CN109917030A - The pre-treating method and detection method of a kind of minodronic acid based on derivatization on solid-phase extraction column in plasma sample - Google Patents
The pre-treating method and detection method of a kind of minodronic acid based on derivatization on solid-phase extraction column in plasma sample Download PDFInfo
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Abstract
The invention discloses pre-treating method and detection method of a kind of minodronic acid based on derivatization on solid-phase extraction column in plasma sample, belong to field of bioanalysis.Provided by the invention detection method includes the following steps: the plasma sample containing minodronic acid first being carried out pre-treatment, again using the concentration of minodronic acid in liquid phase-mass spectrometry combination method detection blood plasma, the pre-treatment include the following steps: will the plasma sample containing minodronic acid to the solid-phase extraction column sample introduction after activation, derivatization reagent is added after elution and carries out column derivatization reaction, after collection reaction solution is dried with nitrogen, is redissolved, supernatant direct injected is taken.This method compensates for minodronic acid can not carry out the defect of liquid phase analysis due to its polarity the reason of, and the dosage of blood plasma is greatly reduced compared with existing derivatization method, simplifies sample pre-treatments step.
Description
Technical field
The invention belongs to field of bioanalysis, and in particular to the pre-treatment of the column derivatization of minodronic acid in a kind of blood plasma
The detection method of minodronic acid concentration in method and blood plasma.
Background technique
Minodronic acid is in a kind of new type heterocycle bis-phosphonic acids compounds, for treating osteoporosis and being dredged by sclerotin
Hypercalcinuria caused by loose disease and malignant tumour, by inhibiting farnesyl pyrophosphate (FPP) synthase activity in osteoclast, suppression
The bone resorption of osteoclast processed reduces bone conversion, plays the effect of prevention and treatment osteoporosis.
Minodronate tablets are as a kind of oral preparation, and dosage is mainly 1mg, and content is low in human plasma, Cmax
For 200pg or so.In biological sample analysis, need to investigate the blood concentration of its half-life period, linear minimum lower limit of quantitation is answered
It should be in Cmax1/10 to 1/20, therefore need to develop it is a kind of efficiently, sensitive detection method detects the minot phosphine in human plasma
Acid content.
Research before reports a kind of column derivatization HPLC-MS/MS method, and the LLOQ of this method is 10pg/ml,
But it is 1mL that plasma volume, which is intended for single use, in this method, and using the WAX solid-phase extraction column of 6cc/150mg, method and step is more, time-consuming, right
For the big bioanalytical assays of sample size and it is not suitable for.
It now needs to develop a kind of highly sensitive, it is easier to which the high-throughput HPLC-MS/MS method of operation is simultaneously used for drug metabolism
Dynamics research.
Summary of the invention
The purpose of the present invention is on the basis of existing technology, provide a kind of minot based on derivatization on solid-phase extraction column
The sensitivity of sample measurement has can be improved in pre-treating method of the phosphonic acids in plasma sample, solves biological sample high throughput
The problem of analysis.
Another mesh of the invention is to provide a kind of using the concentration of minodronic acid in above-mentioned pre-treating method detection blood plasma
Detection method.
Technical scheme is as follows:
The pre-treating method of a kind of minodronic acid based on derivatization on solid-phase extraction column in plasma sample, wherein preceding
Processing include the following steps: will the plasma sample containing minodronic acid to the solid-phase extraction column sample introduction after activation, be added after elution
Derivatization reagent carries out column derivatization reaction, after being dried with nitrogen, redissolving, takes supernatant sample introduction.
Solid-phase extraction column of the present invention is Waters Oasis WAX, may further be preferably solid-phase extraction column
For Waters Oasis WAX (30 μm, 60mg).The solid-phase extraction column that the present invention uses is before sample introduction, first by solid-phase extraction column
It is activated, can successively be activated using methanol, pure water.
