CN106644981B - A kind of method of aureomycin content in quick measurement Chlortetracycline premix - Google Patents
A kind of method of aureomycin content in quick measurement Chlortetracycline premix Download PDFInfo
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- CN106644981B CN106644981B CN201611198775.XA CN201611198775A CN106644981B CN 106644981 B CN106644981 B CN 106644981B CN 201611198775 A CN201611198775 A CN 201611198775A CN 106644981 B CN106644981 B CN 106644981B
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/314—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry with comparison of measurements at specific and non-specific wavelengths
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/286—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/286—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising
- G01N2001/2866—Grinding or homogeneising
Abstract
The present invention is for deficiency existing for microbial method and high performance liquid chromatography, a kind of method quickly measuring aureomycin content in Chlortetracycline premix is provided, only need a spectrophotometer that detection can be completed, instrument and equipment less investment, sample treatment and mensuration program are simple, it is easy to implement quick detection, a large amount of samples can disposably be detected, high performance liquid chromatography is not less than for the accuracy and reproducibility of Chlortetracycline premix detection, it is suitable for the continuous detection of the daily production of enterprise, it is accurate and reliable and economical and practical.
Description
Technical field
The invention belongs to analysis and testing technology fields, and in particular to one kind quickly measures aureomycin in Chlortetracycline premix and contains
The method of amount.
Background technique
Aureomycin (Chlortetracycline, CTC) is a kind of broad-spectrum antibiotic for belonging to Tetracyclines, in medical treatment, poultry
Animal husbandry and agriculturally have been widely used.In recent years, tetracycline antibiotics it is antitumor, inhibit in terms of grind
Study carefully, has widened application range of this kind of product in terms of medicine.Chlortetracycline premix be S. aurefaciens whole beer with
Aureomycin calcium salt made of appropriate calcium carbonate, drug ingedient therein is mainly aureomycin, still, also contains some relevant hairs
Ferment by-product, such as tetracycline.Provided in pharmacopeia at present the detection method of aureomycin content mainly using microbial method and
High performance liquid chromatography.The method of microbiology polymorphism aureomycin content, cumbersome, time-consuming, and influence factor is more, requires
Environmental condition is high, and the accuracy of test is difficult to ensure, the repeatability of test is also relatively poor with reproducibility.
" Chinese Pharmacopoeia " discloses the concrete operations that aureomycin hydrochloride content and related substances are detected using HPLC method, wherein
Provide two different chromatographic conditions: alkaline mobile phase and acid mobile phase, alkaline mobile phase use ammonium oxalate, phosphoric acid hydrogen two
The mixed solution of ammonium and dimethylformamide is 8.3 with ammonia solution tune pH, although this method is more convenient, fast than microbial method,
It is the alkaline mobile phase condition of pH8.3, aureomycin is extremely unstable under alkaline condition, oxidizable degradation, and mobile phase contains
Salt, salt are easy crystallization and are sucked out, cause instrument system accessory to wear, shorten the service life of instrument；Acid mobile phase uses high chlorine
Acid, water and dimethyl sulfoxide mixed solution, perchloric acid are acid with strong oxidizing property, perishable instrument system, shorten chromatographic column and instrument
Service life.
Application number CN201510431875.1, entitled " high effective liquid chromatography for measuring Chlortetracycline premix content and phase
The method for closing substance " patent use instead water, phosphoric acid, n,N-Dimethylformamide mixed solution as mobile phase, acidity is more
Mildly, but this method only alleviates the problem of crystallization salt abrasion is with strong oxidizing property acid corrosion to a certain extent, not root
The mobile phase long-time service of this solution, this phosphoric acid can certainly will also corrode instrument system, shorten device longevity.
In addition, the testing cost of high performance liquid chromatography is expensive, sample pretreatment is cumbersome, is not suitable for largely detecting, because
This needs a kind of quick, easy, accurately and reliably detection method for Chlortetracycline premix production.
Summary of the invention
It is pre- to provide a kind of quickly measurement aureomycin for deficiency existing for microbial method and high performance liquid chromatography by the present invention
The method of aureomycin content in mixture.
