CN108732292A - The rapid detection method and device of sufentanil in blood plasma - Google Patents
The rapid detection method and device of sufentanil in blood plasma Download PDFInfo
- Publication number
- CN108732292A CN108732292A CN201810390239.2A CN201810390239A CN108732292A CN 108732292 A CN108732292 A CN 108732292A CN 201810390239 A CN201810390239 A CN 201810390239A CN 108732292 A CN108732292 A CN 108732292A
- Authority
- CN
- China
- Prior art keywords
- sufentanil
- plasma sample
- blood plasma
- elargol
- sample
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/90—Plate chromatography, e.g. thin layer or paper chromatography
-
- 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
Landscapes
- Health & Medical Sciences (AREA)
- Physics & Mathematics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Abstract
The present invention provides a kind of rapid detection methods of sufentanil in blood plasma, include the following steps：Step S1 pre-processes test plasma to obtain test plasma sample；Step S2 prepares standard solution and simulating blood plasma sample；Step S3 draws test plasma sample, simulating blood plasma sample and standard solution and puts on same silica gel plate respectively；Step S4, test plasma sample, simulating blood plasma sample and the standard solution on silica gel plate are unfolded, dry and iodine cylinder colour developing；The elargol of predetermined amount is added dropwise in step S5 respectively at the sufentanil spot of test plasma sample and simulating blood plasma sample；Step S6 carries out Surface enhanced Raman scattering detection to being added dropwise at the spot of elargol；Step S7, calculate the sufentanil concentration in test plasma sample, wherein the uptake of test plasma sample, simulating blood plasma sample and standard solution is 1 μ L in step S3, elargol predetermined amount is 5 μ L in step S5, includes the nano silver particles that average grain diameter is 50nm in the elargol.
Description
Technical field
The present invention relates to the detection methods of sufentanil, and in particular to the rapid detection method of sufentanil in a kind of blood plasma
And device.
Background technology
Sufentanil (sufentanil) is fentanyl analog derivative, entitled N- [4- (methoxyl methyl) -1-2 [2- of chemistry
(2- thienyls) ethyl] -4- piperidyls]-N- hydrocinnamamides, fat-soluble is 1100 times of morphine, easily penetrates blood-brain barrier,
And effective concentration can be reached in intracerebral rapidly, analgesic activity is about 5-10 times of fentanyl, 300-400 times of morphine, is current
The strongest opium kind analgesics of analgesic activity are clinically widely used in maintenance and postoperative town in general anesthesia induction, art
Pain etc..
Sufentanil is suitble to the anesthesia of each section's operation, but since patient is there are individual difference, the dosage of sufentanil answers root
It is adjusted in time according to patient's reaction, to avoid toxicity, for example can occur for a long time when application large dosage sufentanil
Respiration inhibition (sufentanil pharmacological action and clinical application, Pei Hao, Luo Ailin, medical Leader, in November, 2009,1482
Page), it is therefore desirable to Concentration Testing is carried out to the sufentanil in the biological samples such as blood plasma.In addition, medical personnel is for a long time in disease
It works in room or operating room, fentanyl class drug may be touched by the gas that patient breathes out, and thus cause opium sensitive
Or symptom (Mcauliffe P F, Gold M S, Bajpai L, the et al.Second-hand exposure to of habituation
aerosolized intravenous anesthetics propofol and fentanyl may cause
sensitization and subsequent opiate addiction among anesthesiologists and
surgeons[J].Med Hypotheses.2006,66(5):874-882.).Therefore, to the Shu Fen in the biological samples such as blood plasma
Too Buddhist nun carries out the in-situ check and test method of quick, the sensitive fentanyl class compound in vivo of Concentration Testing, especially exploitation, not only right
There is important directive function in the clinical application of the drug, and for the timely screening of such compound in special circumstances and
Subsequent medical treatment is significant.
Currently, the detection method of sufentanil mainly has high performance liquid chromatography, Liquid Chromatography-Mass Spectrometry (LC-
MS), gas chromatography-mass spectrometry (GC-MS) etc..Above method sensitivity is higher, but instrument complexity, higher operating costs,
Detection speed is slower, thus is not suitable for the quick detection at scene.
Thin-layered chromatography (Thin-Layer Chromatography, TLC) is a kind of method for separating and analyzing of classics,
It is widely used in the qualitative and quantitative analysis of drug.Surface enhanced Raman scattering (Surface enhanced Raman
Scattering, SERS) it is answered in many research fields due to its high sensitivity, the advantage that characteristic is strong, detection time is short
With.TLC-SERS joint technology has analytical cycle is short, characteristic is strong advantage, have been used for environmental pollution Site Detection,
Chinese medicine is mixed pseudo- detection, the detection of Chinese medicine addition chemical drug, illegal drug detection, trace illicit drugs inspection and biochemical war agent detection, is faced
The multiple fields such as bed drug surveillance.In TLC-SERS technologies spectral signal detection may be used handheld Raman spectrometer into
Row so that detection device is whole more portable, thus TLC-SERS is highly suitable for Site Detection.But it has not yet to see
The method that Site Detection is carried out to sufentanil in blood plasma with TLC-SERS technologies.
