CN108732292B - Rapid detection method and device for sufentanil in plasma - Google Patents

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

Rapid detection method and device for sufentanil in plasma

technical field

The invention relates to a detection method of sufentanil, in particular to a rapid detection method and device of sufentanil in plasma.

Background technique

Sufentanil is a fentanyl derivative with the chemical name N-[4-(methoxymethyl)-1-2[2-(2-thienyl)ethyl]-4-piperidine [methyl]-N-phenylpropanamide, its fat solubility is 1100 times that of morphine, it can easily penetrate the blood-brain barrier, and can quickly reach an effective concentration in the brain, and its analgesic effect is about 5-10 times that of fentanyl. It is 300-400 times stronger than morphine and is the opioid analgesic with the strongest analgesic effect. It is widely used in general anesthesia induction, intraoperative maintenance and postoperative analgesia.

Sufentanil is suitable for anesthesia in various surgical procedures, but due to individual differences in patients, the dosage of sufentanil should be adjusted in time according to the patient's response to avoid toxic side effects. For example, large doses of sufentanil may cause growth. Time-dependent respiratory depression (Pharmacological Effects and Clinical Application of Sufentanil, Pei Hao, Luo Ailin, Medical Herald, November 2009, p. 1482), so it is necessary to detect the concentration of sufentanil in biological samples such as plasma. In addition, medical workers who work in the ward or operating room for a long time may be exposed to fentanyls through the patient's exhaled gas, which can lead to symptoms of opioid sensitivity or addiction (Mcauliffe P F, Gold M S, Bajpai L, et al. al. Second-hand exposure to aerosolized intravenous anesthetics propofol and fentanyl may causesensitization and subsequent opiate addiction among anesthesiologists and surgeons [J]. Med Hypotheses. 2006, 66(5): 874-882.). Therefore, the concentration detection of sufentanil in biological samples such as plasma, especially the development of a rapid and sensitive on-site detection method for fentanyl compounds in vivo, not only has an important guiding role for the clinical use of this drug, but also has an important role in guiding the clinical use of the drug. Under special circumstances, timely screening and follow-up medical treatment of such compounds are of great significance.

At present, the detection methods of sufentanil mainly include high performance liquid chromatography, liquid chromatography-mass spectrometry (LC-MS), gas chromatography-mass spectrometry (GC-MS) and so on. The above method has high sensitivity, but the instrument is complex, the use cost is high, and the detection speed is slow, so it is not suitable for rapid detection on site.

Thin-layer chromatography (Thin-Layer Chromatography, TLC) is a classic separation and analysis method, which has been widely used in the qualitative and quantitative detection of drugs. Surface-enhanced Raman Scattering (SERS) has been applied in many research fields due to its advantages of high sensitivity, strong characteristic and short detection time. TLC-SERS combined technology has the advantages of short analysis period and strong characteristics. It has been used for on-site detection of environmental pollution, detection of adulteration of traditional Chinese medicine, detection of added chemical drugs in traditional Chinese medicine, detection of illegal drugs, detection of trace drugs and biochemical War agent detection, clinical drug monitoring and other fields. The spectral signal detection in TLC-SERS technology can be carried out with a hand-held Raman spectrometer, making the detection device more portable as a whole, so TLC-SERS is very suitable for on-site detection. However, there is no method for on-site detection of sufentanil in plasma by TLC-SERS technology.

SUMMARY OF THE INVENTION

In order to solve the above problems, a method and a device capable of rapidly and accurately detecting the content of sufentanil in plasma are provided. The inventor of the present invention proposes the following technical solutions on the basis of exploring relevant detection methods and conditions:

