CN103487539A - Method for determining contents of albendazole and metabolites thereof in hemolymph of Bombyx mori by using ultra-fast liquid chromatography/triple-quadrupole tandem mass spectrometry (UFLC-MS/MS) - Google Patents
Method for determining contents of albendazole and metabolites thereof in hemolymph of Bombyx mori by using ultra-fast liquid chromatography/triple-quadrupole tandem mass spectrometry (UFLC-MS/MS) Download PDFInfo
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Abstract
The invention discloses a method for determining the contents of albendazole and metabolites thereof in hemolymph of Bombyx mori by using UFLC-MS/MS. The method comprises the following steps: (1) extracting hemolymph of Bombyx mori so as to obtain a to-be-measured sample solution; (2) measuring standard analytical data of standard albendazole and metabolites thereof; (3) injecting the to-be-measured sample solution obtained in the step (1) into an ultra-fast liquid chromatograph for separation and analysis according to a method used in the step (2) and detecting substance signals by using a triple-quadrupole tandem mass spectrometer so as to obtain analytical data of a to-be-measured sample; and (4) comparing the analytical data of the to-be-measured sample obtained in the step (3) with chromatographic analysis data of albendazole and metabolites thereof so as to realize fast qualitative and quantitative determination of albendazole and metabolites thereof in hemolymph of Bombyx mori.
Description
Technical field
The present invention relates to the assay method of the content of albendazole and metabolin thereof, relate in particular to a kind of method of utilizing the supper-fast liquid chromatography-triple level Four bar mass spectrums (UFLC-MS/MS) of connecting to measure the content of albendazole and metabolin in silkworm hemolymph.
Background technology
China is the original producton location of silkworm and mulberry, has the history of more than 5 000 year, have the laudatory title of " Dong Fangsiguo ".Silk and silk fabrics thereof are the main export-oriented commodities of China, and its volume of production and marketing accounts for 70% of world's total amount, are the unique exporting of monopolizing world commerce market of China.
Silkworm egg production is the basis of sericulture there.Pebrine disease is that silkworm egg is produced to the destructive infectiousness epidemic disease of tool, because it has the quarantine object that the germinal infection ability becomes world Chan Si state.Nearly more than two decades comes, and pebrine disease is national fashion trend in China, eliminates super malicious silkworm egg amount and reaches millions of, the economic loss heaviness.Drug therapy is one of effectively preventing means of current pebrine disease.At the drug treatment of pebrine disease, domestic scholars has been carried out large quantity research, wherein on producing large scale application be the carbendazim powder that I develop.Pebrine disease medicine material more efficient in order to obtain, low toxicity, we have obtained the desirable drug material albendazole (also claiming Albendazole) of result for the treatment of through screening.
Albendazole (albendazole, ABZ) is that in the benzimidazole medicine, the expelling parasite spectrum is wider, and a kind of expelling parasite high-efficiency low-toxicity medicine that insecticidal action is the strongest, be widely used in a plurality of fields such as human medical, livestock and poultry cultivation, aquaculture.It suppresses the absorption of parasite to glucose after metabolism is sulfoxide type or sulfone class in vivo, causes the polypide glycogen depletion, or suppresses the fumaric reductase system, hinders ATP and produces, and parasite can't be survived and breed.Albendazole rapid metabolism in vivo is albendazole-sulfoxide (albendazole sulphoxide, ABZSO), and it is main metabolites, is also the main active of drug effect; And then be converted into Albendazole sulfone (albendazole sulphone, ABZSO
2), finally generate albendazole-2-amino sulfone (albendazole amino sulphone, ABZSO
2-NH
2).As far back as early 1980s, the external existing report of the research to albendazole in different biomaterials and metabolite residue quantity measuring method thereof, Alvinerie and Galtier(1984) when utilizing the forward high performance liquid chromatography to realize in sheep plasma albendazole and two kinds of metabolins thereof, measure, but it fails to detect ABZSO
2-NH
