CN108732292B - Method and device for rapidly detecting sufentanil in blood plasma - Google Patents
Method and device for rapidly detecting sufentanil in blood plasma Download PDFInfo
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Abstract
The invention provides a method for rapidly detecting sufentanil in plasma, which comprises the following steps of S1, pretreating the plasma to be detected to obtain a plasma sample to be detected, S2, preparing a standard solution and a simulated plasma sample, S3, respectively sucking the plasma sample to be detected, the simulated plasma sample and the standard solution, dotting the plasma sample to be detected, the simulated plasma sample and the standard solution on a silica gel plate, S4, carrying out unfolding, airing and iodine cylinder color development on the plasma sample to be detected, S5, respectively dripping a predetermined amount of silver colloid at sufentanil spots of the plasma sample to be detected and the simulated plasma sample, S6, carrying out surface enhanced Raman scattering detection on the spots dripped with the silver colloid, S7, and calculating the concentration of sufentanil in the plasma sample to be detected, wherein the suction amount of the plasma sample to be detected, the simulated plasma sample and the standard solution in the step S3 is 1 mu L, and the predetermined amount of the silver colloid in the step S5 is 5 mu L, and the silver colloid contains silver particles with the average particle size of 50 nm.
Description
Technical Field
The invention relates to a method for detecting sufentanil, in particular to a method and a device for quickly detecting sufentanil in blood plasma.
Background
Sufentanil (sufentanil) is a fentanyl derivative with the chemical name of N- [4- (methoxymethyl) -1-2[2- (2-thienyl) ethyl ] -4-piperidyl ] -N-phenylacrylamide, the lipid solubility of which is 1100 times that of morphine, the sufentanil can easily penetrate through a blood brain barrier and quickly reach an effective concentration in the brain, the analgesic effect is 5-10 times that of fentanil and 400 times that of morphine, the sufentanil is an opioid analgesic with the strongest analgesic effect at present, and the sufentanil is widely applied to general anesthesia induction, maintenance in operation, postoperative analgesia and the like in clinic.
Sufentanil is suitable for anesthesia in various surgical procedures, but due to individual differences among patients, the amount of sufentanil should be adjusted in time according to patient response to avoid toxic side effects, such as respiratory depression over the time that a large dose of sufentanil can be administered (sufentanil pharmacology vs. clinical use, by fisher, loelin, medical guidance, P. 2009, 11, 1482), so concentration detection of sufentanil in biological samples such as plasma is required in addition to the long-term work of medical workers in hospital or operating rooms, there is a possibility that gas exhaled from patients will come into contact with fentanil and thereby cause symptoms of opioid sensitivity or addiction (mcaulie P F, Gold M S, bajp L, et al. second-hand ex situ inhalation, intravenous inhalation, and therapy, 2006, calcium usage and therapy, respectively, and for rapid in vivo detection of specific compounds such as 90, 882, and so there is significant in vivo clinical tests for such compounds, especially for rapid in vivo medical use of sufentanil.
At present, the sufentanil detection method mainly comprises a high performance liquid chromatography, a liquid chromatography-mass spectrometry combined method (L C-MS), a gas chromatography-mass spectrometry combined method (GC-MS) and the like.
The Thin-layer Chromatography (Thin-L a layer Chromatography, T L C) is a classical separation analysis method, and is widely applied to qualitative and quantitative detection of drugs, and Surface Enhanced Raman Scattering (SERS) is applied to many research fields due to the advantages of high sensitivity, strong characteristics and short detection time.T L C-SERS combined technology has the advantages of short analysis period and strong characteristics, and is used for field detection of environmental pollution, detection of adulteration of traditional Chinese medicines, detection of chemical drugs added in traditional Chinese medicines, detection of forbidden medicines, detection of trace drugs and biochemical warfare agents, and clinical drug monitoring, etc. the spectral signal detection in the T L C-SERS technology can be carried out by using a handheld spectrometer, so that the detection device is more portable as a whole, and T L C-SERS is very suitable for field detection, but the SERS technology is not used for field detection of sufentanil in plasma by using T L C-SERS technology at present.
