CN103190500A - 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 property of medicine is cold and cool and that can clear up in the human body heat fries in shallow oil water and cook beverage and drink, eliminating the summer heat in the human body in summer, or the illness such as throat pain that cause for the treatment of winter drying.Except clearing heat and detoxicating, cold tea also can dry promote the production of body fluid, relieve inflammation or internal heat, make eye bright, dissipating bind, detumescence etc., also can control hot eyes headache, dizziness and tinnitus, furuncle swelling toxin and hypertension, can work as cold drink fully summer and drink.

Cordate houttuynia is the dry herb of saururaceae plant houttuynia cordata, and heat-clearing, detoxifcation, diuresis, detumescence effect are arranged, and exploitation is that the antipyretic beverage of major ingredient will produce remarkable social benefit and economic benefit with the cordate houttuynia.

The relevant patent of cold tea about cordate houttuynia has:

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

Chinese patent application 200910103303.5 discloses a kind of herbal tea, and it is to be that feedstock production forms 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 feedstock production forms by Momordica grosvenori, honeysuckle, the sterculia seed, cordate houttuynia, purple perilla, Buddha's hand, mulberry leaf.

At present, the cold tea kind that with the cordate houttuynia is major ingredient is abundant, especially at partial heat in summer how wet weathers, gastrointestinal disorder easily, add the 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, symptom such as be off one's feed.

Summary of the invention

The purpose 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 provides 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 active component of described cold tea contains the composition 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.

Further preferred, 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 the 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 the 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, also can be its multiple, as 1/10,1/100,10 times, 100 times etc.

Described auxiliary material refers in the cold tea preparation process to add for flavoring required supplies.

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

The present invention also provides a kind of method for preparing cold tea, and this method may further comprise the steps:

Take by weighing each composition according to proportioning, the 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 behind the merging filtrate, filter, add sweetener, add water, high-temperature instantaneous sterilization, packing, namely.

Preferably, the preparation method of cold tea provided by the invention, this method may further comprise the steps:

Take by weighing each composition according to proportioning, with the active component mixing, 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 decocting temperature is 70-80 ℃, filters, merging filtrate, in 4 ℃ of environment cold heavy 8-16 hour, with the centrifugal 20-40min of the speed of 3000-5000r, filter, add sweetener, stir, add water, 120-140 ℃ of sterilization 1-20 second, treat soup be cooled to 0-95 ℃ after can, be inverted cooling, packing, namely.

The present invention also provide cold tea clearing heat and detoxicating in the 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 be packed in pop can or the 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.Flavor is hot, 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 the lung carbuncle pyemesis, the phlegm heat panting is coughed, hot dysentery, and heat is drenched, carbuncle sore tumefacting virus.

Peppermint is the dry aerial parts of labiate peppermint.Flavor is hot, and is cool in nature.Has 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 expands vexed.

Chrysanthemum is the dry capitulum of feverfew chrysanthemum.It is sweet, bitter to distinguish the flavor of, cold nature.Have diffusing wind heat-clearing, flat liver makes eye bright, clearing heat and detoxicating effect.Be used for the treatment of anemopyretic cold, it is dizzy to have a headache, red eye, swell pain, and eyes is dim-sighted, the sore pyogenic infections.

Emblic is the dry mature fruit of euphorbia plant emblic.It is sweet, sour, puckery, cool in nature to distinguish the flavor of.Has 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 the blood-head blood stasis, indigestion, abdominal distension, cough, laryngalgia, dry.

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

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

2, in the cold tea provided by the invention, the volatile oil material of fragrance can significantly be covered the fishlike smell of cordate houttuynia in peppermint, chrysanthemum and the sterculia seed, 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 to extract and 120-140 ℃ of high-temperature short-time sterilization, has extracted the active ingredient in the medicinal material fully, and it is refrigerant, sweet and sweet mouthfeel arranged back to have kept cold tea.

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

The specific embodiment

Following examples are used for explanation 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:

Get cordate houttuynia 10kg, chrysanthemum 2kg, peppermint 2kg, emblic 2kg, the sterculia seed 2kg of wash clean, mixing, the decocting that adds 8 times of amounts of weight summation boils 3 times, each decocting time is 20 minutes, and decocting temperature is 70 ℃, filters, merging filtrate, cold sinking 8 hours in 4 ℃ of environment is with the centrifugal 20min of the speed of 3000r, filter, add the aqueous solution that contains the 800kg white granulated sugar, stir, add water to 4500L, 125 ℃ of sterilizations 5 seconds, treat soup be cooled to 95 ℃ after can, be inverted cooling, packing, namely.