In the present invention, using solid-phase extraction column WAX (Oasis, 30 μm of (60mg) Waters) so that minodronic acid with fill out
Filling substance can sufficiently be bonded, when avoiding using other solid-phase extraction columns the excessive blood plasma dosage in pretreatment process, reaction
Between it is too long the problems such as, and then make minodronic acid occur derivative reaction after eluted from filler.This method pair simultaneously
Than different types of solid-phase extraction column WAX (Oasis, 30 μm of (60mg) Waters), WAX (Oasis, 6cc/150 mg
Waters) and solid-phase extraction column Oasis HLB, 30 μm of Waters, as a result, it has been found that solid-phase extraction column Oasis HLB, 30 μm
Column derivatizationization reaction cannot occur for Waters, and the derivatization efficiency of WAX (Oasis, 30 μm of (60mg) Waters) is WAX
40 times of (Oasis, 6cc/150mg Waters)！
The present invention selects trimethyl silicone hydride diazomethane (TMS-DAM) as derivatization reagent.
Because minodronic acid is the very strong diphosphonate of a kind of polarity, the pre-treating method referred in currently available technology can not
So that it is reached retention in chromatography, the detection of concentration can not be carried out, it is necessary to reaction is performed the derivatization, before derivative reaction
Processing method is complicated and cumbersome, while operation difficulty is larger.The present inventor refers to that pre-treatment is special with reference to other correlations
Benefit or article, discovery only provides pre-treating method of the invention, while retaining the detection side that existing related patents or article refer to
Method still can obtain beneficial detection effect.Therefore the inventor of this patent uses the detection side referred in the prior art
Method, and carry out methodology validation.After the completion of minodronic acid pre-treatment derivative reaction, concentration mensuration is opposite to be easier to, and unmatched
Treatment process can not carry out Concentration Testing.
The derivative reaction that the present invention refers to is schematically as follows:
In a preferred embodiment, the plasma sample containing minodronic acid is mixed with dilution and is diluted, then to work
Solid-phase extraction column sample introduction after change, wherein the volume ratio of plasma sample and dilution containing minodronic acid is 1:1.
In a kind of more preferable scheme, dilution is that the mixing of 10mM ammonium acetate-aqueous solution and 10ng/mL inner mark solution is molten
Liquid.The present invention detect blood plasma in minodronic acid concentration when, using minodronic acid-d4 as internal standard.As further preferred
The volume ratio of scheme, above-mentioned ammonium acetate-aqueous solution and inner mark solution is 5:1 to 6:1.
Further, it is successively used after the plasma sample containing minodronic acid to after the solid-phase extraction column sample introduction after activation
Methanol, pure water elute solid-phase extraction column.
After solid-phase extraction column elutes, sequentially add derivatization reagent with volume ratio to solid-phase extraction column is the present invention
The mixed solution of the methanol-water solution of 12:1, performs the derivatization reaction, after repetitive operation 1 to 3 time, collects reaction solution.
Further, 60min is reacted under greenhouse experiment, regathers reaction solution.
Further, the volume ratio for the methanol water mixed solution that derivatization reagent and volume ratio are 12:1 is 1:3.
In a preferred embodiment, the reaction solution of collection is placed in extraction plate and carries out loading to solid-phase extraction column again,
Continue derivative reaction.In the case where not influencing effect of the present invention, can it is appropriate selection reaction temperature and when
Between.For example, reacting 50-60min under greenhouse experiment.
Further, it after the reaction solution wait collect carries out derivative reaction on the solid-phase extraction column, sequentially adds
The reaction was continued for the methanol-water solution that a certain amount of derivatization reagent and volume ratio are 12:1.
Further, the volume ratio for the methanol water mixed solution that derivatization reagent and volume ratio are 12:1 is 1:3.
Further, 50-60min is reacted under greenhouse experiment.
Further, after derivative reaction, collection reaction solution keeps the temperature 60min at 65~75 DEG C and is dried with nitrogen again.It is excellent
Select reaction solution heat preservation at 70 DEG C.