Disclosure of Invention
In order to solve the problems, the invention provides a method and a device for rapidly and accurately detecting the content of sufentanil in blood plasma, and the inventor of the invention provides the following technical scheme on the basis of searching related detection methods and conditions:
the invention provides a method for rapidly detecting sufentanil in plasma, which is used for detecting the content of sufentanil in plasma to be detected and is characterized by comprising the following steps of S1, preprocessing the plasma to be detected to obtain a plasma sample to be detected, S2, preparing a sufentanil solution as a standard solution, adding sufentanil into a blank plasma sample to prepare a simulated plasma sample containing sufentanil, S3, respectively sucking the plasma sample to be detected, the simulated plasma sample and the standard solution and dotting the plasma sample and the standard solution on the same silica gel plate, S4, using dichloromethane-methanol as a developing agent to develop, air-dry and iodine cylinder color the plasma sample to be detected, the positions of sufentanil spots in the plasma sample to be detected and the simulated plasma sample to be detected are determined according to the positions of sufentanil in the standard solution, respectively dripping a predetermined amount of silver colloid at the sufentanil spots in the plasma sample to be detected and the simulated plasma sample to be detected, S6, performing surface enhanced scattering detection on the positions of the sufentanil spots in the plasma sample to be detected, respectively, detecting the sufentanil spots in the plasma sample to obtain a simulated plasma sample containing a simulated silver colloid concentration peak, and a simulated plasma sample, wherein the peak of the silver colloid is calculated according to be detected, the peak intensity of the sample before step S3526, the sample to be detected, the sample is calculated by using a spectrum of the sample to be detected, the sample to be detected sample, the peak of the sample to be detected, the sample to be detected sample, the sample is calculated by using a spectrum of.
The method for rapidly detecting sufentanil in blood plasma provided by the invention can also have the technical characteristics that the silver colloid in the step S5 is prepared by the following steps of weighing 34mg of silver nitrate and dissolving in 200m L water to obtain a silver nitrate solution, heating, refluxing and condensing the silver nitrate solution to slight boiling, adding a trisodium citrate solution with the mass percentage of 6m L of 1% to form a mixed solution, keeping heating until the color of the mixed solution is changed from colorless transparency to light yellow and further changed into light gray and slightly green, continuing to heat for 30min after the color is changed, stopping heating, and carrying out water bath cooling on the mixed solution to obtain the silver colloid.
The method for rapidly detecting sufentanil in blood plasma provided by the invention can also have the technical characteristics that the conditions for surface enhanced Raman scattering detection in the step S6 are that the laser power is 100mW, the integration time is 5S and the magnification of a microscope system is 20.
The method for rapidly detecting sufentanil in blood plasma provided by the invention can also have the technical characteristics that the analysis processing on the spectrum signal in the step S6 is as follows: selecting 300-1700cm of spectral signal-1Smoothing, baseline correction and normalization processing are carried out, the processed spectrum signals are plotted to obtain an SERS spectrum corresponding to the spectrum signals, and the sufentanil characteristic peak intensity is 1004cm in the SERS spectrum-1The intensity of the characteristic peak at (a).
The method for rapidly detecting sufentanil in blood plasma provided by the invention can also have the technical characteristics that the pretreatment method in the step S2 comprises the following steps: taking to-be-detected plasma or simulated plasma as to-be-processed plasma, adding acetonitrile according to a volume ratio of 1:2, carrying out vortex oscillation for 1min, then centrifuging for 10min at 13000rpm, taking centrifuged supernatant, drying the supernatant at 25 ℃ of a nitrogen blowing instrument, and then adding methanol with the same volume as the to-be-processed plasma for redissolution to obtain a corresponding plasma sample.