The present invention provides a rapid detection method for sufentanil in plasma, which is used for detecting the content of sufentanil in plasma to be tested. Plasma sample to be tested; step S2, prepare a sufentanil solution as a standard solution, and add sufentanil to a blank plasma sample to prepare a simulated plasma sample containing sufentanil; step S3, draw the plasma sample to be tested, The simulated plasma sample and the standard solution are placed on the same silica gel plate; in step S4, dichloromethane-methanol is used as a developing agent to develop, air dry and iodine the plasma sample to be tested, the simulated plasma sample and the standard solution on the silica gel plate. Color development; step S5, with reference to the position of the sufentanil spot in the standard solution to determine the spot position of sufentanil in the plasma sample to be tested and the simulated plasma sample, and in the sufentanil spot of the plasma sample to be tested and the simulated plasma sample A predetermined amount of silver glue is added dropwise to the spots; step S6, surface-enhanced Raman scattering detection is performed on the spots where the silver glue has been dropped, and the spectral signals obtained by the detection are analyzed and processed to obtain the plasma samples to be tested and the simulated plasma samples. The characteristic peak of sufentanil and its intensity; step S7, according to the relationship between the characteristic peak intensity of sufentanil in the simulated plasma sample and the concentration of sufentanil, the concentration of sufentanil in the plasma sample to be tested is calculated, wherein In step S3, the absorption amount of the plasma sample to be tested, the simulated plasma sample and the sufentanil standard solution is 1 μL, and the predetermined amount of the silver glue in step S5 is 5 μL, and the silver glue contains nano-silver particles with an average particle size of 50 nm. .

The method for rapid detection of sufentanil in plasma provided by the present invention may also have such technical features, wherein, the silver glue in step S5 is prepared by the following method: 34 mg of silver nitrate is weighed and dissolved in 200 mL of water to obtain a silver nitrate solution; The silver nitrate solution was heated and refluxed and condensed to slightly boiling, and then 6 mL of trisodium citrate solution with a mass percentage of 1% was added to form a mixed solution; the heating was maintained until the color of the mixed solution changed from colorless and transparent to light yellow and further changed to light gray at the same time. Slightly green, continue heating for 30 min after discoloration, and stop heating; the mixed solution is cooled in a water bath to obtain silver glue.

The rapid detection method for sufentanil in plasma provided by the present invention may also have such technical features, wherein the conditions for the surface-enhanced Raman scattering detection in step S6 are laser power 100mW, integration time 5S, and microscope system magnification 20.

The method for rapid detection of sufentanil in plasma provided by the present invention may also have such technical features, wherein the analysis and processing of the spectral signal in step S6 is: selecting 300-1700 cm ^-1 of the spectral signal for smoothing, baseline After correction and normalization, the processed spectral signal was plotted to obtain the SERS spectrum corresponding to the spectral signal. The intensity of the characteristic peak of sufentanil was the intensity of the characteristic peak at 1004 cm ^-1 in the SERS spectrum.

The rapid detection method of sufentanil in the plasma provided by the invention can also have such technical features, wherein, the pretreatment method of step S2 is: take the plasma to be tested or the simulated plasma as the plasma to be processed, according to the volume ratio 1: 2 Add acetonitrile, vortex for 1 min, centrifuge at 13,000 rpm for 10 min, take the centrifuged supernatant and blow dry at 25°C in a nitrogen blower, then add the same volume of methanol as the plasma to be treated for reconstitution to obtain the corresponding plasma samples.

The method for rapid detection of sufentanil in plasma provided by the present invention may also have such technical features, wherein the simulated plasma is a plurality of sufentanil containing different concentrations of sufentanil respectively, and the simulated plasma contains sufentanil of different concentrations. The content range is 0.85-85.00μg/mL.

The method for rapid detection of sufentanil in plasma provided by the present invention may also have such technical features, wherein, the calculation method in step S7 is: establishing the SERS spectrum of sufentanil in the simulated plasma sample at 1004 cm ^-1 The linear relationship between the intensity of the characteristic peak and the concentration of sufentanil was calculated, and the concentration of sufentanil in the plasma sample to be tested was calculated accordingly.

The invention also provides a rapid detection device for sufentanil in plasma, which is used to detect the content of sufentanil in the plasma to be tested, and is characterized in that it includes: a pretreatment unit, which is used for pre-processing the plasma to be tested. Processed to obtain a plasma sample to be tested; a detection kit, containing a sufentanil solution used as a standard solution, a simulated plasma sample used as a content calculation control and containing sufentanil, a standard solution, simulated plasma Silica gel plate for spotting samples and plasma samples to be tested, a developing cylinder for developing the spotted silica gel plate, an iodine cylinder for color development, and a spectrum for developing sufentanil spots Enhanced silver glue; Raman spectrum detection unit, used for surface-enhanced Raman scattering detection on the sufentanil spots dripped with silver glue, and analyzed and processed the detected spectral signals to obtain simulated plasma samples and samples to be tested. sufentanil characteristic peaks and their intensities in the plasma samples; and a spectral analysis unit for calculating the sufentanil characteristic peaks in the plasma samples to be tested according to the intensities of the sufentanil characteristic peaks in the simulated plasma samples and the plasma samples to be tested Ni concentration, in which, the silver glue contains nano-silver particles with an average particle size of 50nm, and the detection kit also includes a standard aspirator for drawing the standard solution during spotting, and a sampler for drawing the plasma sample to be tested during spotting. The sample aspirator and the silver glue aspirator used to absorb the silver glue when dripping the silver glue, the suction volume of the standard aspirator and the sample aspirator is 1 μL, and the suction volume of the silver glue suction device is 5 μL.

