CN108562661A - A method of turmeric Contained Serum is measured using Thermo UPLC-ITQ-Orbitrap high resolution mass spectrums - Google Patents
A method of turmeric Contained Serum is measured using Thermo UPLC-ITQ-Orbitrap high resolution mass spectrums Download PDFInfo
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Abstract
This application involves a kind of methods for analyzing turmeric Contained Serum using Thermo UPLC ITQ Orbitrap high resolution mass spectrums.The application uses the chemical composition in Thermo UPLC ITQ Orbitrap high resolution mass spectrums analysis hand turmeric Contained Serum to lay the first stone for next step research pharmacokinetics.
Description
Technical field：Turmeric P.E Contained Serum is measured using UPLC-LTQ-Orbitrap this application involves a kind of
Method belongs to Analysis of Chinese Traditional Medicine and measures field.
Background technology：
" treating cold with hot drugs, treating heat with cold drug " is understanding most simple to herbal nature during Chinese medicine disease.Face
Turmeric on bed, the most important medicine difference of Radix Curcumae are that the two is respectively used to treatment YING jaundice (raw yellow subcutaneous ulcer) and yang jaundice card (damp and hot Huang
Subcutaneous ulcer).In order to study the two to cold, medicine of a warm nature sex differernce, multinomial research has been carried out early period.Turmeric clinic is usually used in treating liver and gall
Disease, turmeric first recorded in《Tang materia medica》, " for blade root all like Radix Curcumae, Hua Chun is born in root, and seedling and goes out, and summer flower is rotten, and s.m.p, root has
Yellow, green, Bai Sanse ", the method done is with Radix Curcumae with you." the lotus medicine of western army people meaning ".This section records explanation Turmeric at that time
It should be turmeric various plants ".《The Illustrated Book on Plants》Carry " turmeric,《Tang materia medica》Begin to record.... the full root like canna of its shape
Such as ginger, color and Huang, gas micro-pungent ", it is described to be consistent with modern turmeric (Curcuma.LongaL.), illustrate that Qing Dynasty's Turmeric is
CurcumaLongaL. rhizome ", 2010 editions States Pharmacopoeia specifications, turmeric is the drying of zingiberaceous plant turmeric (Curcum/longaL.)
Rhizome, winter cauline leaf are excavated when withered, are cleaned, are boiled or steam to the saturating heart, dry, and fibrous root is removed ".Chinese medicine thinks that turmeric is warm-natured, taste
Pungent, bitter, returns spleen, Liver Channel, which has blood-breaking, inducing meastruation to relieve menalgia effect, " is clinically used for chest side of body shouting pain, amenorrhoea, rheumatism shoulder arm pain
Bitterly, tumbling and swelling mainly originates in the ground such as China Sichuan, Guangdong, Fujian, Jiangxi, Guangxi.By early-stage study：Turmeric water
The cold Wen Yuqi turmerics cellulose content of extract, turmeric alcohol extract, Turmeric P.E has certain positive correlation, that is, there is certain " amount
One property " relationship.In order to further analyze, the relationship of turmeric and fever and chills, it would be desirable to enter the substance base in animal body to turmeric
Plinth, i.e. chemical composition are accurately analyzed.
Invention content：
Turmeric is the rhizome of zingiberaceous plant turmeric curcumalongaL., warm-natured, has blood-breaking, inducing meastruation to relieve menalgia work(
Energy.In order to study its pharmacological property, on the basis of early-stage study, the effective component group that further accurate analysis turmeric is warm-natured is needed,
The chemical composition in Contained Serum after selecting Thermo UPLC-ITQ-Orbitrap high resolution mass spectrums to give animal to turmeric
It is analyzed, and is determined through experimentation best analysis method, finally analyzed each ingredient in turmeric Contained Serum, be
Research pharmacokinetics lays the first stone in next step.
