CN108072691A - A kind of method of Etomidate in quick, sensitive detection blood - Google Patents
A kind of method of Etomidate in quick, sensitive detection blood Download PDFInfo
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- CN108072691A CN108072691A CN201611033876.1A CN201611033876A CN108072691A CN 108072691 A CN108072691 A CN 108072691A CN 201611033876 A CN201611033876 A CN 201611033876A CN 108072691 A CN108072691 A CN 108072691A
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- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
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- G—PHYSICS
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Abstract
The invention discloses a kind of methods of Etomidate in quick, sensitive detection blood.This method is using Ion mobility spectrometry as basic detection technique, by the use of ionic migration spectrometer as detection means, using cation high pressure mode, with reference to traditional liquid-liquid extraction pre-treating method, high temperature pyrolysis analysis input mode, sample introduction process addition internal standard calibrating reagent.Device in invention is stable, reliable, and method is easy, quick, efficient.The single sample detection and analysis time is less than 30S.Detection sensitivity is high, and measurement detection limit can reach 0.1ng/ μ l；Quantitative analysis concentration range is in 0.5 20ng/ μ l.The qualitative and quantitative analysis method established meets doctor to human body administration concentration analyst coverage.Detection method in the present invention can be widely used for clinical administration depth analysis, instruct the clinical accurate medication of doctor.
Description
Technical field
The invention discloses a kind of methods of Etomidate in quick, sensitive detection blood.This method is with Ion transfer
Spectral technology is basic detection technique, by the use of ionic migration spectrometer as detection means, using cation high pressure mode, with reference to tradition
Liquid-liquid extraction pre-treating method, high temperature pyrolysis analysis input mode.Device in invention is stable, reliable, method is easy, it is quick,
Efficiently.The qualitative and quantitative analysis method established meets doctor to anaesthesia patient clinical administration concentration analysis scope.In the present invention
Detection method can be widely used for clinical administration depth analysis, instruct the accurate medication of clinician.
Background technology
Anesthesia is to impose calmness, analgesia, hypnosis, flesh pine to patient by anaesthetic to ensure being smoothed out for operation
Deng many-sided effectiveness, to eliminate perception and reaction of the patient to various stimulations in operation.The clinical implementation of general anesthesia includes inhaling
Enter two kinds of anesthesia and total intra-venous anesthesia.It is suitble to the requirement for the compound for doing anesthetic,general：It is stable in physicochemical property, nontoxic, fast
Speed distribution, metabolizable and excretion enable anaesthesia process to reach " fast fiber crops are fast to wake up "；Since these features cause the quick of anesthetic
Dynamic monitoring has difficulties.During real-time Monitored anesthesia, the concentration of anesthetic, which is that Anesthetist is with high safety, effectively anaesthetizes
Foundation.Main anesthetic when implementing total-intravenous anesthesia is Propofol and Etomidate, there is no real-time Monitored anesthesia both at home and abroad
The equipment of concentration.
Etomidate is the derivative of imidazoles, and chemical name is R- (+)-ethyl -1- (1- phenethyls) -1- hydrogen-imidazoles -5-
Carboxylate.Etomidate molecular weight is 244, and some adverse reactions to reduce Etomidate introduce the new of liplid emulsions and match somebody with somebody
Side.The Clinical practice dosage and method recommended both at home and abroad at present are all specific due to nothing quickly and effectively blood concentration assay method
Using there is mistake often.
The method without the blood concentration of monitoring Etomidate in real time domestic and international at present.In operation 75% Etomidate with
Protein binding.When the state of an illness (such as hepatopathy, nephrosis) influences haemocyanin, different journeys can occur for the ratio of free (unbonded) drug
The variation of degree may enhance its pharmacological action.Etomidate increases by 1 times in the distribution volume of liver cirrhosis patient, but it is clear
Except rate is normal, therefore it eliminates half-life period as normal 2 times.Its initial distribution half-life and clinical drug effect may be constant.Age
The initial distribution volume reducing of Etomidate can be made by increasing, and clearance rate declines.
