Emulsion containing spectinomycin hydrochloride and lincomycin hydrochloride and preparation method thereof

Abstract

The invention relates to an emulsion containing spectinomycin hydrochloride and lincomycin hydrochloride and a preparation method thereof, and belongs to the field of animal medicine. The emulsion is prepared by jointly combining spectinomycin hydrochloride, lincomycin hydrochloride, tea polyphenol, lipoic acid, polyvinyl alcohol (PVA) and the like and matching with a specific ultrasonic treatment process, and the method remarkably improves the stability of the emulsion, reduces drug degradation and improves the bioavailability of the drug. Animal experiments show that the emulsion prepared by the invention can be quickly and completely absorbed after being injected, the antibody level after vaccination in an animal body can be remarkably improved, and the validity period of the antibody is prolonged. The emulsion disclosed by the invention is simple in preparation method and suitable for large-scale production, and has a relatively good application prospect.

Description

Technical Field

[0001] The invention belongs to the technical field of animal medicine, and in particular relates to an emulsion containing spectinomycin hydrochloride and lincomycin hydrochloride and a preparation method thereof. Background Art

[0002] Lincomycin hydrochloride and spectinomycin hydrochloride are commonly used antibiotics in clinical practice, and are widely used in the prevention and treatment of infectious diseases in livestock and poultry. Lincomycin hydrochloride is a macrolide antibiotic that mainly works by inhibiting bacterial protein synthesis. It has a broad-spectrum antibacterial activity, especially against Gram-positive bacteria and some anaerobic bacteria. Spectinomycin hydrochloride belongs to the aminoglycoside antibiotics, which mainly binds to bacterial ribosomes and interferes with their protein synthesis, thereby exerting a bactericidal effect. The two are often used in combination to enhance the antibacterial effect and expand the antibacterial spectrum.

[0003] However, when lincomycin hydrochloride and spectinomycin hydrochloride are made into emulsion preparations, stability problems are often faced. Emulsion is a thermodynamically unstable system, which is prone to stratification, aggregation, precipitation and other phenomena. More importantly, lincomycin hydrochloride and spectinomycin hydrochloride are prone to degradation reactions in emulsion systems, resulting in reduced efficacy or even failure. This degradation may be related to a variety of factors, including the pH value in the emulsion, the choice of emulsifier, storage conditions, and environmental factors such as light and heat.

[0004] In the prior art, although there are some methods that attempt to improve the stability of the emulsion by adjusting the emulsion formula or adding antioxidants, the effect is still unsatisfactory, especially during long-term storage, the degradation problem of the drug is still prominent. Therefore, it is urgent to develop new technical means or improve the existing formula to improve the stability of lincomycin hydrochloride and spectinomycin hydrochloride emulsions, extend their shelf life, and thus ensure the clinical efficacy of the drugs. Summary of the invention

[0005] The invention provides an emulsion containing spectinomycin hydrochloride and lincomycin hydrochloride and a preparation method thereof, thereby realizing long-term stable storage of the high-hydrochloric acid lincomycin and spectinomycin hydrochloride emulsions.

[0006] First, the present invention provides an emulsion containing spectinomycin hydrochloride and lincomycin hydrochloride with long-term stability, wherein each 25 ml of the emulsion is composed of the following components in terms of mass percentage:

[0007] 5wt% lincomycin hydrochloride;

[0008] 10wt% of spectinomycin hydrochloride;

[0009] 6 wt% of a mixture of Span 40 emulsifier and Tween 20 emulsifier;

[0010] 0.2wt% tea polyphenols;

[0011] 0.1 wt% lipoic acid;

[0012] 0.2 wt% polyvinyl alcohol;

[0013] 10wt% white oil for injection;

[0014] The remainder is water for injection.

[0015] Preferably, in the mixture of Span40 emulsifier and Tween20 emulsifier, the mass ratio of Span40 emulsifier to Tween20 emulsifier is 1:1.

[0016] Preferably, the emulsion is prepared by the following steps:

[0017] 1) Under a certain temperature and nitrogen protection, a mixture of 6 wt% of Span 40 emulsifier and Tween 20 emulsifier is dissolved in 10 wt% of white oil for injection to form an oil phase;

[0018] 2) dissolving lincomycin hydrochloride, spectinomycin hydrochloride, tea polyphenols, lipoic acid and polyvinyl alcohol in water for injection, and subjecting to ultrasonic treatment at 400 W for 30 minutes to obtain an aqueous phase;

[0019] 3) mixing the oil phase with the water phase and emulsifying to obtain a primary emulsion;

[0020] 4) Emulsifying again to obtain a uniform emulsion;

[0021] 5) sterilizing by a microporous filter membrane to obtain the emulsion.

