CN106018593A - HPLC-MS/MS (high performance liquid chromatography-tandem mass spectrometry) detection method for dauricine concentration - Google Patents
HPLC-MS/MS (high performance liquid chromatography-tandem mass spectrometry) detection method for dauricine concentration Download PDFInfo
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Abstract
The invention belongs to the technical field of pharmaceutical analysis, and relates to an HPLC-MS/MS (high performance liquid chromatography-tandem mass spectrometry) detection method for dauricine concentration, in particular to a method for detecting the dauricine concentration of a plasma sample with HPLC-MS/MS. The method adopts an internal standard method, and tetrandrine is taken as an internal standard substance. The method has the advantages of high specificity, high sensitivity and good accuracy, the limit of quantification of the dauricine in plasma is 2.12 ng/ml, and the method can be widely applied to a test for the dauricine concentration of a biological sample, particularly a test for the dauricine concentration of the plasma sample.
Description
Technical field
The invention belongs to pharmaceutical analysis technical field, the method relating to high performance liquid chromatography-tandem mass detection Dauricine concentration, it particularly relates to use the method for Dauricine concentration in high performance liquid chromatography-tandem mass detection plasma sample.
Background technology
Dauricine (Dauricine, Dau) it is a kind of bisbenzylisoquinoline alkaloid of isolated from the subterraneous stem of menispermaceous plants Caulis menispermi, study and shown that it has the effect of arrhythmia and antiinflammatory, in addition, the newest research also shows that it has anticarcinogenic effect, including suppression growth of tumour cell, alleviates drug-resistant effect and promotes the apoptosis etc. of cancerous cell.Having two alicyclic ring tertiary amine groups in Dauricine molecular structure, have stronger fat-soluble, water solublity is poor.Less about the research that Dauricine pharmacokinetics is relevant the most both at home and abroad, but existing research display, after the research of rat Internal pharmacokinetics shows its gastric infusion, absolute bioavailability is about 15-19%.Rat is in the research display of body unidirectional intestinal perfusion model, and Dauricine is in intestinal permeable membrane ability, and the barrier action of gastrointestinal tract epithelial cell is the key factor affecting its oral administration biaavailability.Therefore, the research emphasis of novel Dauricine drug-supplying system is placed on the bioavailability improving Dauricine oral administration by the most increasing researcher.
And in recent years, self-micro-emulsification medicine-releasing system (self-(micro) emulsifying drug delivery systems, SMEDDS), owing to can effectively facilitate the oral absorption of the medicine of poorly water-soluble, has obtained studying widely.SMEDDS is made up of oil phase, surfactant and cosurfactant, is scattered in aqueous medium and forms oil-in-water diameter and less than the microemulsion drop of 100 nanometers thus improve the water solublity of medicine.Additionally, SMEDDS is conducive to protecting the medicine of unstable chemcial property, controlling the release of medicine, in stomach, quick self emulsifying promotes that medicine in gastrointestinal dispersion and quickly absorbs, and SMEDDS can be administered by different route of administration.Therefore, for improving the oral administration biaavailability of Dauricine, present inventor's early stage is prepared for Dauricine SMEDDS.But, how to evaluate whether SMEDDS has the effect improving Dauricine bioavailability？The mensuration of internal Dauricine concentration must be carried out, but how to realize the accurate detection of Dauricine concentration in Dauricine concentration especially plasma sample？At present, prior art lacks a kind of effective ways quantitative determining Dauricine concentration.
Summary of the invention
In view of this, the method that the invention provides a kind of high performance liquid chromatography-tandem mass detection Dauricine concentration, the method is to use high performance liquid chromatography-tandem mass to detect, and can realize the accurate detection of Dauricine concentration.
For achieving the above object, the technical scheme is that
The method of high performance liquid chromatography-tandem mass detection Dauricine concentration, described method is to use internal standard method, with tetrandrine as internal standard substance.
High performance liquid chromatography-tandem mass technology (LC-MS/MS) is using liquid chromatograph as piece-rate system, and mass spectrum is detecting system.Sample is separated by flowing on a column, after being ionized, through mass spectrographic mass analyzer by ion isolation, smashes, then is separated by mass number by fragment, and device obtains chromatogram after testing.High performance liquid chromatography and mass spectrographic advantage are carried out complementation by LC-MS technology, by the chromatograph high separating power to complex sample, combine with mass spectrographic high sensitivity, realize the quantitative analysis to complex sample, and simplify the pretreatment process of sample, make sample analysis easier, be therefore widely used in many fields such as pharmaceutical analysis, food analysis and environmental analysis.
