CN102375040A - Method for determining content of simimtecan and chimmitecan in human or animal whole blood - Google Patents
Method for determining content of simimtecan and chimmitecan in human or animal whole blood Download PDFInfo
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- CN102375040A CN102375040A CN2010102607580A CN201010260758A CN102375040A CN 102375040 A CN102375040 A CN 102375040A CN 2010102607580 A CN2010102607580 A CN 2010102607580A CN 201010260758 A CN201010260758 A CN 201010260758A CN 102375040 A CN102375040 A CN 102375040A
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Abstract
The invention provides a method for determining the content of simimtecan and chimmitecan in human or animal whole blood. The method comprises the steps of pretreating human or animal whole blood, carrying out separation on blood plasma and separating and detecting simimtecan and chimmitecan in the blood plasma by using the method of separation and detection; the invention is characterized in that the human or animal whole blood contacts with one carboxylesterase inhibitor or more carboxylesterase inhibitors in the process of pretreatment. According to the invention, the carboxylesterase inhibitors are utilized to block in vitro hydrolysis of simimtecan into chimmitecan, which guarantees that the result of in vivo exposure level inspection on the prodrug of simimtecan and the metabolite chimmitecan and the result of efficiency of conversion of the prodrug into the metabolite are reliable.
Description
Technical field
The invention belongs to the Pharmaceutical Analysis field, relate to a kind of bright method of replacing health and lucky miaow to replace Kang Hanliang of happiness of measuring.More specifically, relate to the bright method of replacing health and lucky miaow to replace Kang Hanliang of happiness in a kind of human or animal's of mensuration whole blood.
Background technology
Liking bright is the camptothecin derivant of newly developing for health (9-allyl-10-(4-piperidinyl piperidine) formyloxy camptothecine), is used to treat malignant tumour.This compound can be hydrolyzed into active metabolite-Ji miaow through carboxy-lesterase in vivo and produce antineoplastic curative effect for health (reaction equation 1).Carry out clinical before with CLINICAL PHARMACOKINETIS STUDY ON in, need investigate exposure level in its body for health and lucky miaow for the Kang Jinhang quantitative test to the happiness in animal and human's body plasma sample is bright.
According to the Literature Consult result, no matter like animal blood samples such as bright rat, mouse in the existing analytical approach for health in processing, still when bright human blood sample for health is liked in processing, all do not take the active method of any blocking-up carboxy-lesterase.Wherein, some bibliographical informations adopts after the blood sampling immediately centrifugal separation plasma and cryopreservation until the method for analyzing
[1-7], some bibliographical information adopts whole blood to gather rearmounted method of preserving on ice
[2,4,7], some document is not mentioned related measure when gathering whole blood sample
[8-11]
It is extremely unstable in rat plasma to like the bright health of replacing, even also can be generated lucky miaow by the carboxy-lesterase hydrolysis in the blood plasma for health external.If inhibition is timely and effectively liked the bright Kang Jixu of replacing and is hydrolyzed to lucky miaow for health when zoopery is taken a blood sample, can cause the bright interior exposure level miaow on the low side and lucky of health body that replaces of happiness to replace the higher illusion of exposure level in the health body, cause the inaccuracy of experimental result.
The present invention is intended to like the bright ubiquitous false positive results proposition of the conversion ratio rational solution that is converted into active metabolite for health to current camptothecin cancer therapy drug Ji miaow for exposure level in the health body and prodrug: promptly in analytical approach, adopt optionally carboxy-lesterase suppressant-BNPP; External carboxy-lesterase in the whole blood is suppressed; Make the bright Kang Buzai of replacing of prodrug happiness continue to be converted into lucky miaow for health, thereby guaranteed the accuracy of analytical approach effectively external.
List of references:
1.Escoriaza?J，Aldaz?A，Castellanos?C，Calvo?E，Giráldez?J.(2000)Simple?and?rapid?determination?of?irinotecan?and?its?metabolite?SN-38?in?plasma?by?high-performance?liquid-chromatography：application?to?clinical?pharmacokinetic?studies.J.Chromatogr.B，740：159-168.