It is special this application involves a kind of method for analyzing Turmeric P.E Contained Serum using UPLC-LTQ-Orbitrap
Sign is to take sample to be tested (Turmeric P.E Contained Serum), carries out pre-treatment, divided using UPLC-LTQ-Orbitrap
Analysis, identifies the group in Contained Serum and is divided into Curcumin (curcumin), Dihydrocurcumin (double hydrogen curcumins), 5-
Hydroxy-1,7-bis (4-hydroxyphenyl) -3-heptanone (5- hydroxyls -1,7- bis- (4- hydroxy phenyls) -3- heptan
Ketone), Tetrahydrobisdemethoxycurcumin (tetrahydrochysene Bisdemethoxycurcumin).
A method of Turmeric P.E Contained Serum being analyzed using UPLC-LTQ-Orbitrap, which is characterized in that take and wait for
Sample (Turmeric P.E Contained Serum) is carried out pre-treatment, is analyzed using UPLC-LTQ-Orbitrap, identify and contain
Group in medicine serum is divided into Curcumin (curcumin), Dihydrocurcumin (double hydrogen curcumins), 5-Hydroxy-1,7-
Bis (4-hydroxyphenyl) -3-heptanone (bis- (4- the hydroxy phenyls) -3- heptanone of 5- hydroxyls -1,7-),
Tetrahydrobisdemethoxycurcumin (tetrahydrochysene Bisdemethoxycurcumin), wherein Turmeric P.E Contained Serum sample
Product are：Give Turmeric P.E gavage to rat, successive administration 16 days, anesthesia, abdominal aortic blood, centrifugation, collection serum.Its
Middle Turmeric P.E preparation method is：Turmeric is taken, is crushed, 40 mesh sieve is crossed, adds 10 times of amount water, first impregnates 0.5h, decocts 2 times, often
Secondary 45min, filtration, merge decocting liquid, concentration to get.
A method of Turmeric P.E Contained Serum being analyzed using UPLC-LTQ-Orbitrap, which is characterized in that take and wait for
Sample (Turmeric P.E Contained Serum) is carried out pre-treatment, is analyzed using UPLC-LTQ-Orbitrap, identify and contain
Group in medicine serum is divided into Curcumin (curcumin), Dihydrocurcumin (double hydrogen curcumins), 5-Hydroxy-1,7-
Bis (4-hydroxyphenyl) -3-heptanone (bis- (4- the hydroxy phenyls) -3- heptanone of 5- hydroxyls -1,7-),
Tetrahydrobisdemethoxycurcumin (tetrahydrochysene Bisdemethoxycurcumin), wherein Turmeric P.E Contained Serum sample
Product are：Give Turmeric P.E gavage to rat, successive administration 16 days, anesthesia, abdominal aortic blood, centrifugation, collection serum.Its
Middle Turmeric P.E preparation method is：Turmeric is taken, is crushed, 40 mesh sieve is crossed, adds 10 times of amount water, first impregnates 0.5h, decocts 2 times, often
Secondary 45min, filtration, merge decocting liquid, concentration to get；The pre-treating method of wherein sample to be tested is：Take Turmeric P.E drug containing
1800 μ L of chromatography methanol are added in 600 μ L of serum, and fully shaking 50 seconds carries out albumen precipitation, centrifuge (12000r/min,
10min), nitrogen dries up, and adds 100 μ L of methanol, centrifuges (15000r/min, 15 seconds), takes supernatant to obtain the final product.
