CN103278466A - Quality control method for toad venom injection - Google Patents
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- CN103278466A CN103278466A CN2013101571867A CN201310157186A CN103278466A CN 103278466 A CN103278466 A CN 103278466A CN 2013101571867 A CN2013101571867 A CN 2013101571867A CN 201310157186 A CN201310157186 A CN 201310157186A CN 103278466 A CN103278466 A CN 103278466A
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Abstract
The invention relates to a quality control method for measuring the content of toad venom in a toad venom injection based on ultraviolet spectrophotometry. The method comprises preparing a standard curve and obtaining a calculation formula of the standard curve, and is characterized by also comprising a sample pretreatment, adding a masking agent which is acetone or methyl aldehyde or a mixture of acetone and methyl aldehyde to produce a masking effect with sodium hydrogen sulfite, undergoing a nucleophilic addition reaction with the masking agent and the sodium hydrogen sulfite, then adding dimethylamino benzene formaldehyde/hydrochloric acid solution to undergo a chromogenic reaction, detecting a light absorption value A of the reaction solution at a wavelength of 540 to 560 nm, substituting the light absorption value A into the calculation formula of the stander curve to calculate, and obtaining the content of the toad venom in the toad venom injection sample. Through the method provided by the invention, sodium hydrogen sulfite is masked by using the masking agent of sodium hydrogen sulfite which is an antioxidant, and antioxidant sodium hydrogen sulfite components are prevented from affecting measuring results. The quality control method has the good effects of good stability and high accuracy.
Description
Technical field
The invention belongs to the drug quality analysis technical field, relate to a kind of Venenum Bufonis Injection method of quality control based on ultraviolet spectrophotometry.
Background technology
Venenum Bufonis Injection is made up of compositions such as the dried venom of toads, sodium bisulfite, phenmethylol, Tween-80s, and the dried venom of toads is main active drug, and other compositions are auxiliary material.Indole derivatives in the total composition of toad cake extract (water-soluble indoles total alkaloids) has certain physiologically active, so adopt indoles alkali as the control index.The quality of the dried venom of toads is seldom controlled in standard, the HPLC method can be measured the content of the dried venom of toads, but the requirement height to instrument, present most of animal pharmaceutical factory difficulty carries out, adopt ultraviolet spectrophotometry can control the quality that intermediate is water-soluble indoles total alkaloids aborning, fast and convenient, effect is better.Ultraviolet spectrophotometry is main to generate coloring matter according to water-soluble indoles total alkaloids and paradime thylaminobenzaldehyde solution reaction, controls dried venom of toads quality by the absorption photometric of measuring this coloring matter.And in the Venenum Bufonis Injection quality control process of reality, during for the detection of the water-soluble indoles total alkaloids of dried venom of toads effective constituent, sodium bisulfite in the Venenum Bufonis Injection is antioxidant, has very strong reductibility, can disturb the chromogenic reaction (redox reaction) of Venenum Bufonis Injection content assaying method, thereby influence the assay result of Venenum Bufonis Injection, measurement result is lower more than 30% than actual content, cause quality control to produce error, the accuracy of method can not get guaranteeing.
Also the dried venom of toads method of quality control in the relevant parenteral solution that contains dried venom of toads composition has been carried out some researchs through retrieval discovery domestic scholars, its research contents is as follows:
Chinese patent CN200510119552.5, applicant Shenyang Pharmaceutical University, disclose a kind of Senso-like efficient liquid-phase chromatograph finger print atlas identifying method, adopted high performance liquid chromatography, drawn the distinctive bufotenine class of toad venom and bufotalin class chemical fingerprint; Utilize high performance liquid chromatograph, UV detecting device, chromatographic work station, get dried venom of toads sample 25mg to be measured, put in the tool plug conical flask, precision adds methyl alcohol 20ml, claims to decide weight, add hot reflux 1h, put coldly, claim again to decide weight, supply the weight that subtracts mistake with methyl alcohol, shake up, cross 0.2 μ m filter membrane and namely get the toad venom need testing solution.With dried venom of toads need testing solution to be measured, at analytical column, phase flows: methyl alcohol (A)-damping fluid, carry out gradient elution, sample size 20 μ l, retention time 90min, namely obtain toad venom distinctive chemical fingerprint separately, and to all characteristic peaks all or major part carry out chemistry and belong to.This method can be measured the efficient liquid-phase chromatograph finger print atlas of toad venom exactly, thereby for the true and false of differentiating the dried venom of toads with formulate dried venom of toads medicinal material and the Venenum Bufonis Injection quality standard provides reliable scientific basis.But this HPLC method can be measured the content of the dried venom of toads, but to the requirement height of instrument, causes detecting cost, and present most of animal pharmaceutical factory difficulty carries out.