The method for rapidly detecting sufentanil in blood plasma provided by the invention can also have the technical characteristics that the plurality of simulated blood plasma respectively contain sufentanil with different concentrations, and the content range of the sufentanil contained in the simulated blood plasma is 0.85-85.00 mu g/m L.
The method for rapidly detecting sufentanil in blood plasma provided by the invention can also have the technical characteristics that the calculation method in the step S7 is as follows: establishing 1004cm in sufentanil SERS spectrum in simulated plasma sample-1The intensity of the characteristic peak is in linear relation with the concentration of sufentanil, and the concentration of sufentanil in the blood plasma sample to be tested is calculated according to the linear relation.
The invention also provides a device for rapidly detecting sufentanil in blood plasma, which is used for detecting the content of sufentanil in blood plasma to be detected and is characterized by comprising a pretreatment unit, a detection kit, a Raman spectrum detection unit and a spectrum analysis unit, wherein the pretreatment unit is used for pretreating the blood plasma to be detected to obtain a blood plasma sample to be detected, the detection kit comprises a sufentanil solution used as a standard solution, a simulated blood plasma sample used as a content calculation control and containing sufentanil, a silica gel plate used for spotting the standard solution, the simulated blood plasma sample and the blood plasma sample to be detected, an expansion cylinder used for expanding the spotted silica gel plate, an iodine cylinder used for developing color, silver gel used for performing spectrum enhancement on the expanded sufentanil spots, the Raman spectrum detection unit is used for performing surface enhanced Raman scattering detection on the sufentanil spots dripped with the silver gel and analyzing a detected spectrum signal, a characteristic peak and the intensity of the sufentanil in the simulated blood plasma sample and the blood plasma sample to be detected are obtained, the spectrum analysis unit is used for obtaining the sufentanil concentration in the simulated blood plasma sample and the blood plasma sample to be detected, the sufentanil concentration in the simulated blood plasma sample and the blood plasma sample to be detected is obtained by the sufentanil analyzer, the sample to be detected comprises 355 mu silver gel, the silver gel absorption sample is absorbed by the silver gel absorption unit, the absorption unit is used for absorbing.
Action and Effect of the invention
According to the method for detecting sufentanil in blood plasma provided by the invention, the silver colloid containing the nano silver particles with the average particle size of 50nm is used as the enhancing reagent, the silver colloid has a good signal enhancing effect on the characteristic peak of sufentanil, and no signal is generated at the characteristic peak of sufentanil, so that the T L C-SERS method can be applied to the rapid detection of sufentanil in blood plasma.
Drawings
FIG. 1 is a UV-Vis spectrum of a silver paste according to an embodiment of the present invention;
FIG. 2 is a scanning electron microscope image of silver paste according to an embodiment of the present invention;
FIG. 3 is a diagram showing the development results of thin layer chromatography according to the embodiment of the present invention;
FIG. 4 is a schematic diagram of a sample detection result according to an embodiment of the present invention;
FIG. 5 is a graph of the results of the surface enhanced Raman spectroscopy detection limits of plasma samples containing different concentrations of sufentanil according to an example of the present invention;
figure 6 is a sufentanil concentration-signal intensity standard curve for an example of the invention.
Detailed Description
The following examples are given to illustrate specific embodiments of the present invention. In the following embodiments, the Raman spectrometer used is a BWS415-785H type portable Raman spectrometer with an excitation light source of 785nm and a resolution of 3.5cm-1@912nm, spectral range 175--1Mixing ofA BAC151 video micro-Raman detection system (eyepiece × 20) was prepared, and the thin-layer plate was a silica gel plate having a coating thickness of 0.2mm to 0.25mm and a silica gel powder particle size of (8. + -.2). mu.m ≧ 80%, and the reagents used in the examples were obtained from general commercial sources unless otherwise specified, and the experimental conditions not specified were determined according to the conventional experimental conditions or the conditions recommended by the supplier.