2.Owens?TS，Dodds?H，Fricke?K，Hanna?SK，Crews?KR.(2003)High-performance?liquid?chromatographic?assay?with?fluorescence?detection?for?the?simultaneous?measurement?of?carboxylate?and?lactone?forms?of?irinotecan?and?three?metabolites?in?human?plasma.J.Chromatogr.B，788：65-74.
3.Escoriaza?J，Aldaz?A，Castellanos?C，Calvo?E，Giráldez?J.(2000)Simple?and?rapid?determination?of?irinotecan?and?its?metabolite?SN-38?in?plasma?by?high-performance?liquid-chromatography：application?to?clinical?pharmacokinetic?studies.J.Chromatogr.B，740：159-168.
4.de?Jong?FA，Mathijssen?RH，de?Bruijn?P，Loos?WJ，Verweij?J，Sparreboom?A.(2003)Determination?of?irinotecan?(CPT-11)and?SN-38?in?human?whole?blood?and?red?blood?cells?by?liquid?chromatography?with?fluorescence?detection.J.Chromatogr.B，795：383-388.
5.Poujol?S，Pinguet?F，Malosse?F，Astre?C，Ychou?M，Culine?S，Bressolle?F.(2003)Sensitive?HPLC-fluorescence?method?for?irinotecan?and?four?major?metabolites?in?human?plasma?and?saliva：application?to?pharmacokinetic?studies.Clin.Chem.，49：1900-1908.
6.Yang?X，Hu?Z，Chan?SY，Goh?BC，Duan?W，Chan?E，Zhou?S.(2005)Simultaneous?determination?of?the?lactone?and?carboxylate?forms?of?irinotecan?(CPT-11)and?its?active?metabolite?SN-38?by?high-performance?liquid?chromatography：application?to?plasma?pharmacokinetic?studies?in?the?rat.J.Chromatogr.B，821：221-228.
7.Khan?S，Ahmad?A，Guo?W，Wang?YF，Abu-Qare?A，Ahmad?I.(2005)A?simple?and?sensitive?LC/MS/MS?assay?for?7-ethyl-10-hydroxycamptothecin?(SN-38)in?mouse?plasma?and?tissues：application?to?pharmacokinetic?study?of?liposome?entrapped?SN-38(LE-SN38).J.Pharm.Biomed.Anal.，37：135-142.
8.Loos?WJ，de?Bruijn?P，Verweij?J，Sparreboom?A.(2000)Determination?of?camptothecin?analogs?in?biological?matrices?by?high-performance?liquid?chromatography.Anticancer?Drugs，11：315-324.
9.Vali?AM，Shafaghi?B，Dadashzadeh?S.(2005)Simple?and?sensitive?high?performance?liquid?chromatographic?method?for?the?simultaneous?quantitation?ofthe?lactone?and?carboxylate?forms?of?topotecan?in?human?plasma.J.Chromatogr.B，818：205-212.
10.D′Esposito?F，Tattam?BN，Ramzan?I，Murray?M.(2008)A?liquid?chromatography/electrospray?ionization?mass?spectrometry?(LC-MS/MS)assay?for?the?determination?of?irinotecan?(CPT-11)and?its?two?major?metabolites?in?human?liver?microsomal?incubations?and?human?plasma?samples.J.Chromatogr.B，875：522-530.
11.Bardin?S，Guo?W，Johnson?JL，Khan?S，Ahmad?A，Duggan?JX，Ayoub?J，Ahmad?I.(2005)Liquid?chromatographic-tandem?mass?spectrometric?assay?for?the?simultaneous?quantification?of?Camptosar?and?its?metabolite?SN-38?in?mouse?plasma?and?tissues.J.Chromatogr.A，1073：249-255.