Invention action and effect

According to the method for detecting sufentanil in plasma provided by the present invention, since silver glue containing nano-silver particles with an average particle size of 50 nm is used as an enhancing reagent, the silver glue has a good signal for the characteristic peaks of sufentanil Enhanced effect, and no signal is generated at the characteristic peak of sufentanil, so TLC-SERS method can be applied to the rapid detection of sufentanil in plasma. In the present invention, since the spotting amount used is 1 μL, the silver glue containing nano-silver particles with an average particle size of 50 nm is used, and the drop amount of the silver glue is 5 μL. Appropriate amount of nano-silver particles can play a more ideal signal enhancement effect.

Description of drawings

Fig. 1 is the ultraviolet-visible spectrogram of the silver glue of the embodiment of the present invention;

Fig. 2 is the scanning electron microscope picture of the silver glue of the embodiment of the present invention;

Fig. 3 is the TLC development result diagram of the embodiment of the present invention;

4 is a schematic diagram of a sample detection result according to an embodiment of the present invention;

5 is a graph showing the detection limit results of surface-enhanced Raman spectroscopy of plasma samples containing different concentrations of sufentanil according to an embodiment of the present invention;

FIG. 6 is a standard curve of sufentanil concentration-signal intensity according to an embodiment of the present invention.

Detailed ways

The specific embodiments of the present invention will be described below with reference to the examples. In the following examples, the Raman spectrometer used is a BWS415-785H portable Raman spectrometer with an excitation light source of 785nm, a resolution of ^{3.5cm-1@912nm} , a spectral range of 175-2700cm ^-1 , and a BAC151 video microscope. Mann detection system (eyepiece×20); the thin-layer plate is a silica gel plate with a coating thickness of 0.2mm-0.25mm and a silica gel powder particle size of (8±2) μm≧80%. In addition, the reagents used in the examples were obtained from general commercial sources unless otherwise specified, and the unspecified experimental conditions were in accordance with conventional experimental conditions or conditions suggested by suppliers.

<Example>

1. Reagent preparation and preparation

1.1 Preparation of silver glue

The silver glue adopted in the present embodiment contains nano-silver particles with an average particle size of 50nm, and the silver glue is obtained with reference to the prior art (that is, the Lee method), and the preparation method is as follows:

34 mg of silver nitrate was weighed and dissolved in 200 mL of water to obtain a silver nitrate solution. The silver nitrate solution was heated under reflux and condensed to a slight boil, and then 6 mL of 1% by mass trisodium citrate solution was added to form a mixed solution. Keep heating the mixed solution until the color of the mixed solution changes from colorless and transparent to light yellow, and further changes to light gray and slightly green. The silver glue of silver particles was stored in a 250mL brown bottle at 4°C until use.

FIG. 1 is an ultraviolet-visible spectrum diagram of the silver paste according to the embodiment of the present invention, and FIG. 2 is a scanning electron microscope diagram of the silver paste according to the embodiment of the present invention.

As shown in Figure 1, the maximum absorption peak of the silver glue is at 414 nm, and the half-peak width is narrow, indicating that the silver glue particles are uniformly dispersed. As shown in Figure 2, the nano-silver particles are full in shape, uniform in size, without stacking or adhesion, and the average particle size is about 50 nm.

1.2 Preparation of stock solutions and standard solutions

An appropriate amount of sufentanil reference substance was precisely weighed and prepared into a stock solution with a concentration of 3.4 mg/mL in methanol. Take an appropriate amount of the above stock solution and dilute it with methanol into a series of concentrations of 1.7 mg/mL, 850.0 μg/mL, 425.0 μg/mL, 212.5 μg/mL, 85.0 μg/mL, 42.5 μg/mL, 8.5 μg/mL. The solution was used as a standard solution and stored at 4°C for later use.