2005 the 36th volumes of guangdong agricultural science the 2nd phase 158-160 page or leaf, Duan Yali has introduced ultraviolet spectrophotometry and has controlled dried venom of toads quality in the crisp yellow parenteral solution, it is regulated by effective pH value, remove the interference of yellow Cen glycosides component, generate the principle of coloring matter according to water-soluble indoles total alkaloids and paradime thylaminobenzaldehyde solution reaction, measuring substance that show color after chromogenic reaction is finished is the absorbance at 555nm place at wavelength, and this detection method is effective, accuracy, stability, repeated higher.This method can be controlled dried venom of toads content indirectly, controls its quality as an inspection item in quality standard.Yet the dried venom of toads method of quality control that they introduce is mainly used in dried venom of toads quality control in the crisp yellow parenteral solution, owing to mainly contain the pure traditional Chinese medicine that Chinese crude drugs such as the dried venom of toads, Huang Cen, cornu bubali are processed in the crisp yellow parenteral solution, main interference component is yellow Cen; And the effective constituent in the Venenum Bufonis Injection comprises compositions compositions such as the dried venom of toads, sodium bisulfite, phenmethylol, Tween-80, sodium bisulfite in the Venenum Bufonis Injection is antioxidant, has very strong reductibility, can disturb the chromogenic reaction (redox reaction) of Venenum Bufonis Injection content assaying method, cause the instability of dried venom of toads method of quality control, testing result occurs than mistake, influences the accuracy of control method.
Summary of the invention
Purpose of the present invention: exist control method to be subjected to the influence of antioxidant in the dried venom of toads quality for overcoming in the existing Venenum Bufonis Injection, the detection method instability, the testing result error is bigger, shortcomings such as method of quality control cost height, time-consuming length provide a kind of Venenum Bufonis Injection method of quality control that adds anti-oxidant screening agent based on ultraviolet spectrophotometry.This method is sheltered sodium bisulfite with the screening agent of the sodium bisulfite antioxidant of utilization, avoids other components to the influence of measurement result, and this quality checking and controlling method reaches good stability, the high excellent results of accuracy.
Technical scheme of the present invention is as follows: a kind of Venenum Bufonis Injection method of quality control, this method comprise that typical curve preparation, typical curve computing formula obtain, and this method is further comprising the steps of:
The step 1) pre-service: get Venenum Bufonis Injection sample 5 ~ 10ml and place test tube, ultrasonic processing 5 ~ 10min under 20 ~ 25 ℃ of conditions obtains pretreated dried venom of toads injected sample.
Step 2) masking action: get pretreated dried venom of toads injected sample 5 ~ 6ml that step 1) obtains and place tool plug test tube, add anti-oxidant screening agent according to the ratio that dried venom of toads injected sample and anti-oxidant screening agent volume ratio are 5ml:1 ~ 4ml, mixing is placed under 20～25 ℃ of conditions and carries out masking action 1 ~ 15min, sodium bisulfite generation nucleophilic addition in screening agent and the dried venom of toads injected sample obtains sample after the masking action.
Step 3) chromogenic reaction: toward step 2) add paradime thylaminobenzaldehyde hydrochloric acid (2 → 3) solution 4.0 ~ 6.0ml after the masking action that obtains in the sample, mixing is placed on chromogenic reaction 30 ~ 40min takes place under 20～25 ℃ of conditions, obtains solution after the chromogenic reaction.
Step 4) is measured absorbance: solution 3.5 ~ 5ml places the ultraviolet spectrophotometer glass dish after getting the chromogenic reaction that step 3) obtains, and measures the light absorption value A at 540 ~ 560nm wavelength place, reading and recording light absorption value A.
