CN103412082B - HPLC (High Performance Liquid Chromatography) technique-based detection method for contents of jasmonic acid compounds in lycoris radiate - Google Patents
HPLC (High Performance Liquid Chromatography) technique-based detection method for contents of jasmonic acid compounds in lycoris radiate Download PDFInfo
- Publication number
- CN103412082B CN103412082B CN201310370131.4A CN201310370131A CN103412082B CN 103412082 B CN103412082 B CN 103412082B CN 201310370131 A CN201310370131 A CN 201310370131A CN 103412082 B CN103412082 B CN 103412082B
- Authority
- CN
- China
- Prior art keywords
- sample
- jasmonic
- add
- solution
- dihydro
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
- 238000001514 detection method Methods 0.000 title claims abstract description 24
- 241001633628 Lycoris Species 0.000 title claims abstract description 19
- 238000004128 high performance liquid chromatography Methods 0.000 title claims abstract description 15
- 238000000034 method Methods 0.000 title abstract description 14
- ZNJFBWYDHIGLCU-HWKXXFMVSA-N jasmonic acid Chemical class CC\C=C/C[C@@H]1[C@@H](CC(O)=O)CCC1=O ZNJFBWYDHIGLCU-HWKXXFMVSA-N 0.000 title abstract 11
- 241000196324 Embryophyta Species 0.000 claims abstract description 18
- 229940088597 hormone Drugs 0.000 claims abstract description 17
- 239000005556 hormone Substances 0.000 claims abstract description 17
- 150000001875 compounds Chemical class 0.000 claims abstract description 13
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 57
- GEWDNTWNSAZUDX-UHFFFAOYSA-N methyl 7-epi-jasmonate Natural products CCC=CCC1C(CC(=O)OC)CCC1=O GEWDNTWNSAZUDX-UHFFFAOYSA-N 0.000 claims description 36
- GEWDNTWNSAZUDX-WQMVXFAESA-N (-)-methyl jasmonate Chemical compound CC\C=C/C[C@@H]1[C@@H](CC(=O)OC)CCC1=O GEWDNTWNSAZUDX-WQMVXFAESA-N 0.000 claims description 27
- 239000000523 sample Substances 0.000 claims description 26
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 15
- 239000000243 solution Substances 0.000 claims description 13
- 239000008346 aqueous phase Substances 0.000 claims description 11
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 9
- 238000000605 extraction Methods 0.000 claims description 8
- 238000002390 rotary evaporation Methods 0.000 claims description 8
- 230000014759 maintenance of location Effects 0.000 claims description 7
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 6
- 238000005374 membrane filtration Methods 0.000 claims description 6
- 239000012071 phase Substances 0.000 claims description 6
- 238000000746 purification Methods 0.000 claims description 6
- 239000012086 standard solution Substances 0.000 claims description 6
- 150000002148 esters Chemical class 0.000 claims description 5
- 239000000706 filtrate Substances 0.000 claims description 5
- 239000008363 phosphate buffer Substances 0.000 claims description 5
- 239000012488 sample solution Substances 0.000 claims description 5
- 239000006228 supernatant Substances 0.000 claims description 5
- 239000007788 liquid Substances 0.000 claims description 4
- -1 polyethylene pyrrolidone Polymers 0.000 claims description 4
- 239000007864 aqueous solution Substances 0.000 claims description 3
- 238000004132 cross linking Methods 0.000 claims description 3
- 238000010790 dilution Methods 0.000 claims description 3
- 239000012895 dilution Substances 0.000 claims description 3
- 230000010354 integration Effects 0.000 claims description 3
- 238000010829 isocratic elution Methods 0.000 claims description 3
- 235000013809 polyvinylpolypyrrolidone Nutrition 0.000 claims description 3
- 229920000523 polyvinylpolypyrrolidone Polymers 0.000 claims description 3
- 230000001105 regulatory effect Effects 0.000 claims description 3
- 239000003463 adsorbent Substances 0.000 claims description 2
- 238000000926 separation method Methods 0.000 abstract description 3
- 230000037353 metabolic pathway Effects 0.000 abstract description 2
- 230000024053 secondary metabolic process Effects 0.000 abstract description 2
- ZNJFBWYDHIGLCU-UHFFFAOYSA-N jasmonic acid Natural products CCC=CCC1C(CC(O)=O)CCC1=O ZNJFBWYDHIGLCU-UHFFFAOYSA-N 0.000 abstract 3
- 239000000126 substance Substances 0.000 abstract 1
