A method and system for calculating the environmental behavior activity of fluoroquinolone antibiotics

Abstract

This invention discloses a method and system for calculating the environmental behavioral activity of fluoroquinolone antibiotics, relating to the field of activity detection technology. The method includes: acquiring a sample of the fluoroquinolone antibiotic to be tested; pretreating the acquired sample; injecting the pretreated sample into a high-performance liquid chromatograph (HPLC) for qualitative analysis of different fluoroquinolone antibiotics to obtain the residence time of the sample in the HPLC mobile phase; and constructing an activity calculation model based on the residence time of the sample in the HPLC mobile phase using a stepwise analysis method to evaluate the activity of the sample under environmental behavior. This invention uses the residence time of fluoroquinolone drugs in the HPLC mobile phase as a bioaccumulation evaluation index for bioactivity calculation, and the constructed activity calculation model for the residence time of fluoroquinolone drugs in the mobile phase meets the requirements of goodness of fit, robustness, and predictive power.

Description

Technical Field

[0001] The present invention relates to the technical field of activity detection, and in particular to a method and system for calculating the environmental behavior activity of fluoroquinolone antibiotics. Background Art

[0002] Antibiotics are a class of secondary metabolites produced by microorganisms (including bacteria, fungi and actinomycetes) or higher animals and plants in the course of their lives that have anti-pathogen or other activities. According to their chemical structure and properties, antibiotics can be divided into sulfonamides, quinolones, tetracyclines, macrolides, polyethers and B-lactams.

[0003] Fluoroquinolone drugs are the most representative class of homologues in drugs and personal care products. In addition to their remarkable medicinal properties such as broad bactericidal spectrum, low toxicity and side effects, and moderate price, as a type of "new pollutants", they also pose direct and potential hazards to microorganisms, animals, plants and human health, and exhibit the characteristics of persistent organic pollutants.

[0004] Due to the wide variety of fluoroquinolone drugs and the fact that new drugs are still being synthesized and developed, there is a lack of experimental test values ​​for the physical and chemical properties of fluoroquinolone drugs, especially those for environmental hazards, which makes it difficult to systematically analyze, calculate and evaluate the environmental behavior activity of fluoroquinolone drugs.

[0005] Therefore, it is an urgent problem for those skilled in the art to propose a method and system for calculating the environmental behavioral activity of fluoroquinolone antibiotics to solve the difficulties existing in the prior art. Summary of the invention

[0006] In view of this, the present invention provides a method and system for calculating the environmental behavior activity of fluoroquinolone antibiotics. For biological activity detection calculation, the residence time of fluoroquinolone drugs in liquid chromatography mobile phase is used as a bio-enrichment evaluation index, and the constructed activity calculation model of the residence time of fluoroquinolone drugs in mobile phase meets the requirements of fitting, robustness and predictability.

[0007] In order to achieve the above object, the present invention adopts the following technical solution:

[0008] A method for calculating the environmental behavior activity of a fluoroquinolone antibiotic drug comprises the following steps:

[0009] S1. Obtaining samples: Obtaining fluoroquinolone antibiotic drug samples to be tested;

[0010] S2. Sample pretreatment: pretreatment of the obtained samples to be tested;

[0011] S3. Activity analysis: inject the pretreated sample into the active liquid chromatograph, conduct qualitative analysis on different fluoroquinolone antibiotics, and obtain the residence time of the sample in the liquid chromatography mobile phase;

[0012] S4. Evaluation calculation: A step-by-step analysis method is used to construct an activity calculation model for the residence time of the sample to be tested in the liquid chromatography mobile phase to evaluate the activity of the sample to be tested under environmental behavior.

[0013] In the above method, optionally, the specific content of preprocessing the obtained sample to be tested in S2 is:

[0014] Grind the fluoroquinolone antibiotic drug sample to be tested, and sieve it, and then sieve out the fertilizer fine powder for later use;

[0015] The fine powder is added to deionized water, heated and stirred in a water bath, the upper layer of liquid is taken and placed in a centrifuge tube for ultrasonic extraction and centrifugation, and the supernatant is collected;

[0016] The supernatant was passed through a microfiltration membrane to collect the filtrate;

[0017] Place the adsorbed particles in a bottle, add methanol and water to activate, add metal salt solution, heat and stir in a water bath, separate with a magnet, pour off the supernatant, wash the solid with phosphate to remove metal ions, and add the filtrate;

[0018] Ultrasonic extraction under UV light;

[0019] The adsorbed particles were taken out, eluted with acetic acid and methanol, and the eluate was fixed to a certain volume and stored as a sample to be tested.