2, specification: the 500ml/ plastic bottle contains 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, mixing, the decocting that adds 20 times of amounts of weight summation boils 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 the speed of 4000r, filtered, add the Aspartame aqueous solution that contains 250kg, stir, add water to 4800L, 130 ℃ of sterilizations 15 seconds, treat soup be cooled to 0 ℃ after can, be inverted cooling, packing, namely.

2, specification: the 300ml/ plastic bottle contains 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, mixing, the decocting that adds 10 times of amounts of weight summation boils 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 the speed of 4000r, filtered, add the stevioside sweet solution that contains 100kg, stir, add water to 1250L, 140 ℃ of sterilizations 1 second, treat soup be cooled to 45 ℃ after can, be inverted cooling, packing, namely.

2, specification: the bottling of 330ml/ metal can contains 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, mixing, the decocting that adds 18 times of amounts of weight summation boils 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 the speed of 5000r, filtered, add the fructose water solution that contains 200kg, stir, add water to 4250L, 120 ℃ of sterilizations 20 seconds, treat soup be cooled to 60 ℃ after can, be inverted cooling, packing, namely.

2, specification: the 500ml/ plastic bottle contains 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, mixing, the decocting that adds 12 times of amounts of weight summation boils 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 the speed of 4000r, filtered, add the aqueous solution that contains the 120kg white granulated sugar, stir, add water to 1800L, 140 ℃ of sterilizations 5 seconds, treat soup be cooled to 20 ℃ after can, be inverted cooling, packing, namely.

2, specification: the bottling of 300ml/ metal can contains 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, mixing, the decocting that adds 15 times of amounts of weight summation boils 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 the speed of 5000r, filtered, add the aqueous solution that contains the 180kg xylitol, stir, add water to 3000L, 130 ℃ of sterilizations 10 seconds, treat soup be cooled to 40 ℃ after can, be inverted cooling, packing, namely.

2, specification: the 1.5L/ plastic bottle contains 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, mixing, the decocting that adds 12 times of amounts of weight summation boils 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 the speed of 4500r, filtered, add the aqueous solution that contains the 156kg maltitol, stir, add water to 2400L, 130 ℃ of sterilizations 12 seconds, treat soup be cooled to 40 ℃ after can, be inverted cooling, packing, namely.

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

Comparative Examples 1: with reference to the embodiment 1 preparation cold tea among the CN200910044662.8

Comparative Examples 2: with reference to the embodiment 2 preparation cold teas among the CN200810170826.7

Experimental example 1: product detects

To embodiment 1-7 and Comparative Examples 1 detect its soluble solid amount, total acid content, contain 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, mould and yeast count, concrete detection method is:

1, the amount of soluble solid: be numbered GB/T12143-2008 according to standard, standard name is the amount that the assay method of soluble solid in the 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. sample is measured:

By specification is proofreaied and correct refractometer before measuring, and counts example with the Abbe refractive power, other refractometer by specification operations; Separately refractometer two and prism dip in ether or ethanol is cleaned with absorbent cotton; Dip in the glass bar of the molten circle of end and to get 2-3 of test solutions, drip in refractometer prism facets central authorities (attention does not make glass bar touch minute surface); The rapid closing prism leaves standstill 1mh, makes test solution evenly not have bubble, and is full of the visual field; Alignment light source is observed objective lens by eyepiece.Regulate alidade, make the visual field be divided into two ones of light and shades, the rotary fine adjustment spiral makes terminator clear again, and makes 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 numbered GB/T12456-2008 according to standard, standard name is that the method in the mensuration of total acid in the food is measured total acid content, and concrete detection method is:

1. reagent and solution:

0.1mol/L NaOH standard titration solution

0.01mol/L NaOH standard titration solution

0.05mol/L NaOH standard titration solution

1% phenolphthalein solution

2. instrument:

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

3. the preparation of test solution

Sample is filtered with quick filter paper, collect filtrate, be used for measuring.

Take by weighing the 10g-50g sample, be accurate to 0.001g, place the 100ml beaker, with about 80 ℃ of water that boiled the content in the beaker is transferred in the 250ml volumetric flask.Place boiling water bath to boil 30min, take out, be cooled to room temperature, be settled to 250ml with the water that boiled.Filter with quick filter paper.Collect filtrate, be used for measuring.

4. the mensuration of sample

Take by weighing the 25.000g-50.000g test solution, make it to contain 0.035-0.07g acid, place the 250ml triangular flask.Add 40ml-60ml water and 0.2ml1% phenolphthalein indicator, do not fade to blush 30s with the titration of 0.1mol/L NaOH standard titration solution.Record consumes the numerical value (V of the volume of 0.1mol/L NaOH standard titration solution ₁).