Invention content
To solve the above problems, provide it is a kind of can quickly and accurately detect in blood plasma the method for sufentanil content and
Device, the present inventor is on the basis of exploring related detecting method and condition, it is proposed that following technical solution：
The present invention provides a kind of rapid detection methods of sufentanil in blood plasma, for the Shu Fentai in test plasma
Buddhist nun's content is detected, which is characterized in that is included the following steps：Step S1 pre-processes test plasma to obtain blood to be measured
Slurry samples；Step S2 prepares sufentanil solution as standard solution, and sufentanil addition blank plasma samples are prepared into
To the simulating blood plasma sample containing sufentanil；It is molten to draw test plasma sample, simulating blood plasma sample and standard respectively by step S3
Liquid is simultaneously put on same silica gel plate；Step S4, using methylene chloride-methanol as solvent to the test plasma sample on silica gel plate
Product, simulating blood plasma sample and standard solution be unfolded, is dried and the colour developing of iodine cylinder；Step S5, Shu Fentai in reference standard solution
The speckle displacement of sufentanil in the location determination test plasma sample and simulating blood plasma sample of Buddhist nun's spot, in test plasma sample
With the elargol that predetermined amount is added dropwise at the sufentanil spot of simulating blood plasma sample respectively；Step S6, to the spot of elargol has been added dropwise
Place carries out Surface enhanced Raman scattering detection and carries out analyzing processing to the spectral signal that detection obtains, and obtains test plasma sample
With the sufentanil characteristic peak and its intensity in simulating blood plasma sample；Step S7, according to sufentanil in simulating blood plasma sample
The sufentanil concentration in test plasma sample is calculated in feature peak intensity and the relationship of sufentanil concentration, wherein step
The uptake of test plasma sample, simulating blood plasma sample and sufentanil standard solution in S3 is 1 μ L, elargol in step S5
Predetermined amount be 5 μ L, the nano silver particles that average grain diameter is 50nm are included in the elargol.
The rapid detection method of sufentanil in blood plasma provided by the invention can also have such technical characteristic,
In, the elargol of step S5 is prepared with the following method：It weighs silver nitrate 34mg and is dissolved in 200mL water that obtain silver nitrate molten
Liquid；Silver nitrate solution is heated to reflux condensation to slightly boiling, the citric acid three sodium solution that 6mL mass percents are 1% is then added
Form mixed solution；Heating is kept until the color of mixed solution is become pale yellow from water white transparency and is further changed to light grey same
When show slightly green, continue after discoloration heat 30min, stop heating；Water-bath cooling is carried out to mixed solution, obtains elargol.
The rapid detection method of sufentanil in blood plasma provided by the invention can also have such technical characteristic,
In, the condition of Surface enhanced Raman scattering detection is laser power 100mW in step S6, time of integration 5S, microscopic system are amplified
Multiple 20.
The rapid detection method of sufentanil in blood plasma provided by the invention can also have such technical characteristic,
In, the analyzing processing carried out to spectral signal in step S6 is：The 300-1700cm of chosen spectrum signal-1Carry out smooth, baseline
Correction and normalized, to treated, spectral signal is mapped to obtain SERS collection of illustrative plates corresponding with spectral signal, is relaxed
Fentanyl feature peak intensity is 1004cm in SERS collection of illustrative plates-1The intensity of the characteristic peak at place.
The rapid detection method of sufentanil in blood plasma provided by the invention can also have such technical characteristic,
In, the preprocess method of step S2 is：Take test plasma or simulating blood plasma as pending blood plasma, according to volume ratio 1:2 are added
Acetonitrile centrifuges 10min after vortex oscillation 1min under the conditions of 13000rpm, takes the supernatant after centrifugation in 25 DEG C of conditions of nitrogen evaporator
Then lower drying is added and is redissolved with the methanol of pending blood plasma same volume, obtains corresponding plasma sample.
The rapid detection method of sufentanil in blood plasma provided by the invention can also have such technical characteristic,
In, simulating blood plasma is the multiple and sufentanil containing various concentration respectively, the sufentanil content model contained in simulating blood plasma
It encloses for 0.85-85.00 μ g/mL.
The rapid detection method of sufentanil in blood plasma provided by the invention can also have such technical characteristic,
In, the computational methods in step S7 are：Establish in simulating blood plasma sample 1004cm in sufentanil SERS collection of illustrative plates-1The characteristic peak at place
Intensity and sufentanil concentration linear relationship, and the sufentanil concentration being calculated accordingly in test plasma sample.
The present invention also provides a kind of device for fast detecting of sufentanil in blood plasma, for the Shu Fen in test plasma
Too Buddhist nun's content is detected, which is characterized in that including：Pretreatment unit, for being pre-processed to obtain to test plasma
Test plasma sample；Detection kit, containing be useful for the sufentanil solution as standard solution, for be used as content calculation pair
According to and containing sufentanil simulating blood plasma sample, for allow standard solution, simulating blood plasma sample and test plasma sample into
The silica gel plate of row point sample, the expansion cylinder for the silica gel plate after point sample to be unfolded and the iodine cylinder for colour developing, and be used for
The elargol of spectrum enhancing is carried out to the sufentanil spot after expansion；Raman spectrum detection unit, for elargol has been added dropwise
Sufentanil spot carries out Surface enhanced Raman scattering detection and carries out analyzing processing to the spectral signal that detection obtains, and obtains mould
Quasi- plasma sample and the sufentanil characteristic peak in test plasma sample and its intensity；And spectroscopy unit, it is used for basis
Sufentanil characteristic peak Strength co-mputation in simulating blood plasma sample and test plasma sample obtains the Shu Fen in test plasma sample
Too Buddhist nun's concentration, wherein comprising the nano silver particles that average grain diameter is 50nm in elargol, detection kit also includes in point sample
When draw standard solution standard extractor, in point sample draw test plasma sample sample extractor and for
The uptake of the elargol extractor of absorption elargol when elargol is added dropwise, standard extractor and sample extractor is 1 μ L, and elargol is drawn
The uptake of device is 5 μ L.