Step 5) result calculates: calculate in the light absorption value A substitution typical curve computing formula that step 4) is obtained, calculate dried venom of toads content in the Venenum Bufonis Injection sample.
As the further restriction of Venenum Bufonis Injection method of quality control of the present invention, described anti-oxidant screening agent is a kind of or their potpourri in acetone, the formaldehyde.
As the further restriction of Venenum Bufonis Injection method of quality control of the present invention, described its ultrasonic power of ultrasonic processing is 50 ~ 80W.
Further restriction as Venenum Bufonis Injection method of quality control of the present invention, described paradime thylaminobenzaldehyde hydrochloric acid (2 → 3) its component of solution and parts by weight thereof are: 10 ~ 20 parts of paradime thylaminobenzaldehyde hydrochloric acid, 2 ~ 5 parts of 10% hydrochloric acid solutions, 80 ~ 90 parts in water.
Further restriction as Venenum Bufonis Injection method of quality control of the present invention, the composition that contains in the described Venenum Bufonis Injection comprises the dried venom of toads, sodium bisulfite, phenmethylol, Tween-80, Venenum Bufonis Injection is the Venenum Bufonis Injection of type for animals, and every 1ml Venenum Bufonis Injection contains indoles total alkaloids amount with serotonin (C
10H
12N
2O) meter 〉=14.0 μ g.
Abundant for the present invention is disclosed, Venenum Bufonis Injection method of quality control of the present invention comprises that the step specific implementation process of typical curve preparation, the acquisition of typical curve computing formula is as follows:
(1) preparation of reference substance solution: precision takes by weighing serotonin hydrochloride reference substance 18mg, puts in the 100ml measuring bottle, adds water and makes dissolving, and be diluted to scale, shake up, precision is measured 10ml, puts in the 50ml measuring bottle, thin up shakes up to scale, namely gets to contain serotonin 0.030mg among every lml.
(2) preparation of typical curve: precision is measured reference substance solution 0.5,1.0,2.0,3.0,4.0,5.0 ml, put respectively in the 10ml measuring bottle, each accurate paradime thylaminobenzaldehyde hydrochloric acid (2 → 3) solution 5.0ml that adds adds water to scale, shake up, chromogenic reaction 20 ~ 40min takes place under 20～25 ℃, is blank with water, according to UV-VIS spectrophotometry, measure absorbance at 540 ~ 560nm wavelength place, be ordinate with the absorbance, concentration is horizontal ordinate, the drawing standard curve.
The cardinal principle that a kind of Venenum Bufonis Injection method of quality control of the present invention is realized is: the feature effective constituent according to Venenum Bufonis Injection is contained water-soluble indoles total alkaloids, and this water-soluble indoles total alkaloids mainly comprises and mainly contains tryptamines indoles, monoterpene indoles, dimeric indole class, utilize indoles alkaloid to generate coloring matter with the paradime thylaminobenzaldehyde solution reaction, and coloring matter is absorption value and coloring matter relevant at 540 ~ 560nm place at wavelength; In the specimen preparation process, by adding anti-oxidant screening agent (a kind of or their potpourri in acetone, the formaldehyde), utilize acetone (or formaldehyde) and sodium bisulfite generation nucleophilic addition, sulphur is as the center of nucleophilic addition, connect a hydroxyl on the generation carbonyl carbon, connect the nucleophilic addition of a sodium sulfonate again, realize sheltering having than the sodium bisulfite of strong antioxidant action, get rid of this antioxidant to the interference of chromogenic reaction, make testing result good stability, accuracy height.
Venenum Bufonis Injection method of quality control of the present invention has produced following good result:
(1) Venenum Bufonis Injection of the present invention adds the stability test result who develops the color behind the developer and shows: do not add acetone, formaldehyde screening agent chromogenic reaction is unstable and react insufficient, so absorbance log is lower; After adding the acetone screening agent antioxidant sodium bisulfite being sheltered, chromogenic reaction is stable and fully, the colour developing effect after 30 minutes absorbance log reach mxm., just descend gradually after 50 minutes, with respect to the colour developing effect that does not add screening agent, good reproducibility.