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Summary of the invention
In order to improve the bright accuracy of replacing health and lucky miaow to replace the Kangding component analysis of happiness in animal and human's body whole blood/plasma sample; Carboxy-lesterase suppressant blocking-up happiness is bright to generate lucky miaow for health for health in external continuation hydrolysis through adopting in the present invention, thereby has guaranteed the bright reliability that is converted into the transformation efficiency result of metabolic product for the lucky miaow of health and metabolic product for exposure level investigation and prodrug in the health body of prodrug happiness.This technical scheme is equally applicable to the plasma sample analysis of other camptothecine compounds (one-tenth ester prodrugs).
Therefore, the purpose of this invention is to provide the bright method of replacing health and lucky miaow to replace Kang Hanliang of happiness in a kind of human or animal's of mensuration whole blood.
Happiness is bright for health and the lucky miaow method for Kang Hanliang in a kind of human or animal's of mensuration whole blood that provides according to the object of the invention; After this method comprises that the whole blood to the human or animal carries out pre-service; Separated plasma adopts method for separating and detecting to separate for Kang Jinhang and detect for health and lucky miaow happiness in the blood plasma is bright, wherein again; Carry out in the preprocessing process at whole blood, human or animal's whole blood is contacted with one or more carboxy-lesterase suppressant the human or animal.
Even because external, the happiness in human or animal's whole blood is bright also can be generated lucky miaow by carboxy-lesterase hydrolysis wherein for health for health.Therefore, for the bright hydrolysis for health of inhibition happiness more early, the human or animal's of collection whole blood should contact with one or more carboxy-lesterase suppressant as early as possible.For example, one or more carboxy-lesterase suppressant can preexist in the heparin tube; Perhaps the blood sampling back adds the carboxy-lesterase suppressant rapidly in human or animal's whole blood.
Preferably, said one or more carboxy-lesterase suppressant can preexist in the heparin tube.Thus, the human or animal's of collection whole blood can contact with these one or more carboxy-lesterase suppressant in time, is hydrolyzed to lucky miaow for health thereby suppress the bright health of replacing of happiness more effectively.
For the human or animal's that make to gather whole blood contacts with one or more carboxy-lesterase suppressant fully, preferably these one or more carboxy-lesterase suppressant are added in the heparin tube with form of powder, and when contacting, shake up rapidly.
Described carboxy-lesterase suppressant can be two (p-nitrophenyl) phosphates (BNPP, bis-p-nitrophenyl-phosphate), TTA (thenoyltrifluoroacetone) or dibenzoyl (benzil).
Preferably, described carboxy-lesterase suppressant is BNPP.BNPP is a species specific carboxy-lesterase suppressant, and its inhibition to carboxy-lesterase is a nonreversibility.Wadkins etc. carry out people's small intestine carboxy-lesterase suppressant synthetic during with activity identification with BNPP as the positive control compound
[12]The inventor investigates carboxy-lesterase inhibiting effect in the rat plasma of BNPP, and its carboxy-lesterase maximum inhibition can reach more than 98%, therefore selects the carboxy-lesterase suppressant of BNPP as the rat blood sample for use.
(for example, the ratio of ml BNPP): mg is 1: 5～1: 10, is preferably 1: 5, can suppress to like bright hydrolysis for health so basically fully for said human or animal's whole blood and said carboxy-lesterase suppressant.
For the rapid blood coagulation of whole blood that prevents the human or animal, the inwall of said heparin tube can be coated with anticoagulant, for example liquaemin.
In said assay method, the process of separated plasma comprises and (for example, 3000rpm) obtains blood plasma with centrifugal with human or animal's after one or more carboxy-lesterase suppressant fully contact whole blood.Further blood plasma is carried out albumen precipitation and centrifugally (for example, 13000rpm), obtain supernatant and be used for separation detection with acetonitrile.
Described method for separating and detecting can adopt well known by persons skilled in the art can reaching brightly to separate for Kang Jinhang and any method of testing goal for health and lucky miaow liking.
Preferably, described separation method is a high performance liquid chromatography.
Said high performance liquid chromatography is used highly effective liquid phase chromatographic system, and actual conditions those skilled in the art of this system just can obtain through simple experiment.Also can be referring to document 1-11.