Invention effect
According to the detection method of sufentanil in blood plasma provided by the invention, as a result of being 50nm containing average grain diameter
Nano silver particles elargol as Contrast agent, which makees the characteristic peak of sufentanil with good signal enhancing
With, and signal is not generated at the characteristic peak of sufentanil, so that TLC-SERS methods can be applied to Shu Fen in blood plasma
The too quick detection of Buddhist nun.In the present invention, the point sample amount due to using uses for 1 μ L containing the nanometer that average grain diameter is 50nm
The elargol of silver particles and the infusion volume of the elargol are 5 μ L, therefore enable to the nanometer silver granuel to interact with determinand
The amount of son is appropriate, plays the role of more preferably signal enhancing.
Description of the drawings
Fig. 1 is the UV-visible spectrum of the elargol of the embodiment of the present invention；
Fig. 2 is the scanning electron microscope diagram of the elargol of the embodiment of the present invention；
Fig. 3 is the Development of Thin-Layer Chromatography result figure of the embodiment of the present invention；
Fig. 4 is the sample detection result schematic diagram of the embodiment of the present invention；
Fig. 5 is the Surface enhanced Raman spectroscopy detection of the plasma sample of the sufentanil containing various concentration of the embodiment of the present invention
Limit result figure；
Fig. 6 is sufentanil concentration-signal strength standard curve of the embodiment of the present invention.
Specific implementation mode
With reference to embodiments come illustrate the present invention specific implementation mode.In following embodiments, used Raman light
Spectrometer is BWS415-785H type Portable Raman spectrometers, excitation light source 785nm, resolution ratio 3.5cm-1@912nm, spectrum model
Enclose 175-2700cm-1, it is equipped with BAC151 video microscopic Raman detecting systems (eyepiece × 20)；Lamellae is coating layer thickness 0.2mm-
The silica gel plate of 0.25mm, silica gel Powder Particle Size (8 ± 2) μ m≤80%.In addition, the reagent employed in embodiment is unless otherwise specified
It is obtained from general commercial sources, the experiment condition being not specified is with reference to conventional laboratory conditions or the item suggested in accordance with supplier
Part.
<Embodiment>
1. reagent is prepared and is prepared
It is prepared by 1.1 elargol
Containing the nano silver particles that average grain diameter is 50nm in elargol employed in the present embodiment, the elargol is with reference to existing
Technology (i.e. Lee methods) is made, and preparation method is specific as follows：
It weighs silver nitrate 34mg and is dissolved in 200mL water and obtain silver nitrate solution, which is heated to reflux condensation
To slightly boiling, the citric acid three sodium solution that 6mL mass percents are 1% is then added and forms mixed solution.It keeps to mixed solution
Heating, until the color of mixed solution is become pale yellow from water white transparency and is further changed to light grey while showing slightly green, discoloration
After continue to heat 30min, stop heating, water-bath cooling then carried out to get the elargol containing nano silver particles to mixed solution,
It sets in 250mL brown bottles and is preserved for use in 4 DEG C.
Fig. 1 is the UV-visible spectrum of the elargol of the embodiment of the present invention, and Fig. 2 is sweeping for the elargol of the embodiment of the present invention
Retouch electron microscope picture.
As shown in Figure 1, the maximum absorption band of elargol is at 414nm, and half-peak breadth is relatively narrow, illustrates that the dispersion of elargol particle is equal
It is even.It is no in heaps or be adhered as shown in Fig. 2, the form of nano silver particles is full, size is uniform, and average grain diameter is about 50nm
Left and right.
The preparation of 1.2 storing solutions and standard solution
Precision weighing sufentanil reference substance is appropriate, and the storing solution of a concentration of 3.4mg/mL is configured to methanol.It takes above-mentioned
Appropriate storing solution, be diluted to respectively with methanol a concentration of 1.7mg/mL, 850.0 μ g/mL, 425.0 μ g/mL, 212.5 μ g/mL,
85.0 μ g/mL, 42.5 μ g/mL, 8.5 μ g/mL a series of solution as standard solution, saved backup in 4 DEG C.
2. detection method
Detection method in the present embodiment is carried out using TLC-SERS, is mainly included the following steps.
Step S1 pre-processes test plasma to obtain test plasma sample；Step S2 prepares sufentanil solution work
For standard solution, and the simulating blood plasma sample containing sufentanil is prepared in sufentanil addition blank plasma samples.
The present embodiment is pre-processed using blank plasma (the rat blank plasma for not containing sufentanil), and pre-
Sufentanil standard solution is added in treated blank plasma samples to obtain simulating blood plasma sample.In addition, in embodiment
Test plasma sample also use simulating blood plasma sample replacement.