[0022] Preferably, the certain temperature is 30-45°C; the emulsification method is 3000-4000rpm for 2-5 minutes; the re-emulsification method is 6000-8000rpm for 8-10 minutes; the microporous membrane sterilization method is filtering using 0.45μm and 0.22μm filter membranes in turn.

[0023] Preferably, the certain temperature is 40° C., the emulsification method is emulsification at 3500 rpm for 4 minutes, and the re-emulsification method is emulsification at 7000 rpm for 10 minutes.

[0024] Secondly, the present invention provides a method for preparing an emulsion containing spectinomycin hydrochloride and lincomycin hydrochloride with long-term stability, characterized in that the preparation method comprises the following steps:

[0025] 1) Under a certain temperature and nitrogen protection, a mixture of 6 wt% of Span 40 emulsifier and Tween 20 emulsifier is dissolved in 10 wt% of white oil for injection to form an oil phase;

[0026] 2) dissolving lincomycin hydrochloride, spectinomycin hydrochloride, tea polyphenols, lipoic acid and polyvinyl alcohol in water for injection, and subjecting to ultrasonic treatment at 400 W for 30 minutes to obtain an aqueous phase;

[0027] 3) mixing the oil phase with the water phase and emulsifying to obtain a primary emulsion;

[0028] 4) Emulsifying again to obtain a uniform emulsion;

[0029] 5) sterilizing by a microporous filter membrane to obtain the emulsion.

[0030] Preferably, the certain temperature is 30-45°C; the emulsification method is 3000-4000rpm for 2-5 minutes; the re-emulsification method is 6000-8000rpm for 8-10 minutes; the microporous membrane sterilization method is filtering using 0.45μm and 0.22μm filter membranes in turn.

[0031] Preferably, the certain temperature is 40° C., the emulsification method is emulsification at 3500 rpm for 4 minutes, and the re-emulsification method is emulsification at 7000 rpm for 10 minutes.

[0032] Secondly, the present invention provides an application of an emulsion containing spectinomycin hydrochloride and lincomycin hydrochloride in preparing a preparation for improving the antibody titer level of poultry vaccines, wherein the emulsion is prepared by the following preparation method:

[0033] 1) at 37° C. and under nitrogen protection, dissolving a mixture of 6 wt % of Span 40 emulsifier and Tween 20 emulsifier in 10 wt % of white oil for injection to form an oil phase;

[0034] 2) dissolving lincomycin hydrochloride, spectinomycin hydrochloride, tea polyphenols, lipoic acid and polyvinyl alcohol in water for injection, and subjecting to ultrasonic treatment at 400 W for 30 minutes to obtain an aqueous phase;

[0035] 3) mixing the oil phase with the water phase and emulsifying at 3500 rpm for 4 minutes to obtain a primary emulsion;

[0036] 4) Emulsification at 7000 rpm for 8 minutes to obtain a uniform emulsion;

[0037] 5) Filter and sterilize using 0.45 μm and 0.22 μm filter membranes to obtain the emulsion.

[0038] Preferably, the poultry vaccines are H9-ZGS vaccine and NDV-Lasota.

[0039] The beneficial effects of the present invention are:

[0040] The present invention effectively inhibits the degradation of spectinomycin hydrochloride and lincomycin hydrochloride in an emulsion system by adding antioxidants such as tea polyphenols and lipoic acid, combined with polyvinyl alcohol (PVA) and optimized ultrasonic treatment conditions, and significantly improves the long-term stability of the emulsion. In particular, under the ultrasonic treatment conditions of 400W and 30 minutes, the degradation rate of the emulsion is significantly reduced, ensuring that the drug remains efficient and stable during storage, preventing discoloration, and extending its shelf life.

[0041] The invention controls the preparation conditions of the emulsion reasonably, so that the average particle size of the emulsion is significantly reduced and the particle distribution is more uniform. The reduction of the particle size not only improves the stability of the emulsion, but also increases the bioavailability of the drug in the body, and promotes faster and more complete absorption of the drug.