The present invention is 35~45 DEG C using above-mentioned reaction solution, temperature is dried with nitrogen, it is preferable that the temperature being dried with nitrogen is 40
℃.It after sample drying, is redissolved with water, supernatant sample introduction is taken to carry out liquid phase-mass spectrometry combination method detection.
The detection method of the concentration of minodronic acid in a kind of detection blood plasma, first carries out the plasma sample containing minodronic acid
Pre-treatment, then the concentration using minodronic acid in liquid phase-mass spectrometry combination method detection blood plasma, wherein pre-treatment includes following step
It is rapid: derivatization reagent is added by the plasma sample containing minodronic acid to the solid-phase extraction column sample introduction after activation, after elution and carries out
Column derivatizationization reaction takes supernatant direct injected after collection reaction solution is dried with nitrogen, is redissolved.
When the present invention detects the detection of the concentration of minodronic acid in blood plasma, it can use and be able to detect blood plasma in the prior art
The detection method of the LC-MS of the concentration of middle minodronic acid.
In a preferred embodiment, the chromatographic condition of LC-MS is as follows:
Liquid-phase condition are as follows: chromatographic column: Eclipse XDB phenyl analytical column (150mm × 4.6mm, 5 μm)；Column temperature: 40 DEG C；
Sample injector temperature: 8 DEG C；Mobile phase: acetonitrile；0.15% formic acid-water (ammonium acetate containing 10mM)；Elution: gradient elution；When detection
Between: 4.30min；Flow velocity: 1.0mL/min；Sample volume: 10 μ L.
Mass Spectrometry Conditions: ion detection mode: multiple-reaction monitoring (MRM)；Ionization mode: Aeroassisted electron spray ion
Change (ESI)；Ion polarity: cation (Positive)；Test object: m/z 393.1 → 267.2 (pentamethyl minodronic acid)
With m/z 397.1 → 271.2 (internal standard)；Mass spectrometry parameters: IonSpray Voltage:5500V；CUR:10psi；CAD:5psi；
TEM:500 DEG C；GS1:55；GS2:50.
Sample dilution: adding the 500 μ L of plasma sample containing minodronic acid into 1.5mL centrifuge tube, and 500 μ L of dilution is added,
Dilution is 10mM ammonium acetate-aqueous solution: minodronic acid-d4 (10ng/mL)=5:1 to 6:1 mixing, vortex mixed.
The activation of solid-phase extraction column: solid-phase extraction column WAX (Oasis, 30 μm of (60mg) Waters) uses 1mL methanol respectively,
1mL pure water is activated in low flow velocity.
Loading: the plasma sample in centrifuge tube is transferred to solid-phase extraction column and carries out loading, with low flow velocity loading.
Elution: it is eluted with 1mL pure water, 1mL methanol, is eluted using low flow velocity respectively.
Derivative reaction: being added 225 μ L methanol-water solutions (12:1) in solid-phase extraction plate, 75 μ L TMS-DAM, fastly
Speed is repeated twice, and reacts at room temperature 1h.Then by collection liquid again loading into solid-phase extraction plate, 50min is reacted at room temperature.Reaction
After add 225 μ L methanol-waters (12:1), 75 μ L TMS-DAM react at room temperature 1h.
Heating reaction: all collection liquids are placed in 70 DEG C of baking ovens, reaction 1h to fully reacting.
Drying: the sample after taking out reaction is placed in 40 DEG C of nitrogen evaporator dryings.
It redissolves: 100 μ L pure water being added after drying, be vortexed and mix.Cation is carried out with the triple level four bars mass spectrums of electron spray one
Qualitative and quantitative analysis.
Using technical solution of the present invention, advantage is as follows:
1. in the present invention, doing internal standard compound, deuterated internal standard and determinand retention time having the same with deuterated minodronic acid
And chemical property.