(2) Venenum Bufonis Injection method of quality control of the present invention, can eliminate interference after adopting macking technique that its sodium bisulfite is sheltered, test findings shows, this law is easy, accurate for dried venom of toads assay process, overcome the measurement result error of traditional detection method (not adding screening agent), low, the time-consuming weak point of cost of determination.
Description of drawings
The nucleophilic addition process of Fig. 1 screening agent and sodium bisulfite.
Embodiment
Below in conjunction with Fig. 1 and embodiment Venenum Bufonis Injection method of quality control of the present invention is described.
Embodiment 1
Paradime thylaminobenzaldehyde hydrochloric acid (2 → 3) its component of solution and the parts by weight thereof used in the present embodiment are: 15 parts of paradime thylaminobenzaldehyde hydrochloric acid, 2 parts of 10% hydrochloric acid solutions, 83 parts in water.
One, the making of typical curve
(1) preparation of reference substance solution: precision takes by weighing serotonin hydrochloride reference substance 18mg, puts in the 100ml measuring bottle, adds water and makes dissolving, and be diluted to scale, shake up, precision is measured 10ml, puts in the 50ml measuring bottle, thin up shakes up to scale, namely gets to contain serotonin 0.030mg among every lml.
(2) preparation of typical curve: precision is measured reference substance solution 0.5,1.0,2.0,3.0,4.0,5.0 ml, put respectively in the 10ml measuring bottle, each accurate paradime thylaminobenzaldehyde hydrochloric acid (2 → 3) solution 5.0ml that adds adds water to scale, shake up, chromogenic reaction 30min takes place under 20 ℃, is blank with water, according to UV-VIS spectrophotometry, measure absorbance at 555nm wavelength place, be ordinate with the absorbance, concentration is horizontal ordinate, the drawing standard curve.
Two, the detection of sample
The step 1) pre-service: get Venenum Bufonis Injection sample 5ml and place test tube, the ultrasonic processing of 50W 7min obtains pretreated dried venom of toads injected sample under 20 ℃ of conditions.
Step 2) masking action: get the pretreated dried venom of toads injected sample 5ml that step 1) obtains and place tool plug test tube, according to dried venom of toads injected sample: the acetone volume ratio is that the ratio of 5ml:2ml is added anti-oxidant screening agent acetone, mixing is placed under 20 ℃ of conditions and carries out masking action 10min, masking action is the nucleophilic addition of screening agent acetone and sodium bisulfite, its course of reaction obtains sample after the masking action as shown in Figure 1.
Step 3) chromogenic reaction: toward step 2) add paradime thylaminobenzaldehyde hydrochloric acid (2 → 3) solution 4.0ml after the masking action that obtains in the sample, mixing is placed on chromogenic reaction 30min takes place under 20 ℃ of conditions, obtains solution after the chromogenic reaction.
Step 4) is measured absorbance: solution 3.5ml places the ultraviolet spectrophotometer glass dish after getting the chromogenic reaction that step 3) obtains, and measures the light absorption value A at 555nm wavelength place, reading and recording light absorption value A.
Step 5) result calculates: calculate in the light absorption value A substitution typical curve computing formula that step 4) is obtained, calculate dried venom of toads content in the Venenum Bufonis Injection sample.
The 20.4ug/ml as a result that the present embodiment sample determination obtains, with the testing result 20.1ug/ml error range of efficient liquid phase HPLC method 1.5%, and the measurement result that the ultraviolet spectrophotometry (the same present embodiment of all the other determination steps) of not adding screening agent obtains is 14.5ug/ml, with respect to efficient liquid phase HPLC method error 27.8%.
Embodiment 2
Paradime thylaminobenzaldehyde hydrochloric acid (2 → 3) its component of solution and the parts by weight thereof used in the present embodiment are: 10 parts of paradime thylaminobenzaldehyde hydrochloric acid, 5 parts of 10% hydrochloric acid solutions, 85 parts in water.
One, the preparation of typical curve
In the present embodiment, chromogenic reaction 35min takes place down at 25 ℃ in reference substance solution, is blank with water, according to UV-VIS spectrophotometry, measures absorbance at 550nm wavelength place, and all the other steps are identical with embodiment 1.