Blank test: water replaces test solution, and record consumes the numerical value (V of the volume of 0.1mol/L NaOH standard titration solution ₂).

Computing formula:

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

In the formula:

X is total acid content (g/kg);

C is the accurate values (mol/L) of NaOH standard titration solution concentration;

V ₁Consume the numerical value (ml) of the volume of NaOH standard titration solution during for burette test;

V ₂Consume the numerical value (ml) of the volume of NaOH standard titration solution during for blank test;

K is the conversion coefficient of acid: malic acid, 0.067; Acetic acid, 0.060; Tartaric acid, 0.075; Citric acid, 0.064; Citric acid, 0.070(contain 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, contain female amount: be numbered GB/T5009.11-2003 according to standard, standard name is that the method in the mensuration of total arsenic and inorganic arsenic in the food 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 by weighing the 4.0g silver nitrate in the 500ml beaker, add suitable quantity of water dissolving back and add 30ml nitric acid, add water to 500ml, store in the brown bottle.

Poly-vinyl alcohol solution (4g/L): take by weighing 0.4g polyvinyl alcohol (degree of polymerization 1500-1800) in small beaker, add 100ml water, heat in the boiling water bath, be stirred to dissolving, insulation 10min takes out and puts cold standby.

Absorption liquid: get each portion of liquor argenti nitratis ophthalmicus (8g/L) poly-vinyl alcohol solution (4g/L), add two parts of volume of ethanol (95%), mixing is as absorption liquid.Now join during use.

The potassium borohydride sheet: potassium borohydride and sodium chloride are mixed levigate by the 1:4 mass ratio, fully make diameter 10mm at tablet press machine behind the mixing, 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 after several minutes and go redundant solution, spread cotton out, after 80 ℃ of oven dry, store in the wide-mouth vial.

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

Arsenic standard reserving solution: take by weighing through 105 ℃ of dry 1h and place drier to be cooled to the arsenic trioxide 0.1320g of room temperature in the 100ml beaker, add 10ml sodium hydroxide solution (2.5mol/L), wait to dissolve the back and add 5ml perchloric acid, 5ml sulfuric acid, put to be heated on the electric hot plate and emit white cigarette, after the cooling, change in the 1000ml volumetric flask, and dilute with water is settled to scale.This solution contains arsenic (pentavalent) 0.100mg for every milliliter.

Arsenic standard application liquid: draw 1.00ml arsenic standard reserving solution in the 100ml volumetric flask, thin up is to scale.This solution contains arsenic (pentavalent) 1.00 μ g for every milliliter.

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

2. instrument: spectrophotometer, arsenic hydride generating means

3. sample is handled:

Draw the 10ml sample in the 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, place (or spending the night) after a few hours, put on the electric hot plate and heat, if solution becomes brown, should add nitric acid decomposes fully organic matter, take off and put coldly, add 15ml water, be heated to again and emit white cigarette, take off, for several times digestive juice is quantitatively changed over to the 100ml arsenic hydride with 20ml moisture and take place in the bottle.Make reagent blank simultaneously.

The preparation of standard series: take place to add arsenic standard application liquid 0,0.25,0.5,1.0,2.0,3.0ml successively in the bottle in 6 100ml arsenic hydrides, add water to 3ml respectively, add 2.0ml sulfuric acid (1+1) again.

4. the mensuration of sample

Take place in the bottle in sample and standard arsenic hydride, add the 0.1g ascorbic acid respectively, 2.0ml KI (500g/L) thiourea solution (50g/L), putting in the boiling water bath in the heating 5min(bottle this moment temperature must not be above 80 ℃), taking-up is put cold, add 1 of methyl red indicator, add about 3.5ml sodium hydroxide solution, transfer to solution with sodium hydroxide solution and just be yellow, add 1.5ml citric acid-ammonium citrate solution, add water to 40ml, add a potassium borohydride tablet, be connected with the absorption tube that fills the 4.0ml absorption liquid by the conduit that is plugged with lead acetate cotton immediately, shake arsenic hydride frequently bottle takes place, react and add a potassium borohydride tablet after 5 minutes again, continuation reaction 5min.Take off absorption tube, use the 1cm cuvette, at the 400nm wavelength, transferring absorbance with standard pipe zero pipe is zero, measures and respectively manages absorbance.Standard series is respectively managed arsenic content to extinction

Degree drawing standard curve or calculating regression equation.