In above-mentioned each method, chromatographic condition is：Chromatographic condition：Chromatographic column：Thermo Scientific Syncronic
(C18,100mm × 2.1mm, 1.7 μm), mobile phase：A be the methanol containing 0.1% formic acid (percent by volume), B be containing
The water of 0.1% formic acid (percent by volume), gradient elution program：0-20min, 5%A, 95%B；15-20min, 100%A, 0%
B；20-25min, 20%A, 80%B.Flow velocity 0.25mL/min, column temperature：55 DEG C, sample size：1 μ L, Detection wavelength：265nm；
In above-mentioned each method, Mass Spectrometry Conditions：Mass spectral analysis using be equipped with the quadrupole bar track of thermoelectricity esi ion source from
Sub- trap mass spectrograph.Negative ions source voltage is respectively 3.7kv and 3.5kv；Capillary heating temperature is：320℃；Sheath atmospheric pressure：
30psi；Assist gas pressure power：10psi.300 DEG C of volume heating evaporation temperature.Sheath rises and auxiliary gas is nitrogen.Collision gas is nitrogen
Gas, pressure 1.5mTorr.Level-one sweep parameter is：Resolution ratio 7000, automatic growth control target are 1 × 106, maximum isolation
Time 50ms, mass-to-charge ratio scanning range 150-1500.Compound component identification uses data dependence scan pattern, design parameter
For：Resolution ratio 17500, automatic growth control target are 1 × 105, maximum isolation time 50ms, maximum 10 ion scan two levels
Fragment, dynamic exclude.
In above-mentioned each processing method, liquid matter system is controlled by 2.2 SP1.48 softwares of Xcalibur, data acquisition and group
Quantitative Treatment is divided to be operated by the software.
A method of Turmeric P.E Contained Serum is analyzed using UPLC-LTQ-Orbitrap high resolution mass spectrums, it is special
Sign is, takes 600 μ L of Turmeric P.E Contained Serum, and 1800 μ L of chromatography methanol are added, and it is heavy to carry out albumen for fully shaking 50 seconds
It forms sediment, centrifuges (12000r/min, 10min), nitrogen drying adds 100 μ L of acetonitrile, centrifuges (15000r/min, 15 seconds), take supernatant
Sample introduction.Wherein, chromatographic condition is：Chromatographic condition：Chromatographic column：Thermo Scientific Syncronic (C18,100mm ×
2.1mm, 1.7 μm), mobile phase：A is the methanol containing 0.1% formic acid (percent by volume), and B is to contain 0.1% formic acid (volume
Percentage) water, gradient elution program：0-20min, 5%A, 95%B；15-20min, 100%A, 0%B：20-25min,
20%A, 80%B.Flow velocity 0.25mL/min, column temperature：55 DEG C, sample size：1 μ L, Detection wavelength：265nm.Mass Spectrometry Conditions：Mass spectrum
Analysis is using the quadrupole rod orbit ion trap mass spectrograph for being equipped with thermoelectricity esi ion source.Negative ions source voltage is respectively
3.7kv and 3.5kv；Capillary heating temperature is：320℃；Sheath atmospheric pressure：30psi；Assist gas pressure power：10psi.Volume heats
300 DEG C of evaporating temperature.Sheath rises and auxiliary gas is nitrogen.Collision gas is nitrogen, pressure 1.5mTorr.Level-one sweep parameter
For：Resolution ratio 7000, automatic growth control target are 1 × 106, maximum isolation time 50ms, mass-to-charge ratio scanning range 150-
1500.Using data dependence scan pattern, design parameter is for compound component identification：Resolution ratio 17500, automatic growth control mesh
It is designated as 1 × 105, maximum isolation time 50ms, maximum 10 ion scan secondary fragments, dynamic exclusion.Liquid matter system by
2.2 SP1.48 softwares of Xcalibur control, and data acquisition and measured portions processing are operated by the software.
A method of turmeric Contained Serum is analyzed using Thermo UPLC-ITQ-Orbitrap high resolution mass spectrums, it is special
Sign is, takes blank serum, 600 μ L of turmeric Contained Serum respectively, and 1800 μ L of chromatography methanol are added, centrifuge (12000r/min,
10min), N2 is dried up, and adds 100 μ L of methanol, is centrifuged (15000r/min, 15min), is taken supernatant sample introduction.