Preferably, the condition of said highly effective liquid phase chromatographic system is:
Chromatographic column: C
18(50mm * 2.1mm; 5 μ m);
Column temperature: room temperature;
Moving phase: solvent orange 2 A: CH
3CN-H
2O (v/v, 1: 99; 0.4 ‰ HCOOH; 0.05 ‰ HCOONH
4); Solvent B:CH
3CN-H
2O (v/v, 99: 1; 0.4 ‰ HCOOH; 0.05 ‰ HCOONH
4);
Condition of gradient elution: 0～0.5min is with 4% solvent B wash-out, and 0.5～2min solvent B at the uniform velocity increases to 100%, 2～3min with 100% solvent B wash-out from 4%, and 3～4min carries out column equilibration with 4% solvent B;
Flow velocity: 0.40mL/min;
Sample size: 5 μ L.
Said detection method is (series connection) Mass Spectrometer Method, is specially: adopt positive ion electron spray ionisation pattern (ESI+), under the collision energy of MRM pattern and 49V, detect bright m/z 599 → 124 ion pairs for health of happiness; Under the collision energy of 35V, detect m/z 405 → 361 ion pairs of lucky miaow for health.
It will be understood by those skilled in the art that for happiness is bright and replace the health Determination on content for health and lucky miaow, can adopt separation or detection method under the different condition to carry out, happiness is bright replaces health for health or lucky miaow as long as these methods are fit to.
Certainly, for ease, preferably use separation or detection method under the same terms, can reach the purpose of fast measuring like this.
Happiness is bright in the mensuration of the present invention human or animal whole blood is specially for health and the lucky miaow method for Kang Hanliang: be coated with liquaemin and add the whole blood that adds the human or animal in the heparin tube of carboxy-lesterase suppressant at inwall in advance; Shake up gently; Centrifuging obtains blood plasma; In this blood plasma, add acetonitrile and carry out albumen precipitation, obtain supernatant after centrifugal; Said supernatant is separated in the LC-MS appearance and detects.
It will be understood by those skilled in the art that in assay, also need the preparation standard curve.
Happiness is bright in mensuration human or animal whole blood of the present invention replaces in the method for Kang Hanliang for health and lucky miaow; When carrying out the standard curve making of quantitative test; Take preprocess method preparation happiness same as described above bright: promptly in the heparin tube that has the carboxy-lesterase suppressant in advance, to add blank plasma or in blank plasma, (for example add a certain amount of carboxy-lesterase suppressant for health standard plasma solutions; BNPP) behind the powder; Add again that happiness is bright to be mixed with for the health standard items for health and lucky miaow that to contain the finite concentration happiness bright for health and the lucky miaow standard plasma solutions for health; And dilute with the blank plasma that contains the carboxy-lesterase suppressant, be mixed with the happiness of a series of concentration bright for health and lucky miaow for health standard plasma solutions.Carry out albumen precipitation for health and lucky miaow for adding acetonitrile in the health standard plasma solutions to this happiness is bright then, the supernatant that obtains after centrifugal is used for LC-MS and separates and detect.
In the assay method of the present invention, adopt specific carboxy-lesterase suppressant (for example, BNPP) carboxy-lesterase in the blood plasma to be suppressed effectively, be hydrolyzed into lucky miaow for health for health in external continuing thereby the blocking-up happiness is bright.Thereby guaranteed the bright reliability that is converted into the transformation efficiency result of metabolic product for the lucky miaow of health and metabolic product for exposure level investigation and prodrug in the health body of prodrug happiness.
Being appreciated that assay method of the present invention can be applied to can be by in the assay of any compound of carboxy-lesterase hydrolysis.As long as being present in the medium (for example, blood plasma or serum) simultaneously, compound of measuring and carboxy-lesterase get final product.
Description of drawings
Fig. 1 is bright for after the health for rat vein gives (dosage is 2.0mg/kg) happiness, and it is bright for health (left scale) and the lucky miaow of the metabolic product blood concentration-time plot for health (right scale) to adopt the method for prior art to measure happiness.
