CN105388058A - Method for simultaneously and quickly extracting multi-target drugs and toxicants in detected material - Google Patents
Method for simultaneously and quickly extracting multi-target drugs and toxicants in detected material Download PDFInfo
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Abstract
The invention provides a method for simultaneously and quickly extracting multi-target drugs and toxicants in a detected material. The method comprises the steps of adopting a buffer solution with the pH of 2-14 to dilute the detected material, then transferring the detected material to a solid phase extraction column, sequentially using ether and dichloromethane to perform elution and mixing and concentrating eluants for inspection, wherein the solid phase extraction column is filled with a filler with specific composition and specific performance. The extracting method can separate and extract out of all drugs and toxicants in the detected material within 5 minutes at a time and has the advantages of being good in universality, convenient to operate, small in working amount, good in impurity removing capacity and high in extraction efficiency, and the extraction rate of all common drugs and toxicants is basically above 90%.
Description
Technical field
The present invention relates to the analytical control technical field of chemicals, be specifically related to multiple goal medicine poisonous substance rapid extracting method simultaneously in a kind of sample.
Background technology
Medicine in sample, the analytical control of poisonous substance are divided into extraction and detect two links, due to making rapid progress of science and technology, instrumental analysis detection technique is made to achieve tremendous development, but extract link still consuming time, loaded down with trivial details, extraction ratio is low and cannot carry out examination to unknown material, it is the bottleneck restricting the technical development of medicine toxicological analysis at present, therefore, set up simple and efficient, extraction ratio is high, Impurity removal is thorough, can be suitable for multi-medicament, method that poisonous substance extracts simultaneously seems particularly important.
Under normal circumstances, the extracting method be most widely used surely belongs to classical liquid-liquid extraction, but the shortcomings such as the method exists complex operation, solvent consumption is large, medicine extraction ratio is low, Impurity removal is thorough, poor reproducibility, easily emulsification.Until phase early 1990s, China just begins one's study the extracting method based on solid phase extraction techniques, solid phase extractions material conventional is at present lipophilic materials and ion exchange resin material, although above-mentioned solid phase abstraction technique obviously can reduce solvent load, improve medicine extraction ratio, avoid emulsification, but apply it to public security when solving a case in process, single extraction generally can only be separated and obtain single or single class medicine, and for Common drugs, the separation and Extraction of poisonous substance often needs to prepare multiple sample, multiple solid phase material, carry out several times, the disposable extraction compared with drug poisonous substance cannot be realized, still there is complex operation, time-consuming (blood examination material extracts generally needs 120 minutes), the problem such as undetected.
In order to overcome the above-mentioned defect of prior art, Chinese patent literature CN102517793A discloses the method that an employing blood detects institute's toxicity fast: get two parts of blood to be checked, portion adds the dilution of pH9 damping fluid, another part of thin up, pour into respectively and be equipped with in diatomaceous chromatographic column, use ether wash-out after the blood adding the dilution of pH9 damping fluid crosses chromatographic column, eluent to obtain extract A in concentrated the doing to the greatest extent of water-bath of 45 DEG C; The blood of thin up is crossed with dichloromethane eluent after chromatographic column, and eluent dryly to the greatest extent to obtain extract B in the water-bath of 50 DEG C is concentrated, utilizes the detecting instrument of Conventional compounds to detect respectively extract A and extract B, can detect as which kind of toxic poisoning.Above-mentioned technology only needs two parts of samples, just extraction can be completed in 5 minutes, significantly simplify experimental implementation, shorten extraction time, but this technology can only be applicable to barbiturates, Benzodiazepines, methaqualone, alkaloids medicament, phenothiazines and metabolin thereof, indandione class, Hydroxycoumarin raticide, Tetramine, organic phosphates, carbamate insecticides, and the systematicness of the common drugs such as methamphetamine is extracted simultaneously, and for other Common drugs, poisonous substance is as designer drug, respiration stimulant, pyrethroid insecticides etc., but can not from sample, separation and Extraction be out with said medicine, that is, above-mentioned technology still cannot cover for all common lethal medicines, the disposable extraction of poisonous substance, still need separately to get sample, the extracting method that extra exploitation is new, effectively can not solve complex operation, workload is large, length consuming time, undetected problem.
Therefore, research and development a kind of simple and efficient, extraction ratio is high, Impurity removal is thorough, can be suitable for the disposable extracting method of all Common drugs, poisonous substance simultaneously, is the still unsolved technical barrier in this area.