[0042] Secondly, when the emulsion prepared by the present invention is used together with a vaccine, the antibody titer is significantly higher than that of the control group, and the antibody is maintained for a longer time, thereby improving the immune effect of the vaccine. BRIEF DESCRIPTION OF THE DRAWINGS

[0043] Figure 1 The color changes of different emulsions containing spectinomycin hydrochloride and lincomycin hydrochloride at 0 months;

[0044] Figure 2 The color changes of different emulsions containing spectinomycin hydrochloride and lincomycin hydrochloride at 1 month;

[0045] Figure 3 The color changes of different emulsions containing spectinomycin hydrochloride and lincomycin hydrochloride at 3 months;

[0046] Figure 4 The color changes of different emulsions containing spectinomycin hydrochloride and lincomycin hydrochloride at 6 months;

[0047] Figure 5 The color changes of different emulsions containing spectinomycin hydrochloride and lincomycin hydrochloride at 12 months;

[0048] Figure 6 This is the absorption of the Example 1 group at 9 hours;

[0049] Figure 7 This is the absorption situation of the Example 8 group after 9 hours. DETAILED DESCRIPTION

[0050] The embodiments of the present invention are for better explanation of the present invention, but the content of the present invention is not limited to the embodiments. Therefore, those skilled in the art can make non-essential improvements and adjustments to the implementation scheme according to the above invention content, which still fall within the protection scope of the present invention.

[0051] Example 1

[0052] Emulsion A containing spectinomycin hydrochloride and lincomycin hydrochloride

[0053] 1) At 40° C. and under nitrogen protection, 6 wt % of Span 40 and tween 20 (mass fraction ratio 1:1) emulsifiers were added to 10 wt % of white oil for injection, and after complete dissolution, an oil phase was obtained;

[0054] 2) dissolving 0.2wt% tea polyphenols and 0.1wt% lipoic acid, 5wt% lincomycin hydrochloride and 10% spectinomycin hydrochloride (the corresponding emulsion volume is 25ml, similar to the subsequent embodiments) in 68.7wt% water for injection, and obtaining an aqueous phase after complete dissolution;

[0055] 3) After mixing the oil phase and the water phase, emulsify at 3500 rpm for 4 min to obtain a primary emulsion;

[0056] 4) emulsifying the primary emulsion again at 7000 rpm for 10 min to finally obtain a uniform emulsion;

[0057] 5) The homogenized emulsion was sterilized by filtering through a 0.45 μm + 0.22 μm combined microporous filter membrane to obtain an emulsion A containing spectinomycin hydrochloride and lincomycin hydrochloride.

[0058] Example 2

[0059] Emulsion B containing spectinomycin hydrochloride and lincomycin hydrochloride

[0060] 1) At 40° C. and under nitrogen protection, 6 wt % of Span 40 and tween 20 (mass fraction ratio 1:1) emulsifiers were added to 10 wt % of white oil for injection, and after complete dissolution, an oil phase was obtained;

[0061] 2) dissolving 0.3wt% tea polyphenols, 0.2wt% polyvinyl alcohol, 5wt% lincomycin hydrochloride and 10% spectinomycin hydrochloride in 68.5wt% water for injection, and after complete dissolution, ultrasonically treating at 400W for 30min to obtain an aqueous phase;

[0062] 3) After mixing the oil phase and the water phase, emulsify at 3500 rpm for 4 min to obtain a primary emulsion;

[0063] 4) emulsifying the primary emulsion again at 7000 rpm for 10 min to finally obtain a uniform emulsion;

[0064] 5) The homogenized emulsion was sterilized by filtering through a 0.45 μm + 0.22 μm combined microporous filter membrane to obtain an emulsion B containing spectinomycin hydrochloride and lincomycin hydrochloride.

[0065] Example 3

[0066] Emulsion C containing spectinomycin hydrochloride and lincomycin hydrochloride

[0067] 1) At 40° C. and under nitrogen protection, 6 wt % of Span 40 and tween 20 (mass fraction ratio 1:1) emulsifiers were added to 10 wt % of white oil for injection, and after complete dissolution, an oil phase was obtained;

[0068] 2) dissolving 0.3 wt % lipoic acid, 0.2 wt % polyvinyl alcohol, 5 wt % lincomycin hydrochloride and 10 wt % spectinomycin hydrochloride in 68.5 wt % water for injection, and after complete dissolution, ultrasonically treating at 400 W for 30 min to obtain an aqueous phase;

[0069] 3) After mixing the oil phase and the water phase, emulsify at 3500 rpm for 4 min to obtain a primary emulsion;

[0070] 4) emulsifying the primary emulsion again at 7000 rpm for 10 min to finally obtain a uniform emulsion;

[0071] 5) The homogenized emulsion was sterilized by filtering through a 0.45 μm + 0.22 μm combined microporous filter membrane to obtain an emulsion C containing spectinomycin hydrochloride and lincomycin hydrochloride.