Wherein, chromatographic condition is chromatographic condition：Chromatographic column：Thermo Scientific Syncronic (C18,100 ×
2.1mm, 1.7um), mobile phase：A is acetonitrile, and B is 0.1% formic acid water, gradient elution program：0-20min, 10%A；20-
45min, 10%~60%；45-50min, 60%~99%；50-60min, 99~10%.Flow velocity 0.2mL/min, column temperature：35
DEG C, sample size：3uL, Detection wavelength：265nm；Mass Spectrometry Conditions：Scan mode：Negative ions scan respectively.Electron spray voltage：+
5KV (cation), -5KV (anion)；Capillary temperature：350℃；Sheath gas：35psi；Secondary air speed：10psi；It sweeps
Tail gas flow velocity：2psi.Scanning of the mass spectrum mode：Mode (is swept) entirely using FT Full Scan to be scanned, scanning range 50-
1500Da；Resolution ratio：30000.MSN：It is scanned using FT modes, resolution ratio：15000.It is set using what data dependence scanned
Fixed to carry out collision induced dissociation (CID) to most strong ion, CID resolution widths are 2Da, and cracking energy is 35%.Wherein turmeric water
Decocting liquid Contained Serum is that rat continuous gavage is taken to be administered 2 days, gives turmeric routine decocting liquid 5ml every time, 2 hours after the last administration,
Blood is taken, centrifuges, supernatant is taken to obtain Contained Serum.
A method of turmeric is analyzed using Thermo UPLC-ITQ-Orbitrap high resolution mass spectrums, which is characterized in that
Turmeric Root 1g is taken respectively, is placed in conical flask with stopper, 50ml methanol is added, and ultrasonic 30min crosses the miillpore filter mistake of 0.22um
After filter, direct injection analysis
Wherein, chromatographic condition is chromatographic condition：Chromatographic column：Thermo Scientific Syncronic (C18,100 ×
2.1mm, 1.7um), mobile phase：A is acetonitrile, and B is 0.1% formic acid water, gradient elution program：0-20min, 10%A；20-
45min, 10%~60%；45-50min, 60%~99%；50-60min, 99~10%.Flow velocity 0.2mL/min, column temperature：35
DEG C, sample size：3uL, Detection wavelength：265nm；Mass Spectrometry Conditions：Scan mode：Negative ions scan respectively.Electron spray voltage：+
5KV (cation), -5KV (anion)；Capillary temperature：350℃；Sheath gas：35psi；Secondary air speed：10psi；It sweeps
Tail gas flow velocity：2psi.Scanning of the mass spectrum mode：Mode (is swept) entirely using FT Full Scan to be scanned, scanning range 50-
1500Da；Resolution ratio：30000.MSN：It is scanned using FT modes, resolution ratio：15000.It is set using what data dependence scanned
Fixed to carry out collision induced dissociation (CID) to most strong ion, CID resolution widths are 2Da, and cracking energy is 35%.
Description of the drawings：
1 Turmeric P.E Contained Serum of attached drawing detects chromatogram-A1：Turmeric P.E radical anion peak figure (BPC), A2：
Turmeric P.E Contained Serum radical anion peak figure (BPC)；
2 Turmeric P.E Contained Serum of attached drawing detects chromatogram-B1：Turmeric P.E radical cation peak figure (BPC)；B2：
Turmeric P.E Contained Serum radical anion peak figure (BPC)
Specific implementation mode：
Using UPLC-LTQ-Orbitrap high resolution mass spectrums, constituent analysis has been carried out to the Contained Serum of turmeric decocting liquid,
And analysis is carried out using turmeric medicinal substances extract and is compareed as retinue.For curcumin ingredient, by accurate molecular weight compare with
And second order ms parsing.
One, laboratory apparatus, reagent and method
1, instrument and reagent：UPLC-ITQ-Orbitrap (Thermo companies of the U.S.), supper-fast liquid chromatogram instrument
(Thermo Fisher Scientific, model DionexTMUltiMateTM300), quadrupole rod orbit ion trap mass spectrograph, first
Alcohol (chromatographic grade), acetonitrile (mass spectrum grade), high purity water, formic acid, nitrogen.