Summary of the invention
The technical problem to be solved in the present invention be to overcome prior art for the Common drugs in sample, poisonous substance extracting method existing for complex operation, large, the consuming time length of workload, the defect of the disposable extraction of all medicine poisonous substances cannot be realized, thus provide a kind of simple and efficient, extraction ratio is high, removal of impurities is thorough and can be suitable for the method for all Common drugs, the disposable extraction of poisonous substance simultaneously.
For this reason, the present invention realizes the technical scheme of above-mentioned purpose and is:
Multiple goal medicine poisonous substance rapid extracting method simultaneously in a kind of sample, comprising:
Adopt the damping fluid of pH=2-14 to carry out equal-volume dilution to sample, mixing, form mixed liquor, described mixed liquor is transferred in solid-phase extraction column, after treating that it infiltrates completely to the filler of described solid-phase extraction column, adopts ether, dichloromethane eluent successively, merge eluent, concentrated for inspection;
Wherein, the composition of the filler of described solid-phase extraction column comprises, and in the gross mass of described filler, the content of silicon dioxide is 80-94wt%, and the content of aluminium oxide is not more than 3wt%, and the content of iron oxide is not more than 2wt%; The bulk density of described filler is not more than 0.4g/cm
3.
Preferably, for the formation of the pH=5-10 of the damping fluid of described mixed liquor.
Preferably, the volume of described mixed liquor and the mass ratio of described filler are 1: (1.15-1.6), wherein the pass of volume/mass is mL/g.
Preferably, in the gross mass of described filler, the content of silicon dioxide is 85-91wt%, and the content of aluminium oxide is 0.4-2.8wt%, and the content of iron oxide is 0.6-2wt%; The bulk density of described filler is 0.2-0.4g/cm
3.
Preferably, the ignition residue of described filler is not more than 2.5%, and loss on drying is not more than 2.5%.
Preferably, the ignition residue of described filler is not more than 2%, and loss on drying is not more than 2%.
Preferably, the volume ratio of described ether and mixed liquor is (3-8): 1.
Preferably, the volume ratio of described methylene chloride and mixed liquor is (3-8): 1.
Preferably, described sample is one or more in blood, urine, animal viscera and tissue, food or washing product.
Multiple goal medicine poisonous substance rapid extracting method simultaneously in sample of the present invention, in the filler of its solid-phase extraction column, the content of silicon dioxide is 80-94wt%, the content of aluminium oxide is not more than 3wt%, the content of iron oxide is not more than 2wt% and the bulk density of filler is not more than 0.4g/cm
3time, the protein in sample can be made, the large polar impurity molecule such as amino acid is adsorbed on the top layer of filler molecule more firmly by the polar group in its structure, the non-polar group of impurity molecule is consistent outwardly with bound drug simultaneously, poisonous substance molecule, like this in the process of organic solvent wash-out, be conducive on the one hand destroying impurity and medicine, the intermolecular combination of poisonous substance, strengthen medicine, the desorption effect of poisonous substance molecule, and then guarantee very high extraction ratio, also be not easy to impurity molecule from the desorb filler on the other hand, improve the removal effect to impurity, make disposable extracting method of the present invention can reduce the interference of impurity in sample.Preferably, dioxide-containing silica is 85-91wt%, and alumina content is 0.4-2.8wt%, and iron oxide content is 0.6-2wt%, and filler bulk density is 0.2-0.4g/cm
3time, more contribute to the impurity interference thoroughly removed in sample, improve extraction ratio of the present invention.
Eluent after the present invention concentrates can adopt separately the instruments such as ultraviolet spectrum, gas chromatography, high performance liquid chromatography or the instrument such as above-mentioned instrument and mass spectrographic combined instrument to carry out analysis and resolution.
Technical scheme tool of the present invention has the following advantages:
1, multiple goal medicine poisonous substance rapid extracting method simultaneously in sample provided by the invention, select the potpourri with specific composition and particular characteristic as the filler of solid-phase extraction column, and with ether, methylene chloride is that eluant, eluent carries out wash-out to sample respectively, just can by whole medicines contained in sample within 5 minutes, the disposable complete separation and Extraction of poisonous substance is out for detection, thus it is strong to make extracting method of the present invention possess universality, easy and simple to handle, workload is little, purification capacity is strong, the advantage that extraction efficiency is high, to all common lethal medicines, the extraction ratio of poisonous substance is substantially all more than 90%.
