CN104111284A - Method for rapidly sensitively detecting sodium valproate in blood - Google Patents
Method for rapidly sensitively detecting sodium valproate in blood Download PDFInfo
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Abstract
The invention discloses a method for rapidly sensitively detecting sodium valproate in blood. An ion migration spectrum technology is taken as a basic detection technology, an ion migration spectrometer is utilized as a detection means, a positive-ion high-voltage mode is employed, a halogen-lamp thermal desorption sampling manner is employed, and the method does not comprise any solvent extraction pretreatment. An apparatus employed in the method is stable and reliable, and the method is simple, rapid and efficient. The detection analysis time of a single sample is less than 20 S. The detection sensitivity is high, the measure detection limit can reach 1 mg/L, and the quantitative analysis concentration scope is 5-500 mg/L. The established qualitative quantitative analysis method satisfies the human-body medicine-administration concentration analysis scope (50-100 mg/L) of a doctor to an epilepsy patient. The detection method is widely applicable to clinical medicine-administration deep analysis, and helps to guide a doctor to clinically precise administrate a medicine.
Description
Technical field
The invention discloses the method for sodium vedproate in a kind of quick, sensitive detection blood.This method, taking Ion mobility spectrometry as basic detection technique, utilizes ionic migration spectrometer as detection means, adopts negative ion high pressure mode, and input mode is analysed in Halogen lamp LED pyrolysis, without any solvent extraction pre-treating method.Device in invention is stable, reliable, and method is easy, speed is fast, efficiently.The qualitative and quantitative analysis method of setting up meets the human body administration concentration analyst coverage (50-100mg/L) of doctor to epileptic patient.Detection method in the present invention can be widely used in clinical administration depth analysis, instructs the accurate medication of clinician.
Background technology
Sodium vedproate is antiepileptic, and its mechanism of action is illustrated not yet completely.Experiment is shown in that this product can increase the synthetic of GABA and reduce the degraded of GABA, thereby the concentration of rising inhibitory neurotransmitter γ-aminobutyric acid (GABA) reduces neuronic excitability and suppresses outbreak.In electric Physiological Experiment, see that this product can produce the inhibition Na similar to phenytoinum naticum
+the effect of passage.Liver is had to infringement.Oral stomach and intestine absorb rapidly and completely, approximately 1～4 hour blood concentration peaking, and bioavilability is nearly 100%, and effective blood drug concentration is 50～100 μ g/mL.Plasma protein binding rate approximately 94% when blood concentration is about 50 μ g/mL; When blood concentration is about 100 μ g/mL, plasma protein binding rate is about 80～85%.When exceeding 120 μ g/mL, blood concentration can there is obvious adverse reaction.Along with blood concentration increases, free fraction increases, thereby increases the gradient (10～20% that the concentration in brain liquid is Plasma) that enters brain tissue, T
1/2it is 7～10 hours.Mainly be distributed in extracellular fluid and liver, kidney, intestines and brain tissue etc.Most of by liver metabolism, comprise and being combined with glucuronic acid and some oxidizing process, mainly discharged by kidney, discharge with ight soil on a small quantity and breathe out.Can, by placenta, can be secreted into milk.
Adult's usual amounts: every day is by body weight 15mg/kg or 2～3 clothes of 600～1200mg gradation every day.When beginning, by 5～10mg/kg, after one week, increase progressively, till controlling outbreak.In the time that exceeding 2500mg, every consumption per day answers part vic, to reduce gastrointestinal irritation.Every daily maximum for by body weight be no more than 30mg/kg or every day 1.8～2.4g.Children's's usual amounts: press weighing scale identical with adult, also can every day 20～30mg/kg, point take for 2～3 times or every day 15mg/kg, increase week about as required by 5～10mg/kg, till effectively maybe can not tolerating.
