CN110609027A - Method for rapidly detecting chlorpromazine hydrochloride in feed - Google Patents
Method for rapidly detecting chlorpromazine hydrochloride in feed Download PDFInfo
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- CN110609027A CN110609027A CN201910850436.2A CN201910850436A CN110609027A CN 110609027 A CN110609027 A CN 110609027A CN 201910850436 A CN201910850436 A CN 201910850436A CN 110609027 A CN110609027 A CN 110609027A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/65—Raman scattering
- G01N21/658—Raman scattering enhancement Raman, e.g. surface plasmons
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Abstract
The invention discloses a method for rapidly detecting chlorpromazine hydrochloride in feed, which comprises the following steps: (1) extracting a sample to obtain a solution to be detected; (2) adding the solution to be detected into the surface reinforcing agent, adding the coagulant, and uniformly mixing to obtain a mixed solution; (3) performing Raman spectrum measurement on a blank sample containing a standard substance by using a Raman spectrometer, and establishing a chlorpromazine hydrochloride standard substance database; (4) performing Raman spectrum measurement on the mixed solution by using a Raman spectrometer; (5) the raman spectrometer automatically identifies negative and positive samples. The method of the invention can simplify the pretreatment step of the analysis process, shorten the detection period, improve the detection sensitivity and reduce the detection cost.
Description
Technical Field
The invention relates to a detection method of chemical substances, in particular to a method for rapidly detecting chlorpromazine hydrochloride in feed.
Background
Chlorpromazine hydrochloride is a representative medicine of thiazine, is a common hypnotic, tranquilizer and hypnotic medicine, is a blocker of central dopamine receptors, acts on the central nervous system, and is mainly used as a tranquilizer in veterinary clinic. The feed added with the medicine can indirectly play a fattening role and reduce the death rate of animals in the transportation process. If the chlorpromazine hydrochloride enters a human body through animal food, poisoning can be caused after long-term accumulation, pathological changes of the liver and the kidney of the human body can be caused in large dose, and even life and health are damaged.
At present, the chlorpromazine hydrochloride in the feed is detected mainly by a High Performance Liquid Chromatography (HPLC) and liquid chromatography-mass spectrometry (LC-MS) combined method. Wherein the lowest detection concentration of the HPLC method is 1mg/kg, and the lowest detection concentration of the LC-MS method is 0.2 mg/kg. The two methods have the biggest defects of complex pretreatment, long test period and large-scale instrument and equipment, so that the two methods are not suitable for real-time and rapid field detection, and therefore, the identification method with high sensitivity, simplicity, accuracy and low cost is necessary.
Disclosure of Invention
The invention aims to solve the technical problem of providing a method for quickly detecting chlorpromazine hydrochloride in feed, which has the advantages of simple structure, low cost and simple and convenient operation.
The Raman spectrum is the same as the infrared spectrum, belongs to a molecular vibration spectrum, can reflect the characteristic structure of molecules, and has higher sensitivity and can be used for single molecule detection compared with optical analysis methods such as infrared, near infrared, mass spectrum and the like; the Raman response of water is very weak, and the method can be used for detecting the water-containing sample. Compared with HPLC and the like, the method has the advantages of less sample demand and low purity requirement, can reduce the pretreatment process of the sample, saves time and realizes the rapid nondestructive detection of the sample.
A method for rapidly detecting chlorpromazine hydrochloride in feed comprises the following steps:
(1) extracting a sample to obtain a solution to be detected;
(2) adding the solution to be detected into the surface reinforcing agent, adding the coagulant, and uniformly mixing to obtain a mixed solution;
(3) performing Raman spectrum measurement on a blank sample containing a standard substance by using a Raman spectrometer, and establishing a chlorpromazine hydrochloride standard substance database;
(4) performing Raman spectroscopy on the mixed solution by using a Raman spectrometer at 786 +/-5 cm-1，1091±5cm-1，1165±5cm-1，1234±5cm-1，1304±5cm-1，1562±5cm-1The spectrum is used as the qualitative characteristic peak shift of chlorpromazine hydrochloride in the feed;
(5) the raman spectrometer automatically identifies negative and positive samples.
The invention discloses a method for rapidly detecting chlorpromazine hydrochloride in feed, which comprises the following steps of (1): weighing 1g of feed, crushing, placing in a centrifuge tube, adding 2mL of 0.1% sodium hydroxide ethyl acetate solution, ultrasonically vibrating for 1min, centrifuging at 4000rpm for 1min, taking supernatant, adding 1mL of distilled water, vibrating, centrifuging at 4000rpm for 1min, and obtaining supernatant as to-be-detected liquid.
The method for rapidly detecting chlorpromazine hydrochloride in the feed comprises the following steps of (2) taking 200 mu L of liquid to be detected into a test bottle, adding 100 mu L of surface reinforcing agent, adding 100 mu L of coagulant, and vibrating for 5s to obtain a mixed solution.
