CN102262010A - Method for testing chemical titer of chlorotetracycline fermentation liquor - Google Patents
Method for testing chemical titer of chlorotetracycline fermentation liquor Download PDFInfo
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- CN102262010A CN102262010A CN 201110212108 CN201110212108A CN102262010A CN 102262010 A CN102262010 A CN 102262010A CN 201110212108 CN201110212108 CN 201110212108 CN 201110212108 A CN201110212108 A CN 201110212108A CN 102262010 A CN102262010 A CN 102262010A
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- fermentation liquor
- acidifying
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- oxalic acid
- aureomycin
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Abstract
The invention discloses a method for testing the chemical titer of chlorotetracycline fermentation liquor, which can improve the working efficiency of a laboratory and enhance the titer precision. The method comprises the following steps: carrying out acidification pretreatment on the chlorotetracycline fermentation liquor, and detecting the chemical titer of the chlorotetracycline fermentation liquor. The method is characterized in that the acidification pretreatment is implemented by adding fermentation liquor per unit volume into an oxalic acid solution with a certain concentration to acidify the fermentation liquor, so that acidified liquor is obtained, and the PH value of the acidified liquor is controlled to range between 0.9 and 1.25. By using the method disclosed by the invention, the working efficiency of a laboratory can be improved, and the labor intensity of the lab technician can be reduced; meanwhile, because the oxalic acid is fully used, the consumption of the oxalic acid is saved; and the method mainly can provide accurate titer data for production analysis and control, and provide accurate economic technical indexes for enterprise production and assessment.
Description
Technical field
The present invention relates to a kind of chemical titer method of testing, particularly a kind of aureomycin fermentation liquor chemical titer method of testing that relates to aureomycin fermentation liquor chemical titer.
Background technology
In general, in the general aureomycin fermentation liquor, except that aureomycin, all contain a large amount of mycelium, unspent nutrient culture media; Range protein jelly and pigment; Heavy metal ion and aureomycin produce other metabolic products of bacterium etc., and these impurity are soluble a bit, and some is insoluble, so.When surveying chemical titer,, must carry out pre-service to fermentation liquor in order to separate effectively and to extract aureomycin.Promptly carry out acidifying, make that the unit in the mycelia is discharged fully, change aqueous phase over to, after the filtration, get filtrate survey and tire, if the pre-service acidifying is not thorough, aureomycin is tired and can not be discharged fully, remains in the filter residue, then can cause measurement result on the low side.
Disposal route commonly used at present is: adopt solid oxalic acid that fermentation liquor is carried out acidification, since fermentation liquor be carry disease germs the silk viscous fluid, so solid oxalic acid is difficult to dissolving, must fully shake up during acidifying, wasted the oxalic acid that can't dissolve of most costliness when wasting time and energy again, and acidifying is not thorough, some is tired remain in the filter residue, data are on the low side, and prior art also just is acidified to PH1.5-1.8 in the requirement of acidifying simultaneously, potential of hydrogen control is higher, acidifying is not thorough, and is unfavorable to discharging unit, makes part tire and remains in the filter residue, be to cause to tire on the low sidely, it is higher to refine yield.
Summary of the invention
The object of the present invention is to provide a kind of work efficiency that improves the laboratory, improve the aureomycin fermentation liquor chemical titer method of testing of the precision of tiring.
The object of the present invention is achieved like this: it comprises that the acidifying pre-service of aureomycin fermentation liquor and aureomycin fermentation liquor chemical titer detect, and is characterized in that: described acidifying pre-service is carried out the PH scope that acidifying obtains acidifying solution and acidifying solution and is controlled between 0.9 ~ 1.25 for the fermentation liquor with unit volume adds certain density oxalic acid solution.Specifically, the acidifying pre-service obtains acidifying solution for the oxalic acid solution that splashes into 0.1 ~ 0.5mol/L of 1 ~ 3 unit volume in the fermentation liquor with 1 unit volume carries out acidifying.
The present invention has the following advantages: 1, improve the work efficiency in laboratory, alleviate lab technician's labour intensity.2, make full use of oxalic acid, saved the use of oxalic acid; 3, conventional art improves the most yield of refining in back all below 100% because the liquid data of tiring are on the low side, and it is higher then to cause dry product to tire, and refines all un-reasonable phenomenons about 120% of yield for a long time, has correctly reflected production level.The data of tiring accurately are provided for production analysis and control.Provide economic and technical norms accurately to enterprise's production and examination.Summary of the invention
It comprises that the acidifying pre-service of aureomycin fermentation liquor and aureomycin fermentation liquor chemical titer detect, and described acidifying pre-service is carried out acidifying and obtained the PH scope of acidifying solution and acidifying solution between 0.9 ~ 1.25 for the fermentation liquor with unit volume adds certain density oxalic acid solution.