< example >
1. Preparation and formulation of reagents
1.1 silver colloid preparation
The silver colloid used in the present example contains nano silver particles with an average particle size of 50nm, and is prepared by referring to the prior art (i.e., L ee method), and the preparation method specifically is as follows:
weighing 34mg of silver nitrate, dissolving in 200m L water to obtain a silver nitrate solution, heating, refluxing and condensing the silver nitrate solution to slightly boil, adding a trisodium citrate solution with the mass percent of 6m L to form a mixed solution, keeping heating the mixed solution until the color of the mixed solution is changed from colorless and transparent to light yellow and further to light gray and slightly green, continuing heating for 30min after color change, stopping heating, then cooling the mixed solution in water bath to obtain the silver colloid containing the nano silver particles, and storing the silver colloid in a 250m L brown bottle at 4 ℃ for later use.
Fig. 1 is a uv-vis spectrum of a silver paste according to an embodiment of the present invention, and fig. 2 is a scanning electron microscope image of a silver paste according to an embodiment of the present invention.
As shown in FIG. 1, the maximum absorption peak of the silver colloid is 414nm, and the half-peak width is narrow, which indicates that the silver colloid particles are uniformly dispersed. As shown in fig. 2, the silver nanoparticles are full and uniform in size, have no stacking or adhesion, and have an average particle size of about 50 nm.
1.2 preparation of stock solutions and Standard solutions
An appropriate amount of sufentanil reference substance is precisely weighed, methanol is used for preparing stock solution with the concentration of 3.4mg/m L, an appropriate amount of the stock solution is taken, and the stock solution is respectively diluted into a series of solutions with the concentrations of 1.7mg/m L, 850.0 mu g/m L, 425.0 mu g/m L, 212.5 mu g/m L, 85.0 mu g/m L, 42.5 mu g/m L and 8.5 mu g/m L by using the methanol as standard solutions, and the solutions are stored at 4 ℃ for later use.
2. Detection method
The detection method in the embodiment is performed by using T L C-SERS, and mainly includes the following steps.
Step S1, preprocessing the blood plasma to be detected to obtain a blood plasma sample to be detected; step S2, preparing sufentanil solution as standard solution, and adding sufentanil to the blank plasma sample to prepare a simulated plasma sample containing sufentanil.
This example uses a blank plasma (i.e., a sufentanil-free rat blank plasma) for pretreatment, and a sufentanil standard solution is added to the pretreated blank plasma sample to obtain a mock plasma sample. In addition, the plasma sample to be tested in the examples is also replaced by a simulated plasma sample.
The pretreatment method of blank plasma comprises the following steps: taking a proper amount of blood plasma, and mixing the blood plasma with the blood plasma according to a volume ratio of 1:2 adding acetonitrile, carrying out vortex oscillation for 1min, centrifuging at 13000rpm for 10min, removing protein, taking all supernatant, drying at 25 ℃ under a nitrogen blowing instrument, and adding methanol with the same volume as that of the blank plasma for redissolution to obtain a blank plasma sample.
Then, a proper amount of sufentanil standard solution is respectively added into the blank plasma samples to prepare a series of solutions with final sufentanil concentration of 340.00 mu g/m L, 170.00 mu g/m L, 85.00 mu g/m L, 42.50 mu g/m L, 21.25 mu g/m L, 8.50 mu g/m L, 4.25 mu g/m L and 0.85 mu g/m L as simulated plasma samples with different concentrations, and the simulated plasma samples are stored at 4 ℃ for later use.
And step S3, respectively sucking the simulated plasma sample and the standard solution, and spotting on the same silica gel plate.
And step S4, developing the sufentanil standard solution and the simulated plasma sample on the silica gel plate by using dichloromethane-methanol as a developing agent, taking out the silica gel plate after development, drying the silica gel plate, and placing the silica gel plate in an iodine jar for color development.
And step S5, determining the positions of the sufentanil spots in the simulated plasma samples according to the positions of the sufentanil spots in the standard solution, and then respectively dripping a preset amount of silver colloid at the sufentanil spots of the simulated plasma samples.
