure with sample is same) and eliminate and disturb.Also the matrix modifier ammonium dihydrogen phosphate of equivalent in the time of will adding with Specimen Determination while drawing plumbous calibration curve.

Computing formula:

In formula: $X=\frac{(c_1-c_0)\×V\×1000}{m\×1000\×1000}$

X is lead content in sample (mg/kg or mg/L);

C ₁for measuring lead content (ng/ml) in sample liquid;

C ₀for lead content in blank solution (ng/ml);

V is the quantitative cumulative volume of sample digestive juice (ml);

M is sample mass or volume (g or ml).

5, copper content: be GB/T5009.13-2003 according to standard number, standard name is that the method in the mensuration of copper in food is measured copper content:

1. reagent and solution:

Nitric acid, benzinum, nitric acid (10%), nitric acid (0.5%), nitric acid (1+4), nitric acid (1+6), copper titer, copper standard solution I, copper standard solution II;

2. instrument:

Muffle furnace, bruisher, atomic absorption spectrophotometer

3. the mensuration of sample

Absorption 0,1.0,2.0,4.0,6.0,8.0,10.0ml copper standard solution I (1.0 μ g/ml), be placed in respectively 10ml volumetric flask, adds nitric acid (0.5%) and be diluted to scale, mixes.

Condition determination: lamp current 3mA-6mA, wavelength 324.8nm, spectral band-width 0..5nm, air mass flow 9L/min, acetylene flow 2L/min, lamp holder height 6mm, deuterium lamp background correction.With copper standard liquid content and corresponding absorbance, drawing standard curve or calculated line regression equation, absorption of sample value and curve relatively or substitution Solving Equations obtain content.

(1ml=0.10 μ g), is placed in respectively 10ml volumetric flask, adds nitric acid (0.5%) and is diluted to scale, shakes up to draw 0,1.0,2.0,4.0,6.0,8.0,10.0ml copper standard solution II.Copper titer 10-20 μ L in sample liquid after treatment, reagent blank liquid and each volumetric flask is imported respectively and is adjusted to optimum condition graphite furnace atomizer and measures.Reference conditions: lamp current 3-6mA, wavelength 324.8nm, spectral band-width 0.5nm, protective gas 1.5L/min(atomized stage is stopped the supple of gas or steam).Operating parameter: dry 90 ℃, 20s; Ashing, 20s; Be raised to 800 ℃, 20s; 2300 ℃ of atomization, 4s.With copper standard liquid II series content and corresponding absorbance, drawing standard curve or calculated line regression equation, absorption of sample value and curve relatively or substitution Solving Equations obtain content.

Computing formula:

$X=\frac{(A_1-A_2)\×V\×1000}{m\×1000}$

In formula:

X ₁for the content (mg/Kg or mg/L) of copper in sample;

A ₁for the content (μ g/ml) of copper in test sample;

A ₂for the content (μ g/ml) of copper in reagent blank liquid;

V ₁for the cumulative volume after sample treatment (ml);

M ₁for sample quality (volume) (g or ml).

6, zinc content: be GB/T5009.14-2003 according to standard number, the method in the mensuration that standard name is Zinc in Foods is measured zinc content, and concrete grammar is:

1. reagent and solution:

Sodium acetate solution (2mol/L), acetic acid (2mol/L), acetic acid-acetate buffer, ammoniacal liquor (1+1), hydrochloric acid (0.02mol/L), hydroxylamine hydrochloride solution (200g/L), hypo solution (250g/L), dithizone-carbon tetrachloride solution (0.1g/L), dithizone are used liquid, Zinc standard solution, zinc standard solution, phenol red indicator solution (1g/L)

2. instrument: spectrophotometer

3. sample digestion:

Draw 10ml or 20ml sample, be placed in 250ml-500ml nitrogen fixing bottle, addend grain bead, first removes ethanol or carbon dioxide with little fire heating, then adds 5ml-10ml nitric acid-perchloric acid mixed liquor, and after mixing, little fire slowly heats, and the effect for the treatment of relaxes, and lets cool.Along a bottle wall, add 5ml or 10ml sulfuric acid, then heat, start to become when brown to liquid in bottle, constantly along bottle wall, drip nitric acid-perchloric acid mixed liquor complete to organic matter decomposition.Add high flame, to producing white cigarette, until mouthful white cigarette of bottle emit clean after, in bottle liquid produce again white cigarette for digestion completely, this solution is should be clear and bright colourless or be with micro-yellow, lets cool.Add 20ml water boil, then go remaining nitric acid to till producing white cigarette, so process twice, let cool.Solution after cold is moved in 50ml or 100ml volumetric flask, wash nitrogen fixing bottle with water, washing lotion is incorporated in volumetric flask, lets cool, and adds water to scale, mixes.The every 10ml of solution after constant volume is equivalent to 1g sample, quite adds sulfuric acid amount 1ml.Get and the nitric acid-perchloric acid mixed liquor and the sulfuric acid that digest sample same amount, by Same Way, do reagent blank test.

4. the mensuration of sample

Draw the rear sample solution of constant volume of 5.0ml-10.0ml digestion and the reagent blank liquid of same amount, be placed in respectively 125m1 separatory funnel, add 5m1 water, 0.5ml hydroxylamine hydrochloride solution (200g/L), shake up, add again 2 phenol red indicator, be adjusted to redness with ammoniacal liquor (1+1), then add 2.Add again 5ml dithizone-carbon tetrachloride solution (0.1g/L), violent jolting 2min, stratification.Carbon tetrachloride layer is moved in another separatory funnel, and water layer extracts with a small amount of dithizone-carbon tetrachloride solution jolting, and each 2ml-3ml, until dithizone-carbon tetrachloride solution is green constant.Merge extract, use 5m1 water washing, hydrochloric acid carbon tetrachloride layer for (0.02mol/L) extracts 2 times, at every turn 10ml, and violent jolting 2min during extraction, merging hydrochloric acid (0.02mol/L) extract, and wash away residual dithizone with a small amount of carbon tetrachloride.

Absorption 0.0,1.0,2.0,3.0,4.0,5.0ml zinc standard solution, be placed in respectively 125ml separatory funnel, respectively adds 0.02mol/L hydrochloric acid to 20ml.In funnel, respectively add hydrochloric acid to 20m1.Then add respectively 10ml acetic acid-acetate buffer, 1m125% hypo solution, shake up, respectively add 10.0ml dithizone to use liquid, violent jolting 2min.After stratification, carbon tetrachloride layer is filtered in lcm cuvette through absorbent cotton, with zero pipe, regulate zero point, in wavelength 530nm place, measure absorbance, drawing standard curve or obtain regression equation, according to recording absorbance, from calibration curve, check in the content that is equivalent to zinc, or absorbance substitution regression equation is tried to achieve to the content of zinc.

7, stanniferous amount: be GB/T5009.16-2003 according to standard number, the method in the mensuration that standard name is Tin in Food is measured stanniferous amount, and concrete grammar is:

1. reagent and solution:

Tartaric acid solution (100g/L), ascorbic acid (10g/L), animal glue (5g/L), instructions phenolphthalein solution (10g/L), ammoniacal liquor (1+1), sulfuric acid (1+9), phenylfluorone solution (0.1g/L), tin titer, tin standard solution

2. sample digestion:

Draw 10ml or 20ml sample, be placed in 250ml-500ml nitrogen fixing bottle, addend grain bead, first removes ethanol or carbon dioxide with little fire heating, then adds 5ml-10ml nitric acid-perchloric acid mixed liquor, and after mixing, little fire slowly heats, and the effect for the treatment of relaxes, and lets cool.Along a bottle wall, add 5ml or 10ml sulfuric acid, then heat, start to become when brown to liquid in bottle, constantly along bottle wall, drip nitric acid-perchloric acid mixed liquor complete to organic matter decomposition.Add high flame, to producing white cigarette, until mouthful white cigarette of bottle emit clean after, in bottle liquid produce again white cigarette for digestion completely, this solution is should be clear and bright colourless or be with micro-yellow, lets cool.Add 20ml water boil, then go remaining nitric acid to till producing white cigarette, so process twice, let cool.Solution after cold is moved in 50ml or 100ml volumetric flask, wash nitrogen fixing bottle with water, washing lotion is incorporated in volumetric flask, lets cool, and adds water to scale, mixes.The every 10ml of solution after constant volume is equivalent to 1g sample, quite adds sulfuric acid amount 1ml.Get and the nitric acid-perchloric acid mixed liquor and the sulfuric acid that digest sample same amount, by Same Way, do reagent blank test.

3. the mensuration of sample

Draw respectively 1.00-5.00ml sample digestive juice and the reagent blank with amount, be placed in respectively 25ml colorimetric cylinder.Absorption 0,0.20,0.40,0.60,0.80,1.00ml tin standard solution, be placed in respectively 25ml colorimetric cylinder.

Respectively add 0.5ml tartaric acid solution and 1 instructions phenolphthalein solution, mix, respectively add ammoniacal liquor and be neutralized to pale red.Add 3ml H ₂sO ₄, 1ml gelatin solution and 2.5ml ascorbic acid solution, add water to again 25ml, mix, respectively add 2ml phenylfluorone solution, mix, after 1h, measure, with water, regulate zero point with 2cm cuvette, survey absorbance in wavelength 490nm place, standard each point deducts after zero pipe light absorption value, drawing standard curve or calculated line regression equation, sample light absorption value and curve relatively or substitution equation obtain content.

Computing formula:

$X=\frac{(m_1-m_2)\×1000}{m_3\×(V_2/V_1)\×1000}$

In formula:

X is tin content in sample (mg/kg or mg/L);

M ₁in order to measure the quality of tin in sample digestive juice, (μ g);

M ₂for the quality of tin in reagent blank liquid, (μ g);

M ₃for sample mass (g);

V ₁for the cumulative volume (ml) of sample digestive juice;

V ₂for measuring the volume (ml) with sample digestive juice.

8, iron-holder: be GB/T5009.90-2003 according to standard number, standard name is that the method in the mensuration of iron in food, magnesium, manganese is measured iron-holder, and concrete grammar is:

1. reagent and solution: hydrochloric acid, nitric acid, perchloric acid, mixed acid digestion liquid (nitric acid+perchloric acid is pressed 4:1 and mixed), 0.5mol/L salpeter solution, standard liquid (iron, magnesium, copper standard liquid), Standard Applying Solution

2. instrument: atomic absorption spectrophotometer

3. Specimen Determination:

Materials 5.0-10.0g in 250ml beaker in tall form, add mixed sour digestive juice 20-30ml, upper cover surface plate.Be placed in hot digestion in electric hot plate or electric sand-bath, as sample does not digest, can add again several milliliters of nitration mixture, continue hot digestion, until water white transparency.Add several ml waters, heating is to remove unnecessary nitric acid again.When the liquid of beaker approaches 2-3ml, take off cooling.Deionization is washed and is shifted in 10ml scale test tube, adds water and is settled to scale.Get with the nitration mixture that digests sample same amount and close sample digestive juice, by aforesaid operations, make reagent blank and measure.In wavelength 248.3nm place, survey absorbance.

Computing formula:

$X=\frac{(c-c_0)\×V\×f\×100}{m\×1000}$

In formula:

X is constituent content in sample (mg/100g);

C is for measuring constituent content (μ g/ml) in sample liquid;

C ₀for constituent content in blank solution (μ g/ml);

V is sample constant volume (ml);

F is extension rate;

M is the quality (g) of sample.

9, BHC and DDT residual quantity: be GB/T5009.19-2008 according to standard number, standard name is that the method in the mensuration of organo-chlorine pesticide multicomponent residual quantity in food is measured BHC and DDT residual quantity, and its concrete grammar is:

1. reagent and solution:

Acetone, n-hexane, 30 ℃-60 ℃ of benzinum boiling ranges, benzene, sulfuric acid, anhydrous sodium sulfate, metabisulfite solution (20g/L), pesticide standard storing solution, agricultural chemicals hybrid standard working solution;

Standard sample of pesticide: BHC (α-HCH, β-HCH, γ-HCH and δ-HCH) purity >99%; DDT (ρ, ρ '-DDE, o, ρ '-DDT, ρ, ρ '-DDD, ρ, ρ '-DDT) purity >99%.

2. instrument:

Gas chromatograph, rotary evaporator, N-evaporimeter, refiner, the multiplex oscillator of speed governing, centrifuge

3. sample pretreating:

Sample 0.5g, in 10ml scale test tube, is settled to scale with petroleum ether dissolution.Add 1mol concentrated sulfuric acid purification, jolting 0.5min, in the centrifugal I5min of 3000r/min.Get supernatant and carry out GC analysis.

4. the mensuration of sample

Packed column gas chromatography condition: chromatographic column: internal diameter 3mm, the glass column of long 2m; The in-built 80 order-100 order diatomite that are coated with 1.5%OV-17 and 2%QF-1 mixed stationary liquid.

Carrier gas: High Purity Nitrogen, flow velocity 110ml/min; Column temperature: 185 ℃; 225 ℃ of detector temperatures; 195 ℃ of injector temperatures.Sample size is 1 μ L-10 μ L.External standard method is quantitative.

Computing formula: $X=\frac{A_1}{A_2}\×\frac{m_1}{m_2}\×\frac{V_1}{V_2}\×\frac{1000}{1000}$

In formula:

X is the single content (mg/kg) of BHC, DDT and isomers thereof or metabolin in sample;

A ₁for the peak value (peak height or area) of the each component of determined sample;

A ₂for the peak value (peak height or area) of each pesticide composition standard;

M ₁for the content (ng) of single pesticide standard solution;

M ₂for the sampling amount (g) of determined sample;

V ₁for the dilution volume (ml) of determined sample;

V ₂for the sampling volume (μ L) of determined sample.

10, methamidophos residue amount: be GB/T5009.103-2003 according to standard number, standard name is that the method in the mensuration of acephatemet and orthene persticide residue in vegetable food is measured methamidophos residue amount, and its concrete grammar is:

1. reagent and solution: acetone, carrene, anhydrous sodium sulfate, active carbon, acephatemet, acephatemet standard liquid

2. instrument: gas chromatograph (thering is flame photometric detector), electric agitator, K-D inspissator or rotary evaporator, centrifuge

3. sample pretreating:

The 5g that materialses, washes in the centrifuge tube of 50ml with the gradation of 45ml acetone, adds 5ml water, mix, centrifugal 5min under 3000r/min, draws supernatant, oil reservoir adds 10ml water and l0ml acetone more below, and centrifugal 5min draws supernatant, merge supernatant twice, with concentrated concentrated near the doing of clamoring of K-D, residue and water add 40g anhydrous sodium sulfate, grind and are thousand powderies, pour in tool plug conical flask, add 0.3g active carbon, 60ml chloroform; Vibration 0.5h, suction filtration, is settled to 5ml, treats gas chromatographic analysis.

4. the mensuration chromatographic condition of sample:

Chromatographic column: glass column, internal diameter 3mm, long 0.5m, in-built 2%DEGS/ChromosorbW AW-DMCS, 80 order-100 orders.Air-flow: carrier gas: nitrogen 70ml/min, air 0.7kg/cm ², hydrogen 1.2kg/cm ².Temperature: 200 ℃ of injection ports, 180 ℃ of column temperatures.Qualitative: the retention time with methamidophos pesticide standard specimen is qualitative.Quantitative: quantitative by external standard method, with the standard specimen solution of acephatemet and ether methamidophos pesticide concentration known, make external standard, quantitative by peak height.

Computing formula: $X=\frac{A_1}{A_2}\×\frac{m_1}{m_2}\×\frac{V_1}{V_2}\×\frac{1000}{1000}$

In formula:

X _i---component organophosphor content (mg/kg) in sample;

Esi---inject sample component organophosphor content (ng);

The peak height (mm) of hi---sample;

Hsi---the peak height of component (mm) in standard specimen;

Vi---concentrated constant volume (ml);

V ₂---inject the volume (μ L) of chromatograph test sample;

The quality (g) of m---sample.

11, potassium sorbate amount: be GB/T5009.29-2003 according to standard number, standard name is that the method in sorbic acid in food, benzoic mensuration is measured potassium sorbate amount, and its concrete grammar is:

1. reagent and solution:

Methyl alcohol, weak aqua ammonia (1+1), acetic acid are pressed solution (0.02mol/L), sodium bicarbonate solution (20g/L), sorbic acid standard inventory solution, sorbic acid standard mixing use solution

2. instrument:

High performance liquid chromatograph

3. Specimen Determination

Take 5.00g-10.0g sample, with ammoniacal liquor (1+1), adjust pH approximately 7, add water and be settled to proper volume, centrifugation, supernatant is through 0.45 μ m membrane filtration.

High performance liquid chromatography reference conditions: YWG-C18,4.6mm × 250mm, the stainless steel column of 10 μ m.Mobile phase: methyl alcohol-ammonium acetate solution (0.02mol/L) (5:95), flow velocity: 1ml/min, sample size: 10 μ L, detector: UV-detector, 230nm wavelength, 0.2AUFS, qualitative according to retention time, external standard peak area method is quantitative.

Computing formula: $X=\frac{A\×1000}{m\×\frac{V_2}{V_1}\×1000}$

In formula:

The content of sorbic acid (g/kg) in X-sample;

The quality of sorbic acid (mg) in A-sampling volume;

V ₂-sampling volume (ml);

V ₁-sample dilution cumulative volume (ml);

M-sample mass (g).

12, sulfur dioxide residual quantity: be GB/T5009.34-2003 according to standard number, the method in the mensuration that standard name is sulphite in foods is measured sulfur dioxide residual quantity, and its concrete grammar is:

1. reagent and solution:

Tetrachloro mercury sodium absorption liquid: take 13.6g mercury bichloride and 6.0g sodium chloride, soluble in water and be diluted to 1000ml, placement is spent the night, standby after filtering.

Sulfamic acid ammonium salt solution (12g/L)

Formalin (2g/L): draw the formaldehyde (36%) of 0.55ml without polymerization precipitation, be diluted with water to 100ml, mix.

Starch indicator solution: take 1g soluble starch, with little water furnishing pasty state, slowly, in impouring 100ml boiling water, with adding with stirring, boil, let cool standbyly, this solution faces the used time and now joins.

Potassium ferrocyanide solution: take 10.6g potassium ferrocyanide ［ K ₄fe(CN) ₆3H ₂o ］, be dissolved in water and be diluted to 100ml;

Acetic acid zinc solution: take 22g zinc acetate ［ Zn(CH ₃cOO) ₂2H ₂o ］ be dissolved in a small amount of water, add 3ml glacial acetic acid, be diluted with water to 100ml.

Hydrochloric acid Pararosaniline solution: take 0.1g hydrochloric acid Pararosaniline (C19H18N2Cl4H2O; Prosanilinen hydrochlo-ride) in mortar, add a small amount of water grinding and make to dissolve and be diluted to 100ml.Take out 20ml, be placed in 100ml volumetric flask, add hydrochloric acid (1+1), after fully shaking up, make solution by red stain Huang, if not flavescence drips a small amount of hydrochloric acid again to occurring yellow, then be diluted with water to scale, mix standby (as replaced with hydrochloric acid is pinkish red without hydrochloric acid Pararosaniline).

Iodine solution ［ c(1/2I ₂)=0.1mol/L ］.

Sodium thiosulfate standard solution ［ c(Na ₂s ₂o ₃5H ₂o)=0.1mol/L ］.

Sulfur dioxide standard liquid: take 0.5g sodium hydrogensulfite, be dissolved in 200ml tetrachloro mercury sodium absorption liquid, placement is spent the night, supernatant filters standby with quantitative filter paper.

2. instrument: spectrophotometer.

3. the mensuration of sample:

Draw 10.0ml sodium hydrogensulfite one tetrachloro mercury sodium solution in 250ml iodine flask, add 100ml water, accurately add 20.00ml iodine solution (0.1mol/L), 5ml glacial acetic acid, shake up, be positioned over and with sodium thiosulfate (0.1mol/L) standard liquid, be titrated to rapidly after the 2min of dark place faint yellowly, add 0.5ml starch indicator solution, continue to drop to colourless.Separately get 100ml water, accurately add iodine solution 20.0ml(0.1mol/L), 5ml glacial acetic acid, by Same Way, do reagent blank test.

Computing formula: $X=\frac{(V_2-V_1)\×c\×32.03}{10}$

In formula:

X ₁---sulfur dioxide concentration of standard solution

V ₁---measure with sodium hydrogensulfite-tetrachloro mercury sodium solution consumption sodium thiosulfate standard solution volume (ml);

V ₂---reagent blank consumes sodium thiosulfate standard solution volume (ml);

The molar concentration (mol/L) of c---sodium thiosulfate standard solution.

13, salmonella: be GB/T4789.31-2003 according to standard number, standard name is that " the enterobacteriaceae bacteriophage method of inspection of microbiological test of food hygiene salmonella, Shigella and Diarrheogenil Escherichia coli " measures salmonella, and its concrete grammar is:

1. culture medium and reagent: bacteriophage is used in nutrient agar, nutrient broth, peptone water, indole reagent, triple sugar iron agar, enterobacteriaceae diagnosis

2. operating procedure:

Will colony inoculation be tried in nutrient broth pipe, in 36 ℃ of overnight incubation.The full ring of this broth culture of picking one, is diluted in peptone water that a pipe fills 1-2ml in vitro, makes to become 1: 200-1: 400 dilution bacterium liquid, bacteria containing amount is about 1 × 10 ⁶/ ml.

With quantitative nipple dropper, in a bacterial plaque, drip one, bacteriophage, be followed successively by O-I, C, Sh, E, CE, E-4 and Ent.Syringe needle is installed on dropper No. 4, and every milliliter is about 100.Every dropper only drips a kind of bacteriophage, and needle point definitely can not contact planar surface, strictly prevents cross pollution.Dropper of every use, the bacterial plaque position that whole bacterium to be tried should be able to be dripped to bacteriophage all dropwises.While dripping bacteriophage, agar plate must be placed on level table.After 7 kinds of bacteriophages all dropwise, summary etc. several minutes, treat that phagocytosis body fluid is dry.Upset is dull and stereotyped, and put 36 ℃ and cultivate 5-6h, and the once result of each observation of spending the night.

If only there is one or two strain culture to do bacteriophage test, the oese picking bacteriophage that available diameter is 3mm, drips successively in bacterial plaque.Scorching hot oese can picking bacteriophage after must be completely cooling.

Spot semar technique is divided into quarter by agar plate surface, and every equal portions can be for smearing a strain bacterial cultures.Every strain culture is smeared 3 bacterial plaques, 2 of outer rings, 1 of inner ring.Cotton swab semar technique can be smeared test organisms liquid to become band on agar plate, and each flat board approximately can be smeared 5 cingulas.Summaries etc. several minutes, the bacterial plaque of waiting is dry.

Drip successively above-mentioned 3 kinds of bacteriophages, at 36 ℃, cultivate 5-6h, and the once result of each observation of spending the night.

14, staphylococcus aureus: be GB4789.10-2010 according to standard number, standard name is that the method in the check of food microbiological analysis staphylococcus aureus is measured staphylococcus aureus, and concrete detection method is:

1. equipment and material:

Constant incubator, refrigerator, constant water bath box, balance, homogenizer, oscillator, aseptic straw, aseptic conical flask, sterile petri dish, syringe, pH meter or pH colorimetric cylinder or accurate pH test paper.

2. culture medium and reagent:

10% sodium chloride trypticase soybean broth, 7.5% sodium chloride meat soup, blood agar plate, Baird-Parker agar plate, brain heart leachate meat soup (BHI), rabbit plasma, phosphate buffer, the little inclined-plane of nutrient agar, gram staining liquid, SPSS.

3. operating procedure:

With aseptic straw, draw 25ml sample and put in the aseptic conical flask (sterile glass beads of preset right quantity in bottle) that fills 225ml phosphate buffer or physiological saline, fully mix, make the even liquid of sample of 1:10.

With 1ml aseptic straw or the even liquid 1ml of micropipettor absorption 1:10 sample, along tube wall, slowly note in the sterile test tube that fills 9ml dilution (dilution liquid level is not touched at attention suction pipe or suction nozzle tip), jolting test tube or use 1 1ml aseptic straw instead and repeatedly blow and beat it is mixed, makes the even liquid of sample of 1:100.Prepare 10 times of even liquid of series of diluted samples.Often increase progressively dilution once, use 1 1ml aseptic straw or suction nozzle instead.

According to the estimation to sample contamination situation, select 2-3 the even liquid of suitable dilution sample (fluid sample can comprise stoste), increase progressively when dilution carrying out 10 times, each dilution factor is drawn respectively the even liquid of 1ml sample and is added respectively with 0.3ml, 0.3ml, 0.4ml inoculum concentration

10 times of serial dilutions are selected 2-3 the continuous even liquid of suitable dilution sample, inoculation Baird-Parker plate count and coagulase test of blood plasma.Enter three Baird-Parker flat boards, then, with the whole flat board of aseptic L rod coating, note not touching plate edge.Before use, as Baird-Parker planar surface has the globule, can be placed in the incubator of 25 ℃-50 ℃ dry, until the globule of planar surface disappears.

Cultivate: under normal conditions, after coating, by standing flat board 10min, as sample liquid is difficult for absorbing, flat board can be placed on to 36 ℃ ± 1 ℃ of incubator and cultivates 1h; Deng the even liquid of sample, absorb rear upset plate, be inverted in incubator, 36 ℃ ± 1 ℃ cultivation, 45h-48h.

Selection has the flat board of typical staphylococcus aureus bacterium colony, and 3 all clump counts of flat board of same dilution factor are aggregated in the flat board between 20CFU-200CFU, counting colonies typical number.

4. computing formula:

$T=\frac{\mathrm{AB}}{\mathrm{Cd}}$

In formula:

T---Gold Samples staphylococcus aureus clump count;

A---the sum of a certain dilution factor colonies typical;

B---the clump count of a certain dilution factor clotting of plasma enzyme positive;

C---a certain dilution factor is for the clump count of coagulase test of blood plasma;

D---dilution gfactor.

15, Shigella: be GB/T4789.31-2003 according to standard number, standard name be microbiological test of food hygiene salmonella, Shigella and Diarrheogenil Escherichia coli the enterobacteriaceae bacteriophage method of inspection measure Shigella, according to 13) method measure.

16, total plate count: be GB4789.2-2010 according to standard number, standard name is that the method during food security national standard microbiological Test total plate count is measured is measured total plate count, and its concrete grammar is:

1. equipment and material:

Constant incubator, refrigerator, constant water bath box, balance, homogenizer, oscillator, aseptic straw, aseptic conical flask, sterile petri dish, pH meter or pH colorimetric cylinder or accurate pH test paper, magnifying glass are or/and colonometer.

2. culture medium and reagent:

Plate count agar culture medium, phosphate buffer, SPSS

3. operating procedure:

With aseptic straw, draw 25ml sample and put in the aseptic conical flask (sterile glass beads of preset right quantity in bottle) that fills 225ml phosphate buffer or physiological saline, fully mix, make the even liquid of sample of 1:10.

With 1ml aseptic straw or the even liquid 1ml of micropipettor absorption 1:10 sample, along tube wall, slowly note in the sterile test tube that fills 9ml dilution (dilution liquid level is not touched at attention suction pipe or suction nozzle tip), jolting test tube or use 1 aseptic straw instead and repeatedly blow and beat it is mixed, makes the even liquid of sample of 1:100.According to above method, prepare 10 times of even liquid of series of diluted samples.Often increase progressively dilution once, use 1 1ml aseptic straw or suction nozzle instead.

According to the estimation to sample contamination situation, select 2-3 the even liquid of suitable dilution sample (fluid sample can comprise stoste), increase progressively when dilution carrying out 10 times, draw the even liquid of 1ml sample in aseptic plate, each dilution factor makees two plates.Meanwhile, drawing respectively the blank dilution of 1ml adds in two aseptic plates and makes blank.

In time 15ml-20ml is cooled to plate count agar culture medium (can be positioned in 46 ℃ ± 1 ℃ constant water bath box and the be incubated) pour plate of 46 ℃, and rotates plate it is mixed.

After agar solidifies, by Flat plate turnover, cultivate 48h ± 2h for 36 ℃ ± 1 ℃.30 ℃ ± 1 ℃ cultivation 72h ± 3h of aquatic products.

If may contain in sample while filling the air the bacterium colony of growth on agar medium surface, can cover skim agar medium (about 4ml) by the agar surface after solidifying, solidify rear upset flat board and cultivate.

Colony counting: can detect by an unaided eye, if desired with magnifying glass or colonometer, record extension rate and corresponding colony counts.Colony counting represents with CFU (colony-forming units, CFU).

Choose clump count between 30CFU-300CFU, without the plate count total plate count that spreads colony growth.Lower than the flat board of 30CFU, record concrete clump count, be greater than being recorded as of 300CFU and how can not count.Each dilution clump count should adopt two dull and stereotyped averages.

When one of them flat board has larger sheet colony growth, should not adopt, and should be using the flat board without sheet colony growth as this dilution clump count; If sheet bacterium colony less than dull and stereotyped half, and in all the other half, bacterium colony distributes very evenly, is multiplied by 2 after can calculating half flat board, represents a flat-plate bacterial colony number.

When there is the chain-like growth in the obvious boundary line of nothing between bacterium colony on flat board, using every strand as a colony counting.

4. computing formula: $N=\mathrm{\ΣC}/(n_1+0.1n_2)d$

N---clump count in sample;

Σ C---dull and stereotyped (containing the flat board of optimum range clump count) clump count sum;

N ₁---the dull and stereotyped number of the first dilution factor (low extension rate);

N ₂---the dull and stereotyped number of the second dilution factor (highly diluted multiple);

D---dilution gfactor (the first dilution factor).

17, coli-group number: be GB/T4789.3-2010 according to standard number, standard name is that the method in food security national standard microbiological Test Escherichia coli counting is measured coli-group number;

1. equipment and material:

Refrigerator, constant incubator, homogenizer, constant temperature oscillator, microscope, electronic balance, aseptic conical flask, aseptic wide-mouth bottle, aseptic straw, aseptic plate, colonometer.