[0072] Example 4

[0073] Emulsion D containing spectinomycin hydrochloride and lincomycin hydrochloride

[0074] 1) At 40° C. and under nitrogen protection, 6 wt % of Span 40 and tween 20 (mass fraction ratio 1:1) emulsifiers were added to 10 wt % of white oil for injection, and after complete dissolution, an oil phase was obtained;

[0075] 2) dissolving 0.2wt% tea polyphenols and 0.1wt% lipoic acid, 5wt% lincomycin hydrochloride and 10% spectinomycin hydrochloride in 68.7wt% water for injection, and after complete dissolution, ultrasonically treating at 400W for 30min to obtain an aqueous phase;

[0076] 3) After mixing the oil phase and the water phase, emulsify at 3500 rpm for 4 min to obtain a primary emulsion;

[0077] 4) emulsifying the primary emulsion again at 7000 rpm for 10 min to finally obtain a uniform emulsion;

[0078] 5) The homogenized emulsion was sterilized by filtering through a 0.45 μm + 0.22 μm combined microporous filter membrane to obtain an emulsion D containing spectinomycin hydrochloride and lincomycin hydrochloride.

[0079] Example 5

[0080] Emulsion E containing spectinomycin hydrochloride and lincomycin hydrochloride

[0081] 1) At 40° C. and under nitrogen protection, 6 wt % of Span 40 and tween 20 (mass fraction ratio 1:1) emulsifiers were added to 10 wt % of white oil for injection, and after complete dissolution, an oil phase was obtained;

[0082] 2) dissolving 0.2wt% tea polyphenols, 0.1wt% lipoic acid, 0.2wt% polyvinyl alcohol, 5wt% lincomycin hydrochloride and 10% spectinomycin hydrochloride in 68.5wt% water for injection, and after complete dissolution, ultrasonically controlling at 200W for 30min to obtain an aqueous phase;

[0083] 3) After mixing the oil phase and the water phase, emulsify at 3500 rpm for 4 min to obtain a primary emulsion;

[0084] 4) emulsifying the primary emulsion again at 7000 rpm for 10 min to finally obtain a uniform emulsion;

[0085] 5) The homogenized emulsion was sterilized by filtering through a 0.45 μm + 0.22 μm combined microporous filter membrane to obtain an emulsion E containing spectinomycin hydrochloride and lincomycin hydrochloride.

[0086] Example 6

[0087] Emulsion F containing spectinomycin hydrochloride and lincomycin hydrochloride

[0088] 1) At 40° C. and under nitrogen protection, 6 wt % of Span 40 and tween 20 (mass fraction ratio 1:1) emulsifiers were added to 10 wt % of white oil for injection, and after complete dissolution, an oil phase was obtained;

[0089] 2) dissolving 0.2wt% tea polyphenols, 0.1wt% lipoic acid, 0.2wt% polyvinyl alcohol, 5wt% lincomycin hydrochloride and 10% spectinomycin hydrochloride in 68.5wt% water for injection, and after complete dissolution, ultrasonically treating at 600W for 30min to obtain an aqueous phase;

[0090] 3) After mixing the oil phase and the water phase, emulsify at 3500 rpm for 4 min to obtain a primary emulsion;

[0091] 4) emulsifying the primary emulsion again at 7000 rpm for 10 min to finally obtain a uniform emulsion;

[0092] 5) The homogenized emulsion was sterilized by filtering through a 0.45 μm + 0.22 μm combined microporous filter membrane to obtain an emulsion F containing spectinomycin hydrochloride and lincomycin hydrochloride.

[0093] Example 7

[0094] Emulsion G containing spectinomycin hydrochloride and lincomycin hydrochloride

[0095] 1) At 40° C. and under nitrogen protection, 6 wt % of Span 40 and tween 20 (mass fraction ratio 1:1) emulsifiers were added to 10 wt % of white oil for injection, and after complete dissolution, an oil phase was obtained;

[0096] 2) dissolving 0.2wt% forsythiaside A, 0.1wt% lipoic acid, 0.2wt% polyvinyl alcohol, 5wt% lincomycin hydrochloride and 10% spectinomycin hydrochloride in 68.5wt% water for injection, and after complete dissolution, ultrasonically treating at 400W for 30min to obtain an aqueous phase;

[0097] 3) After mixing the oil phase and the water phase, emulsify at 3500 rpm for 4 min to obtain a primary emulsion;

[0098] 4) emulsifying the primary emulsion again at 7000 rpm for 10 min to finally obtain a uniform emulsion;

[0099] 5) The homogenized emulsion was sterilized by filtering through a 0.45 μm + 0.22 μm combined microporous filter membrane to obtain an emulsion G containing spectinomycin hydrochloride and lincomycin hydrochloride.