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

In the formula:

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

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

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

4, lead tolerance: be numbered GB/T5009.12-2010 according to standard, standard name is that the method in the mensuration plumbous in the food security national standard 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 by weighing 1.000g metallic lead (99.99%), gradation adds a small amount of nitric acid (4.5), heating for dissolving, and total amount is no more than 37ml, moves into the 1000ml volumetric flask, adds water to scale.Mixing.This solution contains 1.0mg lead for every milliliter;

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

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 is handled:

Take by weighing 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, adding a little funnel clears up on electric furnace, if become brownish black, add mixed acid again, until Mao Baiyan, digestive juice is water white transparency or slightly yellow, puts cold, the sample digestive juice is washed or is filtered in (salinity on digestion back sample is decided) 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 the volumetric flask and is settled to scale, and mixing is standby; Make reagent blank simultaneously.

4. the mensuration of sample

Instrument condition: transfer 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 temperature continue 15s-20s, atomization temperature: 1700 ℃-2300 ℃, continue 4s-5s, and background correction is deuterium lamp or Zeemen effect.

Calibration curve is drawn: the plumbous standard that absorption is prepared is above used liquid 10.0ng/ml(or μ g/L), 20.0ng/ml(or μ g/L), 40.0ng/ml(or μ g/L), 60.0ng/ml(or μ g/L), 80.0ng/ml(each 10 μ L or μ g/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.

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

The use of matrix modifier: to the interference sample is arranged, then inject an amount of matrix modifier ammonium dihydrogen phosphate (4.8) (be generally 5 μ L or with sample with amount) eliminate to disturb.The matrix modifier ammonium dihydrogen phosphate that also will add equivalent when measuring with sample when drawing plumbous calibration curve.

Computing formula:

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

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

c ₁For measuring lead content (ng/ml) in the sample liquid;

c ₀Be lead content in the 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 numbered GB/T5009.13-2003 according to standard, standard name is that the method in the mensuration of copper in the 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 use liquid I, copper standard to use the liquid 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 are used liquid I (1.0 μ g/ml), place the 10ml volumetric flask respectively, add nitric acid (0.5%) and are diluted to scale, mixing.

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 ratio or the substitution equation try to achieve content.

Draw 0,1.0,2.0,4.0,6.0,8.0,10.0ml copper standard and use liquid II (1ml=0.10 μ g), place the 10ml volumetric flask respectively, add nitric acid (0.5%) and be diluted to scale, shake up.Copper titer 10-20 μ L in sample liquid, reagent blank liquid and each volumetric flask after handling imported respectively transfer to the optimum condition graphite furnace atomizer and measure.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 ratio than or the substitution equation try to achieve content.

Computing formula:

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

In the formula:

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

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

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

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

m ₁Be sample quality (volume) (g or ml).

6, zinc content: be numbered GB/T5009.14-2003 according to standard, standard name is that the method in the mensuration of zinc in the food 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 use liquid, zinc standard liquid, zinc standard to use liquid, phenol red indicator solution (1g/L)

2. instrument: spectrophotometer

3. sample digestion:

Draw 10ml or 20ml sample, place the 250ml-500ml nitrogen fixing bottle, addend grain bead is removed ethanol or carbon dioxide with little fire heating earlier, adds 5ml-10ml nitric acid-perchloric acid mixed liquor again, and behind the mixing, little fire slowly heats, and the effect for the treatment of relaxes, and puts cold.Add 5ml or a 10ml sulfuric acid along the bottle wall, heating again, liquid begins to become when brown to the bottle, and it is complete to organic matter decomposition constantly to drip nitric acid-perchloric acid mixed liquor along the bottle wall.Strengthen firepower, to producing white cigarette, treat the white cigarette of mouthful bottle emit clean after, in the bottle liquid produce again white cigarette for digestion fully, this solution is should be clear and bright colourless or be with little yellow, puts cold.Add the 20ml water boil, go remaining nitric acid till produce white cigarette again, so handle twice, put cold.Solution after cold is moved in 50ml or the 100ml volumetric flask, wash nitrogen fixing bottle with water, washing lotion is incorporated in the volumetric flask, puts coldly, adds water to scale, mixing.The every 10ml of solution behind the constant volume is equivalent to the 1g sample, quite adds sulfuric acid amount 1ml.Get and the nitric acid that digests the sample same amount-perchloric acid mixed liquor and sulfuric acid, do the reagent blank test by Same Way.

4. the mensuration of sample

Draw the sample solution of 5.0ml-10.0ml digestion back constant volume and the reagent blank liquid of same amount, place the 125m1 separatory funnel respectively, add 5m1 water, 0.5ml hydroxylamine hydrochloride solution (200g/L), shake up, add 2 phenol red indicator again, (1+1) transfers to redness with ammoniacal liquor, adds 2 again.Add 5ml dithizone-carbon tetrachloride solution (0.1g/L) again, violent jolting 2min, standing demix.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 is till dithizone-carbon tetrachloride solution green is constant.Merge extract, use the 5m1 water washing, carbon tetrachloride layer extracts 2 times with hydrochloric acid (0.02mol/L), each 10ml, and violent jolting 2min during extraction, merging hydrochloric acid (0.02mol/L) extract, and with the residual dithizone of a small amount of carbon tetrachloride flush away.