At present, clinically commonly use based on EEG signals detection and the technology analyzed to reflect patient's depth of anesthesia indirectly, such as
BIS, entropy index etc., but these technologies also quite have in accurately assessment patient's depth of anesthesia to reduce the effect in terms of intraoperative diagnosis
Dispute.The realization precisely anaesthetized, it is to be understood that real-time concentration distribution and variation of the anesthetic in effect compartment and blood after administration
Trend, it would therefore be highly desirable to develop the instrument and method of the blood concentration of measurement anesthetic in real time.Etomidate therapeutic drug monitoring
Foreign countries' HPLC methods, the country only some reports, HPLC methods monitor time-consuming and laborious, complex pretreatment, more mainly with HPLC methods
Etomidate therapeutic drug monitoring is especially imprecise, so also using less both at home and abroad, many places are in the scientific research stage.Gas-chromatography
(GC), high performance liquid chromatography (HPLC), gas chromatography mass spectrometry (GC-MS) and LC-MS (HPLC-MS) though blood concentration can be monitored,
Real-time online measuring is unable to, and since equipment is huge, need complicated sample pre-treatments, takes the shortcomings of longer, even if
Laboratory draws the related data of drug, but anesthesia terminates already, therefore Yi Shang equipment is impossible to be applied to clinic.Use ion
It migrates spectrometer and monitors Etomidate blood concentration at home and abroad without report.
One kind of ion mobility spectrometry (Ion Mobility Spectrometry, IMS) technology appearance the 1970s is fast
Fast separation detection technique, compared with traditional mass spectrum, chromatographic apparatus, have it is simple in structure, high sensitivity, analyze speed is fast, knot
The characteristics of fruit is reliable.The IMS that we study at present has been widely used in medical detection, chemical warfare agent, drugs, explosion physical prospecting
The fields such as survey, environmental monitoring, monitoring poisonous gas, fire monitoring, water pollution monitoring and Food Monitoring.The research and development of this method are inevitable
Trigger the innovation of full vein Etomidate anesthesia, promote the innovative technology and innovation theory in Intravenous Anesthesia field, be to realize
Precisely anesthesia, the unique key technology of safe vein anesthesia.
The content of the invention
The present invention relates to a kind of methods of Etomidate in quick, sensitive detection blood, and detection device is Ion transfer
Spectrometer, step are as follows：
1) Etomidate drug standards are dissolved in Blood plasma in vitro or isolated blood, are configured to 5 or more containing difference
The Etomidate pharmaceutical samples of blood concentration, Etomidate drug concentration is in 0.5-20ng/ μ l in sample, by same volume
Organic solvent extraction after, the organic phase of volume A is taken to drop on sample introduction slide glass respectively, while adds the interior calibration of volume A respectively
Quasi- reagent, the scope of volume A is 10-50 μ l；After room temperature (20-30 DEG C) volatilizes solvent, using ionic migration spectrum detection instrument, use
Cation high pressure mode tests and analyzes, and after Thermal desorption sample is ionized, the continuous ion that records reaches ionic migration spectrum detection device
Transit time and signal peak strength ion transfer spectrogram；
2) according to Etomidate and the transit time of internal standard reagent, the maximum signal at two peaks is recorded, then sample
Signal strength seeks ratio with internal standard signal strength；Generation standard curve is fitted with corresponding signal intensity rate according to blood concentration
Equation.
3) the in vitro blood sample to be measured of the human or animal of the injection Etomidate drug of 100-1000 μ L is taken, use is organic
Solvent extraction method extracts object to be measured object；Volume A object to be measured objects is taken to drop on sample introduction slide glass, while add the internal standard of volume A
Calibrating reagent；After room temperature (20-30 DEG C) volatilizes organic solvent, using ionic migration spectrum detection instrument, using cation high pressure mode
It tests and analyzes, after Thermal desorption sample, the continuous transit time and signal peak strength for recording ion and reaching ionic migration spectrum detection device
Ion transfer spectrogram；It calculates sample signal intensity and seeks ratio with internal standard signal strength.
4) obtained signal peak transit time is detected according to Etomidate drug in step 1) and internal standard and determines blood to be measured
The signal peak of Etomidate drug and internal standard peak in liquid sample, to Etomidate quality assurance of drug in blood sample to be measured；By step
3) ratio brings step 2) standard curve equation into, further determines that the content of Etomidate drug in blood.
