CN1959407A - Method for constructing fingerprint HPLC of Shatian pomelo, and normal fingerprint - Google Patents
Method for constructing fingerprint HPLC of Shatian pomelo, and normal fingerprint Download PDFInfo
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- CN1959407A CN1959407A CN 200610122923 CN200610122923A CN1959407A CN 1959407 A CN1959407 A CN 1959407A CN 200610122923 CN200610122923 CN 200610122923 CN 200610122923 A CN200610122923 A CN 200610122923A CN 1959407 A CN1959407 A CN 1959407A
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- 238000004128 high performance liquid chromatography Methods 0.000 title claims abstract description 20
- 240000000560 Citrus x paradisi Species 0.000 title claims 7
- 238000000034 method Methods 0.000 title abstract description 9
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims abstract description 12
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 10
- 238000012360 testing method Methods 0.000 claims abstract description 10
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- PQLVXDKIJBQVDF-UHFFFAOYSA-N acetic acid;hydrate Chemical compound O.CC(O)=O PQLVXDKIJBQVDF-UHFFFAOYSA-N 0.000 claims abstract description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 5
- 238000010828 elution Methods 0.000 claims abstract description 4
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- BGEBZHIAGXMEMV-UHFFFAOYSA-N 5-methoxypsoralen Chemical compound O1C(=O)C=CC2=C1C=C1OC=CC1=C2OC BGEBZHIAGXMEMV-UHFFFAOYSA-N 0.000 claims description 8
- KGGUASRIGLRPAX-UHFFFAOYSA-N Meranzin hydrate Chemical compound C1=CC(=O)OC2=C(CC(O)C(C)(C)O)C(OC)=CC=C21 KGGUASRIGLRPAX-UHFFFAOYSA-N 0.000 claims description 8
- 239000000706 filtrate Substances 0.000 claims description 6
- 238000010276 construction Methods 0.000 claims description 5
- FTVWIRXFELQLPI-ZDUSSCGKSA-N (S)-naringenin Chemical compound C1=CC(O)=CC=C1[C@H]1OC2=CC(O)=CC(O)=C2C(=O)C1 FTVWIRXFELQLPI-ZDUSSCGKSA-N 0.000 claims description 4
- LSZONYLDFHGRDP-LBPRGKRZSA-N 8-[[(2s)-3,3-dimethyloxiran-2-yl]methyl]-7-methoxychromen-2-one Chemical compound COC1=CC=C2C=CC(=O)OC2=C1C[C@@H]1OC1(C)C LSZONYLDFHGRDP-LBPRGKRZSA-N 0.000 claims description 4
- DBMJZOMNXBSRED-UHFFFAOYSA-N Bergamottin Natural products O1C(=O)C=CC2=C1C=C1OC=CC1=C2OCC=C(C)CCC=C(C)C DBMJZOMNXBSRED-UHFFFAOYSA-N 0.000 claims description 4
- IGWDEVSBEKYORK-UHFFFAOYSA-N Isoimperatorin Natural products O1C(=O)C=CC2=C1C=C1OC=CC1=C2OCC=C(C)C IGWDEVSBEKYORK-UHFFFAOYSA-N 0.000 claims description 4
- ACUXTFUJXCOKNG-UHFFFAOYSA-N Meransin-hydrat-monoacetat Natural products COc1ccc2ccc(=O)oc2c1CC(OC(C)=O)C(C)(C)O ACUXTFUJXCOKNG-UHFFFAOYSA-N 0.000 claims description 4
- 244000179970 Monarda didyma Species 0.000 claims description 4
- 235000010672 Monarda didyma Nutrition 0.000 claims description 4
- RPMNUQRUHXIGHK-PYXJVEIZSA-N apigenin 7-O-neohesperidoside Chemical compound O[C@@H]1[C@H](O)[C@@H](O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=CC(O)=C2C(=O)C=C(C=3C=CC(O)=CC=3)OC2=C1 RPMNUQRUHXIGHK-PYXJVEIZSA-N 0.000 claims description 4
- 229930034861 apigenin-7-O-neohesperidoside Natural products 0.000 claims description 4
- 229960002045 bergapten Drugs 0.000 claims description 4
- KGZDKFWCIPZMRK-UHFFFAOYSA-N bergapten Natural products COC1C2=C(Cc3ccoc13)C=CC(=O)O2 KGZDKFWCIPZMRK-UHFFFAOYSA-N 0.000 claims description 4
- 230000014759 maintenance of location Effects 0.000 claims description 4
- LSZONYLDFHGRDP-UHFFFAOYSA-N meranzin Natural products COC1=CC=C2C=CC(=O)OC2=C1CC1OC1(C)C LSZONYLDFHGRDP-UHFFFAOYSA-N 0.000 claims description 4
- 229940117954 naringenin Drugs 0.000 claims description 4
- WGEYAGZBLYNDFV-UHFFFAOYSA-N naringenin Natural products C1(=O)C2=C(O)C=C(O)C=C2OC(C1)C1=CC=C(CC1)O WGEYAGZBLYNDFV-UHFFFAOYSA-N 0.000 claims description 4