Wherein, the preprocess method of blank plasma is：It takes blood plasma appropriate, is by volume 1：2 are added acetonitrile, vortex oscillation
1min, 13000rpm centrifuge 10min, go after removing protein to take whole supernatants in the lower 25 DEG C of dryings of nitrogen evaporator, add and blank
The methanol of blood plasma same volume redissolves, and obtains blank plasma samples.
Appropriate sufentanil standard solution is added into blank plasma samples respectively later, is configured to sufentanil final concentration
For 340.00 μ g/mL, 170.00 μ g/mL, 85.00 μ g/mL, 42.50 μ g/mL, 21.25 μ g/mL, 8.50 μ g/mL, 4.25 μ g/
A series of simulating blood plasma sample of solution as various concentration of mL, 0.85 μ g/mL, save backup in 4 DEG C.
Step S3 draws simulating blood plasma sample and standard solution, and puts on same silica gel plate respectively.
Step S4, using methylene chloride-methanol as solvent on silica gel plate sufentanil standard solution and simulation
Plasma sample is unfolded, and is dried silica gel plate taking-up after expansion, is placed in iodine cylinder and develops the color.
Step S5, in reference standard solution in the location determination simulating blood plasma sample of sufentanil spot sufentanil spot
Point position, is then added dropwise the elargol of predetermined amount respectively at the sufentanil spot of simulating blood plasma sample.
In the present embodiment, the influence to investigate elargol dripping quantity has carried out the experiment of different dripping quantities, elargol drop respectively
Dosage is respectively 1 μ L, 5 μ L, 10 μ L.
Step S5 carries out Surface enhanced Raman scattering detection (hereinafter referred to as SERS) and right to being added dropwise at the spot of elargol
It detects obtained spectral signal and carries out analyzing processing, obtain sufentanil characteristic peak and its intensity in simulating blood plasma sample.This
In embodiment, above-mentioned SERS is carried out using Portable Raman spectrometer, and testing conditions are laser power 100mW, microscopic system
Amplification factor 20.
In addition, for the spectral signal that Portable Raman spectrometer detects, the present embodiment uses 5.0 Hes of OPUS
13.0 softwares of Matlab handle gained spectrum, chosen spectrum wave band 300-1700cm-1For spectrum analysis (300cm-1
There are nano silver signal interference, 1700cm before-1Afterwards almost without spectral signature), smooth (Sgolay methods), baseline school are carried out to spectrum
8.5 editions software mappings of Origin are used in combination in positive (airPLS methods) and normalization (Min-Max Normalization methods) processing, from
And obtain Raman spectrum corresponding with each standard solution or simulating blood plasma sample.
Step S6 establishes the 1004cm of sufentanil in simulating blood plasma sample-1Feature peak intensity and sufentanil concentration
Linear relationship, and the sufentanil concentration being calculated accordingly in test plasma sample.
3. condition is investigated
The investigation of 3.1 thin-layer chromatography conditions
Using the methylene chloride-methanol of different proportion as the expansion system in step S3, the dichloro of different proportion is compared
The separating effect of methane-methanol show that best methylene chloride-methanol ratio is 9:1.2.
Fig. 3 is the Development of Thin-Layer Chromatography result figure of the embodiment of the present invention.Wherein, band 1 is sufentanil standard solution, item
Band 2 is plasma sample.
As shown in figure 3, being 9 in methylene chloride-methanol ratio:It is unfolded under conditions of 1.2, can get ideal
Separating effect.
The investigation of 3.2 elargol dripping quantities
Influence when the present embodiment has investigated dispensing amount difference to sufentanil Raman signal.That is, being adopted when elargol is added dropwise
With different dripping quantities, and investigate influence of the different dripping quantities to testing result.
When the dripping quantity of elargol is 1 μ L, the characteristic peak that the SERS collection of illustrative plates of sufentanil is shown is less, and signal is relatively low；
When the dripping quantity of elargol is 5 μ L, the characteristic peak appearance that the SERS collection of illustrative plates of sufentanil is shown is more complete, and signal is stronger.More than
The reason of phenomenon may be dispensing amount be 1 μ L when produce " coffee ring effect ", i.e., elargol dropwise addition spot is formed on lamellae,
The edge concentration of its spot is more than centre concentration, and nano silver particles have been concentrated mainly on spot edge so that institute in Raman light path
The enhancing substrate that can be detected is very few, and the signal of determinand is caused not embody all；And when the dripping quantity of elargol is 5 μ L,
The amount of the elargol to interact with determinand after " coffee ring " effect is appropriate, is able to detect that comparatively ideal signal.And
When dispensing amount is 10 μ L, the appearance time of determinand is longer, and the duration is shorter, is unfavorable for the detection of determinand.
Therefore, when the grain size of the nano silver particles contained in the elargol is 50nm, the optimum point of sufentanil SERS detections
5 μ L of glue amount.
The investigation of 3.3 times of integration
Investigated respectively the time of integration be 5s, 10s and 20s when effect.That is, being carried out with Portable Raman spectrometer
The condition of the different times of integration is used when SERS, and investigates influence of the different times of integration to testing result.