The present invention adopts the following technical scheme:
A kind of method of aureomycin content in quick measurement Chlortetracycline premix, comprising the following steps:
The drafting of S1, standard curve
1) drafting of aureomycin hydrochloride standard curve:
It takes the standard items of aureomycin hydrochloride and is configured to a series of standard solution of potency gradients, multiple groups are arranged in each potency
Standard solution, take wherein one group as the first standard control group, reference solution of the first standard control group for correction, the
The processing mode of one standard control group is as follows: the hydrochloric acid of n mole is added；
For remaining each group as the first standard test group, the processing mode of the first standard test group is as follows: being added n moles
Heated after the hydrochloric acid of amount；
It measures the absorbance of each first standard test group and is averaged, then according to mean light absorbency and the first standard
The corresponding relationship of experimental group potency establishes the standard curve of aureomycin hydrochloride；
2) drafting of quadracycline standard curve:
It takes the standard items of quadracycline and is configured to a series of standard solution of potency gradients, multiple groups are arranged in each potency
Standard solution, take wherein one group as the second standard control group, reference solution of the second standard control group for correction, the
The processing mode of two standard control groups is as follows: volume V is addedEDTAEDTA mixed liquor, after standing, add the salt of n mole
Acid；
For remaining each group as the second standard test group, the processing mode of the second standard test group is as follows: volume is added
VEDTAEDTA mixed liquor, after standing, add the hydrochloric acid of n mole, then carry out same S1 1) in heat treatment；
It measures the absorbance of each second standard test group and is averaged, then according to mean light absorbency and the second standard
The corresponding relationship of experimental group potency establishes the standard curve of quadracycline；
The preparation of S2, Chlortetracycline premix sample solution
1) M will be taken after the grinding of Chlortetracycline premix sample, discharges the antibiotic composition in sample sufficiently through dissolution process
Afterwards, filtrate, filtrate volume V are filtered to take0To get V0Chlortetracycline premix sample solution；
The detection of S3, Chlortetracycline premix sample solution total titer
1) the first control group is set and the first experimental group, the first control group and the first experimental group respectively take V1Step S2 matched
The Chlortetracycline premix sample solution of system, the processing mode of the first control group is the same as the first standard control group, the place of the first experimental group
For reason mode with the first standard test group, the first control group and the first experimental group are diluted with water to same volume V11；
2) spectrophotometer detects: using the first control group as reference solution, after correction, measuring the absorbance of the first experimental group
A；
3) calculate: the standard curve of aureomycin hydrochloride obtained by control step S1, absorbance A correspond to potency N1, then
The total titer R of Chlortetracycline premix sample solution1=N1×V11/V1；
Total titer=V of Chlortetracycline premix sample0×R1/M；
The detection of tetracycline activity in S4, Chlortetracycline premix sample solution
1) the second control group is set and the second experimental group, the second control group and the second experimental group respectively take V2Step S2 matched
The Chlortetracycline premix sample solution of system, the processing mode of the second control group is the same as the second standard control group, the place of the second experimental group
For reason mode with the second standard test group, the second control group and the second experimental group are diluted with water to same volume V22；
2) spectrophotometer detects: using the second control group as reference solution, after correction, measuring the absorbance of the second experimental group
B；
3) calculate: the standard curve of quadracycline obtained by control step S1, absorbance B correspond to potency N2, then
Tetracycline activity R in Chlortetracycline premix sample solution2=N2×V22/V2；
Tetracycline activity=V in Chlortetracycline premix sample0×R2/M；
Aureomycin potency R in S5, Chlortetracycline premix sampleCTC=V0×R1/M-V0×R2/M；
It is mould to measure gold by step S1 to S5 respectively by S6, the Chlortetracycline premix sample for taking multiple and different aureomycin contents
Plain potency RCTC, while measuring aureomycin content respectively using high performance liquid chromatography, then by gained aureomycin potency RCTCWith
The aureomycin content of high performance liquid chromatography detection carries out linear fit, obtains regression equation；
S7, Chlortetracycline premix sample to be measured is taken, obtains aureomycin potency R by step S1 to S5CTCAfterwards, it substitutes into and returns
Equation is up to the aureomycin content in the Chlortetracycline premix sample to be measured.