- 238000002965 ELISA Methods 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- 238000001212 derivatisation Methods 0.000 description 3
- 238000002290 gas chromatography-mass spectrometry Methods 0.000 description 3
- 238000000444 liquid chromatography-electrochemical detection Methods 0.000 description 3
- 239000007791 liquid phase Substances 0.000 description 3
- VYFYYTLLBUKUHU-UHFFFAOYSA-N dopamine Chemical compound NCCC1=CC=C(O)C(O)=C1 VYFYYTLLBUKUHU-UHFFFAOYSA-N 0.000 description 2
- 239000003547 immunosorbent Substances 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 229960003638 dopamine Drugs 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000000589 high-performance liquid chromatography-mass spectrometry Methods 0.000 description 1
- 238000004895 liquid chromatography mass spectrometry Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000004885 tandem mass spectrometry Methods 0.000 description 1
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
Abstract
The invention discloses an HPLC (High Performance Liquid Chromatography) technique-based detection method for the contents of jasmonic acid compounds in lycoris radiate. According to the detection method, multiple jasmonic acid hormones in plants can be simultaneously, quickly and accurately detected by extracting and purifying the jasmonic acid compounds in a lycoris radiate sample, establishing a standard substance separation chromatogram, and determining the contents of the jasmonic acid compounds; a detection result is reliable, and the separation degrees of various compounds are good. The detection method can be used for detecting jasmonic acid endogenous hormones in the plants and providing a technical platform for researching the roles played by various jasmonic acid hormones in growth, development and secondary metabolism processes of the plants and simultaneously researching the metabolic pathways of the jasmonic acid compounds in the plants.
Description
Technical field
The present invention relates to a kind of method utilizing high-efficient liquid phase technique HPLC to measure hormone in plant, in particular a kind of detection method based on Jasmonates compounds content in the short-tube lycoris of HPLC technology.
Background technology
Jasmonates compound is the parahormone being extensively present in plant, in plant, content is very micro-, the method that can be used for the Jasmonates hormone detected in plant at present mainly contains: gas chromatography mass spectrometry (GC-MS), LC-MS (HPLC-MS) and enzyme linked immunosorbent detection technology (ELISA), the methods such as UPLC-MS-MS, also liquid chromatography electrochemistry detection method (liquid chromatography-electrochemical detection, HPLC-ECD) is had.
In existing detection technique, independent HPLC detection method can't detect the multiple Jasmonates hormone in plant accurately and quickly.GC-MS detection method, although sensitivity is good, needs first will carry out derivatization when measuring Jasmonates compound.The derivatization adopted is ester derivatization, and jasmonic can be transformed into ester type compound, and endogenous Jasmonates hormone includes the compound such as methyl jasmonate and dihydro methyl jasmonate, so GC-MS method also has certain limitation.Euzymelinked immunosorbent assay (ELISA) detection method (ELISA) and liquid chromatography electrochemistry detection method (HPLC-ECD) have higher specificity.But Jasmonates classes of compounds is more, ELISA method can not detect multiple hormone simultaneously, and repeatability is poor; The HPLC-ECD method reported is reacted at jasmonic and dopamine, and generate the material with electrochemical signals, the method can only detect the content of jasmonic.The step of other detection method is all complicated or higher to the requirement of instrument and equipment, is not easy to the mensuration rapidly and accurately of hormone.
Summary of the invention
The object of the invention is to overcome the deficiencies in the prior art, provide a kind of detection method based on Jasmonates compounds content in the short-tube lycoris of HPLC technology, detect four kinds of Jasmonates hormones in plant comparatively rapidly and accurately simultaneously.