Due to the individual difference of human or animal, before carrying out blood sample detection to be measured, it is true method feasibility should to be carried out first
Recognize：
The quasi- product examine of advance rower is detected to survey：Step 1) experiment will carry out blood plasma or the background signal blank of blood sample is surveyed
Examination, the transit time of blood plasma or blood sample background signal is compared with the transit time that above-mentioned object is obtained, if
It proves that this detection method is feasible without peak is overlapped, following experimentation can be continued；Prove this detection method in this person if having and overlapping peak
Or the blood plasma or blood sample of animal are infeasible.
Organic solvent is ethyl acetate or chloroform, centrifuges more than extraction time 1min；Organic solvent is immiscible with blood plasma, but
It is that drug ingedient in blood plasma is but soluble in extractant；Ensure the extract of ingredient in blood in migration spectrum cation mould simultaneously
Under formula with the non-overlapping interference of target peak.
Internal standard compound belongs to stable titanate ester, and does not have the adjacent benzene two that overlap peak can be completely isolated with blood matrix
Two nonyl ester (DNP) of formic acid or didecyl phthalate (DIDP), solvent are ethyl acetate or chloroform, and concentration is 5-50ng/ μ
l；Instrument difference in the daytime can be excluded by internal standard calibration, ensure data redundancy, realized precisely quantitative.
It is the object maximum signal using blood concentration as abscissa that drug blank, which adds the calibration curve equation to be formed,
It is ordinate with the ratio between internal standard maximum signal (or parsing area ratio)；The Thermal desorption signal acquisition time for 1-30S it
It is interior.
Etomidate drug standards are dissolved in Blood plasma in vitro or isolated blood by step 1), be configured to containing 0.5,1,
2.5th, 5,12.5ng/ μ l Etomidate pharmaceutical samples.
Blood plasma in vitro or blood plasma or blood that isolated blood is human or animal before medication；In vitro blood sample to be measured is medication
The blood of human or animal afterwards；And their sources are identical, i.e. source individual is identical.
Detection device in this method uses cation high pressure mode for ionic migration spectrometer, and instrument mainly includes sample introduction and fills
Put, transference tube (containing ionization source, reaction zone, ion gate, migration area, signal receiving device), signal translating system and gas circuit
Drying system forms, and the sampling device is made of injection port and the carrier gas transfer pipeline being arranged on the outside of injection port, into
Thermal desorption injector pyrolysis eutectoid temperature is connected on the upside of sample mouth and is more than 150-250 DEG C, when sample introduction slide glass sample introduction be placed in into
In sample mouth, sample is sent in ion mobility spectrometry migration tube and carries out analysis detection by carrier gas.
The carrier gas and drift gas source of the gas used in ion mobility spectrometry experiment is independent outer air supply source；Source gas is needed through making a living
Change silica gel, molecular sieve and the activated carbon filtering of processing, remove water, organic matter and dust；Gas source condition is fixed, it is ensured that ion
Migration spectrum background signal is stablized.
Etomidate therapeutic drug monitoring is to ensureing that clinical safety has particularly important meaning.This detection method
Success is developed, and ion mobility spectrometry will be guided into medical field, while be made up China's medical field anaesthetic Etomidate context of detection
Backwardness and gap.
Advantages of the present invention is as follows：
1. compared with traditional liquid-phase chromatography method, have ion mobility spectrometry as the means of Etomidate in analysis blood
It has the advantage that：Research is found that a kind of internal standard reagent of Etomidate ionic migration spectrum detection, establishes a kind of Ion transfer
The standard method of Etomidate concentration in spectrum detection blood.It is realized by internal standard calibration and carries out Etomidate fiber crops with ion mobility spectrometry
Liquor-saturated medicine quantitative analysis, it is poor in the daytime to avoid, reproducible.
2. measuring method simplicity, quick, good reliability, operate without specialty background personnel.Sampling volume is small, simple sample
Product pre-treatment, high sensitivity, qualitative and quantitative analysis are accurate, are very suitable for clinical quick analysis detection.
3. entire instrument is easy to carry；Measuring speed is fast, and the single sample analysis testing time is less than 30S；The operation of instrument
Expense is very low, and consumables are seldom；Instrument it is cost-effective, analyze speed is fast.
Description of the drawings
Fig. 1 is the ion transfer spectrogram that Etomidate is detected under positive ion mode；
Fig. 2 is Etomidate quantitative analysis standard curve under positive ion mode.
Specific embodiment
Fig. 1-2 gives some experiment spectrograms and gives explanation to the present invention.