2. in the present invention, solid-phase extraction column WAX (Oasis, 30 μm of (60mg) Waters) is placed in positive pressure devices, and
96 orifice plates used will be collected to be placed in below extraction column.The nitrogen that this device had both guaranteed that itself derivative reaction generated can make hole
Interior pressure increase is accelerated the speed of derivative reaction, and can suitably be added in insufficient pressure by the nitrogen pore in positive pressure devices
Pressure guarantees the abundant complete of reaction.
3. in the present invention, being tried using derivatization reagent leucoaurin base diazomethane (TMS-DAM) and traditional derivatization
Agent is less compared to toxicity, reduces injury of the derivatization reagent to operator.
4, in the present invention, using solid-phase extraction column WAX (Oasis, 30 μm of (60mg) Waters) so that minodronic acid with
Filler can be sufficiently bonded, and avoid using other solid-phase extraction columns that the blood plasma dosage in pretreatment process is excessive, reaction
The problems such as overlong time, and then eluted from filler after making minodronic acid that derivative reaction occur.
5. in the present invention, derivative reaction is increased the number to methylate on minodronic acid and is determined by the control reaction time
The end-state of reaction finally detects 5- methyl minodronic acid and the deuterated minodronic acid of 5- methyl after derivative reaction
Concentration, for the quantitative analysis of minodronic acid in human plasma, the reaction time is that 60min or so is that effect is best.
6. in the present invention, by controlling the dosage of derivatization reagent, to guarantee that derivatization is anti-within the relatively short time
Should be able to sufficiently completely, 5- methyl-minodronic acid production quantity can satisfy the requirement of minimum quantitative limit.
Detailed description of the invention
Fig. 1 is sample pretreatment process schematic device；
Fig. 2 is mass spectrum response and the pre-treatment efficiency comparative figure of the derivative products of derivatization different time；
Fig. 3 be left figure be 5- methyl minodronic acid, right figure is the deuterated minodronic acid of 5- methyl by liquid phase-mass spectrometry skill
The chromatogram that art detects.
Specific embodiment
Detection method of the invention is further described by following embodiment and attached drawing, but these embodiments are not
Any restrictions are constituted to the present invention.
Instrument used in the present invention: superelevation phase liquid chromatogram (Shimadzu LC30)；Mass spectrum (API5500, Applied
Biosystems/Sciex)；Pure water meter；Microbalance；Centrifuge etc..Solvent used in the present invention: acetonitrile, water, formic acid,
Isopropanol, ammonium acetate, methanol.
The EDTA blood coagulation slurry of blank derives from health volunteer.
Liquid-phase condition: chromatographic column: Eclipse XDB phenyl analytical column (150mm × 4.6mm, 5 μm)；Column temperature: 40 DEG C；Into
Sample device temperature: 8 DEG C；Mobile phase: acetonitrile: 0.15% formic acid-water (ammonium acetate containing 10mM)；Elution: gradient elution；Detection time:
4.30min；Flow velocity: 1.0mL/min；Sample volume: 10 μ L；Autosampler washes needle liquid: 90:10:1=methanol: water: formic acid is (strong
It washes)；10:90=methanol: water (weak to wash)；Autosampler washes needle program: Seal wash:0.5min；Volume: 100 μ L is washed by force；
Weak volume of washing: 100 μ L.
Mass Spectrometry Conditions: ion detection mode: multiple-reaction monitoring (MRM)；Ionization mode: Aeroassisted electron spray ion
Change (ESI)；Ion polarity: cation (Positive)；Test object: m/z 393.1 → 267.2 (pentamethyl minodronic acid)
With m/z 397.1 → 271.2 (internal standard)；Mass spectrometry parameters: IonSpray Voltage:5500V；CUR:10psi；CAD:5psi；
TEM:500 DEG C；GS1:55；GS2:50.
Pretreating device schematic diagram is shown in Fig. 1.In the present invention, by solid-phase extraction column WAX (Oasis, 30 μm (60mg)
Waters it) is placed in positive pressure devices, and be placed in 96 orifice plates used are collected below extraction column.This device had both guaranteed itself to spread out
The nitrogen that biochemical reaction generates can make pressure increase in hole, accelerate the speed of derivative reaction, and can pass through in insufficient pressure
Nitrogen pore in positive pressure devices suitably pressurizes, and guarantees the abundant complete of reaction.