Directly to add the oxalic acid solid at the fermentation liquor of unit volume different with tradition, the present invention is dissolved oxalic acid the back in advance in adding fermentation liquor, can avoid the dissolving of conventional art solid oxalic acid slow, dissolve shortcomings such as insufficient, improved work efficiency on the one hand, alleviate lab technician's labour intensity, can also avoid the waste of oxalic acid on the other hand, also avoided the fermentation liquor acidifying not thorough simultaneously, partly tiring remains in the filter residue shortcoming that data are on the low side.
Also reduce the value of the PH after the acidifying on the other hand relatively, aureomycin is discharged from mycelium fully, further improved the accuracy rate of tiring.
Wherein said acidifying pre-service is carried out acidifying for the oxalic acid solution that the fermentation liquor with 100ml adds 0.1 ~ 0.5mol/L.When the oxalic acid solution of this concentration is poured fermentation liquor into, both can acidifying fermentation liquid, also can dilute simultaneously fermentation liquor, improve the precision of tiring.
The step of aureomycin fermentation liquor chemical titer detection is got interior the concussion to abundant the mixing obtaining dilution of container that the 5ml acidifying solution is packed into and contained dilution water for (1), wherein dilute the water yield=(X5 of acidifying solution extension rate/3-1), the acidifying solution extension rate: tire by estimating, tiring after the dilution is approximately 1000u/mL and determines extension rate; (2) draw dilution 1mL respectively in 2 50mL volumetric flasks, each adds the 5mL hydrochloric acid solution, wherein does blank for one, add purified water rapidly and be diluted to scale and shake up, another was put in 100 ℃ of water-baths water-bath 5 minutes, be cooled to room temperature after, add purified water and be diluted to scale, shake up; (3) with proofreading and correct good spectrophotometer, solution in the above-mentioned volumetric flask is carried out colorimetric, read absorbance; (4) according to the survey absorbance, on the typical curve table, find corresponding unit value, multiply by extension rate, be the aureomycin chemical titer of sample.
Below in conjunction with experimental data present technique is described:
As shown in table 1: as to adopt tiring of different extension rate mensuration
Below adopt PH numerical value and tire numerical value for adopting 0.1mol/L respectively, 0.3mol/l, the mean value of the resulting numerical value of oxalic acid solution of 0.5mol/L.
Table 1
As shown in table 2: the dilution of measuring under the different PH conditions of following employing is tired
(wherein the concentration of oxalic acid of Cai Yonging all is in 0.1 ~ 0.5mol/L scope interval)
Table 2
As shown in table 3: as to adopt the tire variation of increasing degree and yield of different measuring methods
| Reagent number | Measure the u/ml that tires by former method | Yield 1 | Dilute the u/ml that tires | Yield 2 |
| 1 | 15210 | 143 | 19740 | 90 |
| 2 | 15888 | 138 | 20559 | 95 |
| 3 | 16432 | 121 | 21294 | 93 |
| 4 | 15210 | 131 | 19740 | 92 |
| 5 | 17323 | 120 | 22449 | 93 |
| 6 | 16304 | 121 | 21126 | 94 |
| 7 | 16208 | 125 | 21000 | 97 |
| 8 | 16881 | 109 | 21945 | 84 |
| 9 | 16096 | 119 | 20853 | 92 |
| 10 | 15792 | 127 | 20433 | 99 |
| 11 | 16000 | 115 | 20727 | 89 |
| 12 | 16416 | 119 | 21273 | 92 |
| 13 | 16512 | 116 | 21420 | 89 |
| 14 | 16304 | 114 | 21105 | 88 |
| 15 | 16208 | 128 | 21038 | 99 |
| 16 | 16779 | 113 | 21735 | 88 |
| 17 | 16660 | 117 | 21578 | 90 |
| 18 | 16208 | 106 | 21000 | 82 |
| 19 | 17442 | 112 | 22635 | 86 |
| Mean value | 16399 | 118 | 21256 | 91 |
| Rate of growth | ? | ? | +29.62% | -22.75% |
* the rate of growth in the table 3 is to measure potency ratio than gained by former method.
Generally speaking, method originally has about 30% tire and stay in the filter residue during acidifying, can the actual response fermentation level and refinement green with dilution method, and help producing and analyze and control, for my company's examination provides economic and technical norms accurately, improved popularity of enterprise.