In this example, in order to examine the influence of the dropping amount of the silver paste, experiments were carried out with different dropping amounts, and the dropping amounts of the silver paste were 1 μ L, 5 μ L, and 10 μ L, respectively.
And step S5, performing surface enhanced Raman scattering detection (SERS) on the spot to which the silver colloid is dripped, and analyzing and processing the detected spectral signal to obtain the sufentanil characteristic peak and the strength of the sufentanil characteristic peak in the simulated plasma sample. In this embodiment, the SERS is performed by a portable raman spectrometer, and the detection conditions include a laser power of 100mW and a microscope system magnification of 20.
In addition, for the spectrum signal detected by the portable raman spectrometer, the embodiment adopts the OPUS 5.0 and Matlab 13.0 software to process the obtained spectrum, and selects the spectrum band of 300--1For spectral analysis (300 cm)-1Before the interference of nano-silver signal, 1700cm-1The latter with almost no spectral features), the spectra were smoothed (Sgolay method), baseline corrected (airP L S method) and normalized (Min-Max Normalization method) and plotted using Origin version 8.5 software to obtain raman spectra corresponding to each standard solution or simulated plasma sample, respectively.
Step S6, establishing 1004cm of sufentanil in a simulated plasma sample-1And (4) linear relation between the characteristic peak intensity and the sufentanil concentration, and calculating to obtain the sufentanil concentration in the blood plasma sample to be detected.
3. Conditional investigation
3.1 examination of conditions of thin layer chromatography
Different proportions of dichloromethane-methanol are adopted as the development system in the step S3, and compared with the separation effect of different proportions of dichloromethane-methanol, the optimal dichloromethane-methanol proportion is 9: 1.2.
FIG. 3 is a developed diagram of thin layer chromatography according to an embodiment of the present invention. Where, lane 1 is sufentanil standard solution and lane 2 is plasma sample.
As shown in FIG. 3, when the development is carried out at a ratio of dichloromethane to methanol of 9:1.2, a preferable separation effect can be obtained.
3.2 investigation of the amount of dropping of the silver colloid
This example investigates the effect of different amounts of glue on sufentanil raman signals. Namely, different dropping amounts are adopted when the silver colloid is dropped, and the influence of the different dropping amounts on the detection result is examined.
The method is characterized in that when the dropping amount of the silver colloid is 1 mu L, the characteristic peaks displayed by the SERS spectrum of sufentanil are fewer and the signal is lower, when the dropping amount of the silver colloid is 5 mu L, the characteristic peaks displayed by the SERS spectrum of sufentanil are complete and the signal is stronger, the reason of the phenomenon can be that the 'coffee ring effect' is generated when the dropping amount of the silver colloid is 1 mu L, namely the silver colloid is dropped on a thin-layer plate to form spots, the edge concentration of the spots is greater than the central concentration, and the nano silver particles are mainly concentrated on the edges of the spots, so that the enhanced substrate which can be detected in a Raman light path is too small, and the signal of an object to be detected cannot be completely reflected, when the dropping amount of the silver colloid is 5 mu L, the amount of the silver colloid which interacts with the object to be detected after the 'coffee ring' effect is proper, so that an ideal signal can be detected, and when the dropping amount of the silver colloid is 10 mu L, the peak time of the object to be detected is longer, the duration is shorter.
Therefore, when the particle size of the silver nanoparticles contained in the silver colloid is 50nm, the optimal colloid amount detected by sufentanil SERS is 5 μ L.
3.3 examination of integration time
The effect was examined for integration times of 5s, 10s and 20s, respectively. That is, different integration time conditions are adopted when SERS is performed by a portable raman spectrometer, and the influence of different integration times on the detection result is examined.
When the integration time is 5s, the peak intensity of the object to be detected is strong, the number of peaks is large, and 6-8 spectra can be collected; when the integration time is 10s, the peak intensity of the object to be measured is enhanced, the number of the output peaks is not obviously changed, but the thin-layer plate coating is easily scorched due to the lengthened laser irradiation time, the object to be measured is damaged by the laser irradiation, and 2-4 spectra can be collected; when the integration time is 20s, the thin-layer plate coating is quickly scorched and the spectrum cannot be collected.