The present invention is achieved by the following technical solutions, the present invention includes following steps:
(1) extraction and purification of sample
Get the fresh plant leaf 10g of testing sample, add the methanol solution that 20 ~ 30ml volumetric concentration is 80%, add adsorbent, ice bath grinds to form homogenate, gets supernatant rotary evaporation to aqueous phase after centrifugal; Add phosphate buffer, extraction after decolouring, then redissolve, after Spe C18 pillar purifying, use 0.45 μm of membrane filtration, filtrate is as sample solution;
(2) setting of pick-up unit
Chromatographic column is Eclipse XDB-C18,250mm × 4.6mm, 5 μm; Mobile phase A methyl alcohol, the acetic acid aqueous solution of B pH3.6; Isocratic elution A:B=1:1, flow velocity is 1mL/min, determined wavelength 235nm, and column temperature is 25 DEG C, and treat that baseline is steady, sample size is 10 μ L;
(3) Specification Curve of Increasing
Accurately take the standard items of jasmonic JA, dihydro jasmonic DJA, methyl jasmonate MeJA and dihydro methyl jasmonate DMeJA respectively, add methyl alcohol and dissolve, shake up and be mixed with the standard solution that mass concentration is 1.0 ~ 1.2mg/ml; Get each standard solution respectively, add methyl alcohol stepwise dilution, obtained mass concentration is respectively the standard serial solution of 200 μ g/ml, 100 μ g/ml, 50 μ g/ml, 20 μ g/ml, 10 μ g/ml, 5 μ g/ml; Precision measures 10 μ l sample introductions in high performance liquid chromatograph respectively, record chromatogram, respectively with the content x of each standard items for horizontal ordinate, with peak area y for ordinate, drawing standard curve, the content of horizontal ordinate standard items is in 5 ~ 200 mcg/ml, the retention time RT obtaining standard items jasmonic, dihydro jasmonic, methyl jasmonate and dihydro methyl jasmonate is respectively: jasmonic RT is 4.380min, the RT of dihydro jasmonic is 6.256min, the RT of methyl jasmonate is 8.603min, and the RT of dihydro methyl jasmonate is 13.196min;
(4) the Jasmonates content in testing sample short-tube lycoris tissue is detected
Short-tube lycoris sample is carried out HPLC detection according to step (2) and step (3) after step (1) extraction and purification, sample introduction 10 μ l, peak area under the same retention time of integration, disposable detection obtains four Jasmonates hormones and content thereof in short-tube lycoris sample to be measured.
In described step (1), get fresh plant leaf 10g, add the methanol solution that 20 ~ 30ml volumetric concentration is 80%, hold over night at 4 DEG C, add cross-linking polyethylene pyrrolidone PVPP solution, ice bath grinds to form homogenate, gets supernatant rotary evaporation to aqueous phase after centrifugal; Add phosphate buffer, and then add the decolouring of isopyknic sherwood oil, retain aqueous phase, in triplicate, when ether is mutually without the pH to 8.0 regulating aqueous phase during color, isopyknic ethyl acetate is then used to extract, repeat twice, merge ester phase, rotary evaporation, is concentrated into dry, then joining 1ml volumetric concentration is redissolve in 60% methyl alcohol, after Spe C18 pillar purifying, use 0.45 μm of membrane filtration, filtrate is as sample solution.
The present invention has the following advantages compared to existing technology: method of the present invention can detect multiple Jasmonates hormone simultaneously, and testing result is reliable, and various compound separation degree is better.May be used for detecting the Jasmonates endogenous hormones in plant, for studying the effect that various Jasmonates hormone plays in growth and development of plants and secondary metabolism process, simultaneously for the study metabolic pathways of Jasmonates compound in plant provides technology platform.
Accompanying drawing explanation
Fig. 1 is the high-efficient liquid phase chromatogram of jasmonic JA of the present invention, dihydro jasmonic DJA, methyl jasmonate MeJA and dihydro methyl jasmonate DMeJA standard items;
Fig. 2 is the high-efficient liquid phase chromatogram of the present invention's short-tube lycoris sample to be measured.
Embodiment
Elaborate to embodiments of the invention below, the present embodiment is implemented under premised on technical solution of the present invention, give detailed embodiment and concrete operating process, but protection scope of the present invention is not limited to following embodiment.