2, and the UV detector sensitivity of using is lower, only reaches 100,000/rank.Utilize equally high performance liquid chromatography, Lanchote etc. (1998) are only to ABZSO and corresponding body and ABZSO
2detect, adopt ground fluorescence detector the former height of remolding sensitivity an order of magnitude.To at the beginning of 21 century, both at home and abroad human medical, livestock and poultry cultivation, aquaculture etc. multi-field to various biological organization materials in the research report of albendazole or benzimidazoles assay method also be enriched gradually.As immunoassay, capillary electrophoresis, gas-matter coupling method, liquid-matter coupling method etc.Prochazkova etc. (2000) utilize the albendazole content in nonaqueous capillary electrophoresis and common two kinds of method comparative determination blood of human body of HPLC, find that the detection effect of the two is suitable.Kitzman etc. (2002) utilize the C18 solid-phase extraction column to be extracted before machine on HPLC the Human Blood containing albendazole, can effectively dispel impurity, get rid of and disturb.Mottier etc. (2003) also utilize solid-phase extraction column, reverse hplc technology the benzimidazole medicament contg in the tapeworm pin main bodies such as sheep, ox to be measured simultaneously.Batzias and Delis(2004) utilize reverse phase liquid chromatography, fluorescence detector to realize sheep plasma albendazole metabolite content Accurate Determining.Although the high performance liquid chromatography application is comparatively extensive, also there is obviously deficiency, the sample preparation process is more loaded down with trivial details, and length consuming time is subject to impurity and disturbs.Recent years, fast development along with modern chromatographic technique, reported successively the research that utilizes ultra high efficiency liquid phase, mass spectrum and supper-fast LC-MS-MS etc. to measure biological tissue's Pesticide Residues both at home and abroad, make largely the method bring into play on the mensuration of biomaterial pesticide concentration that it is with strong points, the highly sensitive and fast superiority of detection rates.Yet, for people or livestock and poultry animal, the silkworm small volume, the hemolymph sampling is difficulty relatively, and sampling amount is restricted, drug concentration is low, add that the endogenous material such as inorganic salts, lipid, protein, self metabolin disturb, the detection of Tissues of Silkworm Bombyx Moril material is proposed to requirements at the higher level, above method all is not suitable for the detection of Tissues of Silkworm Bombyx Moril Chinese traditional medicine.At present, there is not yet for albendazole in silkworm hemolymph and metabolite content thereof and carry out the relevant report of accurate qualitative and quantitative study simultaneously.
Summary of the invention
The purpose of this invention is to provide a kind of method of utilizing the supper-fast liquid chromatography-triple level Four bar mass spectrums (UFLC-MS/MS) of connecting to measure the content of albendazole and metabolin in silkworm hemolymph, the method can be carried out while fast qualitative, quantitative measurement to albendazole in silkworm hemolymph and metabolin thereof.
The objective of the invention is to be achieved through the following technical solutions: a kind of method of utilizing the supper-fast liquid chromatography-triple level Four bar mass spectrums (UFLC-MS/MS) of connecting to measure the content of albendazole and metabolin in silkworm hemolymph comprises the following steps:
(1) get silkworm hemolymph and extracted, obtain test sample solution;
(2) prepare the albendazole of a series of variable concentrations and the mixed standard solution of metabolin thereof, inject respectively supper-fast liquid chromatograph, carry out compartment analysis, and with triple level Four bar mass spectrometer detection material ion signals, obtain the standard analysis data of albendazole and metabolin thereof;
(3), by step (2) method, the test sample solution of getting in step (1) injects supper-fast liquid chromatograph, carries out compartment analysis, and by triple level Four bar mass spectrometer detection material signals, the analysis data of acquisition test sample;
(4) the analysis data of test sample step (3) obtained and the chromatogram analysis data of albendazole and metabolin thereof are compared, and can carry out quantitative and qualitative analysis to albendazole in silkworm hemolymph and metabolin thereof.