Rapid determination of sodium valprate in serum is greater than 100mgL
-1time, toxic and side effect increase and curative effect without obvious increase.Now report the bad reaction about sodium vedproate, according to mechanism of action classification, mainly contain the following aspects: cause calmness by suppressing adenyl cyclase, infringement cognition and driving ability, cause hypothyroidism, Weight gain, psoriasis and alopecia; Cause digestive tract reaction by intending serotonin; Cause hepatic lesion and hyperammonemia by anti-carnitine effect; Promote bone to absorb by rising parathormone, cause pancreatic stones and bring out pancreatitis with this; Suppress hematopoiesis by suppressing cell reproduction effect, Inhibit sperm is grown and sperm motility; By causing that allergic reaction causes rare lethal hepatonecrosis and eosinophilia's property pleural effusion; Sodium vedproate tool reproduction bad reaction, causes polycystic ovary syndrome and teratogenic effect.
Due to the therapeutic dose narrow range of sodium vedproate, curative effect and toxicity and blood concentration are in close relations, and between patient, individual difference is very large, therefore monitor very necessary to blood concentration in its body.High performance liquid chromatography, sodium vedproate does not have aromatic rings or conjugated double bond structures, and under ultraviolet light, without absorbing, general HPLC method can not directly be monitored through UV-detector.Because lacking effective uv absorption in sodium vedproate molecular structure, while adopting its blood concentration of high effective liquid chromatography for measuring that ultraviolet detects, sensitivity is compared with low and disturbing factor is many, so mostly adopt the method for fluoroscopic examination, but that testing conditions requires is higher.Fluorescence polarization immunoassay, fluorescence polarization immunoassay mainly relies on TDx-FLx fluorescence polarization immunoassay analyzer, get after serum, adopt the sodium vedproate kit of Abbott Laboratories of the U.S. can complete easy, rapidly determination of plasma concentration, but TDxFLx fluorescence polarization immunoassay analyzer cost is higher, basic medical unit is difficult to carry out.Treat being usually used in clinically simultaneously.Detect the medicine (dilantin sodium, carbamazepine, valpromide, stable, sodium phenobarbital) of epileptics by vapor-phase chromatography, get the highest effective blood drug concentration, carry out interference experiment, gained chromatogram and blank serum chromatogram are just the same, any chromatographic peak do not detected.
The one that ion mobility spectrometry (Ion Mobility Spectrometry, IMS) technology 20 century 70s occur separates detection technique fast, compared with traditional mass spectrum, chromatographic apparatus, have simple in structure, highly sensitive, analysis speed is fast, the feature of reliable results.Can in atmospheric environment, detect micro substance, be suitable for on-the-spot use.We IMS of research has been widely used in the fields such as chemical warfare agent, drugs, explosive detection, environmental monitoring, monitoring poisonous gas, fire monitoring, water pollution monitoring and Food Monitoring at present.Just can determine the existence of evaluating objects material by transit time, and application peak area or peak height can be determined the concentration of respective substance.
Summary of the invention
The present invention relates to the method for sodium vedproate in a kind of quick, sensitive detection blood, pick-up unit is ionic migration spectrometer, the blood sample containing sodium vedproate medicine is analysed on sample introduction thin slice in Halogen lamp LED pyrolysis, and carrier gas is sent to sample steam in ion mobility spectrometry migration tube, to carry out analyzing and testing.Specific as follows:
1) by sodium vedproate medicine dissolving in organic solvent, taking 10-1000mg/L sodium vedproate medicine as standard items, getting 10-100 μ L standard items drops on sample introduction thin slice, utilize ion mobility spectrometry detector, adopting positive ion high pressure mode to detect analyzes, pyrolysis is analysed after sample, measures transit time and the signal peak strength of ion arrival ion mobility spectrometry detecting device;
2) extract the injection of 10-100 μ L or take the human or animal's of sodium vedproate medicine blood sample to be measured, drop on sample introduction thin slice, utilize ion mobility spectrometry detector, adopting negative ion high pressure mode to detect analyzes, pyrolysis is analysed after sample, measures transit time and the signal peak strength of ion arrival ion mobility spectrometry detecting device;
3) the signal peak transit time obtaining according to sodium vedproate drug test in step 1) is determined the signal peak of sodium vedproate medicine in blood sample to be measured, records peak intensity simultaneously, further determines the content of sodium vedproate medicine in blood.