2, multiple goal medicine poisonous substance rapid extracting method simultaneously in sample provided by the invention, limiting the volume (mL) of loading mixed liquor with the ratio of the quality (g) of filler is 1: (1.15-1.6), its reason is, the consumption of filler is too small, not only make mixed liquor in loading process can not be adsorbed in filling surface completely, not adding eluant, eluent just has part loading mixed liquor to flow out, add impurity, reduce extraction ratio, and in elution process, be also unfavorable for the desorb of medicine, poisonous substance molecule, reduce extraction ratio equally; But the consumption of filler can not be excessive, can cause that the consumption of eluant, eluent is many, time and effort consuming and extraction ratio has no lifting.
3, multiple goal medicine poisonous substance rapid extracting method simultaneously in sample provided by the invention, inventor is found by long-term scientific research, the ignition residue of the filler of solid-phase extraction column is not more than 2.5%, loss on drying is not more than 2.5%, preferred ignition residue is not more than 2%, when loss on drying is not more than 2%, can to avoid in loading and elution process, because the heat release of filler self, expansion etc. change the harmful effect produced elute effect, making extracting method accuracy of the present invention, favorable reproducibility.
4, multiple goal medicine poisonous substance rapid extracting method simultaneously in sample provided by the invention, also define the consumption of eluant, eluent ether, methylene chloride, guarantee the medicine that filler adsorbs, poisonous substance molecule to elute completely better, and effectively can prevent impurity desorb, minimizing solvent load, shortening extraction time under this prerequisite.In the present invention, the consumption of eluant, eluent is not The more the better, and too much can cause waste of solvent, the more important thing is the impurity content also increased in eluent, extraction ratio also has no improvement.
In addition, extracting method of the present invention can be applicable to all biological materials as blood, urine, animal viscera and tissue, and abiotic sample is as the disposable extraction of contained drug, poisonous substance in food or washing product etc., thus the plurality of advantages such as the inventive method universality is strong, easy and simple to handle, workload is little, purification capacity is strong, extraction efficiency is high are more highlighted.
Embodiment
Below in conjunction with specific embodiment to multiple goal medicine poisonous substance in sample provided by the present invention simultaneously rapid extracting method be described in detail.
Embodiment 1
The blood examination material of sodium hydrogen phosphate-phosphate sodium dihydrogen buffer solution to 1ml of 1ml, pH=6.6 is adopted to dilute, mix to obtain mixed liquor, mixed liquor is transferred in the solid-phase extraction column being filled with 2.3g filler, liquid to be mixed infiltrates after to filler completely, the ether getting 7ml carries out wash-out, get the dichloromethane eluent of 7ml again, merge the eluent of twice, concentrated for inspection.
Filler in the present embodiment consists of: in filler gross mass, and silica 1 .9g, aluminium oxide 69mg, iron oxide 32mg, surplus are calcium oxide, magnesium oxide and organic matter; And the bulk density of this filler is 0.4g/cm
3, ignition residue is 1%, loss on drying is 0.7%.
Embodiment 2
The urine examination material of damping fluid to 0.5ml of 0.5ml, pH=5 is adopted to dilute, mix to obtain mixed liquor, mixed liquor is transferred in the solid-phase extraction column being filled with 1.5g filler, liquid to be mixed infiltrates after to filler completely, the ether getting 5ml carries out wash-out, get the dichloromethane eluent of 5ml again, merge the eluent of twice, concentrated for inspection.
Filler in the present embodiment consists of: in filler gross mass, and silica 1 .36g, aluminium oxide 38mg, iron oxide 26mg, surplus are calcium oxide, magnesium oxide and organic matter; And the bulk density of this filler is 0.28g/cm
3, ignition residue is 0.5%, loss on drying is 0.9%.
Embodiment 3
The blood examination material of damping fluid to 1ml of 1ml, pH=2 is adopted to dilute, mix to obtain mixed liquor, mixed liquor is transferred in the solid-phase extraction column being filled with 2.3g filler, liquid to be mixed infiltrates after to filler completely, the ether getting 5.5ml carries out wash-out, get the dichloromethane eluent of 8ml again, merge the eluent of twice, concentrated for inspection.
Filler in the present embodiment consists of: in filler gross mass, and silica 1 .84g, aluminium oxide 69mg, iron oxide 13.8mg, surplus are calcium oxide, magnesium oxide and organic matter; And the bulk density of this filler is 0.4g/cm
3, ignition residue is 2.5%, loss on drying is 2%.
Embodiment 4
The urine examination material of damping fluid to 0.5ml of 0.5ml, pH=14 is adopted to dilute, mix to obtain mixed liquor, mixed liquor is transferred in the solid-phase extraction column being filled with 1.6g filler, liquid to be mixed infiltrates after to filler completely, the ether getting 3ml carries out wash-out, get the dichloromethane eluent of 5.5ml again, merge the eluent of twice, concentrated for inspection.