2, chromatographic condition：
Chromatographic column：Thermo Scientific Syncronic (C18,100mm × 2.1mm, 1.7 μm), mobile phase：A is
Methanol containing 0.1% formic acid (percent by volume), B are the water containing 0.1% formic acid (percent by volume), gradient elution journey
Sequence：0-20min, 5%A, 95%B；15-20min, 100%A, 0%B；20-25min, 20%A, 80%B.Flow velocity 0.25mL/
Min, column temperature：55 DEG C, sample size：1 μ L, Detection wavelength：265nm.
3, Mass Spectrometry Conditions：
Mass spectral analysis is using the quadrupole rod orbit ion trap mass spectrograph for being equipped with thermoelectricity esi ion source.Negative ions source electricity
Pressure is respectively 3.7kv and 3.5ky；Capillary heating temperature is：320℃；Sheath atmospheric pressure：30psi；Assist gas pressure power：10psi.
300 DEG C of volume heating evaporation temperature.Sheath rises and auxiliary gas is nitrogen.Collision gas is nitrogen, pressure 1.5mTorr.Level-one is swept
Retouching parameter is：Resolution ratio 7000, automatic growth control target are 1 × 106, maximum isolation time 50ms, mass-to-charge ratio scanning range
150-1500.Using data dependence scan pattern, design parameter is for compound component identification：Resolution ratio 17500, automatic gain control
Target processed is 1 × 105, maximum isolation time 50ms, maximum 10 ion scan secondary fragments, dynamic exclusion.Liquid matter system by
2.2 SP1.48 softwares of Xcalibur control, and data acquisition and measured portions processing are operated by the software.
4, sample-pretreating method：
Turmeric medicinal material sample：Turmeric Root 1g is taken, is placed in conical flask with stopper, 50ml methanol, ultrasonic 30min, mistake is added
After the filtering with microporous membrane of 0.22um, direct injection analysis.
Turmeric Contained Serum sample is：Give Turmeric P.E gavage to rat, successive administration 16 days, anesthesia, abdomen is actively
Arteries and veins takes blood, and serum is collected in centrifugation.Wherein Turmeric P.E preparation method is：Turmeric is taken, is crushed, 40 mesh sieve is crossed, adds 10 times of amounts
Water, first impregnates 0.5h, decocts 2 times, each 45min, filtration, merges decocting liquid, concentration to get.
Sample treatment：
600 μ L of turmeric Contained Serum are taken, 1800 μ L of chromatography methanol are added, fully shaking 50 seconds carries out albumen precipitation, centrifugation
(12000r/min, 10min), nitrogen drying, adds 100 μ L of acetonitrile, centrifuges (15000r/min, 15 seconds), takes supernatant to obtain the final product.
5. sample analysis：The above-mentioned sample handled well is taken, according to above-mentioned chromatographic condition, Mass Spectrometry Conditions sample introduction.
Two, analysis result
1. Turmeric P.E and Turmeric P.E Contained Serum analysis result
1 Turmeric P.E of table and Turmeric P.E Contained Serum analysis result
**：Indicate the ingredient detected in Contained Serum
Brief summary：It is respectively Curcumin (turmerics that 4 compounds have been analyzed and identified from the Contained Serum of Turmeric P.E
Element), Dihydrocurcumin (double hydrogen curcumins), 5-Hydroxy-1,7-bis (4-hydroxyphenyl) -3-
Heptanone (bis- (4- the hydroxy phenyls) -3- heptanone of 5- hydroxyls -1,7-), Tetrahydrobisdemethoxycurcumin (four
Hydrogen Bisdemethoxycurcumin).Result above is to carry out turmeric pharmacokinetic in next step to provide basis.
Claims (10)
1. a kind of method for analyzing Turmeric P.E Contained Serum using UPLC-LTQ-Orbitrap, it is characterised in that take to be measured
Sample (Turmeric P.E Contained Serum) is carried out pre-treatment, is analyzed using UPLC-LTQ-Orbi trap.