[0100] Example 8

[0101] Emulsion containing spectinomycin hydrochloride and lincomycin hydrochloride H

[0102] 1) At 40° C. and under nitrogen protection, 6 wt % of Span 40 and tween 20 (mass fraction ratio 1:1) emulsifiers were added to 10 wt % of white oil for injection, and after complete dissolution, an oil phase was obtained;

[0103] 2) dissolving 0.2wt% tea polyphenols, 0.1wt% lipoic acid, 0.2wt% polyvinyl alcohol, 5wt% lincomycin hydrochloride and 10% spectinomycin hydrochloride in 68.5wt% water for injection, and after complete dissolution, ultrasonically treating at 400W for 30min to obtain an aqueous phase;

[0104] 3) After mixing the oil phase and the water phase, emulsify at 3500 rpm for 4 min to obtain a primary emulsion;

[0105] 4) emulsifying the primary emulsion again at 7000 rpm for 10 min to finally obtain a uniform emulsion;

[0106] 5) The homogenized emulsion is sterilized by filtering through a 0.45 μm + 0.22 μm combined microporous filter membrane to obtain an emulsion H containing spectinomycin hydrochloride and lincomycin hydrochloride.

[0107] Example 9

[0108] Emulsion I containing spectinomycin hydrochloride and lincomycin hydrochloride

[0109] 1) At 30° C. and under nitrogen protection, 6 wt % of Span 40 and tween 20 (mass fraction ratio 1:1) emulsifiers were added to 10 wt % of white oil for injection, and after complete dissolution, an oil phase was obtained;

[0110] 2) dissolving 0.2wt% tea polyphenols, 0.1wt% lipoic acid, 0.2wt% polyvinyl alcohol, 5wt% lincomycin hydrochloride and 10% spectinomycin hydrochloride in 68.5wt% water for injection, and after complete dissolution, ultrasonically treating at 400W for 30min to obtain an aqueous phase;

[0111] 3) After mixing the oil phase and the water phase, emulsify at 3000 rpm for 2 min to obtain a primary emulsion;

[0112] 4) The primary emulsion is then emulsified again at 6000 rpm for 8 min to finally obtain a uniform emulsion;

[0113] 5) The homogenized emulsion was sterilized by filtering through a 0.45 μm + 0.22 μm combined microporous filter membrane to obtain an emulsion I containing spectinomycin hydrochloride and lincomycin hydrochloride.

[0114] Example 10

[0115] Emulsion containing spectinomycin hydrochloride and lincomycin hydrochloride

[0116] 1) At 45° C. and under nitrogen protection, 6 wt % of Span 40 and tween 20 (mass fraction ratio 1:1) emulsifiers were added to 10 wt % of white oil for injection, and after complete dissolution, an oil phase was obtained;

[0117] 2) dissolving 0.2wt% tea polyphenols, 0.1wt% lipoic acid, 0.2wt% polyvinyl alcohol, 5wt% lincomycin hydrochloride and 10% spectinomycin hydrochloride in 68.5wt% water for injection, and after complete dissolution, ultrasonically treating at 400W for 30min to obtain an aqueous phase;

[0118] 3) After mixing the oil phase and the water phase, emulsify at 4000 rpm for 5 min to obtain a primary emulsion;

[0119] 4) emulsifying the primary emulsion again at 8000 rpm for 10 min to finally obtain a uniform emulsion;

[0120] 5) The homogenized emulsion was sterilized by filtering through a 0.45 μm + 0.22 μm combined microporous filter membrane to obtain an emulsion J containing spectinomycin hydrochloride and lincomycin hydrochloride.

[0121] Experimental test 1

[0122] Long-term stability test of spectinomycin hydrochloride and lincomycin hydrochloride emulsions prepared by different methods

[0123] The emulsions of spectinomycin hydrochloride and lincomycin hydrochloride prepared according to the method of Examples 1-8 were placed at 25±2°C and relative humidity of 60%±5% for 12 months. The color and content were tested at 1, 3, 6 and 12 months. The results are shown in Table 1. The color change results are shown in Table 1. Figure 1-Figure 5 shown.

[0124] The content is determined according to the method for the content of spectinomycin and lincomycin in the pharmacopoeia, that is, the content of spectinomycin is determined according to the antibiotic microbiological detection method in the national veterinary pharmacopoeia, and the detected bacteria is Klebsiella pneumoniae; the method for determining the content of lincomycin hydrochloride: high performance liquid chromatography (HPLC-UV) is used to determine its content.