The method for rapidly detecting chlorpromazine hydrochloride in the feed, disclosed by the invention, comprises the step (2) of using a nano gold colloidal solution or a nano silver colloidal solution as the surface enhancer and using a sodium chloride solution with the concentration of 0.1g/mL as the coagulant.
The method for rapidly detecting chlorpromazine hydrochloride in the feed, disclosed by the invention, comprises the step (2) of taking a nano gold colloid solution as a surface reinforcing agent.
The invention discloses a method for rapidly detecting chlorpromazine hydrochloride in feed, which comprises the following steps of (3): accurately weighing 1g of blank sample, placing the blank sample in a centrifuge tube, adding chlorpromazine hydrochloride standard solution with total concentration of 100ppm, 10ppm, 1ppm and 100ppb, respectively measuring Raman spectra according to experimental parameters in 2, drawing a standard curve of concentration-Raman spectrum peak intensity, respectively performing Raman spectrum measurement on the blank sample containing standard substances with different concentrations by using a Raman spectrometer, establishing a chlorpromazine hydrochloride standard substance database, forming a standard spectrum library, and using 786 +/-5 cm of the standard curve-1，1091±5cm-1，1165±5cm-1，1234±5cm-1，1304±5cm-1，1562±5cm-1The spectrum is taken as the qualitative characteristic peak shift of chlorpromazine hydrochloride.
The method for rapidly detecting chlorpromazine hydrochloride in the feed, disclosed by the invention, comprises the following steps of (3) and (4) in Raman spectrum measurement: frequency stabilized laser lightThe emission wavelength of the source is 785 +/-1 nm, the line width is less than 0.1nm, and the energy is more than or equal to 250 mW; spectral resolution is less than or equal to 15cm-1(ii) a Spectral response range 500cm-1～2000cm-1。
The method for rapidly detecting the chlorpromazine hydrochloride in the feed comprises the following steps of (5) matching and calculating the test result obtained in the step (4) and the chlorpromazine hydrochloride in the standard spectrogram library obtained in the step (3) by using a Raman spectrometer, judging the result of the chlorpromazine hydrochloride in the sample according to a spectrogram characteristic Raman spectrum and a built-in calibration curve, displaying the test result, and judging whether the sample is negative or positive, wherein the negative represents that the sample does not contain the chlorpromazine hydrochloride or is less than 100ppb, and the positive represents that the sample contains the chlorpromazine hydrochloride and is more than or equal to 100 ppb.
Compared with the prior art, the method for rapidly detecting chlorpromazine hydrochloride in the feed is characterized in that:
the method for rapidly detecting the chlorpromazine hydrochloride in the feed uses the surface enhanced Raman spectroscopy to rapidly detect the chlorpromazine hydrochloride in the feed, adopts the nano gold or nano silver prepared by the chemical reduction method as a substrate, and uses the chlorpromazine hydrochloride as a probe molecule, so that the pretreatment steps of the analysis process can be simplified, the detection period can be shortened, the detection sensitivity can be improved, and the detection cost can be reduced. For example, chlorpromazine hydrochloride is measured in the specification NY/T1458-2007, the pretreatment comprises a series of steps of extraction, blow-drying dissolution, purification, elution and the like, and the method only needs to extract a sample without further purification, so that the pretreatment step is simplified. And compared with the traditional Raman, the surface enhanced Raman can improve the sensitivity. The method introduces nano gold/nano silver as a surface reinforcing agent to realize the rapid detection of chlorpromazine hydrochloride in the feed sample.
The method is characterized in that nanogold/nanosilver is used as a reinforcing agent, a feed sample is pretreated, Raman spectrum detection is carried out, meanwhile, instrument software carries out matching calculation on a test result and chlorpromazine hydrochloride in a standard spectrogram library, and according to a spectrogram characteristic Raman spectrum and a built-in calibration curve, the result judgment is carried out on the chlorpromazine hydrochloride in the sample: and displaying the test result and judging whether the test result is negative or positive. Negative means that the sample does not contain chlorpromazine hydrochloride or is less than 100ppb, and positive means that the sample contains chlorpromazine hydrochloride and is greater than or equal to 100 ppb.
The method for rapidly detecting chlorpromazine hydrochloride in the feed is further described below with reference to the accompanying drawing.
Drawings
FIG. 1 is a surface enhanced Raman spectrum of chlorpromazine hydrochloride in the feed of the invention;
FIG. 2 is a schematic diagram showing the identification of characteristic peaks of chlorpromazine hydrochloride of standard samples of different concentrations in the invention;
FIG. 3 is a graph showing the variation trend of the concentration of chlorpromazine hydrochloride in the feed substrate and the intensity of a characteristic peak.