The concrete operations of described step (1) are: get silkworm hemolymph and carry out the vortex oscillation treatment before extraction, after the vortex oscillation treatment, the mixed liquor of acetonitrile and ethyl acetate of take is extract, by silkworm hemolymph and extracting liquid volume ratio, be that 1 ︰ 5 ~ 1 ︰ 8 are mixed, vortex vibration 3 ~ 10 min, then by the centrifuging supernatant, residue is repeated to extract once, merge supernatant, centrifugal, membrane filtration, obtain test sample solution.
In step of the present invention (2) and (3), the chromatographic condition of supper-fast liquid chromatograph is: chromatographic column: Agilent ZORBAX C18(4.6 * 100 mm, 3.0 μ m); 40 ℃ of column temperatures; Mobile phase: Yi Jing ︰ 0.005 mol/L formic acid (V ︰ V)=80% ︰ 20% ~ 90% ︰ 10%, Gradient elution; Sample size 10 μ L; Flow velocity 0.1000 ~ 0.2 000 mL/min.
In step of the present invention (2) and (3), triple level Four bar mass spectrometer testing conditions are: ionization mode: ESI(+); Multiple reaction monitoring scanning of the mass spectrum pattern (MRM); Atomization gas (NEB): 6.00 ~ 8.00 psi, gas curtain gas (CUR) 23.00 ~ 26.00 psi, collision gas (CAD): 3.00 ~ 6.00 psi, ionization temperature (TEM): 500.00 ~ 600.00 ℃, spray voltage (IS): 5000.00 ~ 6000.00 V, inject voltage (EP): 8.00 ~ 12.00 V, collision cell penetrates voltage (CXP): 8.00 ~ 12.00 V.
As an embodiment of the invention, the albendazole of a series of variable concentrations of described step (2) and metabolin mixed standard solution thereof are respectively albendazole and the metabolin mixed standard solution thereof that albendazole and metabolite concentration thereof are 5 μ g/L, 62.5 μ g/L, 125 μ g/L, 250 μ g/L, 625 μ g/L and 1 250 μ g/L.
Analysis data of the present invention comprise retention time, peak area and the fragment ion Mass Spectrometer Method result etc. of albendazole and metabolin thereof.In described step (2), the peak area obtained of take is ordinate, and concentration is that horizontal ordinate is drawn albendazole and the corresponding typical curve of metabolin thereof, and obtains albendazole and the corresponding regression equation of metabolin thereof: ABZ:PAR=3.7242 * 10
6c+3.4938 * 10
4, R
2=0.9927; ABZSO:PAR=6.4613 * 10
5c-4.8495 * 10
3, R
2=0.9905; ABZSO
2: PAR=1.0269 * 10
6c+89.1365, R
2=0.9972; ABZSO
2-NH
2: PAR=1.5396 * 10
6c-8.4586 * 10
3, R
2=0.9966.In described step (4), retention time and the fragment ion Mass Spectrometer Method result of comparison test sample and standard items, to albendazole in silkworm hemolymph and metabolin thereof, carry out qualitative, by the peak area data substitution of albendazole and metabolin thereof separately in corresponding regression equation with substance for calculation content, can carry out quantitatively albendazole in silkworm hemolymph and metabolin thereof.
the present invention compared with prior art has the following advantages:
The present invention adopts supper-fast liquid-phase chromatographic analysis sample, can complete a need testing solution analysis within 8 min; Adopt series connection level Four bar mass spectrum MRM monitoring mode simultaneously, selectivity is good, highly sensitive, and can get rid of the interference of other composition in complicated background, the error of having avoided pre-treatment to bring, both are in conjunction with the fast qualitative of having realized easily albendazole in the silkworm hemolymph and metabolin thereof, quantitatively detect, and obviously are better than HPLC method and liquid chromatography-iontrap mass spectroscopy.