[0125] Table 1 Color changes and spectinomycin hydrochloride and lincomycin hydrochloride contents of different emulsions at 1, 3, 6 and 12 months

[0126] Note: The test content is per 25ml of Ligao emulsion: Spectinomycin 10g (10 million units) and lincomycin 5g (according to C 18 H 34 N2O6S) was recorded as 100%.

[0127]

[0128] From the results in Table 1, it can be seen that all examples showed no discoloration and stratification at 1 month, showing good initial stability. However, as time went on, Examples 2 and 3 using a single antioxidant began to show discoloration after 3 months, and the drug content decreased significantly, indicating that a single antioxidant was not as effective as a combination of two antioxidants in terms of long-term stability.

[0129] For Example 1, which did not add polyvinyl alcohol (PVA) and was not ultrasonically treated, the drug content decreased the fastest. This shows that the drug stability was poor without PVA, and the lack of ultrasonic treatment further accelerated this instability. In contrast, although Examples 2 and 3 performed better than Example 1, they still showed significant drug content decreases and discoloration over time, indicating that a single antioxidant cannot fully protect the long-term stability of the drug.

[0130] For Example 4, there was no discoloration and no stratification within 12 months, but the rate of decrease of its drug content was significantly faster than that of Example 8, which shows that the addition of PVA has a significant effect on drug stability.

[0131] In Examples 5, 6 and 8, two antioxidants were used in combination with different ultrasonic treatment methods to further improve the stability of the drug. In particular, in Example 8, which was ultrasonically treated for 30 minutes at 400W power, the contents of spectinomycin hydrochloride and lincomycin hydrochloride were still close to the initial values ​​after 12 months, showing a significant effect. However, the examples using ultrasonic treatment for 30 minutes at 200W or 800W power failed to achieve the same stability, indicating that ultrasonic conditions have an important influence on the stability of the drug.

[0132] In addition, the difference between the antioxidant combinations of Example 7 and Example 8 shows that only a specific combination of antioxidants (such as tea polyphenols and lipoic acid) can achieve the best effect. When tea polyphenols are replaced with other antioxidants, the synergistic effect disappears and the drug stability decreases significantly. This further illustrates that the type and combination of antioxidants play a key role in drug formulation design, and a reasonable combination can maximize the long-term stability of the drug.

[0133] Experimental test 2

[0134] Comparison of the degradation rates of spectinomycin hydrochloride and lincomycin hydrochloride emulsions prepared by different methods

[0135] According to the initial content, the degradation rate at different treatment times was calculated.

[0136] The test results are shown in Table 2:

[0137] Table 2 Comparison of degradation rates of spectinomycin hydrochloride and lincomycin hydrochloride emulsions prepared by different methods

[0138]

[0139] As can be seen from Table 2, the degradation rates of the spectinomycin hydrochloride and lincomycin hydrochloride contents of Examples 1, 2 and 3 increase significantly over time, i.e., the degradation rate of the spectinomycin hydrochloride content in 12 months is close to 40%, and the degradation rate of the lincomycin hydrochloride content in 12 months is also very high, close to 30%. This shows that the effects of using antioxidants alone or without ultrasound are not ideal. In contrast, the degradation rates of Examples 5, 6 and 7, and Test 8 are generally low, showing that the addition of antioxidants, PVA and specific ultrasonic conditions have a positive effect on drug stability. The degradation rates of Tests 5, 6 and 7 are better than those of the test during the entire observation period, but the degradation rate reaches about 20% after 12 months, indicating that specific ultrasonic conditions and specific antioxidant combinations are required. Example 8 confirms that the spectinomycin content of the sample prepared by PVA+tea polyphenols (0.2wt%) and lipoic acid (0.1wt%) under the ultrasonic conditions of 400w, 30min is stable. The comprehensive analysis results show that the coordination of specific excipients and antioxidants effectively controls the degradation rate of the main drugs (lincomycin hydrochloride and spectinomycin hydrochloride) in Example 8, showing excellent long-term stability.

[0140] Experimental test 3

[0141] After diluting the emulsions prepared in Example 1, Example 5, Example 6 and Example 8, the particle size of the emulsions was tested at room temperature of 25° C. using a Malvern Mastersizer 2000 / 3000 laser particle size analyzer to characterize the particle size and main distribution range.