Due to human or animal's individual difference, carry out before blood sample detection to be measured, can first carry out the confirmation of method feasibility:
The detection column criterion product that advance detect: first get 10-100 μ L and do not inject or take the blood sample of the human or animal before sodium vedproate medicine, drop on sample introduction thin slice, utilize ion mobility spectrometry detector, adopt negative ion high pressure mode sample introduction to analyze, pyrolysis is analysed after sample, measure the transit time that ion arrives ion mobility spectrometry detecting device, determine the background signal of blood sample;
Transit time in standard and above-mentioned obtained transit time are compared, if prove that without overlapping this detection method is feasible, can be continued following experimentation; Overlap and prove that this detection method is infeasible in this human or animal's blood sample if having.
Pyrolysis is analysed and is adopted pyrolysis to analyse injector and carry out, and injector is analysed in pyrolysis, and to adopt Halogen lamp LED be thermal source; Hot resolving adopts Halogen lamp LED to irradiate sample 2-20S.
Halogen lamp LED pyrolysis is analysed injector and is opened, closes by controlling Halogen lamp LED, regulates Halogen lamp LED irradiation time between 2-20S; Make blood sample signal not disturb the detection of epilepsy medicine sodium vedproate, blood itself becomes skim membrane-like simultaneously.
Pick-up unit is ionic migration spectrometer, and Halogen lamp LED pyrolysis is analysed on sample introduction thin slice containing after the blood sample of sodium vedproate medicine, and carrier gas is sent to sample in ion mobility spectrometry migration tube, to carry out analyzing and testing;
Ionic migration spectrometer adopts positive ion high pressure mode, mainly comprise sampling device, ionization source, reaction zone, ion gate, migration area, signal reception and detection system and gas circuit drying system, described sampling device is made up of injection port and the carrier gas transfer pipeline that is arranged at injection port outside, be connected with Halogen lamp LED pyrolysis at injection port upside and analyse injector, by regulating Halogen lamp LED irradiation time to change pyrolysis eutectoid temperature; When described sample introduction thin slice sample introduction, be placed in injection port.
Described sample introduction sheeting can be metal grid mesh, tetrafluoro plate, teflon high temperature cloth.Sample introduction sheet thickness is less than pyrolysis and analyses injector injection port height (3mm); Sampling place be taking 0.5-1.2cm as radius, the degree of depth is the groove of 0.5-3mm; This bottom portion of groove is coarse surface or fills disposable reticulate texture small pieces (teflon or iron, aluminum metallic material) at groove, makes blood sample be laid in sampling place surface, increases and carrier gas contact area, is more conducive to carrier gas thermal purging.
The carrier gas using in ion mobility spectrometry experiment and float gas source of the gas for the outer air supply source of independence; Source of the gas gas need to be through silica gel, molecular sieve and the activated carbon filtration of activation processing, except anhydrating, organism and dust; Gas source condition is fixed, and can ensure that ion mobility spectrometry background signal is stable.
In test, be methyl alcohol, ethanol, methylene chloride, chloroform, acetone for the organic solvent diluting.
Sodium vedproate is a kind of not containing the aromatic rings wide spectrum antiepileptic of nonnitrogenous atom again, evident in efficacy to absence petit mal and myoclonic seizure especially, is the choice drug of idiopathy and generalized epilepsy outbreak.Rapid determination of sodium valprate in serum is monitored ensureing that clinical safety has very important meaning.The successful exploitation of this detection method, will guide medical circle into ion mobility spectrometry, make up backwardness and the gap of China's medical circle epilepsy medicine sodium vedproate context of detection simultaneously.
Advantage of the present invention is as follows:
1. compared with traditional liquid-phase chromatography method, ion mobility spectrometry means of sodium vedproate in analyzing blood are had the following advantages: whole instruments weight is less than 15kg, easy to carry; Measuring speed is fast, and the single sample test duration is less than 20S; The operating cost of instrument is very low, and consumables are little; The cost performance of instrument is high, and analysis speed is fast.