As shown in Fig. 2, in the present invention, using solid-phase extraction column WAX (Oasis, 30 μm of (60mg) Waters), so that rice
Promise phosphonic acids can be sufficiently bonded with filler, avoid using other solid-phase extraction columns the blood plasma dosage in pretreatment process
Excessive, the problems such as reaction time is too long, and then eluted from filler after making minodronic acid that derivative reaction occur.
Simultaneously this method compared different types of solid-phase extraction column WAX (Oasis, 30 μm of (60mg) Waters), WAX (Oasis,
6cc/150mg Waters) and solid-phase extraction column Oasis HLB, 30 μm of Waters, as a result, it has been found that solid-phase extraction column
Column derivatizationization reaction cannot occur for Oasis HLB, 30 μm of Waters, and WAX (Oasis, 30 μm of (60 mg) Waters') spreads out
Biochemical efficiency is 40 times of WAX (Oasis, 6cc/150mg Waters)！
In the present invention, derivative reaction is increased the number to methylate on minodronic acid and is determined instead by the control reaction time
The end-state answered finally detects the dense of 5- methyl minodronic acid and the deuterated minodronic acid of 5- methyl after derivative reaction
Degree, for the quantitative analysis of minodronic acid in human plasma, the reaction final result comparison diagram of different time is shown in Fig. 2, the results showed that
Reaction time is that 60min or so is that effect is best.
Using the derivatization method in the present invention, measuring method detailed process is established are as follows:
The preparation of plasma sample:
The preparation of minodronic acid sample: it is appropriate that precision weighs minodronic acid reference substance, after mass calibration coefficient correction, uses
After the dissolution of pure water water, the minodronic acid stock solution that final concentration is 50.0 μ g/mL is obtained, stock solution is stored in -20 DEG C of refrigerators.
Minodronic acid stock solution is diluted with 50% methanol-water, and appropriate blood plasma is then added, and preparing a series of concentration is 10,20,50,
100,200,500,1000,2000pg/mL minodronic acid standard curve sample.
The preparation of minodronic acid-d4 working solution: it is appropriate that precision weighs minodronic acid-d4 reference substance, through mass calibration system
It after number correction, is dissolved with pure water, obtains the minodronic acid-d4 stock solution that ultimate density is 50.0 μ g/mL, stock solution is stored in
- 20 DEG C of refrigerators.Precision measures a certain amount of minodronic acid-d4 and is diluted with 50% methanol-water, and being configured to concentration is 10.0ng/mL
Internal standard working solution, a certain amount of 10mM ammonium acetate-aqueous solution, which is added, makes its volume ratio internal standard (10.0ng/mL): 10mM second
Sour ammonium-aqueous solution=1:6.
As procedure described above, after solid-phase extraction column being activated, loading, elution, derivative reaction are carried out: in solid-phase extraction plate
225 μ L methanol-water solutions (12:1) of interior addition, 75 μ L TMS-DAM are quickly repeated twice, and react at room temperature 1h.It will then collect
Again loading reacts at room temperature 50min into solid-phase extraction plate to liquid.225 μ L methanol-waters (12:1), 75 μ L are added after reaction
TMS-DAM reacts at room temperature 1h.Baking oven reacts 1h, dries up under nitrogen.It is analyzed with established liquid phase-Mass Spectrometry detection method.
Product after derivative reaction: 5- methyl minodronic acid and the deuterated minodronic acid of 5- methyl are joined by liquid phase-mass spectrum
The chromatogram that art detects is shown in Fig. 3.