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

7, stanniferous amount: be numbered GB/T5009.16-2003 according to standard, standard name is that the method in the mensuration of tin in the food is measured the 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 are used liquid

2. sample digestion:

Draw 10ml or 20ml sample, place the 250ml-500ml nitrogen fixing bottle, addend grain bead is removed ethanol or carbon dioxide with little fire heating earlier, adds 5ml-10ml nitric acid-perchloric acid mixed liquor again, and behind the mixing, little fire slowly heats, and the effect for the treatment of relaxes, and puts cold.Add 5ml or a 10ml sulfuric acid along the bottle wall, heating again, liquid begins to become when brown to the bottle, and it is complete to organic matter decomposition constantly to drip nitric acid-perchloric acid mixed liquor along the bottle wall.Strengthen firepower, to producing white cigarette, treat the white cigarette of mouthful bottle emit clean after, in the bottle liquid produce again white cigarette for digestion fully, this solution is should be clear and bright colourless or be with little yellow, puts cold.Add the 20ml water boil, go remaining nitric acid till produce white cigarette again, so handle twice, put cold.Solution after cold is moved in 50ml or the 100ml volumetric flask, wash nitrogen fixing bottle with water, washing lotion is incorporated in the volumetric flask, puts coldly, adds water to scale, mixing.The every 10ml of solution behind the constant volume is equivalent to the 1g sample, quite adds sulfuric acid amount 1ml.Get and the nitric acid that digests the sample same amount-perchloric acid mixed liquor and sulfuric acid, do the reagent blank test by Same Way.

3. the mensuration of sample

Draw 1.00-5.00ml sample digestive juice respectively and with the reagent blank of measuring, place the 25ml colorimetric cylinder respectively.Absorption 0,0.20,0.40,0.60,0.80,1.00ml tin standard are used liquid, place the 25ml colorimetric cylinder respectively.

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

Computing formula:

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

In the formula:

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

m ₁For measuring the quality (μ g) of tin in the sample digestive juice;

m ₂Quality (μ g) for tin in the reagent blank liquid;

m ₃Be sample mass (g);

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

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

8, iron-holder: be numbered GB/T5009.90-2003 according to standard, standard name is that the method in the mensuration of iron, magnesium, manganese in the food 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 application liquid

2. instrument: atomic absorption spectrophotometer

3. sample is measured:

Materials 5.0-10.0g in the 250ml beaker in tall form, add mixed sour digestive juice 20-30ml, the loam cake surface plate.Place hot digestion on electric hot plate or the electric sand-bath, do not digest as sample and can add several milliliters of nitration mixture again, continue hot digestion, till water white transparency.Add several ml waters again, heating is to remove unnecessary nitric acid.The liquid for the treatment of beaker takes off cooling during near 2-3ml.The deionization washing is also shifted in 10ml scale test tube, adds water and is settled to scale.Get with the nitration mixture that digests the sample same amount and close the sample digestive juice, make reagent blank by aforesaid operations and measure.Survey absorbance in wavelength 248.3nm place.

Computing formula:

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

In the formula:

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

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

c ₀Be constituent content in the 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 numbered GB/T5009.19-2008 according to standard, standard name is that the method in the mensuration of organo-chlorine pesticide multicomponent residual quantity in the 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 (purity of α-HCH, β-HCH, γ-HCH and δ-HCH)〉99%; DDT (ρ, ρ '-DDE, o, ρ '-DDT, ρ, ρ '-DDD, ρ, the purity of ρ '-DDT)〉99%.

2. instrument:

Oscillator, centrifuge are used in gas chromatograph, rotary evaporator, N-evaporimeter, refiner, speed governing more

3. sample is handled:

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

4. the mensuration of sample

Packed column gas chromatography condition: chromatographic column: internal diameter 3mm, the glass column of long 2m; Interior dress is coated with the 80 orders-100 order diatomite 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 the formula:

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

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

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

m ₁Content (ng) for single pesticide standard solution;

m ₂Sampling amount (g) for determined sample;

V ₁Dilution volume (ml) for determined sample;

V ₂Sampling volume (μ L) for determined sample.