2. easy, quick, the good reliability of this measuring method, operates without specialty background personnel.Sampling volume is little, n.s. pre-treatment, highly sensitive, qualitative and quantitative analysis is accurate, is applicable to very much clinical express-analysis and detects.
Brief description of the drawings
Fig. 1 is the ion mobility spectrometry apparatus structural principle schematic diagram that detects sodium vedproate;
Fig. 2 is that under positive ion mode, background signal ion mobility spectrometry figure (RIP) is analysed in Halogen lamp LED pyrolysis;
Fig. 3 is the ion mobility spectrometry figure of the hot resolved detection 10 μ L of Halogen lamp LED, 1000ppm sodium vedproate standard under positive ion mode;
Fig. 4 is the ion mobility spectrometry figure of the hot resolved detection 10 μ L blood blank of Halogen lamp LED under positive ion mode;
Fig. 5 is the ion mobility spectrometry figure that detects sodium vedproate in 10 μ L, 100ppm blood under positive ion mode.
Embodiment
Fig. 1 is the ion mobility spectrometry apparatus structural principle schematic diagram that detects sodium vedproate in blood, and this instrument mainly comprises following components: sampling device, ionization source, reaction zone, ion gate, migration tube, make-up gas and signal receive and detection system.Sampling device can be that injector is analysed in the Halogen lamp LED pyrolysis that is applicable to solid or liquid, mainly analyses cavity, sample introduction thin slice and carrier gas transfer pipeline by Halogen lamp LED and lampshade and support, pyrolysis and forms.
In Fig. 1,1 is ionized region, and 2 is migration area, and 3 analyse injector for Halogen lamp LED pyrolysis, and 4 is gas-carrier pipeline, and 5 is ion gun, and 6 is gas outlet, and 7 is ion gate, and 8 is migration ring, and 9 is aperture plate, and 10 for floating air pipe, and 11 is detecting device.
Fig. 2-5 provided some experiment spectrograms to the present invention give with explanation.The experiment condition of these spectrograms is: ionization source is radioactivity
63the ion mobility spectrometry in Ni source, positive ion mode high-voltage power supply, injector is analysed in the pyrolysis of stepper motor Halogen lamp LED.When experiment, migration tube temperature remains on 100 DEG C, and Halogen lamp LED irradiation time is set to 8S, carrier gas (air), floats gas (air) air-flow and is respectively 400mL/min, 600mL/min.
Embodiment 1
In Fig. 2, under positive ion mode, Halogen lamp LED irradiation air reagent ion peak appears at about 7.8ms;
By sodium vedproate medicine dissolving in methyl alcohol, preparation 1000mg/L sodium vedproate standard items, getting 10 μ L standard items drops on sample introduction thin slice, utilize ion mobility spectrometry detector, adopting positive ion high pressure mode to detect analyzes, pyrolysis is analysed after sample, measures transit time and the signal peak strength of ion arrival ion mobility spectrometry detecting device;
Fig. 3 is the ion mobility spectrometry figure that detects 10 μ L, 1000ppm sodium vedproate standard under positive ion mode, and sodium vedproate detection signal is at about 11.6ms.This detection signal is for proofreading and correct with blood sodium vedproate detection signal.
Embodiment 2
The blood sample that extracts 10-100 μ L with microsyringe is in sampling place of sample introduction thin slice, and Halogen lamp LED irradiates 8S blood sample detection signal at about 9ms and 11.8ms, as shown in Figure 4; Relatively can find out that with subsequent result endogenous material in blank blood do not disturb the detection of sodium vedproate epilepsy medicine.
Embodiment 3
Sodium vedproate in blood (100ppm) detection signal as shown in Figure 5.Sodium vedproate in blood (100ppm) detection signal is at about 11.64ms, the about 100mv of signal intensity.
Quantitative test test, according to the signal intensity of sodium vedproate in the blood of variable concentrations, can, at the built-in component analysis graticule equation of halting of doctor's administration concentration scope, analyze the blood concentration after patient's administration.Quantitative test concentration range is at 5-500mg/L.The qualitative and quantitative analysis method of setting up meets the human body administration concentration analyst coverage (50-100mg/L) of doctor to epileptic patient.