[0020] In the above method, optionally, in S3, high performance liquid chromatography-electrospray ionization tandem mass spectrometry is used to determine the antibiotic content in the sample to be tested, the chromatographic column is BEH C18, the column temperature is 35° C., the flow rate is 0.3 ml / min, and the injection volume is 5 μL.

[0021] In the above method, optionally, in S4, a stepwise analysis method is used to construct an activity calculation model for the residence time of the sample to be tested in the liquid chromatography mobile phase, and the specific content of evaluating the activity of the sample to be tested under environmental behavior is:

[0022] The samples were sorted according to their residence time in the liquid chromatography mobile phase, and then the homologues were divided into a modeling group and a verification group using the interval-samping method.

[0023] Inputting the modeling group into the constructed activity calculation model to train the activity calculation model; after several trainings, a trained activity calculation model is obtained;

[0024] The validation set was input into the trained activity computational model to evaluate the activity under ambient behavior in the liquid chromatography mobile phase.

[0025] In the above method, optionally, the activity calculation model expression is:

[0026] Residence time = 44.1-2.14q c7 -70.0q o14 +0.265μ+0.0478α+0.00673β yyy

[0027] Among them, o14 and c7 are the charge values ​​of the fluoroquinolone drug molecule.

[0028] The above method optionally constructs an activity calculation model using the simulation coefficient R 2 , Fisher constant value F, interaction test coefficient q 2 Performance evaluation is performed using the predicted sum of squares PRESS:

[0029]

[0030] 0.85≤k≤1.15or0.85≤k'≤1.15

[0031] The expressions for k and k' are:

[0032]

[0033] Among them, k and k' are Tropsha parameters, is the prediction simulation coefficient obtained using the validation group data, y i is the experimental detection value, is the predicted value of residence time obtained by the model, is the mean value of the dependent variable of the modeling group.

[0034] In the above method, optionally, the fluoroquinolone antibiotic is one or more of norfloxacin, ciprofloxacin, levofloxacin, ofloxacin and moxifloxacin.

[0035] A system for calculating the environmental behavior activity of fluoroquinolone antibiotics, using any one of the above methods for calculating the environmental behavior activity of fluoroquinolone antibiotics, comprising: a sample acquisition module, a sample pretreatment module, an activity analysis module and an evaluation calculation module;

[0036] A sample acquisition module is connected to the input end of the sample pretreatment module and is used to acquire a fluoroquinolone antibiotic drug sample to be tested;

[0037] A sample preprocessing module, connected to the input end of the activity analysis module, for preprocessing the acquired sample to be tested;

[0038] The activity analysis module is connected to the input end of the evaluation calculation module and is used to inject the pretreated sample to be tested into the active liquid chromatograph to perform qualitative analysis on different fluoroquinolone antibiotics and obtain the residence time of the sample to be tested in the liquid chromatography mobile phase;

[0039] The evaluation calculation module is used to construct an activity calculation model of the residence time of the sample to be tested in the liquid chromatography mobile phase by using a stepwise analysis method, and to evaluate the activity of the sample to be tested under environmental behavior.

[0040] It can be seen from the above technical solution that, compared with the prior art, the present invention provides a method and system for calculating the environmental behavior activity of fluoroquinolone antibiotics, which has the following beneficial effects:

[0041] (1) The pretreatment step uses heating in a water bath and ultrasonic extraction, which has simple operation steps and low cost;

[0042] (2) For biological activity detection calculations, the residence time of fluoroquinolone drugs in the liquid chromatography mobile phase was used as the bio-enrichment evaluation index, and the constructed activity calculation model of the residence time of fluoroquinolone drugs in the mobile phase met the requirements of fitting, robustness, and predictability. BRIEF DESCRIPTION OF THE DRAWINGS

[0043] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on the provided drawings without paying creative work.