2. Detection method

The detection method in this embodiment adopts TLC-SERS, which mainly includes the following steps.

In step S1, the plasma to be tested is pretreated to obtain a plasma sample to be tested; in step S2, a sufentanil solution is prepared as a standard solution, and sufentanil is added to a blank plasma sample to prepare a simulated plasma sample containing sufentanil.

In this example, blank plasma (ie, blank rat plasma without sufentanil) is used for pretreatment, and a standard solution of sufentanil is added to the blank plasma sample after pretreatment to obtain a simulated plasma sample. In addition, the plasma samples to be tested in the examples were also replaced by simulated plasma samples.

Among them, the pretreatment method of blank plasma is as follows: take an appropriate amount of plasma, add acetonitrile in a volume ratio of 1:2, vortex for 1 min, centrifuge at 13,000 rpm for 10 min, remove the protein and take all the supernatant and dry it at 25°C under a nitrogen blower , and then add the same volume of methanol as the blank plasma to reconstitute to obtain a blank plasma sample.

After that, an appropriate amount of sufentanil standard solution was added to the blank plasma samples respectively, and the final concentrations of sufentanil were 340.00 μg/mL, 170.00 μg/mL, 85.00 μg/mL, 42.50 μg/mL, 21.25 μg/mL, A series of solutions of 8.50 μg/mL, 4.25 μg/mL, and 0.85 μg/mL were used as simulated plasma samples of different concentrations, which were stored at 4°C for future use.

Step S3, draw the simulated plasma sample and the standard solution respectively, and place them on the same silica gel plate.

In step S4, dichloromethane-methanol is used as a developing agent to develop the sufentanil standard solution and the simulated plasma sample on the silica gel plate.

Step S5 , determine the spot positions of sufentanil in the simulated plasma sample with reference to the positions of the sufentanil spots in the standard solution, and then drop a predetermined amount of silver glue on the sufentanil spots of the simulated plasma sample respectively.

In this embodiment, experiments with different drop amounts were carried out to investigate the influence of the drop amount of silver glue, and the drop amounts of silver glue were respectively 1 μL, 5 μL, and 10 μL.

In step S5, surface-enhanced Raman scattering detection (hereinafter referred to as SERS) is performed on the spots on which the silver glue has been added, and the detected spectral signals are analyzed and processed to obtain the characteristic peaks of sufentanil in the simulated plasma sample and their intensities. . In this embodiment, the above-mentioned SERS is carried out by a portable Raman spectrometer, and the detection conditions are a laser power of 100 mW and a magnification of the microscope system of 20.

In addition, for the spectral signal detected by the portable Raman spectrometer, in this example, OPUS 5.0 and Matlab 13.0 software were used to process the obtained spectrum, and the spectral band 300-1700 cm ^-1 was selected for spectral analysis (there was nano silver before 300 cm ^-1 ) Signal interference, almost no spectral features after 1700cm ^-1 ), the spectra were smoothed (Sgolay method), baseline corrected (airPLS method) and normalized (Min-Max Normalization method), and were plotted with Origin version 8.5 software, so that Raman spectra corresponding to each standard solution or simulated plasma sample were obtained.

Step S6, establish a linear relationship between the 1004 cm ^-1 characteristic peak intensity of sufentanil in the simulated plasma sample and the concentration of sufentanil, and calculate the concentration of sufentanil in the plasma sample to be tested accordingly.

3. Condition inspection

3.1 Investigation of thin-layer chromatography conditions

Different ratios of dichloromethane-methanol are used as the development system in step S3, and the separation effects of different ratios of dichloromethane-methanol are compared, and the optimal ratio of dichloromethane-methanol is 9:1.2.

FIG. 3 is a graph showing the results of thin-layer chromatography development in an embodiment of the present invention. Among them, the band 1 is the sufentanil standard solution, and the band 2 is the plasma sample.

As shown in Figure 3, the development is carried out under the condition that the ratio of dichloromethane-methanol is 9:1.2, and a relatively ideal separation effect can be obtained.