Embodiment 1
The experiment condition used in experiment is：The ion mobility spectrometry that ionization source ionizes for lamp, positive ion mode high-voltage power supply,
Stepper motor Thermal desorption injector.Migration tube temperature is maintained at 100 DEG C during experiment, and gas is floated in 200 DEG C of injector, carrier gas (air)
(air) air-flow is respectively 300mL/min, 400mL/min.Internal standard selects DNP, 10ng/ μ l, 10 μ l of sample injection volume, and internal standard takes
The 10 μ l room temperatures (20-30 DEG C) that are mixed are dried.
Fig. 1 gives the ion transfer spectrogram that Etomidate is detected under positive ion mode.
As shown in Figure 1, it is calibrated by ionic mobility.When acetone transit time is in 3.24ms, Etomidate list
Body peak transit time is 4.64ms, and internal standard DNP transit times are located at 7ms.Follow-up test carries out qualitative analysis as standard.
Embodiment 2
The experiment condition used in experiment is：The ion mobility spectrometry that ionization source ionizes for lamp, positive ion mode high-voltage power supply,
Stepper motor Thermal desorption injector.Migration tube temperature is maintained at 100 DEG C during experiment, and gas is floated in 200 DEG C of injector, carrier gas (air)
(air) air-flow is respectively 300mL/min, 400mL/min.Internal standard selects DNP, 10ng/ μ l, 10 μ l of sample injection volume, and internal standard takes
The 10 μ l room temperatures (20-30 DEG C) that are mixed are dried.Etomidate drug is configured to rely on miaow containing 0.5,1,2.5,5,12.5ng/ μ l
The sample of ester drug.
Fig. 2 gives Etomidate quantitative analysis standard curve under positive ion mode.
According to Etomidate and the transit time of internal standard reagent, continuous acquisition ion mobility spectrometry Figure 30 seconds, continuous acquisition is selected
Sample signal peak intensity and the ratio of internal standard signal strength maximum.According to blood concentration (0-12.5ng/ μ l) and peak signal
Intensity rate fitting generation calibration curve equation；Such as Fig. 2, calibration curve equation Y=0.126*X+0.265, coefficient R>
0.98.The qualitative and quantitative analysis method established meets human administration concentration analysis scope of the doctor to anaesthesia patient.
Embodiment 3
The experiment condition used in experiment is：The ion mobility spectrometry that ionization source ionizes for lamp, positive ion mode high-voltage power supply,
Stepper motor Thermal desorption injector.Migration tube temperature is maintained at 100 DEG C during experiment, and gas is floated in 200 DEG C of injector, carrier gas (air)
(air) air-flow is respectively 300mL/min, 400mL/min.Internal standard selects DNP, 10ng/ μ l, 10 μ l of sample injection volume, and internal standard takes
The 10 μ l room temperatures (20-30 DEG C) that are mixed are dried.
According to Etomidate drug and internal standard detect obtained signal peak transit time determine in blood sample to be measured according to
The signal peak of miaow ester drug and internal standard peak are held in the palm, maximum signal ratio is brought into calibration curve equation, it can further really
Determine the content of Etomidate drug in blood.