Filler in the present embodiment consists of: in filler gross mass, and silica 1 .41g, aluminium oxide 42mg, iron oxide 30mg, surplus are calcium oxide, magnesium oxide and organic matter; And the bulk density of this filler is 0.3g/cm
3, ignition residue is 2%, loss on drying is 2.5%.
Embodiment 5
The milk sample of damping fluid to 0.75ml of 0.75ml, pH=8 is adopted to dilute, mix to obtain mixed liquor, mixed liquor is transferred in the solid-phase extraction column being filled with 2g filler, liquid to be mixed infiltrates after to filler completely, the ether getting 8ml carries out wash-out, get the dichloromethane eluent of 3ml again, merge the eluent of twice, concentrated for inspection.
Filler in the present embodiment consists of: in filler gross mass, and silica 1 .7g, aluminium oxide 8mg, iron oxide 26mg, surplus are calcium oxide, magnesium oxide and organic matter; And the bulk density of this filler is 0.3g/cm
3, ignition residue is 0.2%, loss on drying is 1%.
Embodiment 6
The liquid detergent sample of damping fluid to 0.5ml of 0.5ml, pH=10 is adopted to dilute, mix to obtain mixed liquor, mixed liquor is transferred in the solid-phase extraction column being filled with 1.375g filler, liquid to be mixed infiltrates after to filler completely, the ether getting 6ml carries out wash-out, get the dichloromethane eluent of 6ml again, merge the eluent of twice, concentrated for inspection.
Filler in the present embodiment consists of: in filler gross mass, and silica 1 .05g, aluminium oxide 18.4mg, iron oxide 17.2mg, surplus are calcium oxide, magnesium oxide and organic matter; And the bulk density of this filler is 0.25g/cm
3, ignition residue is 0.1%, loss on drying is 0.8%.
Comparative example 1
The blood examination material of sodium hydrogen phosphate-phosphate sodium dihydrogen buffer solution to 1ml of 1ml, pH=6.6 is adopted to dilute, mix to obtain mixed liquor, mixed liquor is transferred in the solid-phase extraction column being filled with 2.3g filler, liquid to be mixed infiltrates after to filler completely, the ether getting 7ml carries out wash-out, collect eluent, concentrated for inspection.
In this comparative example, the composition of filler is identical with embodiment 1.
Comparative example 2
The blood examination material of sodium hydrogen phosphate-phosphate sodium dihydrogen buffer solution to 1ml of 1ml, pH=6.6 is adopted to dilute, mix to obtain mixed liquor, mixed liquor is transferred in the solid-phase extraction column being filled with 2.3g filler, liquid to be mixed infiltrates after to filler completely, the methylene chloride getting 7ml carries out wash-out, collect eluent, concentrated for inspection.
In this comparative example, the composition of filler is identical with embodiment 1.
Comparative example 3
The blood examination material of sodium hydrogen phosphate-phosphate sodium dihydrogen buffer solution to 1ml of 1ml, pH=6.6 is adopted to dilute, mix to obtain mixed liquor, mixed liquor is transferred to and is filled with in the common commercially available diatomaceous solid-phase extraction column of 2.3g, liquid to be mixed infiltrates after to filler completely, the ether getting 7ml carries out wash-out, get the dichloromethane eluent of 7ml again, merge the eluent of twice, concentrated for inspection.
Adopt ultraviolet differential derivative spectrophotomtry, vapor-phase chromatography, high performance liquid chromatography to detect the confession sample product in embodiment 1-2 and comparative example 1-3 respectively, the extraction ratio data obtained are as shown in table 1-3:
The extraction ratio that table 1 adopts ultraviolet differential derivative spectrophotomtry to record
The extraction ratio that table 2 adopts gas-phase spectrum method to record
The extraction ratio that table 3 adopts efficient liquid phase spectroscopic methodology to record
Note: in table 1-3,---represent and do not detect.
As can be seen from table 1-3, extracting method provided by the invention can be applicable to all Common drugs contained in sample, the disposable extraction of poisonous substance, extraction ratio is substantially all more than 90%, said medicine, poisonous substance comprises barbiturates somnifacient, benzodiazepines minor tranquilizer, methaqualone, alkaloids medicament, phenothiazines neuroleptic agent and metabolin thereof, indandione raticide, Hydroxycoumarin raticide, Tetramine rat poison, organophosphorus insecticides, carbamate insecticides, and the common drugs such as methamphetamine, designer drug, respiration stimulant, pyrethroid insecticides, antiparkinsonian drug, antiepileptic, antidepressants and psychoanaleptic, antalgesic etc. 17 class more than totally 80 plants medicine, poisonous substance.Further, for same medicine, the extraction ratio data adopting different detection methods to obtain are also substantially identical.