- 235000007625 naringenin Nutrition 0.000 claims description 4
- DFPMSGMNTNDNHN-ZPHOTFPESA-N naringin Chemical compound O[C@@H]1[C@H](O)[C@@H](O)[C@H](C)O[C@H]1O[C@H]1[C@H](OC=2C=C3O[C@@H](CC(=O)C3=C(O)C=2)C=2C=CC(O)=CC=2)O[C@H](CO)[C@@H](O)[C@@H]1O DFPMSGMNTNDNHN-ZPHOTFPESA-N 0.000 claims description 4
- ARGKVCXINMKCAZ-UZRWAPQLSA-N neohesperidin Chemical compound C1=C(O)C(OC)=CC=C1[C@H]1OC2=CC(O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O3)O[C@H]3[C@@H]([C@H](O)[C@@H](O)[C@H](C)O3)O)=CC(O)=C2C(=O)C1 ARGKVCXINMKCAZ-UZRWAPQLSA-N 0.000 claims description 4
- ARGKVCXINMKCAZ-UHFFFAOYSA-N neohesperidine Natural products C1=C(O)C(OC)=CC=C1C1OC2=CC(OC3C(C(O)C(O)C(CO)O3)OC3C(C(O)C(O)C(C)O3)O)=CC(O)=C2C(=O)C1 ARGKVCXINMKCAZ-UHFFFAOYSA-N 0.000 claims description 4
- RPMNUQRUHXIGHK-SBDOOABHSA-N rhoifolin Natural products O([C@@H]1[C@@H](O)[C@H](O)[C@H](CO)O[C@H]1Oc1cc(O)c2C(=O)C=C(c3ccc(O)cc3)Oc2c1)[C@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@H](C)O1 RPMNUQRUHXIGHK-SBDOOABHSA-N 0.000 claims description 4
- 238000002386 leaching Methods 0.000 claims description 3
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 claims description 3
- 239000012982 microporous membrane Substances 0.000 claims description 3
- 238000002360 preparation method Methods 0.000 claims description 3
- 239000006228 supernatant Substances 0.000 claims description 3
- 238000012546 transfer Methods 0.000 claims description 3
- 238000005303 weighing Methods 0.000 claims description 3
- 244000276331 Citrus maxima Species 0.000 abstract description 30
- 235000001759 Citrus maxima Nutrition 0.000 abstract description 6
- 238000009835 boiling Methods 0.000 abstract 1
- 238000001514 detection method Methods 0.000 abstract 1
- 239000000203 mixture Substances 0.000 abstract 1
- 239000004576 sand Substances 0.000 abstract 1
- 239000013558 reference substance Substances 0.000 description 6
- 239000003153 chemical reaction reagent Substances 0.000 description 4
- 241000196324 Embryophyta Species 0.000 description 3
- 238000010521 absorption reaction Methods 0.000 description 2
- 235000013399 edible fruits Nutrition 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000011156 evaluation Methods 0.000 description 2
- 241000411851 herbal medicine Species 0.000 description 2
- 239000002398 materia medica Substances 0.000 description 2
- 239000012071 phase Substances 0.000 description 2
- 238000003908 quality control method Methods 0.000 description 2
- HNSDLXPSAYFUHK-UHFFFAOYSA-N 1,4-bis(2-ethylhexyl) sulfosuccinate Chemical compound CCCCC(CC)COC(=O)CC(S(O)(=O)=O)C(=O)OCC(CC)CCCC HNSDLXPSAYFUHK-UHFFFAOYSA-N 0.000 description 1
- 241000207199 Citrus Species 0.000 description 1
- 206010011224 Cough Diseases 0.000 description 1
- 206010036790 Productive cough Diseases 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 208000024794 sputum Diseases 0.000 description 1
- 210000003802 sputum Anatomy 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
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- Investigating Or Analysing Biological Materials (AREA)
- Investigating Or Analyzing Non-Biological Materials By The Use Of Chemical Means (AREA)
Abstract
A method for configurating HPLC fingerprint atlas of sand flat shaddock includes taking said shaddock sample powder; water boiling it; filtering; concentrating; adding alcohol and stewing; taking top clear liquid as test sample; carrying out HPLC analysis as using chromatographic column of Merck Lichrospher RP-18e ( f micron m, 250 x 4.0mm), linear gradient elution of 0min-50min, methanol of 30%-60%, acetic acid-water mixture of 36% and detection wavelength of 320nm to obtain HPLC atlas of said shaddock; comparing 18 batches of said atlas to obtain standard fingerprint atlas of said shaddock.