Device in the present invention is stable, reliable, and method is easy, quick, efficient.Single sample detects and is less than 20S analysis time.Detection sensitivity is high, measures detectability and can reach 1mg/L; Quantitative test concentration range is at 5-500mg/L.The qualitative and quantitative analysis method of setting up meets the human body administration concentration analyst coverage (50-100mg/L) of doctor to epileptic patient.Detection method in the present invention can be widely used in clinical administration depth analysis, instructs the clinical accurate medication of doctor.
Claims (8)
1. a method for sodium vedproate in quick, sensitive detection blood, is characterized in that:
1) by sodium vedproate medicine dissolving in organic solvent, taking 10-1000mg/L sodium vedproate medicine as standard items, getting 10-100 μ L standard items drops on sample introduction thin slice, utilize ion mobility spectrometry detector, adopting positive ion high pressure mode to detect analyzes, pyrolysis is analysed after sample, measures transit time and the signal peak strength of ion arrival ion mobility spectrometry detecting device;
2) extract the injection of 10-100 μ L or take the human or animal's of sodium vedproate medicine blood sample to be measured, drop on sample introduction thin slice, utilize ion mobility spectrometry detector, adopting positive ion high pressure mode to detect analyzes, pyrolysis is analysed after sample, measures transit time and the signal peak strength of ion arrival ion mobility spectrometry detecting device;
3) the signal peak transit time obtaining according to sodium vedproate drug test in step 1) is determined the signal peak of sodium vedproate medicine in blood sample to be measured, records peak intensity simultaneously, further determines the content of sodium vedproate medicine in blood.
2. method according to claim 1, is characterized in that:
Due to human or animal's individual difference, carry out before blood sample detection to be measured, can first carry out the confirmation of method feasibility:
The detection column criterion product that advance detect: first get 10-100 μ L and do not inject or take the blood sample of the human or animal before sodium vedproate medicine, drop on sample introduction thin slice, utilize ion mobility spectrometry detector, adopt negative ion high pressure mode sample introduction to analyze, pyrolysis is analysed after sample, measure the transit time that ion arrives ion mobility spectrometry detecting device, determine the background signal of blood sample;
Transit time in claim step 1) and above-mentioned obtained transit time are compared, if prove that without overlapping this detection method is feasible, can be continued following experimentation; Overlap and prove that this detection method is infeasible in this human or animal's blood sample if having.
3. method according to claim 1 and 2, is characterized in that:
It is thermal source that injector employing Halogen lamp LED is analysed in pyrolysis, and hot resolving adopts Halogen lamp LED to irradiate sample 2-20S.
4. method according to claim 3, is characterized in that: Halogen lamp LED pyrolysis is analysed injector and opened, closes by controlling Halogen lamp LED, regulates Halogen lamp LED irradiation time between 2-20S; Make blood sample signal not disturb the detection of sodium vedproate, blood itself becomes skim membrane-like simultaneously.
5. method according to claim 1, is characterized in that:
Pick-up unit is ionic migration spectrometer, and Halogen lamp LED pyrolysis is analysed on sample introduction thin slice containing after the blood sample of sodium vedproate medicine, and carrier gas is sent to sample in ion mobility spectrometry migration tube, to carry out analyzing and testing;
Ionic migration spectrometer adopts positive ion high pressure mode, mainly comprise sampling device, ionization source, reaction zone, ion gate, migration area, signal reception and detection system and gas circuit drying system, described sampling device is made up of injection port and the carrier gas transfer pipeline that is arranged at injection port outside, be connected with Halogen lamp LED pyrolysis at injection port upside and analyse injector, by regulating Halogen lamp LED irradiation time to change pyrolysis eutectoid temperature; When described sample introduction thin slice sample introduction, be placed in injection port.