The present embodiment comprises the following steps:
(1) extraction and purification of sample
Get the fresh plant leaf 10g of testing sample, add the methanol solution that 25ml volumetric concentration is 80%, hold over night at 4 DEG C, add cross-linking polyethylene pyrrolidone PVPP solution, ice bath grinds to form homogenate, gets supernatant rotary evaporation to aqueous phase after centrifugal; Add phosphate buffer, and then add the decolouring of isopyknic sherwood oil, retain aqueous phase, in triplicate, when ether is mutually without the pH to 8.0 regulating aqueous phase during color, isopyknic ethyl acetate is then used to extract, repeat twice, merge ester phase, rotary evaporation, is concentrated into dry, then joining 1ml volumetric concentration is redissolve in 60% methyl alcohol, after Spe C18 pillar purifying, use 0.45 μm of membrane filtration, filtrate is as sample solution;
(2) setting of pick-up unit
The HPLC device of the present embodiment: Agilent1200 high performance liquid chromatograph workstation, Agilent1200 series pump, variable-wavelenght detector, chromatographic column is Eclipse XDB-C18,250mm × 4.6mm, 5 μm; Mobile phase A methyl alcohol, the acetic acid aqueous solution of B pH3.6; Isocratic elution A:B=1:1, flow velocity is 1mL/min, determined wavelength 235nm, and column temperature is 25 DEG C, and treat that baseline is steady, sample size is 10 μ L;
(3) Specification Curve of Increasing
Accurately take the standard items of jasmonic, dihydro jasmonic, methyl jasmonate and dihydro methyl jasmonate respectively, add methyl alcohol and dissolve, shake up and be mixed with the standard solution that mass concentration is 1.0 ~ 1.2mg/ml; Get each standard solution respectively, add methyl alcohol stepwise dilution, obtained mass concentration is respectively the standard serial solution of 200 μ g/ml, 100 μ g/ml, 50 μ g/ml, 20 μ g/ml, 10 μ g/ml, 5 μ g/ml; Precision measures 10 μ l sample introductions in high performance liquid chromatograph respectively, record chromatogram, respectively with the content x of each standard items for horizontal ordinate, with peak area y for ordinate, drawing standard curve, as shown in Figure 1, in figure, JA, DJA, MeJA and DMeJA are jasmonic, dihydro jasmonic, methyl jasmonate and dihydro methyl jasmonate respectively; The content of horizontal ordinate standard items is in 5 ~ 200 mcg/ml, the retention time RT obtaining standard items jasmonic, dihydro jasmonic, methyl jasmonate and dihydro methyl jasmonate is respectively: jasmonic RT is 4.380, the RT of dihydro jasmonic is 6.256, the RT of methyl jasmonate is 8.603, and the RT of dihydro methyl jasmonate is 13.196;
(4) the Jasmonates content in testing sample short-tube lycoris tissue is detected
Short-tube lycoris sample is carried out HPLC detection according to step (2) and step (3) after step (1) extraction and purification, sample introduction 10 μ l, peak area under the same retention time of integration, disposable detection obtains four Jasmonates hormones and content thereof in short-tube lycoris sample to be measured, as shown in Figure 2, can judge according to the retention time of jasmonic, dihydro jasmonic, methyl jasmonate and dihydro methyl jasmonate, draw four Jasmonates hormones in short-tube lycoris sample to be measured, content can judge according to peak area.
Absolute methanol used, acetic acid and deionized water all use 0.45 nanometer micropore membrane filtration above, and absolute methanol used, acetic acid jasmonic, dihydro jasmonic, methyl jasmonate and dihydro methyl jasmonate compound purity are chromatographically pure.