Description
Technical field
The present invention relates to the construction method of shatian pomelo (Citrus grandis (L.) 0sbeck var.shatinyu Hort.) HPLC finger-print, and the shatian pomelo standard finger-print that obtains of method thus.
Background technology
The chromatographic fingerprints of Chinese materia medica Quality Control Technology is an importance of the modernization of Chinese medicine.Chromatographic fingerprints of Chinese materia medica is realizing that the whole controlling party mask of material colony has remarkable advantages, has obtained the approval of international community.British Herbal Pharmacopoeia, India herbal medicine allusion quotation, German medicinal plant association, Canada is medicinal and fragrant plant association all receives the quality control method of finger-print.
Shatian pomelo is the fruit of rue family citrus plant Citrus grandis (L.) Osbeck var.shatinyu Hort., has clearing heat and moistening lung, relieving cough and reducing sputum effect.The present invention has made up the finger-print of shatian pomelo with the HPLC method, but utilizes the quality of HPLC finger-print overall monitor shatian pomelo.
Summary of the invention
The construction method that the purpose of this invention is to provide shatian pomelo (Citrus grandis (L.) Osbeck var.shatinyu Hort.) finger-print, and the shatian pomelo standard finger-print that obtains of method thus.
The construction method of shatian pomelo HPLC finger-print of the present invention comprises the steps:
(1) preparation of need testing solution: precision takes by weighing shatian pomelo sample powder (crossing 40 mesh sieves) 1.0g～5.0g, decoct 1～3 time, add 5～30 times of amounts of water at every turn, decoct 10min～2h at every turn, filter, it is 1.05～1.20 that filtrate is concentrated into relative density, add ethanol and make determining alcohol reach 50%～90%, leave standstill 6～48h, the leaching supernatant, with 0.45 μ m filtering with microporous membrane, filtrate is as need testing solution.
(2) HPLC analyzes: the accurate need testing solution 20 μ l sample introductions of drawing, and chromatographic condition is as follows: chromatographic column is MerckLichrospher RP-18e (5 μ m, 250 * 4.0mm); Moving phase is methyl alcohol-36% acetic acid water mixed liquid (80: 1, transfer pH to 3.0); Adopt the linear gradient elution mode: 0min → 50min, methyl alcohol: 30% → 60%, 36% acetic acid water mixed liquid: 70% → 40%; Flow velocity: 1ml/min; Detect wavelength: 320nm; Obtain shatian pomelo HPLC finger-print.
According to the method for the invention described above, by 18 batches of shatian pomelo samples are made up the HPLC finger-print and analyze comparison, find out its common characteristic peak (totally 13 peaks), obtain the standard finger-print of shatian pomelo.Retention time (the t at 13 total peaks
R) be respectively: 7.28min, 10.38min, 12.11min, 17.11min, 21.74min, 23.05min, 25.91min, 27.95min, 28.85min, 32.36min, 34.51min, 35.95min, 44.25min.They have constituted the fingerprint characteristic of shatian pomelo.
The present invention by absorption curve relatively to reach addition method qualitative, determine that the ownership of main peaks is as follows: t
RThe peak of=10.38min is naringenin, t
RThe peak of=17.11min is aurantiin, t
RThe peak of=21.74min is neohesperidin, t
RThe peak of=23.05min is Rhoifolin, t
RThe peak of=25.91min is meranzin hydrate, t
RThe peak of=27.95min is different meranzin, t
RThe peak of=35.95min is bergapten, t
RThe peak of=44.25min is the bergamot booth.
The present invention has following advantage and purposes: by having or not of total peak in the finger-print, can monitor the quality of shatian pomelo effectively, guarantee stable, the homogeneous, controlled of its quality.