6. method according to claim 1 or 5, is characterized in that:
Described sample introduction sheeting can be metal grid mesh, tetrafluoro plate, teflon high temperature cloth.Sample introduction sheet thickness is less than pyrolysis and analyses injector injection port height (3mm); Sampling place be taking 0.5-1.2cm as radius, the degree of depth is the groove of 0.5-3mm; This bottom portion of groove is coarse surface or fills disposable reticulate texture small pieces (teflon or iron, aluminum metallic material) at groove, makes blood sample be laid in sampling place surface, increases and carrier gas contact area, is more conducive to carrier gas thermal purging.
7. method according to claim 1 or 5, is characterized in that: the carrier gas using in described ion mobility spectrometry experiment and float gas source of the gas for the outer air supply source of independence; Source of the gas gas need to be through silica gel, molecular sieve and the activated carbon filtration of activation processing, except anhydrating, organism and dust; Gas source condition is fixed, and can ensure that ion mobility spectrometry background signal is stable.
8. method according to claim 1, is characterized in that: in experiment, organic solvent is methyl alcohol, ethanol, methylene chloride, chloroform or acetone.
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| CN105738459A (en) * | 2014-12-08 | 2016-07-06 | 中国科学院大连化学物理研究所 | Method for improving blood sodium valproate detection sensitivity |
| CN106198702A (en) * | 2015-05-06 | 2016-12-07 | 中国科学院大连化学物理研究所 | A kind of method of drugs in quick detection saliva |
| CN108132312A (en) * | 2017-12-28 | 2018-06-08 | 北京和合医学诊断技术股份有限公司 | Detect the liquid phase chromatography analytical method of valproic acid medicament contg in blood |
| CN108931516A (en) * | 2018-05-31 | 2018-12-04 | 北京大学 | Save system parameter optimization method and the Serum Elements quantitative analysis method of sample volume |
| CN111105980A (en) * | 2018-10-25 | 2020-05-05 | 中国科学院大连化学物理研究所 | Flash thermal analysis-time-delay purging sample introduction method for drug mixture detection |
| CN112924529A (en) * | 2019-12-06 | 2021-06-08 | 中国科学院大连化学物理研究所 | Trimethylamine oxide detection method based on ion mobility spectrometer and application thereof |
| 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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| CN105738459A (en) * | 2014-12-08 | 2016-07-06 | 中国科学院大连化学物理研究所 | Method for improving blood sodium valproate detection sensitivity |
| CN105738459B (en) * | 2014-12-08 | 2019-09-06 | 中国科学院大连化学物理研究所 | A kind of method of sodium vedproate detection sensitivity in raising blood |
| CN106198702A (en) * | 2015-05-06 | 2016-12-07 | 中国科学院大连化学物理研究所 | A kind of method of drugs in quick detection saliva |
| CN106198702B (en) * | 2015-05-06 | 2019-01-25 | 中国科学院大连化学物理研究所 | A kind of method of drugs in quick detection saliva |
| CN108132312A (en) * | 2017-12-28 | 2018-06-08 | 北京和合医学诊断技术股份有限公司 | Detect the liquid phase chromatography analytical method of valproic acid medicament contg in blood |
| CN108931516A (en) * | 2018-05-31 | 2018-12-04 | 北京大学 | Save system parameter optimization method and the Serum Elements quantitative analysis method of sample volume |
| CN108931516B (en) * | 2018-05-31 | 2020-11-17 | 北京大学 | System parameter optimization method capable of saving sample introduction amount and serum element quantitative analysis method |
| CN111105980A (en) * | 2018-10-25 | 2020-05-05 | 中国科学院大连化学物理研究所 | Flash thermal analysis-time-delay purging sample introduction method for drug mixture detection |
| CN111105980B (en) * | 2018-10-25 | 2021-01-26 | 中国科学院大连化学物理研究所 | Flash thermal analysis-time-delay purging sample introduction method for drug mixture detection |
| CN112924529A (en) * | 2019-12-06 | 2021-06-08 | 中国科学院大连化学物理研究所 | Trimethylamine oxide detection method based on ion mobility spectrometer and application thereof |
| 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 |
| CN114460161B (en) * | 2021-12-27 | 2022-09-27 | 中船重工安谱（湖北）仪器有限公司 | Trace substance detection method based on ion migration time |
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