CN108917292B - Method for drying drug sensitivity test card - Google Patents
Method for drying drug sensitivity test card Download PDFInfo
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- CN108917292B CN108917292B CN201810470281.5A CN201810470281A CN108917292B CN 108917292 B CN108917292 B CN 108917292B CN 201810470281 A CN201810470281 A CN 201810470281A CN 108917292 B CN108917292 B CN 108917292B
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- F—MECHANICAL ENGINEERING; LIGHTING; HEATING; WEAPONS; BLASTING
- F26—DRYING
- F26B—DRYING SOLID MATERIALS OR OBJECTS BY REMOVING LIQUID THEREFROM
- F26B1/00—Preliminary treatment of solid materials or objects to facilitate drying, e.g. mixing or backmixing the materials to be dried with predominantly dry solids
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- F—MECHANICAL ENGINEERING; LIGHTING; HEATING; WEAPONS; BLASTING
- F26—DRYING
- F26B—DRYING SOLID MATERIALS OR OBJECTS BY REMOVING LIQUID THEREFROM
- F26B3/00—Drying solid materials or objects by processes involving the application of heat
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- F—MECHANICAL ENGINEERING; LIGHTING; HEATING; WEAPONS; BLASTING
- F26—DRYING
- F26B—DRYING SOLID MATERIALS OR OBJECTS BY REMOVING LIQUID THEREFROM
- F26B5/00—Drying solid materials or objects by processes not involving the application of heat
- F26B5/04—Drying solid materials or objects by processes not involving the application of heat by evaporation or sublimation of moisture under reduced pressure, e.g. in a vacuum
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Abstract
The invention discloses a method for drying a drug sensitivity test card. The method comprises the following steps: (1) preparing and adding a medicament, namely adding a medicament sensitive solution added with a surfactant into a blank test card hole; (2) pre-cooling a product to be dried; (3) primary drying: sealing the drying equipment in the step (2), vacuumizing, filling nitrogen while vacuumizing, maintaining the vacuum degree, heating the product and preserving heat; (4) drying by desorption, and extracting to below 100pa, and drying by raising the temperature to above 30 deg.C to reduce the water content to below 1%. According to the method, the surfactant is added in the process of preparing the medicine, and nitrogen is filled in the drying process, so that the violent movement in the liquid in the whole process is avoided, the problems of the stability and the hole jumping of the medicine in the drying process are solved, the medicine is effectively adhered in the test hole after the drying is finished, and the hole jumping problem in the transportation process is solved.
Description
Technical Field
The invention belongs to the field of medicines, and particularly relates to a method for drying a drug sensitivity test card.
Background
The drug sensitive test card is a kind of in vitro diagnostic reagent for testing drug sensitivity of microorganism (including bacteria, fungi and mycobacteria), and adopts broth dilution method to test drug sensitivity, and the products sold commercially in China domestic market include ATB series drug sensitive test card of biological Merrier company of France, TDR series biochemical drug sensitive test card of biological science and technology Limited company of Changsha in Hunan, etc. In order to ensure the stability of the drugs in the biochemical drug sensitive test card products in the production process and the storage process and the convenient transportation of the products, the preparation of the products is generally divided into the following steps: weighing the medicine, dissolving and blending the medicine, adding the medicine solution into a blank test card, drying and packaging.
The drying process is a core step in the production process and is a key for ensuring the stability, storage stability, and no hole jumping of the medicine in the drying process and the transportation process in the medicine production process. At present, the common drying processes include freeze-drying, drying under normal pressure, vacuum drying and the like.