10, methamidophos residue amount: be numbered GB/T5009.103-2003 according to standard, standard name is that the method in the mensuration of acephatemet and orthene persticide residue in the vegetable food is measured the 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 (having flame photometric detector), electric agitator, K-D inspissator or rotary evaporator, centrifuge

3. sample is handled:

The 5g that materialses washes in the centrifuge tube of 50ml with the gradation of 45ml acetone, adds 5ml water, mixing, centrifugal 5min under 3000r/min draws supernatant, following oil reservoir adds 10ml water and l0ml acetone again, and centrifugal 5min draws supernatant, merge supernatant twice, concentrate concentrated near the doing of clamoring with K-D, residue and water add the 40g anhydrous sodium sulfate, grind to be thousand powderies, pour in the 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, interior dress 2%DEGS/ChromosorbW AW-DMCS, 80 orders-100 order.Air-flow: carrier gas: nitrogen 70ml/min, air 0.7kg/cm ², hydrogen 1.2kg/cm ²Temperature: 200 ℃ of injection ports, 180 ℃ of column temperatures.Qualitative: the retention time with the methamidophos pesticide standard specimen is qualitative.Quantitatively: quantitative with external standard method, make external standard with the standard specimen solution of acephatemet and ether methamidophos pesticide concentration known, it is quantitative to press peak height.

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

In the formula:

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

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

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

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

Vi---concentrate constant volume (ml);

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

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

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

Take by weighing the 5.00g-10.0g sample, transfer pH about 7 with ammoniacal liquor (1+1), 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.Phase flows: methyl alcohol-ammonium acetate solution (0.02mol/L) (5:95), flow velocity: 1ml/min, sample size: 10 μ L, detector: UV-detector, the 230nm wavelength, 0.2AUFS, qualitative according to retention time, the external standard peak area method is quantitative.

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

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

1. reagent and solution:

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

Sulfamic acid ammonium salt solution (12g/L)

Formalin (2g/L): draw the formaldehyde (36%) that 0.55ml does not have the polymerization precipitation, thin up is to 100ml, mixing.

The starch indicator solution: take by weighing the 1g soluble starch, with little water furnishing pasty state, slowly in the impouring 100ml boiling water, with adding with stirring, boil, put cold standby, this solution faces the time spent and now joins.

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

Acetic acid zinc solution: take by weighing 22g zinc acetate ［ Zn(CH ₃COO) ₂2H ₂O ］ be dissolved in the low amounts of water, add the 3ml glacial acetic acid, thin up is to 100ml.

Hydrochloric acid Pararosaniline solution: take by weighing 0.1g hydrochloric acid Pararosaniline (C19H18N2Cl4H2O; Prosanilinen hydrochlo-ride) in mortar, adds the low amounts of water grinding and make dissolving and be diluted to 100ml.Take out 20ml, place the 100ml volumetric flask, add hydrochloric acid (1+1), make solution by the red stain Huang after fully shaking up, it is yellow to occurring to drip small amount of hydrochloric acid again as not flavescence, and thin up is to scale again, mixing standby (can be with the pinkish red replacement of hydrochloric acid as no 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 ］.

The sulfur dioxide standard liquid: take by weighing the 0.5g sodium hydrogensulfite, be dissolved in the 200ml tetrachloro mercury sodium absorption liquid, placement is spent the night, and 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 the 250ml iodine flask, add 100ml water, accurately add 20.00ml iodine solution (0.1mol/L), the 5ml glacial acetic acid, shake up, be positioned over behind the 2min of dark place rapidly with sodium thiosulfate (0.1mol/L) standard solution titration to faint yellow, add 0.5ml starch indicator solution, continue to drip to colourless.Other gets 100ml water, accurately adds iodine solution 20.0ml(0.1mol/L), the 5ml glacial acetic acid, do reagent blank test by Same Way.

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

In the formula:

X ₁---the sulfur dioxide concentration of standard solution

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

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

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

13, salmonella: be numbered GB/T4789.31-2003 according to standard, standard name is measured salmonella for " microbiological test of food hygiene salmonella, Shigella and cause the enterobacteriaceae bacteriophage method of inspection of rushing down ETEC ", 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:

To wait to try colony inoculation in the nutrient broth pipe, in 36 ℃ of overnight incubation.This broth culture of picking one full ring is diluted in peptone water that a pipe fills 1-2ml in vitro, and making becomes 1: 200-1: 400 dilution bacterium liquid, bacteria containing amount is about 1 * 10 ⁶/ ml.

Drip one in bacteriophage with quantitative nipple dropper a bacterial plaque, be followed successively by O-I, C, Sh, E, CE, E-4 and Ent.No. 4 syringe needles are installed on the dropper, and every milliliter is about 100.Every dropper only drips a kind of bacteriophage, and needle point definitely can not contact planar surface, and strictness prevents cross pollution.Dropper of every usefulness can all dropwise the bacterial plaque position that whole bacterium to be tried should drip bacteriophage.Agar plate must be placed on the level table when dripping bacteriophage.After treating that 7 kinds of bacteriophages all dropwise, phagocytosis body fluid drying is treated in summary etc. several minutes.Upset is dull and stereotyped, puts 36 ℃ and cultivates 5-6h, and spend the night and respectively observe once result.

If only there is one or two strain culture to do the bacteriophage test, then available diameter is the oese picking bacteriophage of 3mm, drips on bacterial plaque successively.Can the picking bacteriophage after scorching hot oese must cool off fully.

The spot semar technique is divided into quarter with 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.The cotton swab semar technique can be smeared test organisms liquid and become band at agar plate, each flat board can be smeared 5 cingulas approximately.Summary etc. several minutes, the bacterial plaque of waiting drying.

Drip above-mentioned 3 kinds of bacteriophages successively, cultivate 5-6h at 36 ℃, and spend the night and respectively observe once result.

14, staphylococcus aureus: be numbered GB4789.10-2010 according to standard, standard name is measured staphylococcus aureus for the method in the check of food microbiological analysis 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:

Draw the 25ml sample with aseptic straw and put in the aseptic conical flask (presetting the sterile glass beads of right quantity in the bottle) that fills 225ml phosphate buffer or physiological saline, fully mixing is made the sample of 1:10 and is spared liquid.

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

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

10 times of serial dilutions are selected the even liquid of 2-3 continuous suitable dilution samples, inoculation Baird-Parker plate count and coagulase test of blood plasma.Go into three Baird-Parker flat boards, with the whole flat board of aseptic L rod coating, note not touching plate edge then.Before the use, as the Baird-Parker planar surface globule is arranged, can be placed in 25 ℃-50 ℃ the incubator dryly, disappear up to the globule of planar surface.

Cultivate: under normal conditions, after the coating, flat board is left standstill 10min, be difficult for absorbing as sample liquid, flat board can be placed on incubator and cultivate 1h for 36 ℃ ± 1 ℃; Absorb back upset plate Deng the even liquid of sample, 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 the 20CFU-200CFU, counting colonies typical number.

4. computing formula:

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

In the formula:

T---staphylococcus aureus clump count in the sample;

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 used for the clump count of coagulase test of blood plasma;

D---dilution gfactor.

15, Shigella: be numbered GB/T4789.31-2003 according to standard, standard name is microbiological test of food hygiene salmonella, Shigella and causes the enterobacteriaceae bacteriophage method of inspection of rushing down ETEC and measure Shigella, according to 13) method measure.

16, total plate count: be numbered GB4789.2-2010 according to standard, 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:

Draw the 25ml sample with aseptic straw and put in the aseptic conical flask (presetting the sterile glass beads of right quantity in the bottle) that fills 225ml phosphate buffer or physiological saline, fully mixing is made the sample of 1:10 and is spared liquid.

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

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

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

After treating that agar solidifies, with the flat board upset, cultivate 48h ± 2h for 36 ℃ ± 1 ℃.Aquatic products are cultivated 72h ± 3h for 30 ℃ ± 1 ℃.

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

Colony counting: can detect by an unaided eye, in case of necessity with magnifying glass or colonometer, record extension rate and corresponding colony counts.Colony counting is with CFU (colony-forming units, CFU) expression.

Choose clump count between 30CFU-300CFU, do not have the plate count total plate count spread colony growth.The flat board that is lower than 30CFU records concrete clump count, can not count greater than being recorded as of 300CFU is many.Each dilution clump count should adopt the average of two flat boards.

When one of them flat board has big sheet colony growth, then should not adopt, and should be with the flat board of no sheet colony growth as this dilution clump count; If the sheet bacterium colony is less than half of flat board, and bacterium colony distributes very evenly in all the other half, multiply by 2 after can calculating half flat board, represents a flat-plate bacterial colony number.

When occurring the chain-like growth in no obvious boundary line between bacterium colony on the flat board, then with every strand as a colony counting.

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

N---clump count in the sample;

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

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

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

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

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

Draw the 25ml sample to the conical flask that fills the 225ml sterile distilled water (can preset the sterile glass beads of right quantity in bottle) with aseptic straw, fully mixing is made the sample of 1:10 and is spared liquid, and the pH value of the even liquid of sample should be between 6.5-7.5.Get the 1ml1:10 dilution and inject the test tube that contains the 9ml sterilized water, other changes 1ml aseptic straw pressure-vaccum repeatedly, and this liquid is the 1:100 dilution.