It is bright for after the health that Fig. 2 is that rat (female) vein (i.v.) gives (dosage is 3.75mg/kg, 7.5mg/kg, 15mg/kg) happiness, and it is bright for health and the lucky miaow blood concentration-time plot for health to adopt method of the present invention to measure happiness.
Whether Fig. 3 is for adding the carboxy-lesterase suppressant to liking bright influence for health blood plasma typical curve result in the rat plasma under the external room temperature different condition.(A) add the bright health blood plasma typical curve that replaces of happiness that carboxy-lesterase inhibitor B NPP obtains; (B) do not add the bright health blood plasma typical curve that replaces of happiness that carboxy-lesterase inhibitor B NPP obtains.
Fig. 4 LC-MS detection method is together measured the bright chromatogram that replaces health and lucky miaow to replace health of happiness in the rat plasma.A, blank rat plasma sample; B has added in the blank rat plasma sample and has liked the bright standard items that replace health and lucky miaow to replace health; C, plasma sample after the rat vein administration.
Embodiment
LC-MS system: the AcQuity that liquid phase-mass spectrometry analytic system (LC/MS/MS) is produced by U.S. Waters company
TMThe API4000 Qtrap mass spectrometer that UPLC series liquid chromatograph appearance and Applied Biosystems company produce is formed, and system works software is EMPOWER Pro (U.S.) and Analysis1.4.2 (U.S.), controls liquid phase and mass spectrometer system respectively.
HPLC level acetonitrile (CH
3CN), methyl alcohol (MeOH), formic acid (HCOOH) and ammonium formate (HCOONH
4) be the reagent that U.S. Merck company produces.Experimental water is prepared by Millipore Direct-Q.Other organic reagent is provided by China Medicine (Group) Shanghai Chemical Reagent Co.,, analyzes pure.
Experimental technique:
1, animal blood specimen collection: add carboxy-lesterase inhibitor B NPP (every 1mL blood adds 5mg approximately) rapidly in animal blood sampling back, shake up back centrifuging and taking blood plasma, blood plasma is to be measured in-70 ℃ of preservations.
2, rat plasma sample pre-treatments: plasma sample (the 50 μ L) room temperature of-70 ℃ of freezing preservations is thawed; Acid acetonitrile solution (containing 0.5% formic acid) the 150 μ L joltings that add the interior mark compound contain 100ng/mL extract (1600rpm, 5min), centrifugal (13; 000rpm; 5min, 4 ℃) after, get supernatant and be used for analyzing.
3, LC-MS analysis condition: chromatographic column: Agilent Eclipse Plus C
18(50mm * 2.1mm; 5 μ m, Agilent, USA); Column temperature: room temperature; Moving phase: solvent orange 2 A: CH
3CN-H
2O (v/v, 1: 99; 0.4 ‰ HCOOH; 0.05 ‰ HCOONH
4); Solvent B:CH
3CN-H
2O (v/v, 99: 1; 0.4 ‰ HCOOH; 0.05 ‰ HCOONH
4); Condition of gradient elution: 0～0.5min is with 4% solvent B wash-out, and 0.5～2min solvent B at the uniform velocity increases to 100%, 2～3min with 100% solvent B wash-out from 4%, and 3～4min carries out column equilibration with 4% solvent B; Flow velocity: 0.40mL/min; Sample size: 5 μ L.Mass Spectrometer Method adopts positive ion electron spray ionisation pattern (ESI+), and under the collision energy of MRM pattern and 49V, it is bright for health m/z 599 → 124 ion pairs to detect happiness; Under the collision energy of 35V, detect m/z 405 → 361 ion pairs of lucky miaow for health, the mass spectrum running parameter is seen table 1.