Internal standard method is to be added in analyzed sample mixture as internal standard substance by certain density pure material, then the sample containing internal standard substance is carried out Treatment Analysis, measure internal standard substance and the peak area of component to be measured, tested component concentration in the sample can be obtained by standard curve method.Internal standard substance typically should meet following requirement: close with determinand structure, has the chemical property of acquaintance, chemical reaction does not occur.
Therefore, suitable internal standard substance particular importance is selected.Present inventor is through repeatedly studying and screening, it was found that a kind of LC-MS/MS method that is particularly suitable for measures the internal standard material of Dauricine concentration: the structural formula of tetrandrine, Dauricine and tetrandrine and mass spectrum are as shown in Figure 1.
High performance liquid chromatography-tandem mass is using liquid chromatograph as piece-rate system, and mass spectrum is detecting system.
Further, described method, the chromatographic column that described high performance liquid chromatography uses is the chromatographic column that octadecyl silane is filled, and the flowing of employing is the mixed liquor of acetonitrile and ammonium formate aqueous solution mutually.The specification of preferred column is 150 × 2.1mm, 5 μm.
Further, described method, the concentration of described ammonium formate aqueous solution is 1.5-2.5mM, and the preferably concentration of ammonium formate aqueous solution is 2mM.
Further, described method, described acetonitrile is 65:35 with the volume ratio that volume ratio is 60-80:20-40, preferably acetonitrile and ammonium formate aqueous solution of ammonium formate aqueous solution.
Further, described method, described acetonitrile is the acetonitrile containing ammonium formate, the concentration of ammonium formate be the concentration of 1.5-2.5mM, preferably ammonium formate be 2mM.
Further, the sample size of described high performance liquid chromatography is 5-50 μ l, preferably 10 μ l.
Further, the column temperature of described high performance liquid chromatography is 20-40 DEG C, preferably 30 DEG C.
Further, the flow velocity of described high performance liquid chromatography is 0.1-0.5ml/min, and preferable flow rate is 0.2ml/min.
Further, described method, described mass spectrum uses multiple reaction monitoring pattern, and detection ion is that cation mode, Dauricine and tetrandrine are respectively adopted the combination of at least one pair of parent ion and daughter ion and carry out Quantitative Monitoring.
Further, described method, the parent ion of described Dauricine employing and being combined as of daughter ion: parent ion m/z is 625 ± 1, and daughter ion m/z is 206 ± 1, and its mass spectrum is as shown in Figure 1A.
Further, described method, the parent ion of described tetrandrine employing and being combined as of daughter ion: parent ion m/z is 623 ± 1, and daughter ion m/z is 174 ± 1, and its mass spectrum is as shown in Figure 1B.
Further, being combined as of described Dauricine uses with tetrandrine parent ion and daughter ion: being combined as of parent ion that described Dauricine uses and daughter ion: parent ion m/z is 625 ± 1, and daughter ion m/z is 206 ± 1；The parent ion of tetrandrine employing and being combined as of daughter ion: parent ion m/z is 623 ± 1, and daughter ion m/z is 174 ± 1.
In a specific embodiment, the invention discloses following liquid chromatograph and Mass Spectrometry Conditions:
The condition that described liquid chromatograph uses includes:
Chromatographic column: watersMS C18Post (150mm × 2.1mm, 5 μm).
Flowing phase: acetonitrile (containing 2mM ammonium formate): 2mM ammonium formate aqueous solution=65:35 (volume ratio).
Sample size: 10 μ l.
Column temperature: 30 DEG C.
Flow velocity: 0.2ml/min.
The condition that described mass spectrum uses includes:
Ion source: electric spray ion source (ESI), capillary voltage is 3.5KV, and taper hole voltage is 35V, and ion source temperature is 110 DEG C, desolventizing gas (N2) flow is 500l/hr, desolventizing temperature is 350 DEG C.Mass spectrum monitoring mode is polyion reaction monitoring (MRM), and detection ion is cation mode, Dauricine, [M+H]+Ion, m/z 625 → 206；Tetrandrine (internal standard), [M+H]+Ion, m/z 623 → 174.
The present invention the most also protect tetrandrine as internal standard substance Dauricine concentration measure in application.