Testing principle of the invention are as follows:
Be added hydrochloric acid in Chlortetracycline premix sample solution, aureomycin and tetracycline therein in acid condition, through adding
A molecular water is sloughed after processed by hot bath and generate yellow dehydration aureomycin and yellow dehydration tetracycline, color depth
(corresponding to the absorbance measured in spectrophotometer) is directly proportional to potency；
EDTA mixed liquor used in the present invention is formulated by sodium hydroxide and disodium ethylene diamine tetraacetate are soluble in water,
Therefore EDTA mixed liquor is added in Chlortetracycline premix sample solution, aureomycin is converted into different under alkaline condition in alkalinity
Aureomycin makes aureomycin lose the property to develop the color in acid condition, and then plus hydrochloric acid heats tetracycline in acid condition
It sloughs 1 molecular water and generates the dehydration tetracycline of yellow, color depth (corresponding to the absorbance measured in spectrophotometer)
It is directly proportional to potency
Based on principles above, standard solution and Chlortetracycline premix sample solution are made and adds hydrochloric acid and/or adds EDTA mixed
The processing of liquid is closed, absorbance is measured, the standard curve of absorbance and potency is established by standard solution, it is pre- by measurement aureomycin
The absorbance of mixture sample solution, reference standard curve obtain aureomycin and tetracycline in Chlortetracycline premix sample solution
The potency of total titer and tetracycline itself, and then aureomycin potency is calculated.
The aureomycin content for finally measuring aureomycin potency obtained by spectrophotometer and high performance liquid chromatography carries out line
Property fitting, obtain regression equation, thereafter for Chlortetracycline premix sample to be measured, only need to measure aureomycin potency, can substitute into
Equation obtains aureomycin content.
Beneficial effects of the present invention are as follows:
(1) instrument and equipment less investment a, it is only necessary to spectrophotometer；
(2) easy to operate, it avoids cumbersome, reduces labor intensity；
(3) reagent dosage is few, has saved detection reagent cost, and agents useful for same alleviates dirt to human and environment nonhazardous
Water process burden；
(4) sample treatment and mensuration program are simple, are easy to implement quick detection, can disposably detect a large amount of samples；
(5) high performance liquid chromatography is not less than for the accuracy and reproducibility of Chlortetracycline premix detection, is suitable for looking forward to
The continuous detection of the daily production of industry, it is accurate and reliable and economical and practical.
Detailed description of the invention
Fig. 1 is the Linear Fit Chart in step S6；
Fig. 2 is the UV-visible absorption spectrum of aureomycin hydrochloride.
Specific embodiment
In order to keep technical purpose of the invention, technical scheme and beneficial effects clearer, with reference to the accompanying drawing and specifically
Embodiment is further illustrated technical solution of the present invention.
A kind of method of aureomycin content in quick measurement Chlortetracycline premix, comprising the following steps:
The drafting of S1, standard curve
1) foundation of aureomycin hydrochloride standard curve
It is accurate draw each 0.20mL, 0.40mL of aureomycin hydrochloride standard solution, 0.60mL that potency is 1000u/mL,
In the volumetric flask of seven groups of 50mL, every group of 3 volumetric flasks are separately added into 2mol/L by 0.80mL, 1.00mL, 1.20mL, 1.40mL
Hydrochloric acid solution 5mL, wherein a volumetric flask add distilled water dilution constant volume, shake up, as reference solution, i.e. the first standard control
Group；Another two carry out the water bath processing of 100 DEG C, 5min, and after cooling plus distilled water dilutes constant volume, shakes up, as the first standard reality
Group is tested, is averaged after test；
Wavelength 440nm is selected on spectrophotometer, is corrected with reference solution, and with 1cm cuvette, blank is made with reference solution
Control measures the absorbance of the first standard test group in above-mentioned 7 groups and is averaged, using potency as ordinate, with mean light absorbency