The freeze-drying process is a drying process at low temperature, and the principle of drying is sublimation of solids. The freeze-drying process includes freezing the product to below eutectic point, and vacuum pumping to sublimate the liquid, so as to reach the aim of dewatering and drying: many thermal substances do not denature or inactivate; when dried at low temperature, some volatile components in the material are lost little; during the freeze-drying process, the growth of microorganisms and the action of enzymes cannot be carried out, so that the original properties can be maintained; because the drying is carried out in a frozen state, the volume is almost unchanged, the original structure is kept, and the concentration phenomenon cannot occur; since the water in the material exists in the form of ice crystals after pre-freezing, the dissolved inorganic salt originally dissolved in water is uniformly distributed in the material. During sublimation, dissolved substances dissolved in water are separated out, so that the phenomenon of surface hardening caused by separation of inorganic salt carried by migration of water in the material to the surface in a common drying method is avoided; because the drying is carried out under vacuum, oxygen is little, and some substances which are easy to oxidize are protected; can eliminate over 95-99% of water and can make the dried product be stored for a long period without deterioration. Chinese patent of CN105087752A discloses a method for preparing a drug sensitive kit, which mainly comprises freezing, sublimating and resolving processes, and the process is a freeze-drying process with obvious characteristics. In order to solve the problem that the medicine is loose and is easy to jump holes in the transportation process after freeze-drying, nutrient substances and antibiotics are added into the test holes at the same time for drying, so that the jump hole rate is reduced, but the medicine is still freeze-dried in nature, and the problem that the medicine is adsorbed in the test holes is not solved fundamentally; and the stability of the product during storage is affected by the moisture absorption of the product during packaging due to the addition of nutrients such as MH broth which are easily hygroscopic.
The normal pressure drying is drying at the temperature lower than the melting point of the product, and the aim of drying is achieved by mainly utilizing the volatilization of the liquid surface. The method is characterized in that: low energy consumption, low equipment requirement and simple operation. Chinese patent of CN101210770A discloses a drying process at normal pressure and temperature, which has the following disadvantages: (1) the drying process is maintained at normal temperature or higher, which is not favorable for drying easily decomposed medicines, such as beta-lactam antibiotics clavulanic acid, imipenem, etc.; (2) the drying process is exposed to air, which is not conducive to easily oxidized drugs, and thus may require the addition of antioxidants and the like.
The vacuum drying mainly utilizes the characteristics of low pressure and low boiling point of liquid, and the vacuum pumping is carried out to reduce the air pressure on the basis of normal-temperature normal-pressure drying so as to achieve the effect of quicker drying. The method is characterized in that: the drying time is shortest, and the operation is simple and convenient; because the vacuum environment is adopted, the oxygen content is low, so that the protective effect is realized on products which are easy to oxidize; the disadvantages are that: it is very easy to bubble during the drying process, causing the medicine to jump the hole. The invention patent of CN1732257A discloses a drying process for forming high-viscosity liquid, which belongs to the vacuum drying range, is suitable for the test hole unit with larger carrier diameter, is not suitable for the test board unit with lower diameter for drying, and is easy to cause bubbling phenomenon and hole jump. This bubbling phenomenon is not caused by the boiling of the liquid, but is caused by the natural dissolved gas inside the liquid being washed out at low pressure, forming bubbles. Meanwhile, when the diameter of the test unit is smaller, the capillary action is more obvious, and the phenomenon of lifting liquid caused by expansion of bubbles due to low air pressure is more prominent, so that hole jumping is easily caused.
Disclosure of Invention
The invention aims to overcome the defects of the background technology, aims to obtain amorphous state of a class of medicines by low-temperature and low-pressure drying, solves the problems that the medicines in the medicine sensitive test card are easy to lose effectiveness, oxidize and jump holes when the medicine sensitive test card is dried by a process, and the dried product is powder or flaky solid which is not beneficial to transportation, and solves the problems of high drying energy consumption and long drying time.
In order to achieve the aim of the invention, the invention provides a method for drying a drug sensitive test card, which comprises the following steps:
(1) preparing and adding a medicament, namely adding a biochemical solution or a medicament sensitive solution added with a surfactant into a blank test card hole;
(2) pre-cooling a product to be dried, and placing the test card subjected to sample adding in the step (1) into drying equipment for pre-cooling;
(3) preliminary drying, sealing the drying equipment in the step (2), vacuumizing, filling nitrogen while vacuumizing, maintaining the vacuum degree, heating the product and preserving heat;
(4) and (4) analyzing and drying, pumping the vacuum degree to be below 100pa, raising the temperature to be above 30 ℃ at the same time, drying until the moisture is reduced to be below 1%, completing drying, and packaging the test card.
Preferably, the HLB value of the nonionic surfactant in step (1) of the present invention is 3.0 to 20.0, for example, 14.0 to 17.0.