And comparative example 1 for the extraction ratio of medicine poisonous substance compared with the present invention, except barbiturates somnifacient, benzodiazepines minor tranquilizer, phenothiazines tranquilizer (not containing metabolin sulfoxide), organophosphorus insecticide, health shaking plasy is wanted, antalgesic, outside antiepileptic, all the other on average reduce more than 31%, and comparative example 1 plants the extraction ratio of medicine poisonous substance all below 80% to more than 30, illustrate for only not obtaining good extraction ratio for the confession sample product of ether wash-out, only have collocation to use the specific eluant, eluent of the present invention to carry out wash-out and could obtain higher extraction ratio.Comparative example 2 for the extraction ratio of medicine poisonous substance compared with the present invention, except most of alkaloid, benzodiazepines minor tranquilizer, carbamate pesticide, Hydroxycoumarin raticide, all the other on average reduce more than 37%, and comparative example 2 plants the extraction ratio of medicine poisonous substance all below 80% to more than 47, illustrate for only with for the confession sample product of dichloromethane eluent, only having collocation to use the specific eluant, eluent of the present invention to carry out wash-out and could obtain higher extraction ratio.In addition, comparative example 3 can only be applicable to the extraction of barbiturates, the sleeping medicine of benzodiazepines, extraction ratio is between 69.8-76.1%, and be not suitable for all the other medicine poisonous substances, and impurity is more, illustrate that the extraction effect using common commercially available zeyssatite as solid phase extraction column stuffing to medicine poisonous substance is poor, the disposable extraction to all medicine poisonous substances in sample cannot be realized at all.In sum, extracting method of the present invention has clear superiority, and comparative example 1-3 is all infeasible.
Obviously, above-described embodiment is only for clearly example being described, and the restriction not to embodiment.For those of ordinary skill in the field, can also make other changes in different forms on the basis of the above description.Here exhaustive without the need to also giving all embodiments.And thus the apparent change of extending out or variation be still among the protection domain of the invention.
Claims (9)
1. a multiple goal medicine poisonous substance rapid extracting method simultaneously in sample, comprising:
Adopt the damping fluid of pH=2-14 to carry out equal-volume dilution to sample, mixing, form mixed liquor, described mixed liquor is transferred in solid-phase extraction column, after treating that it infiltrates completely to the filler of described solid-phase extraction column, adopts ether, dichloromethane eluent successively, merge eluent, concentrated for inspection;
Wherein, the composition of the filler of described solid-phase extraction column comprises, and in the gross mass of described filler, the content of silicon dioxide is 80-94wt%, and the content of aluminium oxide is not more than 3wt%, and the content of iron oxide is not more than 2wt%; The bulk density of described filler is not more than 0.4g/cm
3.
2. extracting method according to claim 1, is characterized in that, for the formation of the pH=5-10 of the damping fluid of described mixed liquor.
3. extracting method according to claim 1 and 2, is characterized in that, the volume of described mixed liquor and the mass ratio of described filler are 1: (1.15-1.6), wherein the pass of volume/mass is mL/g.
4. the extracting method according to any one of claim 1-3, is characterized in that, the content of silicon dioxide is 85-91wt%, and the content of aluminium oxide is 0.4-2.8wt%, and the content of iron oxide is 0.6-2wt%; The bulk density of described filler is 0.2-0.4g/cm
3.
5. the extracting method according to any one of claim 1-4, is characterized in that, the ignition residue of described filler is not more than 2.5%, and loss on drying is not more than 2.5%.
6. extracting method according to claim 5, is characterized in that, the ignition residue of described filler is not more than 2%, and loss on drying is not more than 2%.
7. the extracting method according to any one of claim 1-6, is characterized in that, the volume ratio of described ether and mixed liquor is (3-8): 1.
8. the extracting method according to any one of claim 1-7, is characterized in that, the volume ratio of described methylene chloride and mixed liquor is (3-8): 1.
9. the extracting method according to any one of claim 1-8, is characterized in that, described sample is one or more in blood, urine, animal viscera and histoorgan, food or washing product.
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| JP2007155518A (en) * | 2005-12-06 | 2007-06-21 | Showa Denko Kk | Solid phase extraction cartridge |
| CN102507793A (en) * | 2011-11-24 | 2012-06-20 | 重庆警官职业学院 | Method for detecting poison quickly by using blood |
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