For abscissa, aureomycin hydrochloride standard curve is established；
2) foundation of quadracycline standard curve
It is accurate draw each 0.20mL, 0.40mL of 1000u/mL quadracycline standard solution, 0.60mL, 0.80mL,
In seven groups of 50mL volumetric flasks, every group of 3 volumetric flasks are separately added into EDTA mixed liquor 11mL by 1.00mL, 1.20mL, 1.40mL,
Be stored at room temperature 5min, be then respectively adding the hydrochloric acid solution 5mL of 2mol/L, wherein a volumetric flask add distilled water dilution constant volume,
It shakes up, as reference solution, i.e. the second standard control group；Another two carry out the water bath processing of 100 DEG C, 5min, after cooling plus steam
Distilled water dilution constant volume shakes up, and as the second standard test group, is averaged after test；
Wavelength 440nm is selected on spectrophotometer, is corrected with reference solution, and with 1cm cuvette, blank is made with reference solution
Control measures the absorbance of the second standard test group in above-mentioned 7 groups and is averaged, using potency as ordinate, with mean light absorbency
For abscissa, quadracycline standard curve is established；
The preparation method of EDTA mixed liquor used is following (similarly hereinafter):
Weigh respectively analysis 55 grams of pure cerium hydroxide sodium, 4.55 grams of disodium ethylene diamine tetraacetate in beaker, add 200mL to distill
After water dissolution, it is diluted to 5000mL, is stored in reagent bottle, it is spare；
The preparation of S2, Chlortetracycline premix sample solution
1) Chlortetracycline premix sample is ground into powdery using powder sample device and takes M g in 50mL volumetric flask, Shao Liangduo
After secondary addition soaking agent and yawing, constant volume after bubble-free generates, then ultrasound bath 20min, passes through soaking agent and ultrasonic wave
Water bath processing sufficiently promotes the dissolution of aureomycin and tetracycline, and taking-up is cooled to room temperature, and the effect of above-mentioned dissolution process is to make
Antibiotic composition in sample sufficiently discharges, and then filters to take filtrate, filtrate volume V0=50mL is to get V0The gold of=50mL
Mycin pre-mixing agent sample solution；
The soaking agent is formed by the hydrochloric acid of water, acetone and 5mol/L according to the volume ratio mixed preparing of 1000:500:80；
The concentration of Chlortetracycline premix sample is 0.004~0.007g/mL in gained Chlortetracycline premix sample solution；
The detection of S3, Chlortetracycline premix sample solution total titer
1) the first control group is set and the first experimental group, the first control group and the first experimental group respectively take V1The step of=0.5mL
The rapid prepared Chlortetracycline premix sample solution of S2 is in two 50mL volumetric flasks, and each addition 2mol/L hydrochloric acid 5mL, and first
Control group is diluted with water constant volume, shakes up, cooling after the water bath processing in 100 DEG C of the first experimental group progress, 5min, is diluted with water
Constant volume shakes up, V11=50mL；
2) spectrophotometer detects: using the first control group as reference solution, after correcting at wavelength 440nm, measuring first
The absorbance A of experimental group；
3) calculate: the standard curve of aureomycin hydrochloride obtained by control step S1, absorbance A correspond to potency N1, then
The total titer R of Chlortetracycline premix sample solution1(u/mL)=N1×V11/V1=100N1；
Total titer (u/g)=V of Chlortetracycline premix sample0×R1/ M=50 × R1/M；
The detection of S4, Chlortetracycline premix sample solution tetracycline activity
1) the second control group is set and the second experimental group, the second control group and the second experimental group respectively take V2The step of=1mL
The prepared Chlortetracycline premix sample solution of S2 is in two 50mL volumetric flasks, and each EDTA mixed liquor that 11mL is added, room
After temperature stands 5min, the hydrochloric acid 5mL of 2mol/L is respectively added in the second control group and the second experimental group again, and then the second control group adds water