The present invention is described further below in conjunction with embodiment and accompanying drawing.
Description of drawings:
Fig. 1 is a shatian pomelo standard finger-print of the present invention.Retention time (the t at each the total peak of numeral among the figure
R).
Embodiment
The preferred embodiments of the present invention are as follows:
1 instrument, sample, reagent:
1.1 instrument: DIONEX high performance liquid chromatograph (ASI-100 automatic sampler, ATH-585 column oven, P680 quaternary gradient pump, PDA-100 detecting device); HP1100 high performance liquid chromatograph (quaternary pump, online degasser, column oven, automatic sampler, DAD detecting device).Chromatographic column: Merck Lichrospher RP-18e (5 μ m, 250 * 4.0mm).
1.2 sample: 18 batches of shatian pomelo samples respectively from super flat, the Cangwu, Guangxi in Mei County, Guangdong, Guangxi, Rong County, Guangxi and other places, are the fruit of rue family citrus plant shatian pomelo Citrus grandis (L.) Osbeck var.shatinyu Hort. through evaluation.
1.3 reagent: naringenin reference substance, aurantiin reference substance (Nat'l Pharmaceutical ﹠ Biological Products Control Institute provides); Neohesperidin reference substance, Rhoifolin reference substance, bergapten reference substance (Sigma company provides); Meranzin hydrate, different meranzin, bergamot booth (modern Chinese herbal medicine quality research development centre, Zhongshan University Guangzhou provides, and purity is higher than 99.5%).Liquid-phase chromatographic analysis reagent is chromatographically pure, and it is pure that all the other reagent are analysis, and water is ultrapure water.
2 methods and result
2.1 the preparation of need testing solution: precision takes by weighing shatian pomelo sample powder (crossing 40 mesh sieves) 2.0g, decoct 3 times, add 15 times of amounts of water at every turn, decoct 1h at every turn, filter, it is 1.10～1.12 that filtrate is concentrated into relative density, add ethanol and make determining alcohol reach 80%, leave standstill 24h, the leaching supernatant, with 0.45 μ m filtering with microporous membrane, filtrate is as need testing solution.
2.2HPLC analyze: the accurate need testing solution 20 μ l sample introductions of drawing, chromatographic condition is as follows: chromatographic column is MerckLichrospher RP-18e (5 μ m, 250 * 4.0mm); Moving phase is methyl alcohol-36% acetic acid water mixed liquid (80: 1, transfer pH to 3.0); Adopt the linear gradient elution mode: 0min → 50min, methyl alcohol: 30% → 60%, 36% acetic acid water mixed liquid: 70% → 40%; Flow velocity: 1ml/min; Detect wavelength: 320nm; Obtain shatian pomelo HPLC finger-print.
2.3 determining of total peak: above-mentioned 18 batches of shatian pomelo sample HPLC finger-prints that obtain are analyzed comparison, find out its common characteristic peak (totally 13 peaks), obtain the standard finger-print of shatian pomelo.Retention time (the t at 13 total peaks
R) be respectively: 7.28min, 10.38min, 12.11min, 17.11min, 21.74min, 23.05min, 25.91min, 27.95min, 28.85min, 32.36min, 34.51min, 35.95min, 44.25min.They have constituted the fingerprint characteristic of shatian pomelo.
2.4 the evaluation of main peaks: by absorption curve relatively to reach the reference substance addition method qualitative, determined that the ownership of main peaks is as follows: t
RThe peak of=10.38min is naringenin, t
RThe peak of=17.11min is aurantiin, t
RThe peak of=21.74min is neohesperidin, t
RThe peak of=23.05min is Rhoifolin, t
RThe peak of=25.91min is meranzin hydrate, t
RThe peak of=27.95min is different meranzin, t
RThe peak of=35.95min is bergapten, t
RThe peak of=44.25min is the bergamot booth.