2. culture medium and reagent: lauryl sulfate tryptose broth, BGLB, violet red bile agar, phosphate buffer, SPSS, aseptic 1mol/LNaOH, aseptic 1mol/L HCl

3. operating procedure:

With aseptic straw, draw 25ml sample to filling in the conical flask sterile glass beads of preset right quantity (can in bottle) of 225ml sterile distilled water, fully mix, make the even liquid of sample of 1:10, the pH value of the even liquid of sample should be between 6.5-7.5.Get 1ml1:10 dilution and inject the test tube that contains 9ml sterilized water, separately change 1ml aseptic straw pressure-vaccum repeatedly, this liquid is 1:100 dilution.

Just fermentation test: each sample, select the even liquid of sample (fluid sample can be selected stoste) of 3 suitable serial dilution degree, each dilution factor is inoculated 3 pipe lauryl sulfate tryptone (LST) meat soups, every pipe inoculation 1ml(, as inoculum concentration exceedes 1ml, uses extra quality LST meat soup), cultivate 24h ± 2h for 36 ℃ ± 1 ℃, observe in voltage regulator tube and whether have Bubble formation, 24h ± 2h aerogenesis person carries out multiple fermentation test, and as aerogenesis not continues to be cultured to 48h ± 2h, aerogenesis person carries out multiple fermentation test.Aerogenesis person is not coliform feminine gender.

Multiple fermentation test: get respectively culture 1 with oese from the LST meat soup pipe of aerogenesis and encircle, culture transferring, in BGLB (BGLB) pipe, is cultivated 48h ± 2h for 36 ℃ ± 1 ℃, observes aerogenesis situation.Aerogenesis person, counts coliform-positive pipe.

18, yeast and mold number: be GB/T4789.15-2010 according to standard number, standard name is that the method in food security national standard microbiological Test moulds and yeasts count is measured yeast and mold number, and concrete grammar is:

1. equipment and material:

Refrigerator, constant incubator, homogenizer, constant temperature oscillator, microscope, electronic balance, aseptic conical flask, aseptic wide-mouth bottle, aseptic straw, aseptic plate, sterile test tube, aseptic kraft paper bag, polybag.

2. culture medium and reagent:

Potato-glucose-agar medium, rose bengal medium

3. operating procedure:

With aseptic straw, draw 25ml sample to filling in the conical flask sterile glass beads of preset right quantity (can in bottle) of 225ml sterile distilled water, fully mix, make the even liquid of sample of 1:10.Get 1ml1:10 dilution and inject the test tube that contains 9ml sterilized water, separately change 1ml aseptic straw pressure-vaccum repeatedly, this liquid is 1:100 dilution.Prepare 10 times of even liquid of series of diluted samples.Often increase progressively dilution once, use 1ml aseptic straw instead 1 time.According to the estimation to sample contamination situation, select 2-3 the even liquid of suitable dilution sample (fluid sample can comprise stoste), carry out 10 times increase progressively dilution in, each dilution factor is drawn respectively the even liquid of 1ml sample in 2 aseptic plates.Get respectively 1ml sample diluting liquid adds 2 aseptic plates to make blank simultaneously.In time 15ml-20ml is cooled to potato-glucose-agar or rose bengal medium (can be positioned in 46 ℃ ± 1 ℃ constant water bath box and the be incubated) pour plate of 46 ℃, and rotates plate it is mixed.After agar solidifies, flat board is inverted, cultivate 5d for 28 ℃ ± 1 ℃, observe and record.

The testing result of embodiment 1-7 is in Table 1:

Table 1: the testing result of embodiment 1-7

Note: ^abe only applicable to metal filling.

Table 1 result shows: cold tea prepared by embodiment 1-7, all there is taste fragrance, and free from extraneous odour, physical and chemical index, microbiological indicator etc. all conforms with the regulations.

Experimental example 2: drink mouthfeel investigation

1, investigation method

1) drink crowd: general population

2) drinking method: take embodiments of the invention and comparative example, 1 month, 500ml for each person every day.

3) drink number: every group of 60 people

2, investigation result:

Drink rear fresh and sweet sweet being designated as that have back of obviously feeling;

Drink being designated as of the fresh and sweet micro-hardship of after sensation general;

Drink being designated as of after sensation heavy bitter taste poor.

3, investigation result: in Table 2

Table 2: mouthfeel experimental result

?	Good (number)	Generally (number)	Poor (number)
				Embodiment 1	63.3%（38）	36.7%（22）	0%（0）
Embodiment 2	66.7%（40）	33.3%（20）	0%（0）
				Embodiment 3	70.0%（42）	30.0%（18）	0%（0）
Embodiment 4	70.0%（42）	30.0%（18）	0%（0）
				Embodiment 5	76.7%（46）	23.3.%（14）	0%（0）
Embodiment 6	75.0%（45）	25.0%（15）	0%（0）
				Embodiment 7	78.3.%（47）	21.7%（13）	0%（0）
Comparative example 1	51.7%（31）	48.3%（29）	0%（0）
				Comparative example 2	48.3%（29）	53.3%（31）	0%（0）

Table 2 result shows: the mouthfeel poor (good is less than 52%) of comparative example 1,2, and embodiment 1-7 respectively organizes all higher than 63%.