Therefore, in the detection method of the present invention, the optimal integration time of SERS is 5 s.
From the above examination results, it is understood that in the detection method of the present invention, the optimal dichloromethane-methanol ratio in step S3 is 9:1.2, the optimal dropping amount of silver colloid in step S4 is 5 μ L, and the optimal integration time of SERS in step S5 is 5S.
4. Detecting the effect
4.1 qualitative detection
FIG. 4 is a schematic diagram of a sample detection result according to an embodiment of the present invention. In fig. 4, a is a simulated plasma sample (i.e., a plasma sample with sufentanil added), b is a conventional raman spectrum of sufentanil, c is blank plasma, and d is a silver gel blank control.
As can be seen from FIG. 4, sufentanil is at 600-1500cm-1Multiple characteristic peaks occur within the range, including 655, 1004, 1080, and 1439cm-1(see arrows in the figure). Wherein, 1004cm-1The characteristic peak of (A) is not present in blank plasma, and the silver colloid has obvious enhancement effect on the characteristic peak of (A), so 1004cm-1The characteristic peak can be regarded as sufentanil characteristic peak, namely, the method of the invention can carry out qualitative detection.
4.2 detection Limit
And (3) detecting simulated plasma samples with the sufentanil concentration range of 0.85-340.00 mu g/m L according to the detection method 2 to obtain Raman signals of samples with different concentrations by adopting the optimal conditions obtained by the condition investigation 3.
FIG. 5 is a graph of the detection limit results of Raman spectra of plasma samples of sufentanil at different concentrations according to an example of the present invention, wherein a is 340.00 μ g/m L, b is 170.00 μ g/m L, c is 85.00 μ g/m L, d is 42.50 μ g/m L, e is 21.25 μ g/m L, f is 8.50 μ g/m L, g is 4.25 μ g/m L, and h is 0.85 μ g/m L.
As shown in FIG. 5, when the concentration of sufentanil is in the range of 0.85-85.00. mu.g/m L, the intensity of the characteristic peak gradually increases with the increasing concentration, and the enhancement effect is rather reduced when the concentration is further increased.
The reason for this analysis is probably due to the fact that as the concentration of sufentanil increases, the number of drug molecules increases and the sensitivity of molecular detection correspondingly increases; in the detection process, the dropping amount of the silver colloid is not changed (namely the colloid surface for the drug molecules to adsorb is not changed), the drug molecules may be adsorbed with the silver colloid in a multilayer way or adsorbed with each other, so that the silver colloid can not be uniformly and effectively adsorbed with the surface of the silver nanoparticles, the Raman signal can not be correspondingly enhanced, and even the signal is submerged, so that the effect is not obvious.
In addition, as can be seen from FIG. 5, the minimum detection limit for sufentanil is 0.85 μ g/m L when the signal-to-noise ratio (S/N) is 3.
4.3 plasma sample content analysis
Figure 6 is a sufentanil concentration-signal intensity standard curve for an example of the invention.
As shown in FIG. 6, simulated plasma samples with sufentanil concentrations of 0.85, 4.25, 8.50, 21.25, 42.50, and 85.00 μ g/m L were taken, subjected to T L C-SERS analysis as previously described, and recorded at 1004cm-1And (4) performing linear fitting on the intensity of the characteristic peak and the concentration, and drawing a standard curve. The standard curve equation is: 70.538x +545.71 (r)20.9742), y is 1004cm-1The peak intensity of the sufentanil is shown, x is the concentration of the sufentanil (mu g/m L). The concentration of the sufentanil is equal to 1004cm within the range of 0.85-85.00 mu g/m L-1The characteristic peak intensity has good linearity, which indicates that the method of the invention can carry out the quantitative detection of sufentanil.