In the present invention, the nonionic surfactant in step (1) is 0.001 to 0.1% of a polyol-based surfactant or a polyethylene glycol-based surfactant, such as any one of span 20, span 40, span 60, span 80, polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 80, polysorbate 85, Myrj45, Myrj49, Myrj51, Myrj52, Myrj53, Brij30 and Brij35, for example, 0.01% of span 20.
Preferably, in the present invention, the amount of the sample to be added in step (1) may be 10. mu.l to 200. mu.l, for example, 25. mu.l to 50. mu.l.
Preferably, in the present invention, the pre-cooling temperature for pre-cooling in step (2) may be 2-8 ℃, for example, 2-5 ℃.
In the present invention, the vacuum pumping in step (3) may be performed at a vacuum degree of 2000 pa.
In the present invention, the vacuum degree in step (3) may be maintained at 800-.
In the invention, the product is heated in the step (3), the heat preservation can be heating, and the temperature is controlled within the range of 4-12 ℃.
In the invention, the step (3) is vacuumized, nitrogen is filled in the vacuumized step to maintain the vacuum degree, and meanwhile, the product is heated, and the specific procedure of heat preservation can be as follows:
in the invention, the step (3) is vacuumized, nitrogen is filled in the vacuumized step to maintain the vacuum degree, and meanwhile, the product is heated, and the specific procedure of heat preservation can be as follows:
in the invention, the step (3) is vacuumized, nitrogen is filled in the vacuumized step to maintain the vacuum degree, and meanwhile, the product is heated, and the specific procedure of heat preservation can be as follows:
in the invention, the temperature of the analysis and drying in the step (4) is not more than 40 ℃, and the time is not more than 1 h.
In the invention, the test card is packaged in the light-proof aluminum film bag with low air permeability in the step (4).
Compared with the prior art, the invention has the following advantages:
1. the nonionic surfactant is added, does not participate in chemical reaction, does not influence the microbial drug sensitivity test result, is mainly used for reducing the surface tension of the liquid and reducing the surface energy, has a physical effect, avoids the formation of bubbles in the drying process to cause hole jumping, and simultaneously reduces the surface energy, accelerates the drying rate and reduces the energy consumption;
2. pre-cooling at a low temperature of 2-8 ℃ to ensure the stability of the medicine;
3. the nitrogen is filled, so that the control of the air pressure range is convenient to isolate oxygen, the decomposition of the easily oxidized medicine is avoided, the air flow of the liquid level of the medicine is disturbed, the evaporation rate is increased, and the aim of quick drying is fulfilled;
4. the low pressure reduces the boiling point of the liquid, and the liquid can be dried by programmed temperature and pressure control without freezing the liquid into a solid state, so as to improve the drying rate, reduce the energy consumption, reduce the oxygen content, and avoid the decomposition of the easily oxidized medicine by matching with the nitrogen filling;
5. the invention relates to a closed temperature-controlled and pressure-controlled drying device in a real site, which adopts a freeze dryer (but the adopted drying principle is not freeze drying), ensures that liquid is always in a liquid state in the drying process, breaks a stress structure, and the whole liquid is evaporated on the surface, is filled with nitrogen, maintains relatively stable air pressure, and ensures that no violent movement is generated in the liquid in the whole process, so that excipient/medicine forms an amorphous state after being dried, and has good adhesiveness in holes, thereby solving the problem of hole jumping in the transportation process.
Drawings
FIG. 1 is a flow chart of a drying process of the present invention;
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is described in further detail below with reference to the accompanying drawings and embodiments. Additional aspects and advantages of the invention will be set forth in part in the description which follows and, in part, will be obvious from the description, or may be learned by practice of the invention. It is to be understood that the following description is only illustrative of the present invention and is not to be construed as limiting the present invention.
The terms "comprises," "comprising," "includes," "including," "has," "having," "contains," "containing," or any other variation thereof, as used herein, are intended to cover a non-exclusive inclusion. For example, a composition, process, method, article, or apparatus that comprises a list of elements is not necessarily limited to only those elements but may include other elements not expressly listed or inherent to such composition, process, method, article, or apparatus.