Cold tea mouthfeel that embodiments of the invention provide is fresh and sweet to be had back sweetly, and mouthfeel is better than comparative example 1,2.

Experimental example 3: drink effect research

1, drink crowd: sub-health population.

2, drinking method: take embodiments of the invention and comparative example, 1 month, 500ml for each person every day.

3, drink number: every group of 60 people.

4, index: drink obviously feel clearing heat and detoxicating afterwards, promote the production of body fluid wet one's whistle, relieving restlessness annealing is designated as effective; Drink after sensation clearing heat and detoxicating, promote the production of body fluid wet one's whistle, relieving restlessness annealing is designated as effectively; Drink rear without obvious comfort, be designated as invalid.

5, investigation result: in Table 3

Table 3: drink effect comparison

?	Obvious effective rate (number)	Efficient (number)	Inefficiency (number)
				Embodiment 1	60.0%（36）	40.0%（24）	0%（0）
Embodiment 2	61.7%（37）	38.3%（23）	0%（0）

Embodiment 3	61.7%（37）	38.3%（23）	0%（0）
				Embodiment 4	63.3%（38）	36.7%（22）	0%（0）
Embodiment 5	70.0%（42）	30.0%（18）	0%（0）
				Embodiment 6	66.7%（40）	33.3%（20）	0%（0）
Embodiment 7	70.0%（42）	30.0%（18）	0%（0）
				Comparative example 1	48.3%（29）	51.7%（31）	0%（0）
Comparative example 2	50.0%（30）	50.0%（30）	0%（0）

Table 3 result shows: comparative example 1-2 group obvious effective rate is less than or equal to 50%, and the effect of the each group of embodiment 1-7 is higher than 60%, and obvious effective rate is apparently higher than comparative example 1 and 2.

Result shows, that cold tea of the present invention has is clearing heat and detoxicating, promote the production of body fluid wet one's whistle, the effect of relieving restlessness annealing, effect is better than prior art.

Although, above used general explanation, the specific embodiment and test, the present invention is described in detail, on basis of the present invention, can make some modifications or improvements it, and this will be apparent to those skilled in the art.Therefore, these modifications or improvements without departing from theon the basis of the spirit of the present invention, all belong to the scope of protection of present invention.

Claims (10)

1. the cold tea that heat-clearing wets one's whistle, contains active component, it is characterized in that, the active component of described cold tea is grouped into by the one-tenth of following weight portion: cordate houttuynia 1-12 part, chrysanthemum 0.2-3 part, peppermint 0.2-2 part, emblic 0.2-2 part and sterculia seed 0.2-2 part.

2. cold tea according to claim 1, is characterized in that, the active component of described cold tea is grouped into by the one-tenth of following weight portion: cordate houttuynia 3-10 part, chrysanthemum 0.5-2.5 part, peppermint 0.5-1.5 part, emblic 0.5-1.5 part and sterculia seed 0.5-1.5 part.

3. cold tea according to claim 1, is characterized in that, the active component of described cold tea is grouped into by the one-tenth of following weight portion: cordate houttuynia 4-8 part, chrysanthemum 1-2 part, peppermint 0.8-1.2 part, emblic 0.8-1.2 part and sterculia seed 0.8-1.2 part.

4. according to the cold tea described in claim 1-3 any one, also contain auxiliary material, concrete consumption is 80-250 part.

5. cold tea according to claim 4, is characterized in that, the consumption of auxiliary material is 100-200 part.

6. cold tea according to claim 4, is characterized in that, the consumption of auxiliary material is 120-180 part.

7. cold tea according to claim 4, is characterized in that, described auxiliary material is sweetener, and described sweetener is selected from one or more in white granulated sugar, fructose, stevioside, glycyrrhizin, xylitol, D-sorbite, maltitol, Aspartame.

8. a method of preparing the cold tea described in claim 1-7 any one, is characterized in that, the method comprises the following steps: according to proportioning, take each composition, active component water is decocted 2-3 time, add the water that active component gross weight 8-20 doubly measures, each decocting time is 20-120 minute at every turn, cold heavy, centrifugal after merging filtrate, filter, add sweetener, add water constant volume, high-temperature instantaneous sterilization, packing, obtains.

9. method according to claim 8, it is characterized in that, the method comprises the following steps: according to proportioning, take each composition, active component is mixed, water decocts 2-3 time, add the water that active component gross weight 8-20 doubly measures at every turn, each decocting time is 20-120 minute, decoction temperature is 70-80 ℃, filter, merging filtrate, cold heavy 8-16 hour in 4 ℃ of environment, with the centrifugal 20-40min of speed of 3000-5000r, filter, add sweetener, stir, add water, 120-140 ℃ of sterilization 1-20 second, it is filling after liquid is cooled to 0-95 ℃, be inverted cooling, packing, obtain.

Cold tea described in claim 1-7 clearing heat and detoxicating in preparation treatment, promote the production of body fluid wet one's whistle, the application in the health products of relieving restlessness annealing.