4.4 recovery
Three portions of simulated plasma samples with sufentanil concentrations of 4.25 mu g/m L (low concentration) and 42.50 mu g/m L (high concentration) are prepared as plasma samples to be tested in a manner of adding sufentanil to blank plasma, and then T L C-SERS analysis is carried out on the plasma samples according to a 2. detection method, and 1004cm of the measured plasma samples is measured-1The intensities of the characteristic peaks were substituted into the standard curve to predict the concentrations, and the results are shown in Table 1.
TABLE 1 recovery of sufentanil from plasma by T L C-SERS method
As can be seen from Table 1, the mean recovery of sufentanil from each of the mock plasma samples, which were the plasma samples tested, was 86.00% and 103.45%, respectively, with RSD% < 10%. The result shows that the detection method of the embodiment can be used for rapidly detecting sufentanil in the blood plasma sample to be detected, and the recovery rate is high.
Examples effects and effects
According to the method for detecting sufentanil in blood plasma provided by the embodiment, the silver colloid containing the nano silver particles with the average particle size of 50nm is used as the enhancing reagent, the silver colloid does not generate signals at the characteristic peak of sufentanil, and has a good signal enhancing effect on the characteristic peak of sufentanil, so that the T L C-SERS method can be applied to rapid detection of sufentanil in blood plasma.
In the examples, the preparation of silver colloid is carried out by referring to the L ee method, and the silver nitrate concentration of 34mg/200m L is adopted in the preparation process, and the reaction condition of heating and refluxing for 30min is continued after discoloration, so that nano silver particles with the average particle size of about 50nm can be obtained.
Because the integration time of 5S is adopted during SERS detection, the problem of thin-layer plate scorching caused by overlong integration time can be avoided, and sufficient spectral data can be obtained while the peak intensity of the object to be detected is ensured. In addition, 1004cm is selected-1As the characteristic peak of sufentanil, the interference of other substances in blood plasma can be avoided, and the signal enhancement effect of the silver colloid can be ensured.
The above examples are only intended to illustrate the method of the present invention for rapid detection of sufentanil in plasma, but the manner of detection of sufentanil in plasma according to the method of the present invention can also be in other forms, such as a form of detection device comprising a pretreatment unit, a detection kit, a raman spectroscopy detection unit, and a spectroscopic analysis unit. In this case, the detection kit may contain reagents required in a pre-prepared detection process, such as a standard solution, a simulated plasma sample, a silica gel plate, an expansion cylinder, an iodine cylinder, silver colloid, etc. in the examples; in addition, the test kit may also comprise a standard aspirator for aspirating a standard solution, a sample aspirator for aspirating a plasma sample and a silver colloid aspirator for aspirating silver colloid, which different aspirators each correspond to the respective aspirated amounts of reagents in the examples, respectively, so that a rapid measurement of sufentanil in a plasma sample can be accomplished without the aspirators on site.