[0142] The results are shown in Table 3:

[0143] Table 3 Difference in particle size of spectinomycin hydrochloride and lincomycin hydrochloride emulsions prepared by different methods

[0144] Grouping Example 1 Example 5 Example 6 Example 8 Average particle size 485.6±1.65 445.6±1.35 128.6±1.83 163.6±1.25 PDI 0.292±0.12 0.286±0.10 0.185±0.09 0.172±0.06

[0145] As can be seen from the results in Table 3, different ultrasonic conditions have a significant effect on the particle size and distribution of the spectinomycin hydrochloride and lincomycin hydrochloride emulsions. The average particle sizes of Examples 1 and 5 are 485.6±1.65nm and 445.6±1.35nm, respectively, and the PDI (polydispersity index) is 0.292±0.12 and 0.286±0.10, respectively, indicating that the particle sizes of these emulsions are relatively large and the particle distribution is relatively uneven.

[0146] In contrast, the average particle size of the emulsions prepared under ultrasonic conditions of 400W, 30min and 600W, 30min in Example 8 and Example 6 was significantly reduced, and the PDI was 0.185±0.09 and 0.172±0.06, indicating that the particle size of the emulsion was smaller and the distribution was more uniform. At the same time, combined with the results in Table 1, it can be seen that the stability of Example 8 is significantly better than that of the examples, indicating that the smaller the particle size, the better the stability. The possible reason is that too small a particle size will lead to an excessively large surface area, which will increase the surface free energy in the system, and will easily cause agglomeration or coagulation, resulting in the instability of the emulsion.

[0147] Therefore, in summary, the optimization of ultrasonic treatment power and time, as well as the synergistic effect with PVA and a specific combination of antioxidants, significantly improved the physical properties and stability of spectinomycin hydrochloride and lincomycin hydrochloride emulsions. Appropriate ultrasonic conditions can effectively control the particle size and distribution of the emulsion, improve the stability of the preparation and the bioavailability of the drug, thereby enhancing the overall effect and clinical application value of the preparation.

[0148] Experimental test 4

[0149] Difference in absorption of spectinomycin hydrochloride and lincomycin hydrochloride emulsions prepared by different methods

[0150] 80 white-feathered chickens were randomly divided into Example 1 Group, Example 4 Group, Example 5 Group, and Example 8 Group, with 20 chickens in each group.

[0151] The injection volume was 0.04 mL of subcutaneous injection in the neck; each treatment group was injected at 7 days old, and the drug was administered once in total. After injection, the absorption of the chicken neck was observed (the neck was dissected every 3 hours to observe the absorption), and the stress and mortality were observed at the same time. Seven days after injection, the internal organs and kidneys were dissected to observe whether there were any abnormalities.

[0152] The results obtained are shown in Table 4. The difference between the Example 1 group and the Example 8 group after 9 hours of absorption is as follows: Figure 6 and Figure 7 shown.

[0153] Table 4 Absorption differences of spectinomycin hydrochloride and lincomycin hydrochloride emulsions prepared by different methods

[0154] Example 1 Group Example 4 Group Example 5 Group Example 8 Group Death situation (7 days) 0 0 0 0 Absorption time 24h full absorption 21h full absorption 12h fully absorbed 9h all absorbed liver normal normal normal normal kidney normal normal normal normal

[0155] It can be seen from Table 4 that the emulsions prepared in Examples 1, 4, 5 and 8 of the present invention are all safe. At the same time, it can be seen that the absorption effect of the emulsion prepared in Example 8 when used for injection is significantly better than that of the emulsion prepared in Example 1, 4 or 5.

[0156] Experimental test 5

[0157] A total of 5 groups of experiments were designed, namely, the vaccine group (only vaccine injection), the blank group (none), the Ligo emulsion group (Example 1 group + vaccine), and the Ligo emulsion group (Example 8 group + vaccine), each group having 30 white-feathered broilers;

[0158] After the vaccine injection, blood is drawn at 28 days and 35 days of age for antibody titer testing. The injection ratio of vaccine and Ligo is as follows: 0.3ml vaccine + 0.05ml Ligo. After the injection, blood is drawn at 18 days and 35 days of age to test the titer.

[0159] The results are shown in Table 5:

[0160]

[0161]

[0162] As shown in Table 5, the antibody levels of the vaccinated chickens at 28 and 35 days of age were generally higher than those of the unvaccinated blank group, which indicates that vaccination is effective in increasing the antibody level of chickens.

[0163] The antibody levels of the blank group decreased significantly over time, which may be due to the disappearance of maternal antibodies and the lack of immune protection provided by the vaccine. At the same time, the chickens vaccinated with the vaccine + test group 8 showed higher antibody levels in most cases, which may mean that the test group 8 has a positive effect on improving the antibody level.