Two, the detection of sample
The step 1) pre-service: get Venenum Bufonis Injection sample 7ml for animals and place test tube, the ultrasonic processing of 80W 5min obtains pretreated dried venom of toads injected sample under 25 ℃ of conditions.
Step 2) masking action: get the pretreated dried venom of toads injected sample 5ml that step 1) obtains and place tool plug test tube, according to dried venom of toads injected sample: the formaldehyde volume ratio is that the ratio of 5ml:1ml is added anti-oxidant screening agent formaldehyde, mixing is placed under 25 ℃ of conditions and carries out masking action 5min, masking action is the nucleophilic addition of screening agent formaldehyde and sodium bisulfite, its course of reaction obtains sample after the masking action as shown in Figure 1.
Step 3) chromogenic reaction: toward step 2) add paradime thylaminobenzaldehyde hydrochloric acid (2 → 3) solution 6.0ml after the masking action that obtains in the sample, mixing is placed on chromogenic reaction 35min takes place under 25 ℃ of conditions, obtains solution after the chromogenic reaction.
Step 4) is measured absorbance: solution 5ml places the ultraviolet spectrophotometer glass dish after getting the chromogenic reaction that step 3) obtains, and measures the light absorption value A at 550nm wavelength place, reading and recording light absorption value A.
Step 5) result calculates: calculate in the light absorption value A substitution typical curve computing formula that step 4) is obtained, calculate dried venom of toads content in the Venenum Bufonis Injection sample.
The 21.5ug/ml as a result that the present embodiment sample determination obtains, with the testing result 21.2ug/ml error range of efficient liquid phase HPLC method 1.4%, and the measurement result that the ultraviolet spectrophotometry (the same present embodiment of all the other determination steps) of not adding screening agent obtains is 16.35ug/ml, with respect to efficient liquid phase HPLC method error 23.0%.
Embodiment 3
Paradime thylaminobenzaldehyde hydrochloric acid (2 → 3) its component of solution and the parts by weight thereof used in the present embodiment are: 18 parts of paradime thylaminobenzaldehyde hydrochloric acid, 2 parts of 10% hydrochloric acid solutions, 80 parts in water.
One, the preparation of typical curve
In the present embodiment, chromogenic reaction 40min takes place down at 25 ℃ in reference substance solution, is blank with water, according to UV-VIS spectrophotometry, measures absorbance at 540nm wavelength place, and all the other steps are identical with embodiment 1.
Two, the detection of sample
The step 1) pre-service: get Venenum Bufonis Injection sample 8ml and place test tube, the ultrasonic processing of 60W 10min obtains pretreated dried venom of toads injected sample under 25 ℃ of conditions.
Step 2) masking action: get the pretreated dried venom of toads injected sample 6ml that step 1) obtains and place tool plug test tube, according to dried venom of toads injected sample: acetone: the formaldehyde volume ratio is the potpourri that the ratio of 5ml:2ml:1ml is added anti-oxidant screening agent acetone and formaldehyde, mixing is placed under 25 ℃ of conditions and carries out masking action 1min, masking action is the nucleophilic addition of acetone or formaldehyde screening agent and sodium bisulfite, its course of reaction obtains sample after the masking action as shown in Figure 1.
Step 3) chromogenic reaction: toward step 2) add paradime thylaminobenzaldehyde hydrochloric acid (2 → 3) solution 5.0ml after the masking action that obtains in the sample, mixing is placed on chromogenic reaction 40min takes place under 25 ℃ of conditions, obtains solution after the chromogenic reaction.
Step 4) is measured absorbance: solution 4ml places the ultraviolet spectrophotometer glass dish after getting the chromogenic reaction that step 3) obtains, and measures the light absorption value A at 540nm wavelength place, reading and recording light absorption value A.
Step 5) result calculates: calculate in the light absorption value A substitution typical curve computing formula that step 4) is obtained, calculate dried venom of toads content in the Venenum Bufonis Injection sample.
The 19.4ug/ml as a result that the present embodiment sample determination obtains, with the testing result 20.0ug/ml error range of efficient liquid phase HPLC method 3.0%, and the measurement result that the ultraviolet spectrophotometry (the same present embodiment of all the other determination steps) of not adding screening agent obtains is 13.5ug/ml, with respect to efficient liquid phase HPLC method error 32.5%.