Claims (10)
1. a kind of pre-treating method of minodronic acid based on derivatization on solid-phase extraction column in plasma sample, feature exist
In by the plasma sample containing minodronic acid to the solid-phase extraction column sample introduction after activation, addition derivatization reagent is carried out after elution
Column derivatizationization reaction takes supernatant direct injected after collection reaction solution is dried with nitrogen, is redissolved.
2. preceding processing of the minodronic acid according to claim 1 based on derivatization on solid-phase extraction column in plasma sample
Method, which is characterized in that the solid-phase extraction column is Waters Oasis WAX；The solid-phase extraction column successively use methanol,
Pure water is activated；The derivatization reagent is trimethyl silicone hydride diazomethane.
3. preceding processing of the minodronic acid according to claim 1 based on derivatization on solid-phase extraction column in plasma sample
Method, which is characterized in that the plasma sample containing minodronic acid is mixed with dilution and is diluted, then to the solid phase after activation
The volume ratio of extraction column sample introduction, the plasma sample containing minodronic acid and dilution is 1:1；The dilution is 10mM second
The mixed solution of sour ammonium-aqueous solution and 10ng/mL inner mark solution, it is described in be designated as minodronic acid-d4；It is preferred that the ammonium acetate-
The volume ratio of aqueous solution and inner mark solution is 5:1~6:1.
4. preceding processing of the minodronic acid according to claim 1 based on derivatization on solid-phase extraction column in plasma sample
Method, which is characterized in that the solid-phase extraction column successively uses methanol, pure water to be eluted；After elution, Xiang Suoshu Solid Phase Extraction
The mixed solution for the methanol-water solution that column sequentially adds derivatization reagent and volume ratio is 12:1, performs the derivatization reaction, repeats
After operation twice, reaction solution is collected；It is preferred that reacting 60min under greenhouse experiment, reaction solution is regathered.
5. preceding processing of the minodronic acid according to claim 4 based on derivatization on solid-phase extraction column in plasma sample
Method, which is characterized in that by the reaction solution of collection be placed in extraction plate again to the solid-phase extraction column carry out loading, continue into
Row derivative reaction；It is preferred that reacting 50~60min at room temperature.
6. preceding processing of the minodronic acid according to claim 5 based on derivatization on solid-phase extraction column in plasma sample
Method, which is characterized in that after the reaction solution wait collect carries out derivative reaction on the solid-phase extraction column, sequentially add and spread out
The reaction was continued for the mixed solution for the methanol-water solution that biochemical reagents and volume ratio are 12:1；It is preferred that reacting 50 under greenhouse experiment
~60min.
Before 7. the minodronic acid based on derivatization on solid-phase extraction column according to claim 4 or 6 is in plasma sample
Processing method, which is characterized in that the volume ratio for the methanol water mixed solution that derivatization reagent is 12:1 with volume ratio is 1:3.
8. preceding processing of the minodronic acid according to claim 1 based on derivatization on solid-phase extraction column in plasma sample
Method, which is characterized in that collection reaction solution keeps the temperature 60min at 65~75 DEG C and is dried with nitrogen again；It is preferred that reaction solution is at 70 DEG C
Heat preservation；The temperature being dried with nitrogen is 35~45 DEG C, it is preferable that the temperature being dried with nitrogen is 40 DEG C；It is described to redissolve the molten of use
Agent is water.
9. the method according to claim 1, wherein mass spectrum carries out sample to be tested using multiple-reaction monitoring pattern
Quantitative analysis, it is preferable that sample to be tested quantitative analysis method is established before quantitative analysis.