When the time of integration is 5s, the peak intensity of determinand is stronger, and appearance is more, can acquire 6-8 spectrum；When integral
Between be 10s when, determinand peak intensity enhancing, go out peak number without significant change, but due to laser irradiation time lengthen, thin layer plate coating
It is easily scorched, determinand is irradiated with a laser cause damage, can collect 2-4 spectrum；When the time of integration is 20s, thin layer plate coating
It is scorched quickly, spectrum can not be acquired.
Therefore, in detection method of the invention, the best total of points time of SERS is 5s.
According to above-mentioned investigation result it is found that in the detection method of the present invention, the methylene chloride-methanol in step S3 most preferably compares
Example is 9:The best dripping quantity of elargol in 1.2, step S4 is 5 μ L, and the best total of points time of SERS is 5s in step S5.
4. detection result
4.1 qualitative detection
Fig. 4 is the sample detection result schematic diagram of the embodiment of the present invention.In Fig. 4, a is that simulating blood plasma sample (is added with
The plasma sample of sufentanil), b is the normal Raman collection of illustrative plates of sufentanil, and c is blank plasma, and d is elargol blank control.
Figure 4, it is seen that sufentanil is in 600-1500cm-1The multiple characteristic peaks of appearance in range, including 655,
1004,1080 and 1439cm-1(at arrow in figure).Wherein, 1004cm-1The characteristic peak at place is not present in blank plasma, and
And elargol has apparent humidification, therefore 1004cm to characteristic peak at this-1The characteristic peak at place can be used as sufentanil feature
Peak, that is, method of the invention can carry out qualitative detection.
4.2 detection limits
The optimal conditions obtained using above-mentioned " investigation of 3. conditions " is 0.85-340.00 μ g/ to sufentanil concentration range
The simulating blood plasma sample of mL is detected according to aforementioned " 2. detection method ", obtains the Raman signal of various concentration sample.
Fig. 5 is the Raman spectrogram detection limit result of the plasma sample of the various concentration sufentanil of the embodiment of the present invention
Figure.Wherein, a is 21.25 μ g/ for 42.50 μ g/mL, e for 85.00 μ g/mL, d for 170.00 μ g/mL, c for 340.00 μ g/mL, b
It is 0.85 μ g/mL that mL, f, which are 4.25 μ g/mL, h for 8.50 μ g/mL, g,.
As shown in figure 5, when sufentanil concentration range is 0.85-85.00 μ g/mL, with the gradual increase of concentration, feature
Peak intensity also gradually increases；And if concentration further increases, enhancing effect weakens instead.
Analyze its reason, it may be possible to which, due to the increase with sufentanil concentration, drug molecule quantity increases, Molecular Detection
Sensitivity accordingly increase；And due to constant (the colloid table adsorbed for drug molecule of elargol dripping quantity in detection process
Face is constant), adsorbing mutually between multilayer absorption or drug molecule may occur therewith in drug molecule, thus can not be with silver nanoparticle
Grain surface carries out uniform, effective absorption, cannot Raman signal accordingly be enhanced, or even flood signal, cause effect unknown
It is aobvious.
In addition, as can be drawn from Figure 5, when signal-to-noise ratio (S/N) is 3, the lowest detection of sufentanil is limited to 0.85 μ g/
mL。
4.3 plasma sample content analysis
Fig. 6 is sufentanil concentration-signal strength standard curve of the embodiment of the present invention.
As shown in fig. 6, choosing a concentration of 0.85,4.25,8.50,21.25,42.50 and 85.00 μ g/mL's of sufentanil
Simulating blood plasma sample carries out TLC-SERS analyses, and records 1004cm as previously described-1The feature peak intensity at place is carried out with concentration
Linear fit draws standard curve.Calibration curve equation is：Y=70.538x+545.71 (r2=0.9742), y 1004cm-1
The sufentanil feature peak intensity at place, x are the concentration (μ g/mL) of sufentanil.As it can be seen that in the range of 0.85~85.00 μ g/mL
It is interior, sufentanil concentration and 1004cm-1The feature peak intensity at place has good linear, illustrates that the method for the present invention can carry out
The quantitative detection of sufentanil.
4.4 the rate of recovery
In a manner of adding sufentanil into blank plasma, it is respectively 4.25 μ g/mL (low dense to prepare sufentanil concentration
Degree) and 42.50 μ g/mL (high concentration) each three parts of simulating blood plasma sample as test plasma sample, later according to " 2. detection sides
Method " carries out TLC-SERS analyses to it, the 1004cm that will be measured-1The intensity for locating characteristic peak substitutes into above-mentioned standard curve, predicts it
Concentration, the results are shown in Table 1.
1 TLC-SERS methods of table measure the rate of recovery of sufentanil method in blood plasma
As it can be seen from table 1 the average recycling as sufentanil in each simulating blood plasma sample of test plasma sample
Rate is respectively 86.00% and 103.45%, RSD% < 10%.The result shows that the detection method using the present embodiment can be to be measured
Sufentanil in plasma sample is used for quickly detecting, and the rate of recovery is higher.
Embodiment effect
According to the detection method of sufentanil in blood plasma provided in this embodiment, it is as a result of containing average grain diameter
For the elargol of the nano silver particles of 50nm as Contrast agent, which does not generate signal at the characteristic peak of sufentanil, and
There is good signal enhancing effect to the characteristic peak of sufentanil, so that TLC-SERS methods can be applied to relax in blood plasma
The quick detection of fentanyl.In the present embodiment, for the point sample amount due to using for 1 μ L, it is 50nm's to use containing average grain diameter
The elargol of nano silver particles and the infusion volume of the elargol are 5 μ L, therefore enable to the nanometer to interact with determinand
The amount of silver particles is appropriate, plays the role of more preferably signal enhancing.