3.2 Investigation of the dripping amount of silver glue

In this example, the effects of different dispensing amounts on the Raman signal of sufentanil were investigated. That is, different dripping amounts were used when the silver glue was dripped, and the influence of different dripping amounts on the detection results was investigated.

When the drop amount of silver glue was 1 μL, the SERS pattern of sufentanil showed fewer characteristic peaks and lower signal; when the drop amount of silver glue was 5 μL, the SERS pattern of sufentanil showed The characteristic peaks are more complete and the signal is stronger. The reason for the above phenomenon may be that the "coffee ring effect" occurs when the dispensing volume is 1 μL, that is, the silver glue is dripped on the thin layer board to form spots, and the edge concentration of the spots is greater than the center concentration, and the nano-silver particles are mainly concentrated in the spots. At the edge of the Raman light path, there are too few enhanced substrates that can be detected in the Raman optical path, so that the signal of the object to be tested cannot be fully reflected; and when the drop amount of silver glue is 5 μL, after the "coffee ring" effect, it is closely related to the object to be tested. The amount of silver glue that interacts is appropriate, and an ideal signal can be detected. When the dispensing volume is 10 μL, the peak time of the test object is longer and the duration is short, which is not conducive to the detection of the test object.

Therefore, when the particle size of the silver nanoparticles contained in the silver gel is 50 nm, the optimal amount of gel for SERS detection of sufentanil is 5 μL.

3.3 Investigation of integration time

The effects of integration time of 5s, 10s and 20s were investigated respectively. That is, the conditions of different integration times were used when carrying out SERS with a portable Raman spectrometer, and the influence of different integration times on the detection results was investigated.

When the integration time is 5s, the peak intensity of the analyte is stronger, and there are many peaks, and 6-8 spectra can be collected; when the integration time is 10s, the peak intensity of the analyte is enhanced, and the number of outgoing peaks is not obvious However, due to the prolonged laser irradiation time, the coating of the thin-layer board is easily scorched, and the object to be tested is damaged by the laser irradiation, and 2-4 spectra can be collected; when the integration time is 20s, the coating of the thin-layer board is very Almost burnt, unable to collect spectrum.

Therefore, in the detection method of the present invention, the optimal integration time of SERS is 5s.

According to the above investigation results, in the detection method of the present invention, the optimal ratio of dichloromethane-methanol in step S3 is 9:1.2, the optimal dripping amount of silver glue in step S4 is 5 μL, and the optimal amount of SERS in step S5 is 5 μL. The optimal integration time is 5s.

4. Detection 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 Figure 4, a is a simulated plasma sample (ie, a plasma sample added with sufentanil), b is a conventional Raman spectrum of sufentanil, c is blank plasma, and d is a silver gel blank control.

It can be seen from Fig. 4 that sufentanil has multiple characteristic peaks in the range of 600-1500 cm ^-1 , including 655, 1004, 1080 and 1439 cm ^-1 (see arrows in the figure). Among them, the characteristic peak at 1004cm ^-1 does not exist in blank plasma, and the silver glue has a significant enhancement effect on the characteristic peak, so the characteristic peak at 1004cm ^-1 can be used as the characteristic peak of sufentanil, that is, the present The inventive method enables qualitative detection.

4.2 Detection limit

Using the optimal conditions obtained in the above "3. Condition investigation", the simulated plasma samples with the concentration range of sufentanil ranging from 0.85 to 340.00 μg/mL were tested according to the aforementioned "2. Detection method". Raman signal.

FIG. 5 is a graph showing the detection limit of Raman spectra of plasma samples of different concentrations of sufentanil according to an embodiment of the present invention. where a is 340.00 μg/mL, b is 170.00 μg/mL, c is 85.00 μg/mL, d is 42.50 μg/mL, e is 21.25 μg/mL, f is 8.50 μg/mL, and g is 4.25 μg/mL , h is 0.85μg/mL.

As shown in Figure 5, when the concentration of sufentanil was in the range of 0.85-85.00 μg/mL, the intensity of the characteristic peaks gradually increased as the concentration increased; however, if the concentration further increased, the enhancement effect was weakened.

Analysis of the reasons may be due to the increase in the number of drug molecules with the increase of sufentanil concentration, and the corresponding increase in the sensitivity of molecular detection; The surface remains unchanged), drug molecules may have multi-layer adsorption with them or mutual adsorption between drug molecules, so that uniform and effective adsorption with the surface of silver nanoparticles cannot be carried out, and the Raman signal cannot be correspondingly enhanced, or even drown the signal, resulting in the effect of Not obvious.