Claims (2)
1., based on a detection method for Jasmonates compounds content in the short-tube lycoris of HPLC technology, it is characterized in that, comprise the following steps:
(1) extraction and purification of sample
Get the fresh plant leaf 10g of testing sample, add the methanol solution that 20 ~ 30ml volumetric concentration is 80%, add adsorbent, ice bath grinds to form homogenate, gets supernatant rotary evaporation to aqueous phase after centrifugal; Add phosphate buffer, extraction after decolouring, then redissolve, after Spe C18 pillar purifying, use 0.45 μm of membrane filtration, filtrate is as sample solution;
(2) setting of pick-up unit
Chromatographic column is Eclipse XDB-C18,250mm × 4.6mm, 5 μm; Mobile phase A methyl alcohol, the acetic acid aqueous solution of B pH3.6; Isocratic elution A:B=1:1, flow velocity is 1mL/min, determined wavelength 235nm, and column temperature is 25 DEG C, and treat that baseline is steady, sample size is 10 μ L;
(3) Specification Curve of Increasing
Accurately take the standard items of jasmonic JA, dihydro jasmonic DJA, methyl jasmonate MeJA and dihydro methyl jasmonate DMeJA respectively, add methyl alcohol and dissolve, shake up and be mixed with the standard solution that mass concentration is 1.0 ~ 1.2mg/ml; Get each standard solution respectively, add methyl alcohol stepwise dilution, obtained mass concentration is respectively the standard serial solution of 200 μ g/ml, 100 μ g/ml, 50 μ g/ml, 20 μ g/ml, 10 μ g/ml, 5 μ g/ml; Precision measures 10 μ l sample introductions in high performance liquid chromatograph respectively, record chromatogram, respectively with the content x of each standard items for horizontal ordinate, with peak area y for ordinate, drawing standard curve, the content of horizontal ordinate standard items is in 5 ~ 200 mcg/ml, the retention time RT obtaining standard items jasmonic, dihydro jasmonic, methyl jasmonate and dihydro methyl jasmonate is respectively: jasmonic RT is 4.380min, the RT of dihydro jasmonic is 6.256min, the RT of methyl jasmonate is 8.603min, and the RT of dihydro methyl jasmonate is 13.196min;
(4) the Jasmonates content in testing sample short-tube lycoris tissue is detected
Short-tube lycoris sample is carried out HPLC detection according to step (2) and step (3) after step (1) extraction and purification, sample introduction 10 μ l, peak area under the same retention time of integration, disposable detection obtains four Jasmonates hormones and content thereof in short-tube lycoris sample to be measured.
2. a kind of detection method based on Jasmonates compounds content in the short-tube lycoris of HPLC technology according to claim 1, it is characterized in that, in described step (1), get fresh plant leaf 10g, add the methanol solution that 20 ~ 30ml volumetric concentration is 80%, hold over night at 4 DEG C, add cross-linking polyethylene pyrrolidone PVPP solution, ice bath grinds to form homogenate, gets supernatant rotary evaporation to aqueous phase after centrifugal; Add phosphate buffer, and then add the decolouring of isopyknic sherwood oil, retain aqueous phase, in triplicate, when ether is mutually without the pH to 8.0 regulating aqueous phase during color, isopyknic ethyl acetate is then used to extract, repeat twice, merge ester phase, rotary evaporation, is concentrated into dry, then joining 1ml volumetric concentration is redissolve in 60% methyl alcohol, after Spe C18 pillar purifying, use 0.45 μm of membrane filtration, filtrate is as sample solution.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310370131.4A CN103412082B (en) | 2013-08-22 | 2013-08-22 | HPLC (High Performance Liquid Chromatography) technique-based detection method for contents of jasmonic acid compounds in lycoris radiate |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310370131.4A CN103412082B (en) | 2013-08-22 | 2013-08-22 | HPLC (High Performance Liquid Chromatography) technique-based detection method for contents of jasmonic acid compounds in lycoris radiate |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103412082A CN103412082A (en) | 2013-11-27 |
| CN103412082B true CN103412082B (en) | 2014-12-24 |
Family
ID=49605108
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310370131.4A Expired - Fee Related CN103412082B (en) | 2013-08-22 | 2013-08-22 | HPLC (High Performance Liquid Chromatography) technique-based detection method for contents of jasmonic acid compounds in lycoris radiate |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103412082B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104297410B (en) * | 2014-11-05 | 2016-03-16 | 中国烟草总公司郑州烟草研究院 | A kind of Liquid Chromatography-Tandem Mass Spectrometry detects the method for abscisic acid and jasmonic in fresh tobacco leaves |