Detailed Description
Example 1
A method for rapidly detecting chlorpromazine hydrochloride in feed comprises the following steps:
(1) weighing 1g of feed, crushing, placing in a centrifuge tube, adding 2mL of 0.1% sodium hydroxide ethyl acetate solution, ultrasonically vibrating for 1min, centrifuging at 4000rpm for 1min, taking supernatant, adding 1mL of distilled water, vibrating, centrifuging at 4000rpm for 1min, and obtaining supernatant as to-be-detected liquid.
(2) And (3) putting 200 mu L of liquid to be tested into a test bottle, adding 100 mu L of the surface reinforcing agent, adding 100 mu L of coagulant, and shaking for 5s to obtain a mixed solution. The surface reinforcing agent is a nano gold colloid solution, and the coagulant is a sodium chloride solution with the concentration of 0.1 g/mL.
The preparation method of the nano gold sol comprises the following steps: 200ml of 0.01% chloroauric acid (AuCl) are taken3·HCl·4H2O) aqueous solution to boil, 2.0mL of 1% trisodium citrate (Na) was added with vigorous stirring3C6H5O7) The water solution, golden yellow chloroauric acid water solution, turns red within 2min, boiling for 15min, cooling, and adding 200mL distilled water to 200 mL. The water for preparing the enhancing reagent is primary water specified in GB/T6682.
(3) Establishing a chlorpromazine hydrochloride standard substance database: accurately weighing 1g of blank samples, placing the blank samples into a centrifuge tube, adding chlorpromazine hydrochloride standard solutions with total concentration of 100ppm, 10ppm, 1ppm and 100ppb, respectively measuring Raman spectra of the blank samples according to experimental parameters in 2, drawing a standard curve of concentration-Raman spectrum peak intensity, respectively carrying out Raman spectrum measurement on the blank samples containing standard substances with different concentrations by using a Raman spectrometer, establishing a chlorpromazine hydrochloride standard substance database, and forming a standard spectrum library as shown in figure 2.
At 786 +/-5 cm-1，1091±5cm-1，1165±5cm-1，1234±5cm-1，1304±5cm-1，1562±5cm-1The spectrum is taken as the qualitative characteristic peak shift of chlorpromazine hydrochloride.
(4) Performing Raman spectroscopy on the mixed solution by using a Raman spectrometer at 786 +/-5 cm-1，1091±5cm-1，1165±5cm-1，1234±5cm-1，1304±5cm-1，1562±5cm-1The spectrum is used as the qualitative characteristic peak shift of chlorpromazine hydrochloride in the feed;
(5) and (3) matching and calculating the test result obtained in the step (4) with the chlorpromazine hydrochloride in the standard spectrum library obtained in the step (3) by using the Raman spectrometer, judging the result of the chlorpromazine hydrochloride in the sample according to the spectrum characteristic Raman spectrum and a built-in calibration curve, displaying the test result, and judging whether the sample is negative or positive, wherein the negative represents that the sample does not contain the chlorpromazine hydrochloride or is less than 100ppb, and the positive represents that the sample contains the chlorpromazine hydrochloride and is more than or equal to 100 ppb.
In the step (3) and the step (4), the following test conditions are adopted for Raman spectrum measurement: a frequency stabilized laser source with the emission wavelength of 785 +/-1 nm, the line width of less than 0.1nm and the energy of more than or equal to 250 mW; spectral resolution is less than or equal to 15cm-1(ii) a Spectral response range 500cm-1～2000cm-1。
In the embodiment, the feed is 4% of premix for middle-aged pigs, the adopted raman spectrometer is specifically a raney-shain Via laser raman spectrometer, a surface enhanced raman spectrogram of chlorpromazine hydrochloride in the feed obtained in the step (4) is shown in fig. 1, a characteristic peak identification schematic diagram of chlorpromazine hydrochloride of standard samples with different concentrations in the step (3) is shown in fig. 2, and a concentration and characteristic peak intensity change trend diagram of chlorpromazine hydrochloride in a feed matrix is shown in fig. 3. The result of this example was negative, i.e., chlorpromazine hydrochloride was not contained or was less than 100 ppb.
Example 2
The surface reinforcing agent is a nano silver colloidal solution, and the specific preparation method comprises the following steps:
taking 100mL1.0 mM silver nitrate (AgNO)3) Heating the aqueous solution to boil, and accurately dropwise adding 2.5mL of 1% trisodium citrate (Na) under vigorous stirring3C6H5O7) Boiling the aqueous solution for 1 hr to turn the solution into grey green, cooling, and adding distilled water to 100mL if the solution is less than 100 mL. The water for preparing the enhancing reagent is primary water specified in GB/T6682.
The other steps were the same as in example 1.