[0044] Figure 1 A flow chart of a method for calculating the environmental behavior activity of a fluoroquinolone antibiotic provided by the present invention;

[0045] Figure 2 This is a structural block diagram of the environmental behavior activity calculation system of fluoroquinolone antibiotics provided by the present invention. DETAILED DESCRIPTION

[0046] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0047] Reference Figure 1 As shown, the present invention discloses a method for calculating the environmental behavior activity of fluoroquinolone antibiotics, comprising the following steps:

[0048] S1. Obtaining samples: Obtaining fluoroquinolone antibiotic drug samples to be tested;

[0049] S2. Sample pretreatment: pretreatment of the obtained samples to be tested;

[0050] S3. Activity analysis: inject the pretreated sample into the active liquid chromatograph, conduct qualitative analysis on different fluoroquinolone antibiotics, and obtain the residence time of the sample in the liquid chromatography mobile phase;

[0051] S4. Evaluation calculation: A step-by-step analysis method is used to construct an activity calculation model for the residence time of the sample to be tested in the liquid chromatography mobile phase to evaluate the activity of the sample to be tested under environmental behavior.

[0052] Furthermore, the specific content of preprocessing the obtained sample to be tested in S2 is:

[0053] Grind the fluoroquinolone antibiotic drug sample to be tested, and sieve it, and then sieve out the fertilizer fine powder for later use;

[0054] The fine powder is added to deionized water, heated and stirred in a water bath, the upper layer of liquid is taken and placed in a centrifuge tube for ultrasonic extraction and centrifugation, and the supernatant is collected;

[0055] The supernatant was passed through a microfiltration membrane to collect the filtrate;

[0056] Place the adsorbed particles in a bottle, add methanol and water to activate, add metal salt solution, heat and stir in a water bath, separate with a magnet, pour off the supernatant, wash the solid with phosphate to remove metal ions, and add the filtrate;

[0057] Ultrasonic extraction under UV light;

[0058] The adsorbed particles were taken out, eluted with acetic acid and methanol, and the eluate was fixed to a certain volume and stored as a sample to be tested.

[0059] Furthermore, in S3, high performance liquid chromatography-electrospray ionization tandem mass spectrometry was used to determine the antibiotic content in the sample to be tested, with a BEH C18 column, a column temperature of 35°C, a flow rate of 0.3 ml / min, and an injection volume of 5 μL.

[0060] Furthermore, in S4, a stepwise analysis method is used to construct an activity calculation model for the residence time of the sample to be tested in the liquid chromatography mobile phase, and the specific content of evaluating the activity of the sample to be tested under environmental behavior is:

[0061] The samples were sorted according to their residence time in the liquid chromatography mobile phase, and then the homologues were divided into a modeling group and a verification group using the interval-samping method.

[0062] Inputting the modeling group into the constructed activity calculation model to train the activity calculation model; after several trainings, a trained activity calculation model is obtained;

[0063] The validation set was input into the trained activity computational model to evaluate the activity under ambient behavior in the liquid chromatography mobile phase.

[0064] Furthermore, the activity calculation model expression is:

[0065] Residence time = 44.1-2.14q c7 -70.0q o14 +0.265μ+0.0478α+0.00673β yyy

[0066] Among them, o14 and c7 are the charge values ​​of the fluoroquinolone drug molecule.

[0067] Furthermore, the activity calculation model constructed uses the simulation coefficient R 2 , Fisher constant value F, interaction test coefficient q 2 Performance evaluation is performed using the predicted sum of squares PRESS:

[0068]

[0069] 0.85≤k≤1.15or0.85≤k'≤1.15

[0070] The expressions for k and k' are:

[0071]

[0072] Among them, k and k' are Tropsha parameters, is the prediction simulation coefficient obtained using the validation group data, y i is the experimental detection value, is the predicted value of residence time obtained by the model, is the mean value of the dependent variable of the modeling group.

[0073] Furthermore, the fluoroquinolone antibiotic drug is one or more of norfloxacin, ciprofloxacin, levofloxacin, ofloxacin and moxifloxacin.

[0074] In a specific embodiment, a fluoroquinolone antibiotic drug sample to be tested is obtained; the obtained sample to be tested is pretreated; the pretreated sample to be tested is injected into a high-performance liquid chromatograph, and different fluoroquinolone antibiotics are qualitatively analyzed to obtain the residence time of the sample to be tested in the liquid chromatography mobile phase; a stepwise analysis method is used to construct an activity calculation model for the residence time of the sample to be tested in the liquid chromatography mobile phase, and the activity of the sample to be tested under environmental behavior is evaluated.

[0075] The activity calculation model expression is:

[0076] Residence time = 44.1-2.14q c7 -70.0q o14 +0.265μ+0.0478α+0.00673β yyy

[0077] Among them, o14 and c7 are the charge values ​​of the fluoroquinolone drug molecule.

[0078] The activity calculation model constructed uses the simulation coefficient R 2 , Fisher constant value F, interaction test coefficient q 2 Performance evaluation is performed using the predicted sum of squares PRESS:

[0079]

[0080] 0.85≤k≤1.15or0.85≤k'≤1.15

[0081] The expressions for k and k' are:

[0082]

[0083] Among them, k and k' are Tropsha parameters, is the prediction simulation coefficient obtained using the validation group data, y i is the experimental detection value, is the predicted value of residence time obtained by the model, is the mean value of the dependent variable of the modeling group.

[0084] The model's fitting parameter performance is: R 2 =0.931, F = 24.28, where R 2 The fitting performance is greater than 0.9, and the significance evaluation index is less than the critical value of very significant index 0.01. The model robustness parameter performance is q 2=0.643, a robustness performance index value greater than 0.5, PRESS=14.115, and the residence time of fluoroquinolone drugs in the liquid chromatography mobile phase is used as the bioaccumulation evaluation index. The constructed activity calculation model of the residence time of fluoroquinolone drugs in the mobile phase meets the requirements of fitting, robustness and predictability.

[0085] and Figure 1 Corresponding to the method described above, the embodiment of the present invention also provides a system for calculating the environmental behavior activity of fluoroquinolone antibiotics, for calculating Figure 1 The specific implementation of the method is shown in the following diagram: Figure 2 As shown, it includes: a sample acquisition module, a sample pretreatment module, an activity analysis module and an evaluation calculation module;

[0086] A sample acquisition module is connected to the input end of the sample pretreatment module and is used to acquire a fluoroquinolone antibiotic drug sample to be tested;

[0087] A sample preprocessing module, connected to the input end of the activity analysis module, for preprocessing the acquired sample to be tested;

[0088] The activity analysis module is connected to the input end of the evaluation calculation module and is used to inject the pretreated sample to be tested into the active liquid chromatograph to perform qualitative analysis on different fluoroquinolone antibiotics and obtain the residence time of the sample to be tested in the liquid chromatography mobile phase;

[0089] The evaluation calculation module is used to construct an activity calculation model of the residence time of the sample to be tested in the liquid chromatography mobile phase by using a stepwise analysis method, and to evaluate the activity of the sample to be tested under environmental behavior.

[0090] In this specification, each embodiment is described in a progressive manner, and each embodiment focuses on the differences from other embodiments. The same or similar parts between the embodiments can be referred to each other. For the device disclosed in the embodiment, since it corresponds to the method disclosed in the embodiment, the description is relatively simple, and the relevant parts can be referred to the method part.

[0091] The above description of the disclosed embodiments enables one skilled in the art to implement or use the present invention. Various modifications to these embodiments will be apparent to one skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention will not be limited to the embodiments shown herein, but rather to the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A method for calculating the environmental behavior activity of fluoroquinolone antibiotics, characterized in that: The following steps are involved: S1. Obtaining samples: Obtaining fluoroquinolone antibiotic drug samples to be tested; S2. Sample pretreatment: pretreatment of the obtained samples to be tested; S3. Activity analysis: inject the pretreated sample into the active liquid chromatograph, conduct qualitative analysis on different fluoroquinolone antibiotics, and obtain the residence time of the sample in the liquid chromatography mobile phase; S4. Evaluation calculation: A step-by-step analysis method is used to construct an activity calculation model for the residence time of the sample to be tested in the liquid chromatography mobile phase to evaluate the activity of the sample to be tested under environmental behavior.