The accompanying drawing explanation
Fig. 1-1st, ABZ chromatography of ions figure in hybrid standard product (A);
Fig. 1-2 is ABZ chromatography of ions figure in 2 test samples (A ');
Fig. 2-1st, ABZSO chromatography of ions figure in hybrid standard product (B);
Fig. 2-2nd, ABZSO chromatography of ions figure in test sample (B ');
Fig. 3-1st, ABZSO in hybrid standard product (C)
2chromatography of ions figure;
Fig. 3-2nd, ABZSO in test sample (C ')
2chromatography of ions figure;
Fig. 4-1st, ABZSO in hybrid standard product (D)
2-NH
2chromatography of ions figure;
Fig. 4-2nd, ABZSO in test sample (D ')
2-NH
2chromatography of ions figure;
Fig. 5 is the scanning of the mass spectrum figure of hybrid standard liquid.
Embodiment
materials and methods
1.1 instrument and reagent
Supper-fast liquid chromatography-triple quadrupole bar GC-MS (UFLC-MS/MS) (Japanese Shimadzu, American AB SCIEX company), supersonic wave cleaning machine (SB-5200 DT), Milli-Q ultrapure water apparatus (U.S. Millipore company), Sigma 3k 15 hydro-extractors, electronic analytical balance, vortex mixer (German IKA company), 0.22 μ m micropore
pTFEfilter membrane (experimental facilities company limited is risen in Tianjin, Tianjin), disposable medical asepsis injector etc.
ABZ(99.0%) and ABZSO
2(98.3%) standard items are all purchased from Dr. Ehrenstorfer GmbH, Germany; ABZSO(>=98%) and ABZSO
2-NH
2(98%) respectively purchased from Sigma-Aldrich, USA and Alfa Aesar, USA.Acetonitrile, methyl alcohol etc. are chromatographically pure, respectively purchased from Thermo Fisher Scientific and Oceanpak alexative chemical., and Ltd, all the other reagent are for analyzing pure rank.
standard specimen and test sample are prepared
1.2.1 test is processed
250, the test site of 3 repetitions and ，Mei district, 1 clear area established in test, plays the mulberry leaf that silkworm first feeding feeding soaked with 2000 mg/L albendazole solution five ages and once continues with normal mulberry afterwards, the normal mulberry of clear area feeding.After adding food medicine mulberry, within 2 hours, rise, 30 silkworms are got in each every district at random, get hemolymph from caudal horn and are mixed into a test sample, sample altogether 5 times, are placed in immediately-20 ℃ of refrigerators after every sub-sampling and save backup.
1.2.2 sample preparation
Test sample solution preparation: silkworm hemolymph sample vortex before extracting 1 min that vibrates, extract adopts the mixed liquor of Yi Jing ︰ ethyl acetate=5 ︰ 1.Get 1 mL silkworm hemolymph sample, add 5 mL extracts, vortex 10 min that vibrate; Isolating residue adds 2 mL extracts to repeat to extract once again; Merge supernatant, high speed centrifugation 5 min under 10 000 r/min conditions, cross 0.22 μ m micropore before machine on filtrate
pTFEfilter membrane, obtain test sample solution.
The preparation of negative sample solution: prepared by the hemolymph that blank test district silkworm is obtained as stated above.
1.2.3 standard reserving solution preparation
Test is carried out qualitative, quantitative with external standard method.Accurately take ABZ, ABZSO, ABZSO
2, ABZSO
2-NH
2each 0.01 g of standard items, dissolve with the vibration of acetonitrile lucifuge, makes the hybrid standard storing solution of 20 mg/L, is placed in 4 ℃ of refrigerators standby.During mensuration with the albendazole of variable concentrations gradient, albendazole-sulfoxide, Albendazole sulfone, the amino sulfone standard items of albendazole-2-drawing standard curve.