In embodiment, due to having carried out elargol preparation with reference to Lee methods, and use 34mg/200mL's in preparation process
Continue the reaction condition for being heated to reflux 30min after silver nitrate concentration, discoloration, therefore it is about receiving for 50nm that can obtain average grain diameter
Rice silver particles.
Due to using the time of integration of 5S when SERS is detected, lamellae caused by the time of integration can be avoided long
Burned problem obtains enough spectroscopic datas while ensureing determinand peak intensity.In addition, due to having selected 1004cm-1As
The characteristic peak of sufentanil, therefore the interference of other substances in blood plasma can be avoided, and can ensure the signal enhancing effect of elargol
Fruit.
Above-described embodiment is only used for the rapid detection method of sufentanil in the blood plasma illustrated the present invention, and according to this
The detection method of invention, the detection mode of sufentanil can also be other forms in blood plasma, such as contain pretreatment unit, inspection
The form of the detection device of test agent box, Raman spectrum detection unit and spectroscopy unit.In this case, detection reagent
Box can include required reagent in detection process well prepared in advance, for example, standard solution in embodiment, simulating blood plasma sample,
Silica gel plate, expansion cylinder, iodine cylinder, elargol etc.；In addition, detection kit can also include to be drawn for drawing the standard of standard solution
Device, the sample extractor for drawing plasma sample and the elargol extractor for drawing elargol, these different extractors are equal
The uptake for corresponding respectively to corresponding reagent in embodiment, so as to no extractor at the scene in the case of can also complete blood
The quick measurement of sufentanil in slurry samples.
Claims (8)
1. the rapid detection method of sufentanil in a kind of blood plasma, for being examined to the sufentanil content in test plasma
It surveys, which is characterized in that include the following steps：
Step S1 is pre-processed to obtain test plasma sample to the test plasma；
Step S2 prepares sufentanil solution as standard solution, and sufentanil addition blank plasma samples are prepared
Simulating blood plasma sample containing sufentanil；
Step S3 draws the test plasma sample, the simulating blood plasma sample and the standard solution and puts in same respectively
On silica gel plate；
Step S4, using methylene chloride-methanol as solvent to the test plasma sample, the mould on the silica gel plate
Quasi- plasma sample and the standard solution be unfolded, is dried and the colour developing of iodine cylinder；
Step S5, test plasma sample and the simulation with reference to described in the location determination of sufentanil spot in the standard solution
The speckle displacement of sufentanil in plasma sample, in the sufentanil spot of the test plasma sample and the simulating blood plasma sample
The elargol of predetermined amount is added dropwise at point respectively；
Step S6 carries out Surface enhanced Raman scattering detection and is obtained to detection to being added dropwise at the spot of the elargol
Spectral signal carries out analyzing processing, obtains the sufentanil characteristic peak in the test plasma sample and the simulating blood plasma sample
And its intensity；
Step S7 is calculated according to the relationship of the feature peak intensity of sufentanil in the simulating blood plasma sample and sufentanil concentration
The sufentanil concentration in the test plasma sample is obtained,
Wherein, the uptake of the test plasma sample in step S3, the simulating blood plasma sample and the standard solution
For 1 μ L,
The predetermined amount of elargol described in step S5 is 5 μ L, includes the nano silver particles that average grain diameter is 50nm in the elargol.
2. the rapid detection method of sufentanil in blood plasma according to claim 1, it is characterised in that：
Wherein, the elargol of step S5 is prepared with the following method：
It weighs silver nitrate 34mg and is dissolved in 200mL water and obtain silver nitrate solution；
The silver nitrate solution is heated to reflux condensation to slightly boiling, the trisodium citrate that 6mL mass percents are 1% is then added
Solution forms mixed solution；
Heating is kept until the color of the mixed solution is become pale yellow from water white transparency and is further changed to light grey while omiting
Aobvious green continues to heat 30min after discoloration, stops heating；
Water-bath cooling is carried out to the mixed solution, obtains the elargol.
3. the rapid detection method of sufentanil in blood plasma according to claim 1, it is characterised in that：
Wherein, the condition of the detection of Surface enhanced Raman scattering described in step S6 is laser power 100mW, time of integration 5S, shows
Micro-system amplification factor 20.
4. the rapid detection method of sufentanil in blood plasma according to claim 1, it is characterised in that：
Wherein, it is to the analyzing processing of spectral signal progress in step S6：Choose the 300- of the spectral signal
1700cm-1Smooth, baseline correction and normalized are carried out, spectral signal is mapped to obtain and the spectrum to treated
The corresponding SERS collection of illustrative plates of signal,
The sufentanil feature peak intensity is 1004cm in the SERS collection of illustrative plates-1The intensity of the characteristic peak at place.