2. The method for calculating the environmental behavior activity of a fluoroquinolone antibiotic according to claim 1, characterized in that: The specific content of preprocessing the obtained sample to be tested in S2 is: Grind the fluoroquinolone antibiotic drug sample to be tested, sieve it, and sieve out the fertilizer fine powder for later use; The fine powder is added to deionized water, heated and stirred in a water bath, the upper layer of liquid is taken and placed in a centrifuge tube for ultrasonic extraction and centrifugation, and the supernatant is collected; The supernatant was passed through a microfiltration membrane to collect the filtrate; Place the adsorbed particles in a bottle, add methanol and water to activate, add metal salt solution, heat and stir in a water bath, separate with a magnet, pour off the supernatant, wash the solid with phosphate to remove metal ions, and add the filtrate; Ultrasonic extraction under UV light; The adsorbed particles were taken out, eluted with acetic acid and methanol, and the eluate was fixed to a certain volume and stored as a sample to be tested.

3. The method for calculating the environmental behavior activity of a fluoroquinolone antibiotic according to claim 1, characterized in that: In S3, high performance liquid chromatography-electrospray ionization tandem mass spectrometry was used to determine the antibiotic content in the sample to be tested. The chromatographic column was BEHC18, the column temperature was 35°C, the flow rate was 0.3 ml / min, and the injection volume was 5 μL.

4. The method for calculating the environmental behavior activity of a fluoroquinolone antibiotic according to claim 1, characterized in that: In S4, a stepwise analysis method is used to construct an activity calculation model for the residence time of the sample to be tested in the liquid chromatography mobile phase. The specific content of evaluating the activity of the sample to be tested under environmental behavior is: The samples were sorted according to their residence time in the liquid chromatography mobile phase, and then the homologues were divided into a modeling group and a verification group using the interval-samping method. Inputting the modeling group into the constructed activity calculation model to train the activity calculation model; after several trainings, a trained activity calculation model is obtained; The validation set was input into the trained activity computational model to evaluate the activity under ambient behavior in the liquid chromatography mobile phase.

5. The method for calculating the environmental behavior activity of a fluoroquinolone antibiotic according to claim 4, characterized in that: The activity calculation model expression is: Residence time = 44.1-2.14q c7 -70.0q o14 +0.265μ+0.0478α+0.00673β yyy Among them, o14 and c7 are the charge values ​​of the fluoroquinolone drug molecule.

6. The method for calculating the environmental behavior activity of a fluoroquinolone antibiotic according to claim 1, characterized in that: The activity calculation model constructed uses the simulation coefficient R 2 , Fisher constant value F, interaction test coefficient q 2 Performance evaluation is performed using the predicted sum of squares PRESS: 0.85≤k≤1.15or0.85≤k'≤1.15 The expressions of k and k, are: Among them, k and k' are Tropsha parameters, is the prediction simulation coefficient obtained using the validation group data, y i is the experimental detection value, is the predicted value of residence time obtained by the model, is the mean value of the dependent variable of the modeling group.

7. The method for calculating the environmental behavior activity of a fluoroquinolone antibiotic according to claim 1, characterized in that: The fluoroquinolone antibiotics are one or more of norfloxacin, ciprofloxacin, levofloxacin, ofloxacin and moxifloxacin.

8. A system for calculating the environmental behavior activity of fluoroquinolone antibiotics, characterized in that: A method for calculating the environmental behavior activity of a fluoroquinolone antibiotic drug according to any one of claims 1 to 7, comprising: a sample acquisition module, a sample pretreatment module, an activity analysis module and an evaluation calculation module; A sample acquisition module is connected to the input end of the sample pretreatment module and is used to acquire a fluoroquinolone antibiotic drug sample to be tested; A sample pretreatment module, connected to the input end of the activity analysis module, for pretreatment of the acquired sample to be tested; The activity analysis module is connected to the input end of the evaluation calculation module and is used to inject the pretreated sample to be tested into the active liquid chromatograph to perform qualitative analysis on different fluoroquinolone antibiotics and obtain the residence time of the sample to be tested in the liquid chromatography mobile phase; The evaluation calculation module is used to construct an activity calculation model of the residence time of the sample to be tested in the liquid chromatography mobile phase by using a stepwise analysis method, and to evaluate the activity of the sample to be tested under environmental behavior.

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