It dilutes and constant volume, shakes up；It is cooling after water bath processing in 100 DEG C of second experimental group progress, 5min, it is diluted with water constant volume, shakes
It is even, V22=50mL；
2) spectrophotometer detects: using the second control group as reference solution, after correcting at wavelength 440nm, measuring second
The absorbance B of experimental group；
3) calculate: the standard curve of quadracycline obtained by control step S1, absorbance B correspond to potency N2, then
Tetracycline activity R in Chlortetracycline premix sample solution2(u/mL)=N2×V22/V2=50N2；
Tetracycline activity (u/g)=V in Chlortetracycline premix sample0×R2/ M=50 × R2/M；
Medicine ingredient in S5, Chlortetracycline premix is mainly aureomycin and tetracycline, so in Chlortetracycline premix sample
Aureomycin potency RCTC=V0×R1/M-V0×R2/ M=50 × (R1-R2)/M；
S6, take multiple groups difference aureomycin content and different weight Chlortetracycline premix sample (aureomycin content from 13%,
15%, 20%, 22%, 25% differ), multiple groups absorbance A and B are measured by above-mentioned steps S1 to S5, and correspondence is calculated
R1、R2And RCTC, while the aureomycin content of every group of Chlortetracycline premix sample is measured using high performance liquid chromatography, then will
Gained aureomycin potency RCTCLinear fit is carried out with the aureomycin content of high performance liquid chromatography detection, is examined with spectrophotometer
The aureomycin potency R of surveyCTCFor abscissa, the aureomycin content of high performance liquid chromatography detection is ordinate, do linear regression and
Correlation analysis, draw Linear Fit Chart as shown in Figure 1, and obtain regression equation: aureomycin content=- 6.570+0.9851 ×
RCTC, coefficient R-Sq=99.8%.The detection data of the multiple groups Chlortetracycline premix sample is as shown in table 1.
S7, Chlortetracycline premix sample to be measured is taken, obtains aureomycin potency R by step S1 to S5CTCAfterwards, it substitutes into and returns
Equation is up to the aureomycin content in Chlortetracycline premix sample to be measured.
The detection data of 1 multiple groups Chlortetracycline premix sample of table summarizes
Note: the percentage in sample number into spectrum bracket refers to the mass ratio of Chlortetracycline premix Gold Samples mycin content；
Potency unit g/kg=103u/g。
Present invention selection is corrected wavelength 440nm at, and reason is: detect through ultraviolet-visible spectrophotometer (see Fig. 2,
Table 2), aureomycin hydrochloride has maximum absorption band at 440nm, therefore 440nm is taken to be corrected.
Specific detection method is as follows:
(1) 1000u/mL aureomycin hydrochloride standard solution is prepared, therefrom respectively draws 1mL standard solution in two 50mL capacity
In bottle, it is separately added into the hydrochloric acid of 5mL 2mol/mL, wherein one adds distilled water constant volume, shakes up, as reference solution, another
In 100 DEG C heating water bath 5 minutes, taking-up is cooled to room temperature, and is added water constant volume, is shaken up；
(2) 752N type ultraviolet-visible spectrophotometer is used, booting preheating 20 minutes is surveyed above-mentioned molten at different wave length
The absorbance of liquid, using wavelength as abscissa, absorbance is the absorption spectrogram that ordinate obtains as shown in Fig. 2, absorbing data statistics
It is as shown in table 2:
The ultraviolet-ray visible absorbing data of 2 aureomycin hydrochloride standard solution of table
| Wavelength (nm) | 240 | 400 | 410 | 420 | 430 | 436 | 438 | 440 | 442 | 444 | 446 | 448 | 450 |
| Absorbance | 0.00 | 0.005 | 0.153 | 0.230 | 0.272 | 0.286 | 0.288 | 0.290 | 0.289 | 0.288 | 0.287 | 0.280 | 0.277 |
| Wavelength (nm) | 452 | 454 | 456 | 458 | 460 | 470 | 480 | 490 | 500 | 510 | 520 | 860 | -- |
| Absorbance | 0.273 | 0.268 | 0.259 | 0.254 | 0246 | 0.195 | 0.140 | 0.088 | 0.065 | 0.040 | 0.023 | 0.000 | -- |
It should be noted last that: technical solution of the present invention that the above embodiments are only illustrative and not limiting is any right
The equivalent replacement and do not depart from the modification of spirit and scope of the invention or locally replace that the present invention carries out, should all cover in this hair
Within bright protective scope of the claims.