In addition, it can be concluded from Figure 5 that when the signal-to-noise ratio (S/N) is 3, the minimum detection limit of sufentanil is 0.85 μg/mL.

4.3 Analysis of plasma sample content

FIG. 6 is a standard curve of sufentanil concentration-signal intensity according to an embodiment of the present invention.

As shown in Figure 6, mock plasma samples with sufentanil concentrations of 0.85, 4.25, 8.50, 21.25, 42.50, and 85.00 μg/mL were selected, subjected to TLC-SERS analysis as previously described, and the features at 1004 cm ^-1 were recorded Peak intensities were linearly fitted to concentrations and a standard curve was drawn. The equation of the standard curve is: y=70.538x+545.71 (r ² =0.9742), y is the characteristic peak intensity of sufentanil at 1004 cm ⁻¹ , and x is the concentration of sufentanil (μg/mL). It can be seen that in the range of 0.85-85.00 μg/mL, the sufentanil concentration and the characteristic peak intensity at 1004 cm ⁻¹ have good linearity, indicating that the method of the present invention can perform quantitative detection of sufentanil.

4.4 Recovery rate

By adding sufentanil to blank plasma, three simulated plasma samples with sufentanil concentrations of 4.25 μg/mL (low concentration) and 42.50 μg/mL (high concentration) were prepared as the plasma samples to be tested. , and then perform TLC-SERS analysis according to "2. Detection method", and substitute the measured intensity of the characteristic peak at 1004cm ^-1 into the above standard curve to predict its concentration. The results are shown in Table 1.

Table 1 The recovery of sufentanil in plasma by TLC-SERS method

It can be seen from Table 1 that the average recoveries of sufentanil in the simulated plasma samples used as the plasma samples to be tested were 86.00% and 103.45%, respectively, and the RSD% was less than 10%. The results show that the detection method of this embodiment can rapidly detect sufentanil in the plasma sample to be tested, and the recovery rate is high.

Example function and effect

According to the method for detecting sufentanil in plasma provided in this embodiment, since silver glue containing nano-silver particles with an average particle size of 50 nm is used as the enhancing reagent, the silver glue does not generate at the characteristic peak of sufentanil. It has a good signal enhancement effect on the characteristic peaks of sufentanil, so the TLC-SERS method can be applied to the rapid detection of sufentanil in plasma. In this example, since the spotting volume used is 1 μL, the silver glue containing nano-silver particles with an average particle size of 50 nm is used, and the drop amount of the silver glue is 5 μL, so the interaction with the test object can be made. The appropriate amount of nano-silver particles can play a more ideal signal enhancement effect.

In the embodiment, since the preparation of silver glue was carried out with reference to the Lee method, and the silver nitrate concentration of 34mg/200mL was used in the preparation process, and the reaction conditions of continuing to heat and reflux for 30min after discoloration, nano-silver with an average particle size of about 50nm can be obtained. particle.

Since the integration time of 5S is used in the SERS detection, it can avoid the burning problem of the thin-layer plate caused by the long integration time, and obtain sufficient spectral data while ensuring the peak intensity of the analyte. In addition, since 1004cm ^-1 was selected as the characteristic peak of sufentanil, the interference of other substances in the plasma can be avoided, and the signal enhancement effect of the silver glue can be ensured.

The above embodiment is only used to illustrate the rapid detection method of sufentanil in plasma of the present invention, and according to the detection method of the present invention, the detection method of sufentanil in plasma can also be in other forms, such as containing a pretreatment unit. , a detection kit, a Raman spectrum detection unit, and a detection device for a spectrum analysis unit. In this case, the detection kit may contain pre-prepared reagents required in the detection process, such as standard solutions, simulated plasma samples, silica gel plates, development cylinders, iodine cylinders, silver glue, etc. in the embodiment; in addition, detection reagents The box can also include a standard aspirator for drawing standard solutions, a sample aspirator for drawing plasma samples, and a silver glue aspirator for drawing silver glue, and these different aspirators respectively correspond to the corresponding reagents in the examples. so that the rapid measurement of sufentanil in plasma samples can be accomplished without an aspirator in the field.

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.

Images