On the other hand, after adopting oxalic acid solution, overcome the situation that original solid oxalic acid is difficult to dissolve in dense fermentation liquor, method originally, each sample acidifying concussion needs the time more than 1 minute, after adopting dilution method, just can save this part time, can reduce labor intensity simultaneously, to work in three shifts, measure 4 chemical titers every day, and the sample about 50 is arranged at every turn, and can save the minute about 4 hours average every day.
On the other hand, adopt oxalic acid solution to replace oxalic acid, make expensive oxalic acid effectively to utilize, save oxalic acid about 77% on the one hand, probably can save 1000 yuan of left and right sides expenses in every month, also can avoid the oxalic acid of the reaction of not participating in directly to discharge the environmental pollution that causes on the other hand.
Owing to adopt the oxalic acid solution of concentration known, when acidifying, the scope of PH also compares and better controls, thereby in general the use amount that can save fermentation liquor can save the fermentation liquor of about 16L every day.
From table 3, can learn, in general product has consumption unavoidably in the process of producing, so in general theoretical yield just meets the requirements less than 100%, the yield that traditional method detects is all greater than 100%, and after adopting dilution method and adjusting PH, its yield is more pressed close to actual value substantially all below 100%.
Claims (3)
1. aureomycin fermentation liquor chemical titer method of testing, it comprises that the acidifying pre-service of aureomycin fermentation liquor and aureomycin fermentation liquor chemical titer detect, and is characterized in that: described acidifying pre-service is carried out the PH scope that acidifying obtains acidifying solution and acidifying solution and is controlled between 0.9 ~ 1.25 for the fermentation liquor with unit volume adds certain density oxalic acid solution.
2. aureomycin fermentation liquor chemical titer method of testing according to claim 1 is characterized in that: described acidifying pre-service obtains acidifying solution for the oxalic acid solution that splashes into 0.1 ~ 0.5mol/L of 1 ~ 3 unit volume in the fermentation liquor with 1 unit volume carries out acidifying.
3. aureomycin fermentation liquor chemical titer method of testing according to claim 1, it is characterized in that: the step of aureomycin fermentation liquor chemical titer detection is got interior the concussion to abundant the mixing obtaining dilution of container that the 5ml acidifying solution is packed into and contained dilution water for (1), wherein dilute the water yield=(X5 of acidifying solution extension rate/3-1), the acidifying solution extension rate: tire by estimating, tiring after the dilution is approximately 1000u/mL and determines extension rate; (2) draw dilution 1mL respectively in 2 50mL volumetric flasks, each adds the 5mL hydrochloric acid solution, wherein does blank for one, add purified water rapidly and be diluted to scale and shake up, another was put in 100 ℃ of water-baths water-bath 5 minutes, be cooled to room temperature after, add purified water and be diluted to scale, shake up; (3) with proofreading and correct good spectrophotometer, solution in the above-mentioned volumetric flask is carried out colorimetric, read absorbance; (4) according to the survey absorbance, on the typical curve table, find corresponding unit value, multiply by extension rate, be the aureomycin chemical titer of sample.
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| CN201110212108A CN102262010B (en) | 2011-07-27 | 2011-07-27 | Method for testing chemical titer of chlorotetracycline fermentation liquor |
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| CN201110212108A CN102262010B (en) | 2011-07-27 | 2011-07-27 | Method for testing chemical titer of chlorotetracycline fermentation liquor |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104198418A (en) * | 2014-06-24 | 2014-12-10 | 浦城正大生化有限公司 | High-flux determination method for aureomycin titer |
| CN108168975A (en) * | 2017-12-29 | 2018-06-15 | 浦城正大生化有限公司 | A kind of preprocess method of zymotic fluid containing salinomycin and the assay method of salinomycin chemical titer |
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2011
- 2011-07-27 CN CN201110212108A patent/CN102262010B/en active Active
Non-Patent Citations (1)
| Title |
|---|
| 《河北农业大学学报》 201001 崔文静等人 纳他霉素的提取工艺及效价测定的研究 第128-132页 1-3 第33卷, 第1期 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104198418A (en) * | 2014-06-24 | 2014-12-10 | 浦城正大生化有限公司 | High-flux determination method for aureomycin titer |
| CN108168975A (en) * | 2017-12-29 | 2018-06-15 | 浦城正大生化有限公司 | A kind of preprocess method of zymotic fluid containing salinomycin and the assay method of salinomycin chemical titer |
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| CN102262010B (en) | 2012-07-04 |
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Denomination of invention: Method for testing chemical titer of chlorotetracycline fermentation liquor Effective date of registration: 20170113 Granted publication date: 20120704 Pledgee: Agricultural Bank Chinese Limited by Share Ltd Pucheng County branch Pledgor: Pucheng Chia Tai Biochemistry Co., Ltd. Registration number: 2017350000009 |
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