Fermentation test just: each sample, select the even liquid (fluid sample can be selected stoste) of sample of 3 suitable serial dilution degree, each dilution factor inoculation 3 pipe lauryl sulfate tryptone (LST) meat soup, every pipe inoculation 1ml(such as inoculum concentration surpass 1ml, then use extra quality LST meat soup), cultivate 24h ± 2h for 36 ℃ ± 1 ℃, whether observe has bubble to produce in the voltage regulator tube, 24h ± 2h aerogenesis person is recurred ferment test, then continues to be cultured to 48h ± 2h as aerogenesis not, and the aerogenesis person is recurred the ferment test.The aerogenesis person is not the coliform feminine gender.

The test of recurrence ferment: get culture 1 ring respectively with oese from the LST meat soup pipe of aerogenesis, culture transferring is cultivated 48h ± 2h for 36 ℃ ± 1 ℃ in BGLB (BGLB) pipe, observe the aerogenesis situation.The aerogenesis person counts the coliform-positive pipe.

18, mould and yeast count: be numbered GB/T4789.15-2010 according to standard, standard name is that the method in food security national standard microbiological Test mould and the saccharomycete counting is measured mould and yeast count, 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:

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

The testing result of embodiment 1-7 sees Table 1:

Table 1: the testing result of embodiment 1-7

Annotate: ^aBe only applicable to the metal can.

Table 1 result shows: the cold tea of embodiment 1-7 preparation, taste fragrance is all arranged, and free from extraneous odour, physical and chemical index, microbiological indicator etc. are all up to specification.

Experimental example 2: drink the mouthfeel investigation

1, investigation method

1) drinks the crowd: the general population

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

3) drink number: every group of 60 people

2, investigation result:

Obviously feel fresh and sweet sweet being designated as that have back after drinking;

It is general to drink being designated as of the fresh and sweet little hardship of after sensation;

It is poor to drink then being designated as of after sensation heavy bitter taste.

3, investigation result: see Table 2

Table 2: mouthfeel experimental result

?	Good (number)	Generally (number)	Difference (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 Examples 1	51.7%（31）	48.3%（29）	0%（0）
				Comparative Examples 2	48.3%（29）	53.3%（31）	0%（0）

Table 2 is the result show: Comparative Examples 1,2 mouthfeel relatively poor (good is less than 52%), and each group of embodiment 1-7 all is higher than 63%.

The cold tea mouthfeel that embodiments of the invention provide is fresh and sweet to be had back sweetly, and mouthfeel is better than Comparative Examples 1,2.

Experimental example 3: drink effect research

1, drinks the crowd: sub-health population.

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

3, drink number: every group of 60 people.

4, index: obviously feel clearing heat and detoxicating after drinking, promote the production of body fluid wet one's whistle, relieving restlessness annealing then is designated as produce effects; Drink after sensation clearing heat and detoxicating, promote the production of body fluid wet one's whistle, relieving restlessness annealing then is designated as effectively; Drink the back no obvious comfort then be designated as invalid.

5, investigation result: see Table 3

Table 3: drink effect relatively

?	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 Examples 1	48.3%（29）	51.7%（31）	0%（0）
Comparative Examples 2	50.0%（30）	50.0%（30）	0%（0）

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

The 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 relieving restlessness Effect of annealing, effect is better than prior art.

Though, 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 all belong to the scope of protection of present invention without departing from theon the basis of the spirit of the present invention.

Claims (9)

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 contains following composition: cordate houttuynia, chrysanthemum, peppermint, emblic and the sterculia seed.

2. cold tea according to claim 1 is characterized in that, 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.

3. cold tea according to claim 1 is characterized in that, the active component of described cold tea contains the composition 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.

4. cold tea according to claim 1 is characterized in that, 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.

5. according to each described cold tea of claim 1-3, also contain auxiliary material, concrete consumption is 80-250 part, and preferably, consumption is 100-200 part, more preferably 120-180 part.

6. cold tea according to claim 5 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, the Aspartame.

7. a method for preparing each described cold tea of claim 1-6 is characterized in that, this method may further comprise the steps: take by weighing each composition according to proportioning, the 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, and is cold heavy, centrifugal behind the merging filtrate, filter, add sweetener, add the water constant volume, high-temperature instantaneous sterilization, packing, namely.

8. method according to claim 7 is characterized in that, this method may further comprise the steps: take by weighing each composition according to proportioning, with the active component mixing, water decocts 2-3 time, adds the water that active component gross weight 8-20 doubly measures at every turn, and each decocting time is 20-120 minute, decocting temperature is 70-80 ℃, filter merging filtrate, in 4 ℃ of environment cold heavy 8-16 hour, with the centrifugal 20-40min of the speed of 3000-5000r, filter, add sweetener, stir, add water, 120-140 ℃ of sterilization 1-20 second, treat soup be cooled to 0-95 ℃ after can, be inverted cooling, packing, namely.

The described cold tea of claim 1-6 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.