10. the detection method of the concentration of minodronic acid in a kind of detection blood plasma, which is characterized in that by the blood plasma containing minodronic acid
Sample carries out pre-treatment, then using the concentration of minodronic acid in liquid phase-mass spectrometry combination method detection blood plasma, the pre-treatment includes
Following steps: derivatization examination is added by the plasma sample containing minodronic acid to the solid-phase extraction column sample introduction after activation, after elution
Agent carries out column derivatization reaction and takes supernatant direct injected after collection reaction solution is dried with nitrogen, is redissolved.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111948307A (en) * | 2020-07-27 | 2020-11-17 | 苏州圣临医药科技有限公司 | Liquid chromatography-mass spectrometry azide methane derivatization method |
| CN114113379A (en) * | 2021-11-12 | 2022-03-01 | 四川尚锐分析检测有限公司 | Method for detecting iban sodium phosphate in blood plasma by LC-MS (liquid chromatography-mass spectrometry) |
| CN114384186A (en) * | 2022-01-21 | 2022-04-22 | 王立强 | Method for measuring content of (2E, 4E) -ethyl-4- (pyridine-2-yl imino) -2-ethyl crotonate |
| CN114594170A (en) * | 2020-12-03 | 2022-06-07 | 复旦大学 | In-vivo drug analysis method combining magnetic solid phase extraction with rapid in-situ derivatization |
| CN116482242A (en) * | 2022-12-30 | 2023-07-25 | 杭州百杏生物技术有限公司 | LC-MS/MS method for determining alendronate concentration in biological sample |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2013085344A1 (en) * | 2011-12-08 | 2013-06-13 | 아주대학교 산학협력단 | Method for screening material for prevention or treatment of osteoporosis using leptin secretion of mesenchymal stem cells |
| CN103983709A (en) * | 2014-05-14 | 2014-08-13 | 永信药品工业(昆山)有限公司 | Detection method of minodronic acid by using high performance liquid chromatography |
| CN107543865A (en) * | 2016-06-23 | 2018-01-05 | 天津市汉康医药生物技术有限公司 | The method for separating and detecting minodronic acid |
-
2019
- 2019-01-28 CN CN201910079034.7A patent/CN109917030A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2013085344A1 (en) * | 2011-12-08 | 2013-06-13 | 아주대학교 산학협력단 | Method for screening material for prevention or treatment of osteoporosis using leptin secretion of mesenchymal stem cells |
| CN103983709A (en) * | 2014-05-14 | 2014-08-13 | 永信药品工业(昆山)有限公司 | Detection method of minodronic acid by using high performance liquid chromatography |
| CN107543865A (en) * | 2016-06-23 | 2018-01-05 | 天津市汉康医药生物技术有限公司 | The method for separating and detecting minodronic acid |
Non-Patent Citations (7)
| Title |
|---|
| MEIXIA CHEN ET AL: "Trimethylsilyldiazomethane derivatization coupled with solid-phase extraction for the determination of alendronate in human plasma by LC-MS/MS", 《ANAL BIOANAL CHEM》 * |
| YONG YANG ET AL: "On-cartridge derivatization coupled with solid-phase extraction for the ultra-sensitive determination of minodronic acid in human plasma by LC–MS/MS method", 《JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS》 * |
| 博纳艾杰尔科技: "生物样品分析", 《博纳艾杰尔科技生物样品分析产品介绍》 * |
| 宋伟杰 等: "高效液相色谱法测定米诺膦酸的含量及有关物质", 《中国新药杂志》 * |
| 张琳 等: "不同前处理方法对全血中12种毒品成分检测结果的影响", 《中国法医学杂》 * |
| 田兰 等: "固相萃取结合HPLC-荧光法测定人血浆中维生素B2含量", 《药物分析杂志》 * |
| 邬方宁 等: "高效液相色谱法测定米诺膦酸片中米诺膦酸", 《现代药物与临床》 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111948307A (en) * | 2020-07-27 | 2020-11-17 | 苏州圣临医药科技有限公司 | Liquid chromatography-mass spectrometry azide methane derivatization method |
| CN114594170A (en) * | 2020-12-03 | 2022-06-07 | 复旦大学 | In-vivo drug analysis method combining magnetic solid phase extraction with rapid in-situ derivatization |
| CN114594170B (en) * | 2020-12-03 | 2024-05-17 | 复旦大学 | In-vivo drug analysis method combining magnetic solid phase extraction with rapid in-situ derivatization |
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Application publication date: 20190621 |