Embodiment 4
Paradime thylaminobenzaldehyde hydrochloric acid (2 → 3) its component of solution and the parts by weight thereof used in the present embodiment are: 10 parts of paradime thylaminobenzaldehyde hydrochloric acid, 2 parts of 10% hydrochloric acid solutions, 88 parts in water.
One, the preparation of typical curve
In the present embodiment, chromogenic reaction 35min takes place down at 23 ℃ in reference substance solution, is blank with water, according to UV-VIS spectrophotometry, measures absorbance at 560nm wavelength place, and all the other steps are identical with embodiment 1.
Two, the detection of sample
The step 1) pre-service: get Venenum Bufonis Injection sample 10ml for animals and place test tube, the ultrasonic processing of 70W 8min obtains pretreated dried venom of toads injected sample under 23 ℃ of conditions.
Step 2) masking action: get the pretreated dried venom of toads injected sample 6ml that step 1) obtains and place tool plug test tube, according to dried venom of toads injected sample: the acetone volume ratio is that the ratio of 5ml:4ml is added anti-oxidant screening agent acetone, mixing is placed under 23 ℃ of conditions and carries out masking action 15min, masking action is the nucleophilic addition of screening agent acetone and sodium bisulfite, its course of reaction obtains sample after the masking action as shown in Figure 1.
Step 3) chromogenic reaction: toward step 2) add paradime thylaminobenzaldehyde hydrochloric acid (2 → 3) solution 4.0ml after the masking action that obtains in the sample, mixing is placed on chromogenic reaction 35min takes place under 23 ℃ of conditions, obtains solution after the chromogenic reaction.
Step 4) is measured absorbance: solution 4.5ml places the ultraviolet spectrophotometer glass dish after getting the chromogenic reaction that step 3) obtains, and measures the light absorption value A at 560nm wavelength place, reading and recording light absorption value A.
Step 5) result calculates: calculate in the light absorption value A substitution typical curve computing formula that step 4) is obtained, calculate dried venom of toads content in the Venenum Bufonis Injection sample.
The 22.7ug/ml as a result that the present embodiment sample determination obtains, with the testing result 23.2ug/ml error range of efficient liquid phase HPLC method 2.1%, and the measurement result that the ultraviolet spectrophotometry (the same present embodiment of all the other determination steps) of not adding screening agent obtains is 17.6ug/ml, with respect to efficient liquid phase HPLC method error 24.1%.
It should be noted that at last above embodiment is only unrestricted in order to technical scheme of the present invention to be described.Although the present invention has been described in detail with reference to embodiment, those of ordinary skill in the art is to be understood that, technical scheme of the present invention is made amendment or is equal to replacement, do not break away from the spirit and scope of technical solution of the present invention, it all should be encompassed in the claim of the present invention.The present invention is through dried venom of toads Quality Control Technology personnel Long-Term Scientific Study experience accumulation in the multidigit Venenum Bufonis Injection, and go out by creative work creation, this method is sheltered sodium bisulfite with the screening agent of the sodium bisulfite antioxidant of utilization, avoid antioxidant sodium bisulfite component to the influence of measurement result, this quality checking and controlling method reaches good stability, the high excellent results of accuracy.
Claims (3)
1. Venenum Bufonis Injection method of quality control, this method comprise that typical curve preparation, typical curve computing formula obtain, and it is characterized in that: this method is further comprising the steps of:
The step 1) pre-service: get Venenum Bufonis Injection sample 5 ~ 10ml and place test tube, ultrasonic processing 5 ~ 10min under 20 ~ 25 ℃ of conditions obtains pretreated dried venom of toads injected sample;
Step 2) masking action: get pretreated dried venom of toads injected sample 5 ~ 6ml that step 1) obtains and place tool plug test tube, add anti-oxidant screening agent according to the ratio that dried venom of toads injected sample and anti-oxidant screening agent volume ratio are 5ml:1 ~ 4ml, mixing is placed under 20～25 ℃ of conditions and carries out masking action 1 ~ 15min, sodium bisulfite generation nucleophilic addition in screening agent and the dried venom of toads injected sample obtains sample after the masking action;
Step 3) chromogenic reaction: toward step 2) add paradime thylaminobenzaldehyde hydrochloric acid solution 4.0 ~ 6.0ml after the masking action that obtains in the sample, mixing is placed on chromogenic reaction 30 ~ 40min takes place under 20～25 ℃ of conditions, obtains solution after the chromogenic reaction;
Step 4) is measured absorbance: solution 3.5 ~ 5ml places the ultraviolet spectrophotometer glass dish after getting the chromogenic reaction that step 3) obtains, and measures the light absorption value A at 540 ~ 560nm wavelength place, reading and recording light absorption value A;
Step 5) result calculates: calculate in the light absorption value A substitution typical curve computing formula that step 4) is obtained, calculate dried venom of toads content in the Venenum Bufonis Injection sample.