Claims

1. An emulsion containing spectinomycin hydrochloride and lincomycin hydrochloride with long-term stability, characterized in that: According to mass percentage, every 25ml of emulsion consists of the following components: 5wt% lincomycin hydrochloride; 10wt% of spectinomycin hydrochloride; 6 wt% of a mixture of Span 40 emulsifier and Tween 20 emulsifier; 0.2wt% tea polyphenols; 0.1 wt% lipoic acid; 0.2 wt% polyvinyl alcohol; 10wt% white oil for injection; The remainder is water for injection.

2. The emulsion according to claim 1, characterized in that In the mixture of Span40 emulsifier and Tween20 emulsifier, the mass ratio of Span40 emulsifier to Tween20 emulsifier is 1:

1.

3. The emulsion according to claim 2, characterized in that The emulsion is prepared by the following steps: 1) Under a certain temperature and nitrogen protection, a mixture of 6 wt% of Span 40 emulsifier and Tween 20 emulsifier is dissolved in 10 wt% of white oil for injection to form an oil phase; 2) dissolving lincomycin hydrochloride, spectinomycin hydrochloride, tea polyphenols, lipoic acid and polyvinyl alcohol in water for injection, and subjecting to ultrasonic treatment at 400 W for 30 minutes to obtain an aqueous phase; 3) mixing the oil phase with the water phase and emulsifying to obtain a primary emulsion; 4) Emulsifying again to obtain a uniform emulsion; 5) sterilizing by a microporous filter membrane to obtain the emulsion.

4. The emulsion according to claim 3, characterized in that The certain temperature is 30-45°C; the emulsification method is 3000-4000rpm emulsification for 2-5 minutes; the re-emulsification method is 6000-8000rpm emulsification for 8-10 minutes; the microporous membrane sterilization method is to filter using 0.45μm and 0.22μm filter membranes in turn.

5. The emulsion according to claim 4, characterized in that The certain temperature is 40° C., the emulsification method is emulsification at 3500 rpm for 4 minutes, and the re-emulsification method is emulsification at 7000 rpm for 8 minutes.

6. A method for preparing an emulsion containing spectinomycin hydrochloride and lincomycin hydrochloride with long-term stability, characterized in that: The preparation method comprises the following steps: 1) Under a certain temperature and nitrogen protection, a mixture of 6 wt% of Span 40 emulsifier and Tween 20 emulsifier is dissolved in 10 wt% of white oil for injection to form an oil phase; 2) dissolving lincomycin hydrochloride, spectinomycin hydrochloride, tea polyphenols, lipoic acid and polyvinyl alcohol in water for injection, and subjecting to ultrasonic treatment at 400 W for 30 minutes to obtain an aqueous phase; 3) mixing the oil phase with the water phase and emulsifying to obtain a primary emulsion; 4) Emulsifying again to obtain a uniform emulsion; 5) sterilizing by a microporous filter membrane to obtain the emulsion.

7. The preparation method according to claim 6, characterized in that: The certain temperature is 30-45°C; the emulsification method is 3000-4000rpm emulsification for 2-5 minutes; the re-emulsification method is 6000-8000rpm emulsification for 8-10 minutes; the microporous membrane sterilization method is to filter using 0.45μm and 0.22μm filter membranes in turn.

8. The preparation method according to claim 7, characterized in that: The certain temperature is 40° C., the emulsification method is emulsification at 3500 rpm for 4 minutes, and the re-emulsification method is emulsification at 7000 rpm for 8 minutes.

9. Use of an emulsion containing spectinomycin hydrochloride and lincomycin hydrochloride in the preparation of a preparation for improving the antibody titer level of poultry vaccines, characterized in that: The emulsion is prepared by the following preparation method: 1) at 37° C. and under nitrogen protection, dissolving a mixture of 6 wt % of Span 40 emulsifier and Tween 20 emulsifier in 10 wt % of white oil for injection to form an oil phase; 2) dissolving lincomycin hydrochloride, spectinomycin hydrochloride, tea polyphenols, lipoic acid and polyvinyl alcohol in water for injection, and subjecting to ultrasonic treatment at 400 W for 30 minutes to obtain an aqueous phase; 3) mixing the oil phase with the water phase and emulsifying at 3500 rpm for 4 minutes to obtain a primary emulsion; 4) Emulsification at 7000 rpm for 10 minutes to obtain a uniform emulsion; 5) Filter and sterilize using 0.45 μm and 0.22 μm filter membranes to obtain the emulsion.

10. The use according to claim 9, characterized in that: The poultry vaccines are H9-ZGS vaccine and NDV-Lasota.

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