conditional parameter
1.3.1 liquid phase chromatogram condition
Liquid chromatograph is Shimadzu Prominence UFLC(ULTRA FAST LIQUID CHROMATOGRAPH), chromatographic column: Agilent ZORBAX C18(4.6 * 100 mm, 3.0 μ m); 40 ℃ of column temperatures; Mobile phase: Yi Jing ︰ 0.005 mol/L formic acid (V ︰ V)=(85% ︰ 15%), Gradient elution; Sample size 10 μ L; Flow velocity 0.2 000 mL/min.
the mass spectrum condition
Mass spectrometer is API 4000 MS/MS(AB SCIEX, USA), triple quadrupole bar mass spectrum; Ionization mode: ESI(+); Multiple reaction monitoring scanning of the mass spectrum pattern (MRM); Atomization gas (NEB): 7.00 psi, gas curtain gas (CUR) 25.00 psi, collision gas (CAD): 5.00 psi, ionization temperature (TEM): 550.00 ℃, spray voltage (IS): 5500.00 V, inject voltage (EP): 10.00 V, collision cell penetrates voltage (CXP): 10.00 V.Albendazole and metabolin thereof are at ESI(+) [M+H]+the MRM pattern under the mass spectrum parameter in Table 1.
1.4 data statistic analysis
Utilize automatic integration, the chromatogram mass spectrum map generalization of Analyst analysis software to liquid mass signal data analysis, material peak area; Utilize 2003 pairs of data of Microsoft Office Excel to carry out the statistical study such as calculating of content of material calculating, linear regression analysis, standard relative deviation.
method validation
2.1 qualitative analysis
125 μ g/L mixed standard solutions, silkworm hemolymph test sample solution is sample introduction 10 μ L respectively, and gained UFLC-MS/MS quota ion reconstruct mass of ion chromatogram is shown in Fig. 1-1～Fig. 4-2, and mixed standard solution ion scan mass spectrum is shown in Fig. 5.Under selected chromatographic condition, determinand ABZ, ABZSO, ABZSO
2, ABZSO
2-NH
2the chromatographic peak retention time be respectively 2.45,2.01,2.02,2.27 min, each determinand in test sample and the retention time of hybrid standard product are basically identical.Peak shape is good, and impurity peaks disturbs and can ignore on every side.The signal to noise ratio (S/N ratio) S/N at the reconstruct chromatography of ions peak of the qualitative ion pair of determinand >=3, the signal to noise ratio (S/N ratio) S/N at the reconstruct chromatography of ions peak that quota ion is right >=10; In addition, the relative abundance of ions value of each qualitative ion pair meets the requirement (table 2) (Commission of the European Community, 2002) of the relevant mass spectrometry of European Union, so far can judge in test sample and contain albendazole and metabolin thereof; Otherwise sample is not containing this chemical substance.
quantitative test
To albendazole and the metabolin thereof contained in the silkworm hemolymph test sample, respectively by its quota ion to (
m/z) peak area of (referring to table 1) carries out cubage.Methodological study is as follows:
2.2.1 linear relationship
The hybrid standard storing solution of 20 mg/L is configured to the typical curve sample that concentration is respectively 5 μ g/L, 62.5 μ g/L, 125 μ g/L, 250 μ g/L, 625 μ g/L, 1 250 μ g/L, by measured peak area (peak area ratio, PAR) and concentration (C, μ g/L) carry out linear regression analysis, obtain ABZ, ABZSO, ABZSO
2, ABZSO
2-NH
2the standard solution regression equation is respectively: ABZ:PAR=3.7242 * 10
6c+3.4938 * 10
4, R
2=0.9927; ABZSO:PAR=6.4613 * 10
5c-4.8495 * 10
3, R
2=0.9905; ABZSO
2: PAR=1.0269 * 10
6c+89.1365, R
2=0.9972; ABZSO
2-NH
2: PAR=1.5396 * 10
6c-8.4586 * 10
3, R
2=0.9966, all be good linear relationship.
2.2.2 detectability (LOD) and quantitative limit (LOQ)
When sample size is 10 μ L, during S/N=3, ABZ, ABZSO, ABZSO
2, ABZSO
2-NH
2lOD be respectively 0.39 ng/mL, 5.00 ng/mL, 0.23 ng/mL, 1.78 ng/mL; When sample size is 10 μ L, during S/N=10, LOQ is respectively 1.32 ng/mL, 16.67 ng/mL, 0.76 ng/mL, 5.94 ng/mL.
2.2.3 repeated---precision test
Mixed standard solution and the test sample of 125 μ g/L, 625 μ g/L, tri-kinds of concentration of 1 250 μ g/L are respectively repeated to sample introduction 6 times, and the RSDs of retention time and peak area is in Table 3 separately.Test findings shows, this method has good reappearance.
2.2.4 the recovery---accuracy test
Get 6 parts, the negative silkworm hemolymph sample (each 0.5 mL) made by 1.2.2, each precision adds albendazole and metabolin hybrid standard product 0.5 mL thereof that concentration is 625 μ g/L, and vortex mixes, and crosses the organic miillpore filter of 0.22 μ m, and sample introduction is measured.ABZ, ABZSO, ABZSO
2, ABZSO
2-NH
2the recovery be respectively 81.60%～95.16%, 76.88%～84.31%, 87.39%～99.86%, 73.25%～76.47%; RSD is respectively: 5.26%, 3.61%, 4.65%, 1.67%.Details are in Table 4.
The present invention can summarize with other the concrete form without prejudice to spirit of the present invention or principal character.The above embodiment of the present invention all can only think explanation of the present invention rather than restriction, and any trickle modification, equivalent variations and modification that every foundation essence technology of the present invention is done above embodiment, all belong in the scope of technical solution of the present invention.
Claims (6)
1. a method of utilizing the content of albendazole and metabolin in supper-fast liquid chromatography-triple level Four bar mass spectroscopy silkworm hemolymphs of connecting, is characterized in that, comprises the following steps:
(1) get silkworm hemolymph and extracted, obtain test sample solution;
(2) prepare the albendazole of a series of variable concentrations and the mixed standard solution of metabolin thereof, inject respectively supper-fast liquid chromatograph, carry out compartment analysis, and with triple level Four bar mass spectrometer detection material ion signals, obtain the standard analysis data of albendazole and metabolin thereof;
(3), by step (2) method, the test sample solution of getting in step (1) injects supper-fast liquid chromatograph, carries out compartment analysis, and by triple level Four bar mass spectrometer detection material signals, the analysis data of acquisition test sample;
(4) the analysis data of test sample step (3) obtained and the chromatogram analysis data of albendazole and metabolin are compared, and can carry out quantitative and qualitative analysis to albendazole in silkworm hemolymph and metabolin thereof.
2. the method for utilizing the content of albendazole and metabolin in supper-fast liquid chromatography-triple level Four bar mass spectroscopy silkworm hemolymphs of connecting according to claim 1, it is characterized in that, the concrete operations of described step (1) are: get silkworm hemolymph and carry out the vortex oscillation treatment before extraction, after the vortex oscillation treatment, the mixed liquor of acetonitrile and ethyl acetate of take is extract, by silkworm hemolymph and extracting liquid volume ratio, be that 1 ︰ 5 ~ 1 ︰ 8 are mixed, vortex vibration 3 ~ 10 min, then by the centrifuging supernatant, adopt said method to repeat to extract once to residue, merge supernatant, centrifugal, filter, obtain test sample solution.
3. the method for utilizing the content of albendazole and metabolin in supper-fast liquid chromatography-triple level Four bar mass spectroscopy silkworm hemolymphs of connecting according to claim 1, it is characterized in that, in described step (2) and (3), the chromatographic condition of supper-fast liquid chromatograph is: chromatographic column: Agilent ZORBAX C18,4.6 * 100 mm, 3.0 μ m; 40 ℃ of column temperatures; Mobile phase: Yi Jing ︰ 0.005 mol/L formic acid=80% ︰ 20% ~ 90% ︰ 10%, Gradient elution; Sample size 10 μ L; Flow velocity 0.1000 ~ 0.2 000 mL/min.
4. the method for utilizing the content of albendazole and metabolin in supper-fast liquid chromatography-triple level Four bar mass spectroscopy silkworm hemolymphs of connecting according to claim 1, it is characterized in that, in described step (2) and (3), triple level Four bar mass spectrometer testing conditions are: ionization mode: ESI(+); Multiple reaction monitoring scanning of the mass spectrum pattern; Atomization gas: 6.00 ~ 8.00 psi, gas curtain gas 23.00 ~ 26.00 psi, collision gas: 3.00 ~ 6.00 psi, ionization temperature: 500.00 ~ 600.00 ℃, spray voltage: 5000.00 ~ 6000.00 V, inject voltage: 8.00 ~ 12.00 V, collision cell penetrates voltage: 8.00 ~ 12.00 V.
5. the method for utilizing the content of albendazole and metabolin in supper-fast liquid chromatography-triple level Four bar mass spectroscopy silkworm hemolymphs of connecting according to claim 1, it is characterized in that, the albendazole of a series of variable concentrations of described step (2) and metabolin mixed standard solution thereof are respectively albendazole and the metabolin mixed standard solution thereof that albendazole and metabolite concentration thereof are 5 μ g/L, 62.5 μ g/L, 125 μ g/L, 250 μ g/L, 625 μ g/L and 1 250 μ g/L.
6. the method for utilizing the content of albendazole and metabolin in supper-fast liquid chromatography-triple level Four bar mass spectroscopy silkworm hemolymphs of connecting according to claim 1, it is characterized in that, the analysis data in described step (2) and step (3) comprise retention time, peak area and the fragment ion Mass Spectrometer Method result of albendazole and metabolin thereof; In described step (2), the peak area obtained of take is ordinate, and concentration is that horizontal ordinate is made albendazole and the corresponding typical curve of metabolin thereof, and obtains albendazole and the corresponding regression equation of metabolin thereof; In described step (4), retention time and the fragment ion Mass Spectrometer Method result of comparison test sample and standard items, to albendazole in silkworm hemolymph and metabolin thereof, carry out qualitative, by the peak area data substitution of albendazole and metabolin thereof separately in corresponding regression equation with substance for calculation content, albendazole in silkworm hemolymph and metabolin thereof are carried out quantitatively.
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| CN106770765A (en) * | 2016-12-21 | 2017-05-31 | 潍坊海润华辰检测技术有限公司 | The detection method and application of a kind of albendazole and its metabolin |
| CN113588834A (en) * | 2021-08-04 | 2021-11-02 | 沈阳伟嘉生物技术有限公司 | HPLC (high performance liquid chromatography) detection method and application of albendazole and metabolite thereof in piglet plasma |
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| 陈翠兰: "盐酸氧阿苯达唑及其代谢产物在鲫体内的药物动力学及残留研究", 《中国优秀硕士学位论文全文数据库 医药卫生科技辑》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103969374A (en) * | 2014-05-27 | 2014-08-06 | 广东省农业科学院蚕业与农产品加工研究所 | Method for determining retention capacity of albendazole in mulberry leaves |
| CN106770765A (en) * | 2016-12-21 | 2017-05-31 | 潍坊海润华辰检测技术有限公司 | The detection method and application of a kind of albendazole and its metabolin |
| CN106770765B (en) * | 2016-12-21 | 2019-10-29 | 潍坊海润华辰检测技术有限公司 | The detection method and application of a kind of albendazole and its metabolin |
| CN113588834A (en) * | 2021-08-04 | 2021-11-02 | 沈阳伟嘉生物技术有限公司 | HPLC (high performance liquid chromatography) detection method and application of albendazole and metabolite thereof in piglet plasma |
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| CN103487539B (en) | 2015-06-03 |
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