2. a kind of method for analyzing Turmeric P.E Contained Serum using UPLC-LTQ-Orbitrap, it is characterised in that take to be measured
Sample (Turmeric P.E Contained Serum) is carried out pre-treatment, is analyzed using UPLC-LTQ-Orbitrap, identify drug containing
Group in serum is divided into Curcumin (curcumin), Dihydrocurcumin (double hydrogen curcumins), 5-Hydroxy-1,7-bis
(4-hydroxyphenyl) -3-heptanone (bis- (4- the hydroxy phenyls) -3- heptanone of 5- hydroxyls -1,7-),
Tetrahydrobisdemethoxycurcumin (tetrahydrochysene Bisdemethoxycurcumin).
3. the method that Turmeric P.E Contained Serum is analyzed using UPLC-LTQ-Orbitrap as claimed in claim 1 or 2,
Wherein Turmeric P.E Contained Serum sample is：Give Turmeric P.E gavage to rat, successive administration 16 days, anesthesia, abdomen master
Serum is collected in arterial blood extracting, centrifugation, and wherein Turmeric P.E preparation method is：Turmeric is taken, is crushed, 40 mesh sieve is crossed, adds 10 times of amounts
Water, first impregnates 0.5h, decocts 2 times, each 45min, and filtration merges decocting liquid, be concentrated into 50ml to get.
4. analyzing Turmeric P.E drug containing blood using UPLC-LTQ-Orbitrap as described in any one of claim 1-3
The pre-treating method of clear method, wherein sample to be tested is：600 μ L of Turmeric P.E Contained Serum are taken, chromatography methanol is added
1800 μ L, fully shaking 50 seconds carry out albumen precipitation, centrifuge (12000r/min, 10min), and nitrogen drying adds 100 μ L of methanol,
It centrifuges (15000r/min, 15 seconds), takes supernatant to obtain the final product.
5. analyzing Turmeric P.E drug containing blood using UPLC-LTQ-Orbitrap as described in any one of claim 1-4
Clear method, wherein chromatographic condition are：Chromatographic column：Thermo Scientific Syncronic (C18,100mm × 2.1mm,
1.7 μm), mobile phase：A is the methanol containing 0.1% formic acid (percent by volume), and B is to contain 0.1% formic acid (percent by volume)
Water, gradient elution program：0-20min, 5%A, 95%B；15-20min, 100%A, 0%B；20-25min, 20%A, 80%
B.Flow velocity 0.25mL/min, column temperature：55 DEG C, sample size：1 μ L, Detection wavelength：265nm.
6. analyzing Turmeric P.E drug containing blood using UPLC-LTQ-Orbitrap as described in any one of claim 1-5
Clear method, wherein Mass Spectrometry Conditions are：Mass spectral analysis is using the quadrupole rod orbit ion trap matter for being equipped with thermoelectricity esi ion source
Spectrometer, negative ions source voltage is respectively 3.7kv and 3.5kv；Capillary heating temperature is：320℃；Sheath atmospheric pressure：30psi；
Assist gas pressure power：10psi, 300 DEG C of volume heating evaporation temperature, sheath gas and auxiliary gas are nitrogen, and collision gas is nitrogen, pressure
For 1.5mTorr, level-one sweep parameter is：Resolution ratio 7000, automatic growth control target are 1 × 106, the maximum isolation time
50ms, mass-to-charge ratio scanning range 150-1500, using data dependence scan pattern, design parameter is for compound component identification：Point
Resolution 17500, automatic growth control target are 1 × 105, maximum isolation time 50ms, maximum 10 ion scan secondary fragments,
Dynamic excludes.
7. the side for analyzing turmeric Contained Serum using UPLC-LTQ-Orbitrap as described in any one of claim 1-6
Method, wherein liquid matter system are controlled by 2.2 SP1.48 softwares of Xcalibur, and data acquisition and measured portions processing are soft by this
Part operates.
8. a kind of identifying Curcumin (curcumin) in Turmeric P.E Contained Serum, Dihydrocurcumin (double hydrogen ginger simultaneously
Flavine), 5-Hydroxy-1,7-bis (4-hydroxyphenyl) -3-heptanone (bis- (4- hydroxy benzenes of 5- hydroxyls -1,7-
Base) -3- heptanone), the method for Tetrahydrobisdemethoxycurcumin (tetrahydrochysene Bisdemethoxycurcumin), feature
Be using as described in any one of claim 1-7 using UPLC-LTQ-Orbitrap analysis turmeric Contained Serums
Method.