The bright mass spectrum running parameter that replaces health and lucky miaow to replace health of table 1 application liquid matter coupling technique quantitative test simultaneously happiness
4, the preparation of typical curve: in the blank rat plasma that contains BNPP (blood plasma (ml): BNPP (mg)=1: 5), add the bright acetonitrile standard reserving solution (concentration is 100 μ g/mL) that replaces health and lucky miaow to replace health of happiness; Be made into that to be equivalent to contain happiness bright be the plasma sample of 1000ng/mL for health and lucky miaow for health concentration; Dilute with the blank rat plasma that contains BNPP at interval by 3 times then, the preparation happiness is bright to be the series of standards sample of 1.37～1000ng/mL for health and lucky miaow for the health concentration range.Behind " experimental technique 2 " described method processing standard model (50 μ L), the method by " experimental technique 3 " obtains sample detection response (ratio of test compound and interior mark chromatographic peak peak area) again.(X ng/mL) is independent variable, be dependent variable to detect response (Y), carries out linear regression with least square method, and is weight coefficient with 1/X, tries to achieve the regression equation of mensuration effect and concentration with the sign concentration of above-mentioned standard blood sample.
Adopt bibliographical information
[2]Method (that is: centrifugal separation plasma and cryopreservation immediately after whole blood is gathered; In the analytic process sample is put in the ice bath; And after blood sampling, do not block the carboxy-lesterase activity immediately) measured vein and like bright brightly for health and the lucky miaow concentration for health for happiness in the rat plasma of health (2mg/kg) back, resultant their blood concentration-time curve is seen Fig. 1.Wherein, AUC
Like bright for healthBe 170 ± 6ngh/mL, AUC
Lucky miaow is for healthBe 105 ± 4ngh/mL, AUC
Lucky miaow is for health/ AUC
Like bright for healthRatio be 61.8%.
After adopting blood sampling of the present invention, add the active method of carboxy-lesterase inhibitor B NPP blocking-up carboxy-lesterase immediately; When giving to rat (female) vein (i.v.) that (dosage is 3.75mg/kg, 7.5mg/kg, 15mg/kg) happiness is bright to be analyzed for the blood sample after the health; Like and brightly continue to be converted into lucky miaow and obtained good restraining for health for health is external, resultant happiness is bright sees Fig. 2 for health and lucky miaow for the blood concentration-time curve of health.AUC under three dosage
Lucky miaow is for health/ AUC
Like bright for healthRatio be respectively 6.74% (3.75mg/kg dose groups), 4.51% (7.5mg/kg dose groups) and 3.74% (15mg/kg dose groups).
Add the carboxy-lesterase suppressant under the embodiment 2 external room temperature different conditions in the rat plasma and whether be converted into of the influence of lucky miaow for health to liking the bright health of replacing
Experimental technique:
1, specimen preparation: to the blank rat plasma that contains BNPP (blood plasma (ml): BNPP (mg)=1: 5) or do not contain and add the bright acetonitrile standard reserving solution (concentration is 100 μ g/mL) of happiness in the blank rat plasma of BNPP, be made into that to be equivalent to contain happiness bright be the plasma sample of 333ng/mL and 37.1ng/mL for health concentration for health.
2, rat plasma sample pre-treatments: with embodiment 1.
3, LC-MS analysis condition: with embodiment 1.
The result sees table 2.
Add carboxy-lesterase inhibitor B NPP under the external room temperature different condition of table 2 in the rat plasma and whether be converted into of the influence of lucky miaow for health to liking the bright health of replacing
The result of table 2 shows, in heparin tube, add a certain amount of carboxy-lesterase inhibitor B NPP in advance after, like and brightly continue to be converted into lucky miaow and obtained complete inhibition external for health for health, like does not brightly have conversion for health, does not detect lucky miaow in the sample and replaces health; And in heparin tube, do not add the carboxy-lesterase suppressant, liking bright conversion ratio for health is 63.4-66.0%.
Whether add the carboxy-lesterase suppressant under the embodiment 3 external room temperature different conditions in the rat plasma to liking bright influence for health blood plasma typical curve result
Experimental technique:
1, the preparation of typical curve: to the blank rat plasma that contains BNPP (blood plasma (ml): BNPP (mg)=1: 5) with do not contain and add the bright acetonitrile standard reserving solution (concentration is 100 μ g/mL) of happiness in the blank rat plasma of BNPP for health; Be made into be equivalent to contain the happiness bright for health concentration be the plasma sample of 6000ng/mL; Dilute with the blank rat plasma that contains BNPP at interval by 3 times then, the preparation happiness is bright to be the series of standards sample of 1.37～6000ng/mL for the health concentration range.
2, rat plasma sample pre-treatments: with embodiment 1.
3, LC-MS analysis condition: with embodiment 1.
It is bright for the concentration range internal linear good (r=0.9941) of health typical curve at 1.37-6000ng/mL in blood plasma, to add the latter made happiness of carboxy-lesterase inhibitor B NPP; Do not add the bright health typical curve that replaces of the latter made happiness of carboxy-lesterase suppressant in the blood plasma in same concentration range internal linear relatively poor (r=0.9447), be illustrated in the bright health instability of replacing of happiness in the standard curve making process, have certain conversion (Fig. 3).
Fig. 4 together measures the bright chromatogram that replaces health and lucky miaow to replace health of happiness in the rat plasma for the LC-MS detection method.A, blank rat plasma sample; B has added in the blank rat plasma sample and has liked the bright standard items that replace health and lucky miaow to replace health; C, plasma sample after the rat vein administration.This figure is the representative chromatogram that carries out above-mentioned 3 kinds of embodiment gained.
In sum, adopt and of the present inventionly carry out in the preprocessing process at the whole blood to the human or animal, human or animal's whole blood is contacted with one or more carboxy-lesterase suppressant, promptly one or more carboxy-lesterase suppressant can preexist in the heparin tube; After perhaps the blood sampling back adds the active method of carboxy-lesterase suppressant blocking-up carboxy-lesterase immediately in human or animal's whole blood; Like and brightly continue to be converted into lucky miaow and obtained fully suppressing external for health for health; Resultant happiness is bright replaces the blood concentration of health can reflect its actual conditions in vivo preferably for health and lucky miaow, has guaranteed that the camptothecine compounds prodrug is converted into the transformation efficiency result's of metabolic product reliability.
Claims (9)
1. measure the bright method of replacing health and lucky miaow to replace Kang Hanliang of happiness in human or animal's whole blood for one kind; After this method comprises that the whole blood to the human or animal carries out pre-service; Separated plasma adopts method for separating and detecting to separate for Kang Jinhang and detect for health and lucky miaow happiness in the blood plasma is bright again, it is characterized in that; Carry out in the preprocessing process at whole blood, human or animal's whole blood is contacted with one or more carboxy-lesterase suppressant the human or animal.
2. bright health and the lucky miaow of replacing of happiness is characterized in that for the method for Kang Hanliang said one or more carboxy-lesterase suppressant preexist in the heparin tube in the mensuration human or animal whole blood according to claim 1.
3. bright health and the lucky miaow of replacing of happiness is characterized in that for the method for Kang Hanliang said carboxy-lesterase suppressant is two (p-nitrophenyl) phosphates, TTA or dibenzoyl in the mensuration human or animal whole blood according to claim 1 and 2.
4. the bright method of replacing health and lucky miaow to replace Kang Hanliang of happiness in the mensuration human or animal whole blood according to claim 3, it is characterized in that the ml of human or animal's whole blood and said carboxy-lesterase suppressant: the ratio of mg is 1: 5～1: 10.
5. the bright method of replacing health and lucky miaow to replace Kang Hanliang of happiness in the mensuration human or animal whole blood according to claim 4, it is characterized in that the ml of human or animal's whole blood and said carboxy-lesterase suppressant: the ratio of mg is 1: 5.
6. bright health and the lucky miaow of replacing of happiness is characterized in that for the method for Kang Hanliang the inwall of said heparin tube is coated with liquaemin in the mensuration human or animal whole blood according to claim 2.
7. in the mensuration according to claim 1 human or animal whole blood happiness bright for health and lucky miaow for the method for Kang Hanliang, it is characterized in that, the separated plasma process comprise with human or animal's after one or more carboxy-lesterase suppressant contact the centrifugal blood plasma that obtains of whole blood.
8. bright health and the lucky miaow of replacing of happiness is characterized in that for the method for Kang Hanliang described separation method is a high performance liquid chromatography in the mensuration human or animal whole blood according to claim 1; Used detection method is a Mass Spectrometer Method.
9. measure the bright method of replacing health and lucky miaow to replace Kang Hanliang of happiness in human or animal's whole blood for one kind; This method is specially: be coated with liquaemin and add the whole blood that adds the human or animal in the heparin tube of carboxy-lesterase suppressant at inwall in advance; Shake up; Centrifuging obtains blood plasma, in this blood plasma, adds acetonitrile and carries out albumen precipitation, obtains supernatant after centrifugal; Said supernatant is separated in the LC-MS appearance and detects.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201010260758.0A CN102375040B (en) | 2010-08-23 | 2010-08-23 | Method for determining content of simimtecan and chimmitecan in human or animal whole blood |
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|---|---|---|---|
| CN201010260758.0A CN102375040B (en) | 2010-08-23 | 2010-08-23 | Method for determining content of simimtecan and chimmitecan in human or animal whole blood |
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| CN102375040A true CN102375040A (en) | 2012-03-14 |
| CN102375040B CN102375040B (en) | 2014-02-05 |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102885822A (en) * | 2012-10-08 | 2013-01-23 | 中国科学院大连化学物理研究所 | Carboxylesterase inhibitor with tanshinone skeleton structure and application thereof |
| CN110161159A (en) * | 2019-07-04 | 2019-08-23 | 杭州必益泰得医学科技有限公司 | A kind of biological sample analysis method of Oxcarbazepine bioequivalence test |
| CN110208411A (en) * | 2019-06-10 | 2019-09-06 | 杭州必益泰得医学科技有限公司 | Carboxylesterase inhibitor preparation for drug metabolism detection |
| CN110220757A (en) * | 2019-06-05 | 2019-09-10 | 杭州必益泰得医学科技有限公司 | Blood preseration agent for drug metabolism detection |
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| CN1616460A (en) * | 2003-11-10 | 2005-05-18 | 中国科学院上海药物研究所 | Novel derivative of camptothecine, preparing method and use |
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| WO2003014069A1 (en) * | 2001-08-10 | 2003-02-20 | Pharmacia Italia Spa | Fluoro linkers and their use as linkers for enzyme-activated drug conjugates |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102885822A (en) * | 2012-10-08 | 2013-01-23 | 中国科学院大连化学物理研究所 | Carboxylesterase inhibitor with tanshinone skeleton structure and application thereof |
| CN102885822B (en) * | 2012-10-08 | 2014-09-17 | 中国科学院大连化学物理研究所 | Carboxylesterase inhibitor with tanshinone skeleton structure and application thereof |
| CN110220757A (en) * | 2019-06-05 | 2019-09-10 | 杭州必益泰得医学科技有限公司 | Blood preseration agent for drug metabolism detection |
| CN110220757B (en) * | 2019-06-05 | 2021-08-24 | 浙江龙传生物医药科技有限公司 | Blood preservative for drug metabolism detection |
| CN110208411A (en) * | 2019-06-10 | 2019-09-06 | 杭州必益泰得医学科技有限公司 | Carboxylesterase inhibitor preparation for drug metabolism detection |
| CN110208411B (en) * | 2019-06-10 | 2021-12-24 | 浙江龙传生物医药科技有限公司 | Carboxylesterase inhibitor formulations for drug metabolism detection |
| CN110161159A (en) * | 2019-07-04 | 2019-08-23 | 杭州必益泰得医学科技有限公司 | A kind of biological sample analysis method of Oxcarbazepine bioequivalence test |
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| CN102375040B (en) | 2014-02-05 |
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