Claims (1)
1. a kind of method of aureomycin content in quickly measurement Chlortetracycline premix, which comprises the following steps:
The drafting of S1, standard curve
1) drafting of aureomycin hydrochloride standard curve:
It takes the standard items of aureomycin hydrochloride and is configured to a series of standard solution of potency gradients, multiple groups standard is arranged in each potency
Solution, take wherein one group as the first standard control group, reference solution of the first standard control group for correction, the first mark
The processing mode of quasi- control group is as follows: the hydrochloric acid of n mole is added；
For remaining each group as the first standard test group, the processing mode of the first standard test group is as follows: n mole is added
Heated after hydrochloric acid；
Selection measurement wavelength 440nm, measures the absorbance of each first standard test group and is averaged, and is then inhaled according to average
Luminosity and the corresponding relationship of the first standard test group potency establish the standard curve of aureomycin hydrochloride；
2) drafting of quadracycline standard curve:
It takes the standard items of quadracycline and is configured to a series of standard solution of potency gradients, multiple groups standard is arranged in each potency
Solution, take wherein one group as the second standard control group, reference solution of the second standard control group for correction, the second mark
The processing mode of quasi- control group is as follows: volume V is addedEDTAEDTA mixed liquor, after standing, add the hydrochloric acid of n mole；
For remaining each group as the second standard test group, the processing mode of the second standard test group is as follows: volume V is addedEDTA's
EDTA mixed liquor after standing, adds the hydrochloric acid of n mole, then carries out same S1 1) in heat treatment；
Selection measurement wavelength 440nm, measures the absorbance of each second standard test group and is averaged, and is then inhaled according to average
Luminosity and the corresponding relationship of the second standard test group potency establish the standard curve of quadracycline；
The preparation of S2, Chlortetracycline premix sample solution
1) M will be taken after the grinding of Chlortetracycline premix sample, after dissolution process discharges the antibiotic composition in sample sufficiently, mistake
Leaching filtrate, filtrate volume V0 To get V0 Chlortetracycline premix sample solution；
The detection of S3, Chlortetracycline premix sample solution total titer
1) the first control group is set and the first experimental group, the first control group and the first experimental group respectively take V1Step S2 it is prepared
Chlortetracycline premix sample solution, the processing mode of the first control group is the same as the first standard control group, the processing side of the first experimental group
For formula with the first standard test group, the first control group and the first experimental group are diluted with water to same volume V11；
2) spectrophotometer detects: selection measurement wavelength 440nm after correction, measures first using the first control group as reference solution
The absorbance A of experimental group；
3) calculate: the standard curve of aureomycin hydrochloride obtained by control step S1, absorbance A correspond to potency N1, then
The total titer R of Chlortetracycline premix sample solution1= N1×V11/ V1；
Total titer=V of Chlortetracycline premix sample0×R1/ M；
The detection of tetracycline activity in S4, Chlortetracycline premix sample solution
1) the second control group is set and the second experimental group, the second control group and the second experimental group respectively take V2Step S2 it is prepared
Chlortetracycline premix sample solution, the processing mode of the second control group is the same as the second standard control group, the processing side of the second experimental group
For formula with the second standard test group, the second control group and the second experimental group are diluted with water to same volume V22；
2) spectrophotometer detects: selection measurement wavelength 440nm after correction, measures second using the second control group as reference solution
The absorbance B of experimental group；
3) calculate: the standard curve of quadracycline obtained by control step S1, absorbance B correspond to potency N2, then
Tetracycline activity R in Chlortetracycline premix sample solution2= N2×V22 / V2；
Tetracycline activity=V in Chlortetracycline premix sample0×R2/ M；
Aureomycin potency R in S5, Chlortetracycline premix sampleCTC = V0×R1/ M - V0×R2/ M；
S6, the Chlortetracycline premix sample for taking multiple and different aureomycin contents measure aureomycin effect by step S1 to S5 respectively
Valence RCTC, while measuring aureomycin content respectively using high performance liquid chromatography, then by gained aureomycin potency RCTCWith it is efficient
The aureomycin content of liquid chromatography detection carries out linear fit, obtains regression equation；Wherein, the Chlortetracycline premix sample
In aureomycin content be 13%, 15%, 20%, 22% or 25%；
S7, Chlortetracycline premix sample to be measured is taken, obtains aureomycin potency R by step S1 to S5CTCAfterwards, regression equation is substituted into
Up to the aureomycin content in the Chlortetracycline premix sample to be measured.
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