When an amount, concentration, or other value or parameter is expressed as a range, preferred range, or as a range of upper preferable values and lower preferable values, this is to be understood as specifically disclosing all ranges formed from any pair of any upper range limit or preferred value and any lower range limit or preferred value, regardless of whether ranges are separately disclosed. For example, when a range of "1 to 5" is disclosed, the described range should be interpreted to include the ranges "1 to 4", "1 to 3", "1 to 2 and 4 to 5", "1 to 3 and 5", and the like. When a range of values is described herein, unless otherwise stated, the range is intended to include the endpoints thereof and all integers and fractions within the range.
Further, the technical features of the embodiments of the present invention may be combined with each other as long as they do not conflict with each other.
Example 1
Firstly, preparing a working solution: preparing required antibiotic solutions or biochemical working solutions with different concentrations and types, and adding a nonionic surfactant with the HLB value of 3.0-20.0 in the preparation process;
sample adding: adding the prepared solution into a blank test card, wherein the adding amount of each hole is 25 mu l;
thirdly, precooling: putting the test card after sample adding into a partition plate of a drying box, closing the box door, refrigerating the partition plate, and reducing the temperature of the product to 2-4 ℃;
fourthly, primary drying: the vacuum pump and the nitrogen bleed switch were turned on and the program parameters were set according to the following program.
Analyzing and drying, closing the nitrogen control electromagnetic valve, starting the vacuum pump, pumping the vacuum to be less than 100Pa, and maintaining for 1 h.
Sixthly, packaging: and packaging the product in a vacuum manner by adopting an aluminum foil bag.
The whole drying time is not longer than 14h, wherein the preliminary drying time is not longer than 10 h.
Example 2
Firstly, preparing a working solution: preparing required antibiotic solutions or biochemical working solutions with different concentrations and types, and adding a nonionic surfactant with the HLB value of 3.0-20.0 in the preparation process;
sample adding: adding the prepared solution into a blank test card, wherein the adding amount of each hole is 25 mu l;
thirdly, precooling: putting the test card after sample adding into a partition plate of a drying box, closing the box door, refrigerating the partition plate, and reducing the temperature of the product to 2-4 ℃;
fourthly, primary drying: the vacuum pump and the nitrogen bleed switch were turned on and the program parameters were set according to the following program.
Analyzing and drying, closing the nitrogen control electromagnetic valve, starting the vacuum pump, pumping the vacuum to be less than 100Pa, and maintaining for 1 h.
Sixthly, packaging: and packaging the product in a vacuum manner by adopting an aluminum foil bag.
The whole drying time is not longer than 14h, wherein the preliminary drying time is not longer than 10 h.
Example 3
Firstly, preparing a working solution: preparing required antibiotic solutions or biochemical working solutions with different concentrations and types, and adding a nonionic surfactant with the HLB value of 3.0-20.0 in the preparation process;
sample adding: adding the prepared solution into a blank test card, wherein the adding amount of each hole is 25 mu l;
thirdly, precooling: putting the test card after sample adding into a partition plate of a drying box, closing the box door, refrigerating the partition plate, and reducing the temperature of the product to 2-4 ℃;
fourthly, primary drying: the vacuum pump and the nitrogen bleed switch were turned on and the program parameters were set according to the following program.
Analyzing and drying, closing the nitrogen control electromagnetic valve, starting the vacuum pump, pumping the vacuum to be less than 100Pa, and maintaining for 1 h.
Sixthly, packaging: and packaging the product in a vacuum manner by adopting an aluminum foil bag.
The whole drying time is not longer than 14h, wherein the preliminary drying time is not longer than 10 h.