Claims (6)
1. A method for rapidly detecting sufentanil in blood plasma is used for detecting the content of sufentanil in blood plasma to be detected, and is characterized by comprising the following steps:
step S1, preprocessing the blood plasma to be detected to obtain a blood plasma sample to be detected;
step S2, preparing a sufentanil solution as a standard solution, and adding sufentanil into a blank plasma sample to prepare a simulated plasma sample containing sufentanil;
step S3, respectively sucking the plasma sample to be detected, the simulated plasma sample and the standard solution and spotting on the same silica gel plate;
step S4, adopting dichloromethane-methanol with the ratio of 9:1.2 as a developing solvent to develop and dry the to-be-detected plasma sample, the simulated plasma sample and the standard solution on the silica gel plate and develop color in an iodine jar;
step S5, determining the positions of sufentanil spots in the plasma sample to be detected and the simulated plasma sample according to the positions of the sufentanil spots in the standard solution, and respectively dripping a predetermined amount of silver colloid on the sufentanil spots of the plasma sample to be detected and the simulated plasma sample;
step S6, performing surface enhanced Raman scattering detection on the spot point to which the silver colloid is dripped, and analyzing and processing a detected spectrum signal to obtain sufentanil characteristic peaks and intensities thereof in the plasma sample to be detected and the simulated plasma sample;
step S7, calculating the sufentanil concentration in the blood plasma sample to be detected according to the relation between the characteristic peak intensity of sufentanil in the simulated blood plasma sample and the sufentanil concentration,
wherein the absorption amount of the plasma sample to be tested, the simulated plasma sample and the standard solution in the step S3 is 1 mu L,
the predetermined amount of the silver paste in step S5 is 5 μ L, the silver paste including nano silver particles having an average particle diameter of 50nm,
the preprocessing method of step S2 is: taking the plasma to be detected or the simulated plasma as plasma to be processed, adding acetonitrile according to the volume ratio of 1:2, carrying out vortex oscillation for 1min, then centrifuging for 10min at 13000rpm, taking the centrifuged supernatant, drying the supernatant at 25 ℃ of a nitrogen blowing instrument, and then adding methanol with the same volume as the plasma to be processed for redissolution to obtain a corresponding plasma sample.
2. The method for rapid detection of sufentanil in plasma according to claim 1, wherein:
the silver colloid of the step S5 is prepared by the following method:
weighing 34mg of silver nitrate, and dissolving in 200m L water to obtain a silver nitrate solution;
heating, refluxing and condensing the silver nitrate solution to slightly boil, and then adding a trisodium citrate solution with the mass percent of 6m L being 1% to form a mixed solution;
keeping heating until the color of the mixed solution changes from colorless transparency to light yellow and further changes to light gray while showing slight green, continuing heating for 30min after changing color, and stopping heating;
and cooling the mixed solution in water bath to obtain the silver colloid.
3. The method for rapid detection of sufentanil in plasma according to claim 1, wherein:
wherein, the conditions of the surface enhanced raman scattering detection in the step S6 include a laser power of 100mW, an integration time of 5S, and a microscope system magnification of 20.
4. The method for rapid detection of sufentanil in plasma according to claim 1, wherein:
wherein the analyzing process performed on the spectral signal in step S6 is: selecting 300-1700cm of the spectral signal-1Carrying out smoothing, baseline correction and normalization processing, drawing the processed spectral signals to obtain an SERS spectrum corresponding to the spectral signals,
the sufentanil characteristic peak intensity is 1004cm in the SERS spectrum-1The intensity of the characteristic peak at (a).
5. The method for rapid detection of sufentanil in plasma according to claim 1, wherein:
wherein the simulated plasma is a plurality of plasmas respectively containing sufentanil at different concentrations, and the content of the sufentanil contained in the simulated plasma is in a range of 0.85-85.00 mu g/m L.
6. The method for rapid detection of sufentanil in plasma according to claim 5, wherein:
wherein, the calculating method in step S7 is: establishing 1004cm in the sufentanil SERS spectrum in the simulated plasma sample-1The intensity of the characteristic peak is in linear relation with the concentration of sufentanil, and the concentration of sufentanil in the blood plasma sample to be tested is calculated according to the linear relation.
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| CN112240881A (en) * | 2020-09-30 | 2021-01-19 | 合肥国研汉因检测科技有限公司 | Method for rapidly detecting trace fentanyl by using Raman spectroscopy |
| CN112255343B (en) * | 2020-10-16 | 2021-08-06 | 宁波市疾病预防控制中心 | Method and kit for measuring fentanyl drugs in biological sample |
| CN112782305A (en) * | 2020-12-29 | 2021-05-11 | 苏州海科医药技术有限公司 | Method for analyzing sufentanil concentration in blood plasma sensitively and rapidly and suitable for pharmacokinetic research |
| CN115165777A (en) * | 2022-07-01 | 2022-10-11 | 首都医科大学宣武医院 | Fentanyl class management and control narcotic waste liquid recovery verification system |
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