9. a kind of method that turmeric decocting liquid Contained Serum is analyzed using Thermo UPLC-ITQ-Orbitrap high resolution mass spectrums,
It is characterized in that, taking 600 μ L of turmeric decocting liquid Contained Serum, 1800 μ L of chromatography methanol are added, centrifuge (12000r/min,
10min), N2 is dried up, and adds 100 μ L of methanol, is centrifuged (15000r/min, 15min), is taken supernatant sample introduction.Wherein, chromatographic condition is
Chromatographic condition：Chromatographic column：Thermo Scientific Syncronic (C18,100 × 2.1mm, 1.7um), mobile phase：A is
Acetonitrile, B are 0.1% formic acid water, gradient elution program：0-20min, 10%A；20-45min, 10%~60%；45-50min,
60%~99%；50-60min, 99~10%.Flow velocity 0.2mL/min, column temperature：35 DEG C, sample size：3uL, Detection wavelength：
265nm；Mass Spectrometry Conditions：Scan mode：Negative ions scan respectively.Electron spray voltage：+ 5KV (cation), -5KV (bear from
Son)；Capillary temperature：350℃；Sheath gas：35psi；Secondary air speed：10psi；It winds up gas velocity：2psi.Scanning of the mass spectrum
Mode：Mode (is swept) entirely using FT Full Scan to be scanned, scanning range 50-1500Da；Resolution ratio：30000.MSN：It adopts
It is scanned with FT modes, resolution ratio：15000.The setting scanned using data dependence carries out collision-induced solution to most strong ion
From (CID), CID resolution widths are 2Da, and cracking energy is 35%.Wherein turmeric decocting liquid Contained Serum is that rat is taken continuously to fill
Stomach is administered 2 days, gives turmeric routine decocting liquid 5ml every time, 2 hours after the last administration, takes blood, centrifuges, supernatant is taken to obtain drug containing
Serum.
10. a kind of analyzing turmeric method using Thermo UPLC-ITQ-Orbitrap high resolution mass spectrums, which is characterized in that take ginger
600 μ L of yellow sample are added 1800 μ L of chromatography methanol, centrifuge (12000r/min, 10min), and N2 dryings add 100 μ L of methanol, centrifuge
(15000r/min, 15min) takes supernatant sample introduction.Wherein, chromatographic condition is chromatographic condition：Chromatographic column：Thermo
Scientific Syncronic (C18,100 × 2.1mm, 1.7um), mobile phase：A is acetonitrile, and B is 0.1% formic acid water, gradient
Elution program：0-20min, 10%A；20-45min, 10%~60%；45-50min, 60%~99%；50-60min, 99~
10%.Flow velocity 0.2mL/min, column temperature：35 DEG C, sample size：3uL, Detection wavelength：265nm；Mass Spectrometry Conditions：Scan mode：It is positive and negative
Ion scans respectively.Electron spray voltage：+ 5KV (cation), -5KV (anion)；Capillary temperature：350℃；Sheath gas：
35psi；Secondary air speed：10psi；It winds up gas velocity：2psi.Scanning of the mass spectrum mode：Using the sides FT Full Scan (sweeping entirely)
Formula is scanned, scanning range 50-1500Da；Resolution ratio：30000.MSN：It is scanned using FT modes, resolution ratio：
15000.The setting scanned using data dependence carries out collision induced dissociation (CID) to most strong ion, and CID resolution widths are 2Da,
It is 35% to crack energy.Wherein turmeric sample is to be prepared with the following method：Turmeric, Radix Curcumae powder 1g are taken respectively, are placed in
In conical flask with stopper, 50ml methanol is added, ultrasonic 30min is spare after crossing the filtering with microporous membrane of 0.22um.
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