Example 4
And (3) carrying out a comparative test on the jumping hole rate of the added surfactant and the non-added surfactant:
firstly, preparing a working solution: respectively adding purified water and 0.01% span 20, and preparing a series of vancomycin (with concentration of 0.12/0.25/0.5/1/2/4/8/16 μ g/ml respectively), penicillin (with concentration of 0.03/0.06/0.12/0.25/0.5/1/2/4 μ g/ml respectively), gentamicin (with concentration of 0.12/0.25/0.5/1/2/4/8/16 μ g/ml respectively), and ceftazidime (with concentration of 0.06/0.12/0.25/0.5/1/2/4/8 μ g/ml) solutions with different concentrations, wherein the concentration is not less than 100 ml;
sample adding: adding the solution into a blank test card with 96 holes in the sequence of drug concentration from low to high, wherein the adding amount of each hole is 25 mu l, and each formula of each drug is added into 20 holes respectively;
secondly, the test card after sample adding is placed in a clapboard of a drying box, and drying and packaging are carried out according to the method in the embodiment 1, the embodiment 2 or the embodiment 3;
thirdly, testing the vancomycin and penicillin test card by adopting strains of staphylococcus aureus ATCC29213 and enterococcus faecalis ATCC29212 with the age of 16-24 h; the strains of Escherichia coli ATCC25922 and Pseudomonas aeruginosa ATCC27853 with the age of 16-24h are adopted to test the gentamicin and ceftazidime test card;
fourthly, observing the result, counting the MIC (minimum inhibitory concentration) hole jumping situation, and calculating the hole jumping rate (the hole jumping rate is the number of the holes per the number of the test holes);
the test results are shown in the following table (values reported in the table are mean values of the test cards obtained by the method of example 1, example 2 or example 3, and relative standard deviation is less than 2%)
Example 5
The minimum inhibitory concentration MIC accuracy test and the accelerated stability test after the unstable drug is dried are as follows:
firstly, preparing a working solution: adding 0.01% span 20, preparing a series of penicillin (concentration of 0.03/0.06/0.12/0.25/0.5/1/2/4 μ g/ml respectively), amoxicillin-clavulanic acid (0.25/0.12, 0.5/0.25, 1/0.5, 2/1, 4/2, 8/4, 16/8, 32/16 μ g/ml), cefazolin (0.25/0.5/1/2/4/8/16/32 μ g/ml), cefuroxime (0.5/1/2/4/8/16/32/64 μ g/ml), ceftadine (concentration of 0.06/0.12/0.25/0.5/1/2/4/8 μ g/ml respectively), Cefepime (with concentration of 0.06/0.12/0.25/0.5/1/2/4/8 μ g/ml respectively), meropenem (with concentration of 0.12/0.25/0.5/1/2/4/8/16 μ g/ml respectively), imipenem (with concentration of 0.06/0.12/0.25/0.5/1/2/4/8 μ g/ml respectively) solution, no less than 100 ml;
sample adding: adding the solution into a blank test card with 96 holes in the sequence of drug concentration from low to high, wherein the adding amount of each hole is 25 mu l, and each drug is added into 20 holes respectively;
thirdly, the test card after sample adding is placed in a clapboard of a drying box, and drying and packaging are carried out according to the method in the embodiment 1;
fourthly, testing the dried test card by adopting strains with the age of 16 to 24 hours, such as staphylococcus aureus ATCC29213, enterococcus faecalis ATCC2921, escherichia coli ATCC25922, pseudomonas aeruginosa ATCC27853 and escherichia coli ATCC35218, wherein the specific test results are detailed in the following table; all test results meet the requirements of the quality control standard of the strains.
Placing the test card at 37 ℃, taking out the test card after 2/4/6/8/10/12 weeks respectively for testing, and inspecting whether the stability of different medicines meets the storage requirement of the product, wherein the test result is as follows:
it will be understood by those skilled in the art that the foregoing is only a preferred embodiment of the present invention, and is not intended to limit the invention, and that any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the scope of the present invention.