The present invention also protects the application in detection plasma sample in Dauricine concentration of the described method, and the detection method of the present invention, specificity is strong, highly sensitive, accuracy is good.In detecting plasma sample during Dauricine concentration, the processing method of described plasma sample is: take plasma sample, add methanol, internal standard tetrandrine, adding extractant ethyl acetate, centrifuging and taking supernatant after extraction after mixing, nitrogen dries up, the flowing that residue uses with high performance liquid chromatography is redissolved mutually, and last sample introduction is measured.Use the sample treatment of the present invention, it is possible to ensure the stability of sample.To use concentration be 5.30,53, the plasma sample of 212ng/ml carry out study on the stability, investigation project includes the stability after sample treatment, room temperature stability, three stability of freeze thawing and long-term frozen (-20 DEG C) stability.Result shows that sample keeps stable in 8h after at least processing, and room temperature can keep stable in placing 6h, keeps stablizing within freeze thawing three times, and long-term frozen keeps in 5 days stablizing.Stability test result error is all within 9.39%.
In a specific embodiment, the invention further particularly discloses and use the method for Dauricine concentration in high performance liquid chromatography-tandem mass detection plasma sample, particularly as follows:
(1) medicine-feeding test
It is appropriate that precision weighs Dauricine crude drug, and the NaOH adding 0.1mol/l dissolves in right amount, and is 6-7 with the HCl regulation pH value of 0.5mol/l, uses normal saline constant volume, makes the final concentration of 20mg/ml of Dauricine, is administered as test liquid.
Take Dauricine SMEDDS prescription prepared by present inventor's early stage, prepare Dauricine SMEDDS.The Dauricine SMEDDS mean diameter preparing gained is (58.3 ± 9.21) nm, and drug loading is about 49.46mg/g.
Take 12 and be divided into A, B two groups, often group 6 at random.After fasting 12h, ig is administered, A group rat ig Dauricine test liquid 50mg/Kg, B group rat ig Dauricine SMEDDS20mg/Kg, before being administered and after being administered 0.167,0.333,0.5,0.75,1,2,4,8,12,24,36h taken blood about 0.3ml in heparinization centrifuge tube by retroorbital venous clump respectively, after blood specimen collection immediately centrifugal also separated plasma put-70 DEG C of ultra cold storage freezers preserve to be measured.
(2) process of plasma sample
Precision measures blank rat plasma 0.1ml, adds methanol 5 μ l, internal standard 5 μ l, after mixing 10s, adds extractant ethyl acetate 1ml, after vortex mixed extraction 3min, and 1.3 × 104R/min is centrifuged 5min (centrifugal radius 15cm), takes supernatant, and at 40 DEG C, nitrogen dries up, and residue redissolves mutually with 50 μ l flowings, and sample introduction 10 μ l measures.
(3) preparation of solution
Dauricine and the preparation of internal standard storing solution: take Dauricine and internal standard reference substance about 10mg is the most accurately weighed, dissolve constant volume with methanol.
Dauricine and the preparation of internal standard working solution: take Dauricine and internal standard storing solution, become working solution with methanol dilution respectively.Dauricine working solution concentration is respectively 8480,4240,2120,1060,424,106,42.4ng/ml；Inside it is designated as 2036ng/ml.Storing solution and working solution all set to 0～preserve in 4 DEG C of refrigerators.
(4) chromatograph and Mass Spectrometry Conditions
Chromatographic column: watersMS C18Post (150mm × 2.1mm, 5 μm)；Flowing phase: acetonitrile (containing 2mM ammonium formate): 2mM ammonium formate aqueous solution=65:35 (volume ratio)；Column temperature: 30 DEG C；Flow velocity: 0.2ml/min；Sampling volume: 10 μ l.
Mass spectrum monitoring mode is polyion reaction monitoring (MRM), and detection ion is cation mode, Dauricine, [M+H]+Ion, m/z 625 → 206；Tetrandrine (internal standard), [M+H]+Ion, m/z 623 → 174.Using electric spray ion source (ESI), capillary voltage is 3.5KV, and taper hole voltage is 35V, and ion source temperature is 110 DEG C, desolventizing gas (N2) flow is 500l/hr, desolventizing temperature is 350 DEG C.
(5) detection of sample
Measuring Dauricine under conditions of step (4) and internal standard mass spectrum is shown in Fig. 1, measure blank rat plasma, blank rat plasma+Dauricine+internal standard, rat plasma sample+internal standard chromatogram is shown in Fig. 2.
Beneficial effects of the present invention:
The method of the high performance liquid chromatography-tandem mass detection Dauricine of the present invention, specificity is strong, highly sensitive, accuracy is good, can be widely used for the mensuration of Dauricine sample, in particular for measuring organism Dauricine bioavailability.