2. Venenum Bufonis Injection method of quality control according to claim 1, it is characterized in that: described anti-oxidant screening agent is a kind of or their potpourri in acetone, the formaldehyde.
3. Venenum Bufonis Injection method of quality control according to claim 1 and 2, it is characterized in that: described its component of paradime thylaminobenzaldehyde hydrochloric acid solution and parts by weight thereof are: 10 ~ 20 parts of paradime thylaminobenzaldehyde hydrochloric acid, 2 ~ 5 parts of 10% hydrochloric acid solutions, 80 ~ 90 parts in water.
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|---|---|---|---|---|
| CN107607526A (en) * | 2016-07-11 | 2018-01-19 | 福建大北农水产科技有限公司 | A kind of breeding water body total hardness quick detection kit |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1739598A (en) * | 2005-08-23 | 2006-03-01 | 咸阳步长医药科技发展有限公司 | Chinese medicine injection for treating cancer and its prepn process and quality control method |
| CN1776419A (en) * | 2005-11-18 | 2006-05-24 | 沈阳药科大学 | Senso-like efficient liquid-phase chromatograph finger print atlas identifying method |
| CN102106904A (en) * | 2011-01-20 | 2011-06-29 | 贵州拜特制药有限公司 | Method for preparing pharmaceutical preparation for treating palpitation and cardiodynia and quality control method thereof |
| CN102302542A (en) * | 2011-09-19 | 2012-01-04 | 谢媛媛 | Chinese medicinal oral liquid for treating cancers |
-
2013
- 2013-05-02 CN CN2013101571867A patent/CN103278466A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1739598A (en) * | 2005-08-23 | 2006-03-01 | 咸阳步长医药科技发展有限公司 | Chinese medicine injection for treating cancer and its prepn process and quality control method |
| CN1776419A (en) * | 2005-11-18 | 2006-05-24 | 沈阳药科大学 | Senso-like efficient liquid-phase chromatograph finger print atlas identifying method |
| CN102106904A (en) * | 2011-01-20 | 2011-06-29 | 贵州拜特制药有限公司 | Method for preparing pharmaceutical preparation for treating palpitation and cardiodynia and quality control method thereof |
| CN102302542A (en) * | 2011-09-19 | 2012-01-04 | 谢媛媛 | Chinese medicinal oral liquid for treating cancers |
Non-Patent Citations (4)
| Title |
|---|
| 吴乾菁: "羰基化合物与亚硫酸氢钠亲核加成反应实验的改进与探索", 《四川师范大学学报(自然科学版)》, vol. 13, no. 4, 31 December 1990 (1990-12-31), pages 104 - 107 * |
| 朱蔚玮: "蟾酥注射液质量标准的研究", 《中成药》, vol. 21, no. 2, 31 December 1999 (1999-12-31), pages 63 - 65 * |
| 段亚丽: "紫外分光光度法控制酥黄注射液中蟾酥质量", 《广西农业科技》, vol. 36, no. 2, 31 December 2005 (2005-12-31), pages 158 - 159 * |
| 马卫兴: "药物分析中掩蔽问题的讨论", 《时珍国医国药》, vol. 19, no. 8, 31 December 2008 (2008-12-31), pages 1877 - 1878 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107607526A (en) * | 2016-07-11 | 2018-01-19 | 福建大北农水产科技有限公司 | A kind of breeding water body total hardness quick detection kit |
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Application publication date: 20130904 |