Claims (2)
1. the construction method of a shatian pomelo HPLC finger-print may further comprise the steps:
(1) preparation of need testing solution: precision takes by weighing shatian pomelo sample powder (crossing 40 mesh sieves) 1.0g～5.0g, decoct 1～3 time, add 5～30 times of amounts of water at every turn, decoct 10min～2h at every turn, filter, it is 1.05～1.20 that filtrate is concentrated into relative density, add ethanol and make determining alcohol reach 50%～90%, leave standstill 6～48h, the leaching supernatant, with 0.45 μ m filtering with microporous membrane, filtrate is as need testing solution;
(2) HPLC analyzes: the accurate need testing solution 20 μ l sample introductions of drawing, and chromatographic condition is as follows: chromatographic column is MerckLichrospher RP-18e (5 μ m, 250 * 4.0mm); Moving phase is methyl alcohol-36% acetic acid water mixed liquid (80: 1, transfer pH to 3.0); Adopt the linear gradient elution mode: 0min → 50min, methyl alcohol: 30% → 60%, 36% acetic acid water mixed liquid: 70% → 40%; Flow velocity: 1ml/min; Detect wavelength: 320nm; Obtain shatian pomelo HPLC finger-print;
(3) measure 18 batches of shatian pomelo sample HPLC finger-prints and comparing, obtain the shatian pomelo HPLC standard finger-print that constitutes by its total peak, the retention time (t at 13 common characteristic peaks that this finger-print has
R) be respectively: 7.28min, 10.38min, 12.11min, 17.11min, 21.74min, 23.05min, 25.91min, 27.95min, 28.85min, 32.36min, 34.51min, 35.95min, 44.25min.
2. according to the described shatian pomelo HPLC of claim 1 finger-print, it is characterized in that: t
RThe peak of=10.38min is naringenin, t
RThe peak of=17.11min is aurantiin, t
RThe peak of=21.74min is neohesperidin, t
RThe peak of=23.05min is Rhoifolin, t
RThe peak of=25.91min is meranzin hydrate, t
RThe peak of=27.95min is different meranzin, t
RThe peak of=35.95min is bergapten, t
RThe peak of=44.25min is the bergamot booth.
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| Application Number | Priority Date | Filing Date | Title |
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| CNB200610122923XA CN100485386C (en) | 2006-10-20 | 2006-10-20 | Method for constructing HPLC fingerprint graph spectrum of Shatian pomelo |
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| CNB200610122923XA CN100485386C (en) | 2006-10-20 | 2006-10-20 | Method for constructing HPLC fingerprint graph spectrum of Shatian pomelo |
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|---|---|
| CN1959407A true CN1959407A (en) | 2007-05-09 |
| CN100485386C CN100485386C (en) | 2009-05-06 |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103344729A (en) * | 2013-07-15 | 2013-10-09 | 西南大学 | Method for identifying authenticity and evaluating quality of Fengjie navel oranges |
| CN104316635A (en) * | 2014-10-28 | 2015-01-28 | 石河子大学 | Method for rapidly identifying flavor and quality of fruits |
| CN106442777A (en) * | 2016-09-14 | 2017-02-22 | 广西大学 | Method for identifying Fujian Guanxi honey pomelos by chromatography method |
| CN106442778A (en) * | 2016-09-14 | 2017-02-22 | 广西大学 | Method for identifying Shatian pomelo in Rong County of Guangxi by adopting chromatography |
| CN107383040A (en) * | 2017-07-05 | 2017-11-24 | 浙江大学 | A kind of method that bergamot element is isolated and purified from early fragrant shaddock oil vacuole layer |
-
2006
- 2006-10-20 CN CNB200610122923XA patent/CN100485386C/en not_active Expired - Fee Related
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103344729A (en) * | 2013-07-15 | 2013-10-09 | 西南大学 | Method for identifying authenticity and evaluating quality of Fengjie navel oranges |
| CN104316635A (en) * | 2014-10-28 | 2015-01-28 | 石河子大学 | Method for rapidly identifying flavor and quality of fruits |
| CN106442777A (en) * | 2016-09-14 | 2017-02-22 | 广西大学 | Method for identifying Fujian Guanxi honey pomelos by chromatography method |
| CN106442778A (en) * | 2016-09-14 | 2017-02-22 | 广西大学 | Method for identifying Shatian pomelo in Rong County of Guangxi by adopting chromatography |
| CN106442777B (en) * | 2016-09-14 | 2018-11-27 | 广西大学 | A method of Fu Jian Guanxi small stream honey shaddock is identified using chromatography |
| CN106442778B (en) * | 2016-09-14 | 2019-01-29 | 广西大学 | A method of Guangxi Rong County shatian pomelo is identified using chromatography |
| CN107383040A (en) * | 2017-07-05 | 2017-11-24 | 浙江大学 | A kind of method that bergamot element is isolated and purified from early fragrant shaddock oil vacuole layer |
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| CN100485386C (en) | 2009-05-06 |
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