Such as a unknown concentration Etomidate sample, detect that maximum signal is followed successively by：638mv, 662mv, 608mv,
Corresponding internal standard signal strength is followed successively by 1534mv, 1695mv, 1490mv three times；Ratio is followed successively by：0.3989、0.3316、
0.4080, average value 0.3795.It substitutes into above-described embodiment calibration curve equation, obtains Etomidate blood concentration 0.9ng/ μ
l。
Claims (7)
1. a kind of method of Etomidate in quick, sensitive detection blood, detection device is ionic migration spectrum detection instrument, special
Sign is：
1) Etomidate drug standards are dissolved in Blood plasma in vitro or isolated blood, are configured to 5 or more containing different blood medicines
The Etomidate pharmaceutical samples of concentration, Etomidate drug concentration is in 0.5-20ng/ μ l in sample, by having for same volume
After solvent extraction, the organic phase of volume A is taken to drop on sample introduction slide glass respectively, while add the internal standard calibration examination of volume A respectively
Agent, the scope of volume A is 10-50 μ l；After room temperature (20-30 DEG C) volatilizes solvent, using ionic migration spectrum detection instrument, using just from
Sub- high pressure mode detection and analysis, after Thermal desorption sample is ionized, the continuous migration for recording ion and reaching ionic migration spectrum detection device
Time and signal peak strength ion transfer spectrogram；
2) according to Etomidate and the transit time of internal standard reagent, the maximum signal at two peaks is recorded, then sample signal
Intensity seeks ratio with internal standard signal strength；Generation standard curve side is fitted with corresponding signal intensity rate according to blood concentration
Journey；
3) the in vitro blood sample to be measured of the human or animal of the injection Etomidate drug of 100-1000 μ L is taken, using organic solvent
Extraction method extracts object to be measured object；Volume A object to be measured objects is taken to drop on sample introduction slide glass, while add the internal standard calibration of volume A
Reagent；After room temperature (20-30 DEG C) volatilizes organic solvent, using ionic migration spectrum detection instrument, detected using cation high pressure mode
It analyzes, after Thermal desorption sample, the continuous ion that records reaches the transit time of ionic migration spectrum detection device and signal peak strength ion
Migrate spectrogram；It calculates sample signal intensity and seeks ratio with internal standard signal strength；
4) obtained signal peak transit time is detected according to Etomidate drug in step 1) and internal standard and determines blood sample to be measured
The signal peak of Etomidate drug and internal standard peak in product, to Etomidate quality assurance of drug in blood sample to be measured；It will be in step 3)
Ratio brings step 2) standard curve equation into, further determines that the content of Etomidate drug in blood.
2. according to the method described in claim 1, it is characterized in that：Due to the individual difference of human or animal, blood to be measured is carried out
Before sample detection, method feasibility confirmation should be carried out first：
The quasi- product examine of advance rower is detected to survey：Step 1) tests the background signal skip test that carry out blood plasma or blood sample, will
The transit time of blood plasma or blood sample background signal is compared with the transit time that above-mentioned object is obtained, if without coincidence
Peak proves that this detection method is feasible, can continue following experimentation；Prove this detection method in this human or animal if having and overlapping peak
Blood plasma or blood sample it is infeasible.
3. method according to claim 1 or 2, it is characterised in that：Organic solvent is ethyl acetate or chloroform, and centrifugation is extracted
More than time 1min；Organic solvent is immiscible with blood plasma, but the drug ingedient in blood plasma is but soluble in extractant；It protects simultaneously
Demonstrate,prove blood in ingredient extract under migration spectrum positive ion mode with the non-overlapping interference of target peak.
4. method according to claim 1 or 2, it is characterised in that：Internal standard compound belongs to stable titanate ester, and and blood
Matrix does not have the dinonyl phthalate (DNP) or didecyl phthalate that overlap peak can be completely isolated, solvent
For ethyl acetate or chloroform, concentration is 5-50ng/ μ l；Instrument difference in the daytime can be excluded by internal standard calibration, ensure data weight
Renaturation is realized precisely quantitative.
5. according to the method described in claim 1, it is characterized in that：It is with blood that drug blank, which adds the calibration curve equation to be formed,
Concentration is abscissa, and the ratio between object maximum signal and internal standard maximum signal (or parsing area ratio) are vertical
Coordinate；The Thermal desorption signal acquisition time is within 1-30S.
6. according to the method described in claim 1, it is characterized in that：Etomidate drug standards are dissolved in vitro by step 1)
In blood plasma or isolated blood, be configured to containing 0.5,2.5,5,7.5,10,20ng/ μ l Etomidate pharmaceutical samples.
7. according to the method described in claim 1, it is characterized in that：Blood plasma in vitro or isolated blood are human or animal before medication
Blood plasma or blood；In vitro blood sample to be measured is the blood of human or animal after medication；And their sources are identical, i.e. source individual phase
Together.
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| CN112649494A (en) * | 2020-12-15 | 2021-04-13 | 中国科学院大连化学物理研究所 | Multimodal quantification method based on photoionization ion mobility spectrometry |
| CN112903803A (en) * | 2019-11-19 | 2021-06-04 | 中国科学院大连化学物理研究所 | Device and method for improving detection sensitivity of etomidate blood concentration in operation |
| CN114460161A (en) * | 2021-12-27 | 2022-05-10 | 中船重工安谱（湖北）仪器有限公司 | Ion migration time-based trace substance detection method |
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Application publication date: 20180525 |