Claims (11)
1. A method for drying a drug sensitive test card is characterized by comprising the following steps:
(1) preparing and adding a medicament, namely adding a medicament sensitive solution added with a nonionic surfactant into a blank test card hole;
(2) pre-cooling a product to be dried, and placing the test card subjected to sample adding in the step (1) into drying equipment for pre-cooling;
(3) preliminary drying, sealing the drying equipment in the step (2), vacuumizing, filling nitrogen while vacuumizing, maintaining the vacuum degree, heating the product and preserving heat;
(4) analyzing and drying, namely pumping the vacuum degree to be below 100pa, raising the temperature to be above 30 ℃ for drying at the same time, reducing the moisture to be below 1%, completing drying, and packaging the test card;
the precooling temperature for precooling in the step (2) is 2-8 ℃;
vacuumizing in the step (3), filling nitrogen while vacuumizing, maintaining the vacuum degree, and heating the product, wherein the specific procedure of heat preservation is as follows:
firstly, controlling the air pressure within the range of 2000-2300 Pa, controlling the temperature T to be 4 ℃, and maintaining the temperature T to be 2 h;
controlling the air pressure within the range of 2000-2300 Pa, controlling the temperature to be 6 ℃, and maintaining the time T to be 2 h;
thirdly, controlling the air pressure within the range of 2000-2300 Pa, controlling the temperature to be 8 ℃, and maintaining the temperature for 2 h;
controlling the air pressure within the range of 2000-2300 Pa, controlling the temperature T to 10 ℃, and maintaining the temperature T to 2 h;
controlling the air pressure within the range of 2000-2300 Pa, controlling the temperature to 12 ℃ and maintaining the time T for 2 h;
or as follows:
firstly, controlling the air pressure within the range of 2000-2300 Pa, controlling the temperature T to be 6-8 ℃, and maintaining the temperature T for 2 h;
controlling the air pressure within the range of 1700-2000 Pa, controlling the temperature to be 6-8 ℃, and maintaining the temperature for 2 h;
controlling the air pressure within the range of 1400-1700 Pa, controlling the temperature to be 6-8 ℃, and maintaining the temperature for 2 h;
controlling the air pressure within the range of 1100-1400 Pa, controlling the temperature T to be 6-8 ℃, and maintaining the temperature T for 2 h;
controlling the air pressure within the range of 800-1100 Pa, controlling the temperature to be 6-8 ℃, and maintaining the temperature for 2 h;
or as follows:
firstly, controlling the air pressure within the range of 2000-2300 Pa, controlling the temperature T to be 4 ℃, and maintaining the temperature T to be 2 h;
controlling the air pressure within the range of 1700-2000 Pa, controlling the temperature T to be 6 ℃, and maintaining the time T to be 2 h;
controlling the air pressure within the range of 1400-1700 Pa, controlling the temperature to be 8 ℃ and maintaining the time T to be 2 h;
fourthly, controlling the air pressure within the range of 1100-1400 Pa, controlling the temperature T to be 10 ℃, and maintaining the temperature T to be 2 h;
controlling the air pressure within 800-1100 Pa, controlling the temperature T to 12 ℃, and maintaining the time T to 2 h.
2. The method for drying the drug sensitive test card according to claim 1, wherein the HLB value of the nonionic surfactant is 3.0 to 20.0.
3. The method for drying the drug sensitive test card according to claim 1, wherein the HLB value of the nonionic surfactant is 14.0 to 18.0.
4. The method for drying a drug sensitive test card according to claim 1, wherein the non-ionic surfactant is 0.001-0.1% of a polyol surfactant or a polyethylene glycol type surfactant.
5. The method of drying drug sensitivity test card according to claim 1, wherein the non-ionic surfactant is any one of span 20, span 40, span 60, span 80, polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 80, polysorbate 85, Myrj45, Myrj49, Myrj51, Myrj52, Myrj53, Brij30, and Brij 35.
6. The method for drying a drug sensitive test card according to claim 1, wherein the non-ionic surfactant is span 20 at 0.01%.
7. The method for drying the drug sensitive test card according to claim 1, wherein the sample is added in an amount of 10 to 200. mu.l.
8. The method for drying the drug sensitive test card according to claim 1, wherein the sample is added in an amount of 25 to 50 μ l.
9. The method for drying the drug sensitive test card according to claim 1, wherein the pre-cooling temperature in the step (2) is 2-5 ℃.
10. The method for drying the drug sensitive test card according to claim 1, wherein the temperature for desorption drying in the step (4) is not more than 40 ℃ and the time is not more than 1 h.
11. The method for drying drug sensitive test card according to claim 1, wherein the test card is packaged in the aluminum film bag with low air permeability and being protected from light in the step (4).
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| JP2703179B2 (en) * | 1994-04-22 | 1998-01-26 | 日本エフデイ株式会社 | Freeze drying method and vacuum heating vessel for freeze drying equipment |
| CN101818985B (en) * | 2010-04-22 | 2013-05-29 | 康纳新型材料(杭州)有限公司 | Novel drying oven and drying method thereof |
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