5. the rapid detection method of sufentanil in blood plasma according to claim 1, it is characterised in that：
Wherein, the preprocess method of step S2 is：Take the test plasma or the simulating blood plasma as pending blood plasma,
According to volume ratio 1:2 are added acetonitrile, centrifuge 10min under the conditions of 13000rpm after vortex oscillation 1min, take the supernatant after centrifugation
Liquid dries up under the conditions of 25 DEG C of nitrogen evaporator, is then added and the methanol of the pending blood plasma same volume is redissolved, and obtains pair
The plasma sample answered.
6. the rapid detection method of sufentanil in blood plasma according to claim 1, it is characterised in that：
Wherein, the simulating blood plasma is the multiple and sufentanil containing various concentration respectively, is contained in the simulating blood plasma
Sufentanil content range is 0.85-85.00 μ g/mL.
7. the rapid detection method of sufentanil in blood plasma according to claim 6, it is characterised in that：
Wherein, the computational methods in step S7 are：It establishes in the simulating blood plasma sample in sufentanil SERS collection of illustrative plates
1004cm-1The linear relationship of the intensity and sufentanil concentration of the characteristic peak at place, and be calculated in test plasma sample accordingly
Sufentanil concentration.
8. the device for fast detecting of sufentanil in a kind of blood plasma, for being examined to the sufentanil content in test plasma
It surveys, which is characterized in that including：
Pretreatment unit obtains test plasma sample for being pre-processed to the test plasma；
Detection kit, containing be useful for the sufentanil solution as standard solution, for be used as content calculation compare and contain
There is the simulating blood plasma sample of sufentanil, for allowing the standard solution, the simulating blood plasma sample and the test plasma sample
Product carry out silica gel plate, the expansion cylinder for the silica gel plate after point sample to be unfolded and the iodine cylinder for colour developing of point sample,
And the elargol for carrying out spectrum enhancing to the sufentanil spot after expansion；
Raman spectrum detection unit is dissipated for carrying out surface-enhanced Raman to the sufentanil spot that the elargol has been added dropwise
It penetrates detection and analyzing processing is carried out to the spectral signal that detection obtains, obtain the simulating blood plasma sample and the test plasma sample
Sufentanil characteristic peak in product and its intensity；And
Spectroscopy unit, for according to the sufentanil characteristic peak in the simulating blood plasma sample and the test plasma sample
Strength co-mputation obtains the sufentanil concentration in the test plasma sample,
Wherein, it is the nano silver particles of 50nm comprising average grain diameter in the elargol,
The detection kit also includes for drawing the standard extractor of the standard solution in point sample, in point sample
It draws the sample extractor of the test plasma sample and the elargol for drawing the elargol when the elargol is added dropwise is inhaled
Device is taken,
The uptake of the standard extractor and the sample extractor is 1 μ L, and the uptake of the elargol extractor is 5 μ
L。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810390239.2A CN108732292B (en) | 2018-04-27 | 2018-04-27 | Method and device for rapidly detecting sufentanil in blood plasma |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810390239.2A CN108732292B (en) | 2018-04-27 | 2018-04-27 | Method and device for rapidly detecting sufentanil in blood plasma |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108732292A true CN108732292A (en) | 2018-11-02 |
| CN108732292B CN108732292B (en) | 2020-07-14 |
Family
ID=63939311
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810390239.2A Active CN108732292B (en) | 2018-04-27 | 2018-04-27 | Method and device for rapidly detecting sufentanil in blood plasma |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108732292B (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111521597A (en) * | 2019-02-01 | 2020-08-11 | 中国人民解放军军事科学院军事医学研究院 | Method and kit for detecting fentanyl/morphine compound |
| 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 |
| 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 |
| WO2022077808A1 (en) * | 2020-10-16 | 2022-04-21 | 宁波市疾病预防控制中心 | Method and kit for measuring fentanyl drug in biological sample |
| 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 |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20050054942A1 (en) * | 2002-01-22 | 2005-03-10 | Melker Richard J. | System and method for therapeutic drug monitoring |
| WO2005098429A3 (en) * | 2000-11-08 | 2006-05-26 | Univ Florida | System and method for real-time diagnosis, treatment, and therapeutic drug monitoring |
| CN101790529A (en) * | 2007-08-27 | 2010-07-28 | 施万制药 | 8-azabicyclo[3.2.1]octyl-2-hydroxybenzamide compounds as mu opioid receptor antagonists |
| CN202676599U (en) * | 2012-01-11 | 2013-01-16 | 临沂大学 | Chromatography filter paper suitable for raman spectroscopy detection |
| CN103954608A (en) * | 2014-05-20 | 2014-07-30 | 中国人民解放军第二军医大学 | Method for identifying whether blood-glucose-lowering traditional Chinese medicine contains pioglitazone or rosiglitazone or not |
| CN104949955A (en) * | 2014-06-26 | 2015-09-30 | 中国人民解放军第二军医大学 | Method for detecting ephedrine additives doped in weight-losing traditional Chinese medicines |
| CN107064401A (en) * | 2016-09-30 | 2017-08-18 | 中国人民解放军第二军医大学 | A kind of quick method for detecting ten kinds of pigments of illegal addition in beverage simultaneously |