Accompanying drawing explanation
Fig. 1 is Dauricine and interior target mass spectrum, and wherein A is Dauricine, and B is internal standard.
Fig. 2 is blank rat plasma and the testing result figure of Dauricine concentration in test rat plasma sample, wherein letter A represents blank rat plasma, letter b represents blank rat plasma+Dauricine+internal standard, letter C represents test rat plasma sample+internal standard, numeral 1 represents Dauricine, and 2 represent internal standard.
Detailed description of the invention
In order to make the object, technical solutions and advantages of the present invention clearer, below in conjunction with accompanying drawing, the preferred embodiments of the present invention are described in detail, but protection scope of the present invention is not limited to this.
Instrument used in embodiments of the invention and condition, medicine are as follows with reagent:
Instrument: Waters (Quattro Premier XE MircoMass) High Performance Liquid Chromatography/Mass Spectrometry instrument；MassLynx4.0 chromatographic work station；Germany's Sartorius CP225D electronic balance；Germany's Eppendorf pipettor；Germany's IKA vortex mixed instrument；Centrifuge (Shanghai City An Ting tech equipment factory)；HGC-36A Nitrogen evaporator (upper Haiquan island scientific & trading Co., Ltd.).
Chromatographic column: watersMS C18Post (150mm × 2.1mm, 5 μm)；Column temperature: 30 DEG C；Flow velocity: 0.2ml/min；Sampling volume: 10 μ l.
Mass spectrum monitoring mode is polyion reaction monitoring (MRM), and detection ion is cation mode.Using electric spray ion source (ESI), capillary voltage is 3.5KV, and taper hole voltage is 35V, and ion source temperature is 110 DEG C, desolventizing gas (N2) flow is 500l/hr, desolventizing temperature is 350 DEG C.
Medicine and reagent:
Dauricine crude drug (purity >=98%, the room of testing, room is made by oneself)；Dauricine reference substance (Wei Qi bio tech ltd of Shenzhen, lot number: 20120625: purity: 99.6%)；Tetrandrine reference substance (Nat'l Pharmaceutical & Biological Products Control Institute, lot number: 110711-201001；Purity: 99.7%)；NaOH (analytical pure, Chengdu Ke Long chemical reagent factory)；Ethyl acetate and ammonium formate are chromatographically pure (world company of the U.S.)；Acetonitrile and methanol are chromatographically pure (Fisher company of the U.S.)；Water is the homemade deionized water of laboratory；Remaining reagent is analytical pure.
Healthy SD rat 12, ♂, weight (186 ± 21) g, Medical University Of Chongqing's Experimental Animal Center provide, quality certification SCXK (Chongqing) 2014-0002.
Embodiment 1
(1) chromatographic condition
Chromatographic column: watersMS C18Post (150mm × 2.1mm, 5 μm).
Flowing phase: flowing phase: acetonitrile (containing 2mM ammonium formate): 2mM ammonium formate aqueous solution=65:35 (volume ratio).
(2) Mass Spectrometry Conditions
Mass spectrum monitoring mode is polyion reaction monitoring (MRM), and detection ion is cation mode, Dauricine, [M+H]+Ion, m/z 625 → 206；Tetrandrine (internal standard), [M+H]+Ion, m/z 623 → 174.Using electric spray ion source (ESI), capillary voltage is 3.5KV, and taper hole voltage is 35V, and ion source temperature is 110 DEG C, desolventizing gas (N2) flow is 500l/hr, desolventizing temperature is 350 DEG C.
(3) preparation of solution
Dauricine and the preparation of internal standard storing solution: take Dauricine and internal standard reference substance about 10mg is the most accurately weighed, dissolve constant volume with methanol.
Dauricine and the preparation of internal standard working solution: take Dauricine and internal standard storing solution, become working solution with methanol dilution respectively.Dauricine working solution concentration is respectively 8480,4240,2120,1060,424,106,42.4ng/ml；Inside it is designated as 2036ng/ml.Storing solution and working solution all set to 0 in-4 DEG C of refrigerators and preserve.
(4) Dauricine and the detection of interior target
Measuring Dauricine and internal standard under conditions of step (3), mass spectrum is shown in Fig. 1.
Embodiment 2
(1) medicine-feeding test
It is appropriate that precision weighs Dauricine crude drug, and the NaOH adding 0.1mol/l dissolves in right amount, and is 6-7 with the HCl regulation pH value of 0.5mol/l, uses normal saline constant volume, makes the final concentration of 20mg/ml of Dauricine, is administered as test liquid.
Take Dauricine SMEDDS prescription prepared by present inventor's early stage, prepare Dauricine SMEDDS.The Dauricine SMEDDS mean diameter preparing gained is (58.3 ± 9.21) nm, and drug loading is about 49.46mg/g.
Take 12 and be divided into A, B two groups, often group 6 at random.After fasting 12h, ig is administered, A group rat ig Dauricine test liquid 50mg/Kg, B group rat ig Dauricine SMEDDS 20mg/Kg, before being administered and after being administered 0.167,0.333,0.5,0.75,1,2,4,8,12,24,36h taken blood about 0.3ml in heparinization centrifuge tube by retroorbital venous clump respectively, after blood specimen collection immediately centrifugal also separated plasma put-70 DEG C of ultra cold storage freezers preserve to be measured.
(2) chromatograph and Mass Spectrometry Conditions
Chromatographic column: watersMS C18Post (150mm × 2.1mm, 5 μm)；
Flowing phase: flowing phase: acetonitrile (containing 2mM ammonium formate): 2mM ammonium formate aqueous solution=65:35 (volume ratio)；
Column temperature: 30 DEG C；
Flow velocity: 0.2ml/min；
Sampling volume: 10 μ l.
Mass spectrum monitoring mode is polyion reaction monitoring (MRM), and detection ion is cation mode, Dauricine, [M+H]+Ion, m/z 625 → 206；Tetrandrine (internal standard), [M+H]+Ion, m/z 623 → 174.Using electric spray ion source (ESI), capillary voltage is 3.5KV, and taper hole voltage is 35V, and ion source temperature is 110 DEG C, desolventizing gas (N2) flow is 500l/hr, desolventizing temperature is 350 DEG C.
(3) preparation of solution
Dauricine and the preparation of internal standard storing solution: take Dauricine and internal standard reference substance about 10mg is the most accurately weighed, dissolve constant volume with methanol.
Dauricine and the preparation of internal standard working solution: take Dauricine and internal standard storing solution, become working solution with methanol dilution respectively.Dauricine working solution concentration is respectively 8480,4240,2120,1060,424,106,42.4ng/ml；Inside it is designated as 2036ng/ml.Storing solution and working solution all set to 0 in-4 DEG C of refrigerators and preserve.
(4) plasma sample processes
Precision measures blank rat plasma 0.1ml, adds methanol 5 μ l, internal standard 5 μ l, after mixing 10s, adds extractant ethyl acetate 1ml, after vortex mixed extraction 3min, and 1.3 × 104R/min is centrifuged 5min (centrifugal radius 15cm), takes supernatant, and at 40 DEG C, nitrogen dries up, and residue redissolves mutually with 50 μ l flowings, and sample introduction 10 μ l measures.
(5) method specificity
Measuring Dauricine under conditions of step (2) and internal standard mass spectrum is shown in Fig. 1, measure blank rat plasma, blank rat plasma+Dauricine+internal standard, rat plasma sample+internal standard chromatogram is shown in Fig. 2.From Figure 2 it can be seen that Dauricine and interior target retention time respectively may be about 1.9,2.5min, in this method, plasma endogenous material does not affect Dauricine and measures with interior target, meets Determination of Biological Samples specificity requirement.
(6) mensuration of standard curve
Preparation Dauricine concentration is respectively 424,212,106,53,21.20,5.30,2.12ng mL-1The each 0.1ml of rat plasma sample, plasma sample processing method according to step (4) operates, then according to the condition of step (2) detects, record Dauricine peak area and internal standard peak area, calculating standard curve equation by internal standard method is: Y=0.0788X+0.0056 (r=0.9999, W=1/C2), the range of linearity of the method mensuration rat plasma sample Dauricine concentration obtaining the present invention is 2.12-424ng/ml, and it is 2.12ng/ml (RSD=11.22%, n=15) that the method for the present invention measures the minimum quantitative concentrations of Dauricine in rat plasma.
(7) the method response rate and precision test
Preparation Dauricine concentration respectively 5.30,53, the plasma sample of 212ng/ml carry out precision and method recovery test, with measured value divided by the actual value computational methods response rate, within one day, to measure the result calculating withinday precision of 5 times, measured value calculates day to day precision for three days on end, the results are shown in Table 1.
The table 1 method response rate and Precision test result
(8) recovery of extraction and matrix effect test
The investigation method of recovery of extraction and matrix effect, and article that wherein preparation of biological sample and processing method are published in " Third Military Medical University's journal " with reference to the inventor oral administration biaavailability of Artemether rat " self-micro emulsifying medicament delivery system improve ", 2014,36 (14): 1481.
Result shows, concentration is 5.30,53, Dauricine recovery of extraction is (70.62 ± 6.47) % (n=5) in the rat plasma sample of 212ng/ml, internal standard recovery of extraction is (65.48 ± 4.38) %, (n=5)；Dauricine matrix effect is (98.87 ± 6.34) % (n=5), and internal standard matrix effect is (97.55 ± 4.38) %, (n=5).
(9) stability test
The stability of sample is the key factor of high performance liquid chromatography-tandem mass detection Dauricine concentration, and sample does not the most occur the degraded of target substance most important to testing result.
To use concentration be 5.30,53, the plasma sample of 212ng/ml carry out study on the stability, investigation project includes the stability after sample treatment, room temperature stability, three stability of freeze thawing and long-term frozen (-20 DEG C) stability.Result shows that sample keeps stable in 8h after at least processing, and room temperature can keep stable in placing 6h, keeps stablizing within freeze thawing three times, and long-term frozen keeps in 5 days stablizing.Stability test result error is all within 9.39%.
From above-described embodiment, the method for the high performance liquid chromatography-tandem mass detection Dauricine concentration of the present invention, specificity is strong, highly sensitive, accuracy is good, can be widely used for the mensuration of Dauricine sample.
Finally illustrate is, above example is only in order to illustrate technical scheme and unrestricted, although the present invention being described in detail with reference to preferred embodiment, it will be understood by those within the art that, technical scheme can be modified or equivalent, without deviating from objective and the scope of technical solution of the present invention, it all should be contained in the middle of scope of the presently claimed invention.
Claims (10)
1. the method for high performance liquid chromatography-tandem mass detection Dauricine concentration, it is characterised in that described method is to use internal standard method, with tetrandrine as internal standard substance.
Method the most according to claim 1, it is characterised in that the chromatographic column that described high performance liquid chromatography uses is the chromatographic column that octadecyl silane is filled, and the flowing of employing is the mixed liquor of acetonitrile and ammonium formate aqueous solution mutually.
Method the most according to claim 2, it is characterised in that the concentration of described ammonium formate aqueous solution is 1.5-2.5mM.
Method the most according to claim 2, it is characterised in that described acetonitrile is 60-80:20-40 with the volume ratio of ammonium formate aqueous solution.
Method the most according to claim 2, it is characterised in that described acetonitrile is the acetonitrile containing ammonium formate, the concentration of ammonium formate is 1.5-2.5mM.
Method the most according to claim 2, it is characterised in that described mass spectrum uses multiple reaction monitoring pattern, detection ion is that cation mode, Dauricine and tetrandrine are respectively adopted the combination of at least one pair of parent ion and daughter ion and carry out Quantitative Monitoring.
Method the most according to claim 6, it is characterised in that the parent ion of described Dauricine employing and being combined as of daughter ion: parent ion m/z is 625 ± 1, and daughter ion m/z is 206 ± 1.
Method the most according to claim 6, it is characterised in that the parent ion of described tetrandrine employing and being combined as of daughter ion: parent ion m/z is 623 ± 1, and daughter ion m/z is 174 ± 1.
9. the application in Dauricine concentration in detection plasma sample of the method described in any one of claim 1-8, it is characterized in that, the processing method of described plasma sample is: take plasma sample, add methanol, internal standard tetrandrine, after mixing, add extractant ethyl acetate, centrifuging and taking supernatant after extraction, nitrogen dries up, and the flowing that residue uses with high performance liquid chromatography is redissolved mutually, and last sample introduction is measured.
10. tetrandrine as internal standard substance Dauricine concentration measure in application.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112858497A (en) * | 2020-12-31 | 2021-05-28 | 四川新绿色药业科技发展有限公司 | Method for constructing characteristic spectrum of rhizoma Menispermi standard decoction and detecting quality of rhizoma Menispermi standard decoction |
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| F.M. HAN，ET AL: "Identification of dauricine and its metabolites in rat urine by liquid chromatography–tandem mass spectrometry", 《JOURNAL OF CHROMATOGRAPHY B》 * |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112858497A (en) * | 2020-12-31 | 2021-05-28 | 四川新绿色药业科技发展有限公司 | Method for constructing characteristic spectrum of rhizoma Menispermi standard decoction and detecting quality of rhizoma Menispermi standard decoction |
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