| CN108918710B (en) * | 2018-07-16 | 2021-04-23 | 中国烟草总公司郑州烟草研究院 | Method for detecting endogenous phytohormones in fresh tobacco leaves |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP3916144B2 (en) * | 2002-07-02 | 2007-05-16 | 独立行政法人科学技術振興機構 | Method for producing jasmonic acids and theobroxide using microorganisms |
| CN1281114C (en) * | 2004-08-18 | 2006-10-25 | 南京林业大学 | Long tube lycoris fast breeding method |
| CN102135528A (en) * | 2010-01-26 | 2011-07-27 | 湖南农业大学 | Detection method of jasmonic acid and methyl jasmonate in trace plant fresh sample |
-
2013
- 2013-08-22 CN CN201310370131.4A patent/CN103412082B/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| CN103412082A (en) | 2013-11-27 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103713077B (en) | The method of alpha-aminobutyric acid content in high effective liquid chromatography for measuring red colouring agent for food, also used as a Chinese medicine | |
| CA2685358C (en) | Method of detecting blood plasma danshensu and salvianolic acid b after administration of fuzheng huayu (fzhy) | |
| CN103134871B (en) | Preparation method of test solution for detecting aconitine-type alkaloids through high-efficiency liquid phase chromatography | |
| Jiang et al. | Simultaneous determination of adenine nucleotides, creatine phosphate and creatine in rat liver by high performance liquid chromatography–electrospray ionization-tandem mass spectrometry | |
| CN112526051B (en) | Fmoc-lysine high performance liquid chromatography determination method | |
| Yang et al. | Determination of tetrandrine and fangchinoline in plasma samples using hollow fiber liquid-phase microextraction combined with high-performance liquid chromatography | |
| Lopes et al. | Quantification of carbamazepine and its active metabolite by direct injection of human milk serum using liquid chromatography tandem ion trap mass spectrometry | |
| CN103743844B (en) | The assay method of ethanol content in methyl alcohol | |
| CN107037150A (en) | It is a kind of at the same determine food in Determination of Preservatives, sweetener and colouring agent method | |
| Xu et al. | Determination of schizandrin in rat plasma by high-performance liquid chromatography–mass spectrometry and its application in rat pharmacokinetic studies | |
| CN103412082B (en) | HPLC (High Performance Liquid Chromatography) technique-based detection method for contents of jasmonic acid compounds in lycoris radiate | |
| CN103630618A (en) | Method for determining betaine content in wolfberry | |
| CA2685359A1 (en) | Method of detecting blood plasma amygdalin after administration of fuzheng huayu(fzhy) | |
| CN102288687A (en) | Method for analysing and detecting impurities in ornithine aspartate | |
| CN103323419A (en) | Method for testing nicotine alkaloid in tobacco and tobacco products | |
| CN103852531A (en) | Method for detecting malto-oligosaccharide in beer through HPLC-ELSD (High-Performance Liquid Chromatography-Evaporative Light Scattering Detector) | |
| CN105092720A (en) | Method for measuring genotoxic impurities in pradaxa | |
| CN103954705A (en) | Method for measuring content of allantoin contained in Chinese traditional medicine rhizoma dioscoreae and rhizoma dioscoreae-containing preparation | |
| CN102854277B (en) | Determination method for matrine content in gynecological lotion | |
| CN102706971A (en) | A method for analysis of corticosteroids in cosmetics utilizing reverse microemulsion electrokinetic chromatography by taking an ionic liquid as an additive | |
| CN108802243B (en) | Method for simultaneously detecting bentazone, 2,4-D, 2, 4-dichlorophen, 2,4, 6-trichlorophenol and pentachlorophenol in water | |
| CN106290603A (en) | A kind of utilize Vavle switching method to detect the inorganic anion in plant, organic acid and the method for three kinds of phytochemicalss and application simultaneously | |
| CN108072717B (en) | Method for detecting arginine solution | |
| CN104237410A (en) | Detection method of substituted benzene in environmental water | |
| KR101159064B1 (en) | Quantification of active androgens in urine and blood by isotope dilution-mass spectrometry |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20141224 Termination date: 20150822 |
|
| EXPY | Termination of patent right or utility model |