In the embodiment, the feed is 4% of the premix for the middle pigs, and the obtained judgment result is negative, namely the feed does not contain chlorpromazine hydrochloride or is lower than 100 ppb.
The above-mentioned embodiments are merely illustrative of the preferred embodiments of the present invention, and do not limit the scope of the present invention, and various modifications and improvements of the technical solution of the present invention by those skilled in the art should fall within the protection scope defined by the claims of the present invention without departing from the spirit of the present invention.
Claims (8)
1. A method for rapidly detecting chlorpromazine hydrochloride in feed is characterized by comprising the following steps: the method comprises the following steps:
(1) extracting a sample to obtain a solution to be detected;
(2) adding the solution to be detected into the surface reinforcing agent, adding the coagulant, and uniformly mixing to obtain a mixed solution;
(3) performing Raman spectrum measurement on a blank sample containing a standard substance by using a Raman spectrometer, and establishing a chlorpromazine hydrochloride standard substance database;
(4) performing Raman spectroscopy on the mixed solution by using a Raman spectrometer at 786 +/-5 cm-1，1091±5cm-1，1165±5cm-1，1234±5cm-1，1304±5cm-1，1562±5cm-1Spectrum as feedDisplacement of qualitative characteristic peak of chlorpromazine hydrochloride;
(5) the raman spectrometer automatically identifies negative and positive samples.
2. The method for rapidly detecting chlorpromazine hydrochloride in feed according to claim 1, which is characterized in that: the step (1) is as follows: weighing 1g of feed, crushing, placing in a centrifuge tube, adding 2mL of 0.1% sodium hydroxide ethyl acetate solution, ultrasonically vibrating for 1min, centrifuging at 4000rpm for 1min, taking supernatant, adding 1mL of distilled water, vibrating, centrifuging at 4000rpm for 1min, and obtaining supernatant as to-be-detected liquid.
3. The method for rapidly detecting chlorpromazine hydrochloride in feed according to claim 2, characterized in that: and (2) putting 200 mu L of liquid to be tested into a test bottle, adding 100 mu L of the surface reinforcing agent, adding 100 mu L of coagulant, and shaking for 5s to obtain a mixed solution.
4. The method for rapidly detecting chlorpromazine hydrochloride in feed according to claim 3, wherein the method comprises the following steps: in the step (2), the surface reinforcing agent is a nano gold colloidal solution or a nano silver colloidal solution, and the coagulant is a sodium chloride solution with the concentration of 0.1 g/mL.
5. The method for rapidly detecting chlorpromazine hydrochloride in feed according to claim 4, wherein the method comprises the following steps: in the step (2), the surface reinforcing agent is a nano gold colloid solution.
6. The method for rapidly detecting chlorpromazine hydrochloride in feed according to claim 1, which is characterized in that: the step (3) is specifically as follows: accurately weighing 1g of blank sample, placing the blank sample in a centrifuge tube, adding chlorpromazine hydrochloride standard solutions with total concentration of 100ppm, 10ppm, 1ppm and 100ppb, respectively measuring Raman spectra of the blank sample according to experimental parameters in the specification 2, drawing a standard curve of concentration-Raman spectrum peak intensity, respectively carrying out Raman spectrum measurement on the blank sample containing standard substances with different concentrations by using a Raman spectrometer, and establishing the number of chlorpromazine hydrochloride standard substancesDatabase, forming standard spectrum library with standard curve of 786 + -5 cm-1，1091±5cm-1，1165±5cm-1，1234±5cm-1，1304±5cm-1，1562±5cm-1The spectrum is taken as the qualitative characteristic peak shift of chlorpromazine hydrochloride.
7. The method for rapidly detecting chlorpromazine hydrochloride in feed according to claim 6, which is characterized in that: in the step (3) and the step (4), the following test conditions are adopted for Raman spectrum measurement: a frequency stabilized laser source with the emission wavelength of 785 +/-1 nm, the line width of less than 0.1nm and the energy of more than or equal to 250 mW; spectral resolution is less than or equal to 15cm-1(ii) a Spectral response range 500cm-1～2000cm-1。
8. The method for rapidly detecting chlorpromazine hydrochloride in feed according to claim 7, wherein the method comprises the following steps: in the step (5), the Raman spectrometer performs matching calculation on the test result obtained in the step (4) and the chlorpromazine hydrochloride in the standard spectrum library obtained in the step (3), according to the spectrum characteristic Raman spectrum and a built-in calibration curve, the result judgment is performed on the chlorpromazine hydrochloride in the sample, the test result is displayed, and the negative judgment result is negative or positive, the negative judgment result does not contain the chlorpromazine hydrochloride or is lower than 100ppb, and the positive judgment result represents that the sample contains the chlorpromazine hydrochloride and is greater than or equal to 100 ppb.
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