| CN107655873A (en) * | 2016-07-25 | 2018-02-02 | 岛津分析技术研发（上海）有限公司 | Raman spectrum analysis device and method |
-
2018
- 2018-04-27 CN CN201810390239.2A patent/CN108732292B/en active Active
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2005098429A3 (en) * | 2000-11-08 | 2006-05-26 | Univ Florida | System and method for real-time diagnosis, treatment, and therapeutic drug monitoring |
| US20050054942A1 (en) * | 2002-01-22 | 2005-03-10 | Melker Richard J. | System and method for therapeutic drug monitoring |
| CN101790529A (en) * | 2007-08-27 | 2010-07-28 | 施万制药 | 8-azabicyclo[3.2.1]octyl-2-hydroxybenzamide compounds as mu opioid receptor antagonists |
| CN202676599U (en) * | 2012-01-11 | 2013-01-16 | 临沂大学 | Chromatography filter paper suitable for raman spectroscopy detection |
| CN103954608A (en) * | 2014-05-20 | 2014-07-30 | 中国人民解放军第二军医大学 | Method for identifying whether blood-glucose-lowering traditional Chinese medicine contains pioglitazone or rosiglitazone or not |
| CN104949955A (en) * | 2014-06-26 | 2015-09-30 | 中国人民解放军第二军医大学 | Method for detecting ephedrine additives doped in weight-losing traditional Chinese medicines |
| CN107655873A (en) * | 2016-07-25 | 2018-02-02 | 岛津分析技术研发（上海）有限公司 | Raman spectrum analysis device and method |
| CN107064401A (en) * | 2016-09-30 | 2017-08-18 | 中国人民解放军第二军医大学 | A kind of quick method for detecting ten kinds of pigments of illegal addition in beverage simultaneously |
Non-Patent Citations (2)
| Title |
|---|
| TATSUYUKI KANAMORI等: "Simultaneous analysis of 18 compounds of fentanyl and its analogues by TLC，GC/MS and LC/MS", 《J-STAGEトップ》 * |
| 孔令策 等: "表面增强拉曼光谱检测水中微量的芬太尼", 《光散射学报》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111521597A (en) * | 2019-02-01 | 2020-08-11 | 中国人民解放军军事科学院军事医学研究院 | Method and kit for detecting fentanyl/morphine compound |
| 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 |
| WO2022077808A1 (en) * | 2020-10-16 | 2022-04-21 | 宁波市疾病预防控制中心 | Method and kit for measuring fentanyl drug 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 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN108732292B (en) | 2020-07-14 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108732292A (en) | The rapid detection method and device of sufentanil in blood plasma | |
| Yin et al. | Application and development trends of gas chromatography–ion mobility spectrometry for traditional Chinese medicine, clinical, food and environmental analysis | |
| Fini | Applications of Raman spectroscopy to pharmacy | |
| Martino et al. | Counterfeit drugs: analytical techniques for their identification | |
| Mayr et al. | Theae nigrae folium: Comparing the analytical performance of benchtop and handheld near-infrared spectrometers | |
| Lau et al. | Rapid analysis of Radix puerariae by near-infrared spectroscopy | |
| Duan et al. | Study on the destructive effect to inherent quality of Fritillaria thunbergii Miq.(Zhebeimu) by sulfur-fumigated process using chromatographic fingerprinting analysis | |
| CN105467023B (en) | A kind of method that UPLC measures Herba Andrographitis medicinal materials fingerprint and four kinds of diterpenoid-lactones | |
| CN105758957A (en) | High performance liquid chromatography-tandem mass spectrum detection method for 8 quinolones residuals in vegetables and application thereof | |
| CN105717250A (en) | Method for fast screening topoisomerase I inhibitor from natural product | |
| Voelker et al. | Evaluation of four field portable devices for the rapid detection of mitragynine in suspected kratom products | |
| CN102707007B (en) | Quality detection method of five-flavor manna medicine bath preparation | |
| Kong et al. | Development and validation of a UPLC–ELSD method for fast simultaneous determination of five bile acid derivatives in Calculus Bovis and its medicinal preparations | |
| Maiti et al. | Standard operating procedure to reveal prostate cancer specific volatile organic molecules by infrared spectroscopy | |
| Travo et al. | Basis of a FTIR spectroscopy methodology for automated evaluation of Akt kinase inhibitor on leukemic cell lines used as model | |
| Yu et al. | The pharmacokinetics, bioavailability and excretion of bergapten after oral and intravenous administration in rats using high performance liquid chromatography with fluorescence detection | |
| CN110441413A (en) | The construction method and detection method of qianbai biyan tablets HPLC finger-print | |
| CN113866309B (en) | HPLC fingerprint spectrum establishment method and application of 11 bufogenin components in venenum bufonis | |
| CN104931594B (en) | The detection method of content of 5 hydroxymethyl furfural in a kind of Schisandra chinensis | |
| CN113960241A (en) | Method for rapidly screening illegal addition of prohibited chemicals in health care products based on thin-layer chromatography-near infrared spectrum coupling technology | |
| Dhandar et al. | Development and validation of UV spectrophotometric method for simultaneous estimation of Quinfamide and Mebendazole in in-house pharmaceutical formulation | |
| Attia et al. | Simeprevir oxidative degradation product: Molecular modeling, in silico toxicity and resolution by synchronous spectrofluorimetry | |
| CN109596768A (en) | A kind of detection method of Children stasis relieving particle | |
| CN114216980B (en) | Method for establishing HPLC-ELSD fingerprint of starwort root | |
| CN110261518A (en) | A kind of screening of human body effect of low dose radiation molecular injury marker taurine and verification method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |