CN113203806A - Method for analyzing components of chlorophyll and degradation products thereof in tea - Google Patents
Method for analyzing components of chlorophyll and degradation products thereof in tea Download PDFInfo
- Publication number
- CN113203806A CN113203806A CN202110436988.6A CN202110436988A CN113203806A CN 113203806 A CN113203806 A CN 113203806A CN 202110436988 A CN202110436988 A CN 202110436988A CN 113203806 A CN113203806 A CN 113203806A
- Authority
- CN
- China
- Prior art keywords
- chlorophyll
- tea
- analysis
- degradation
- internal standard
- 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.)
- Pending
Links
- ATNHDLDRLWWWCB-AENOIHSZSA-M chlorophyll a Chemical compound C1([C@@H](C(=O)OC)C(=O)C2=C3C)=C2N2C3=CC(C(CC)=C3C)=[N+]4C3=CC3=C(C=C)C(C)=C5N3[Mg-2]42[N+]2=C1[C@@H](CCC(=O)OC\C=C(/C)CCC[C@H](C)CCC[C@H](C)CCCC(C)C)[C@H](C)C2=C5 ATNHDLDRLWWWCB-AENOIHSZSA-M 0.000 title claims abstract description 76
- 229930002875 chlorophyll Natural products 0.000 title claims abstract description 73
- 235000019804 chlorophyll Nutrition 0.000 title claims abstract description 73
- 239000007857 degradation product Substances 0.000 title claims abstract description 44
- 238000000034 method Methods 0.000 title claims abstract description 27
- 241001122767 Theaceae Species 0.000 title claims abstract 17
- 238000004458 analytical method Methods 0.000 claims abstract description 41
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 17
- 238000004895 liquid chromatography mass spectrometry Methods 0.000 claims abstract description 9
- 150000001875 compounds Chemical class 0.000 claims description 53
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 33
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 27
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims description 16
- BZLVMXJERCGZMT-UHFFFAOYSA-N Methyl tert-butyl ether Chemical compound COC(C)(C)C BZLVMXJERCGZMT-UHFFFAOYSA-N 0.000 claims description 16
- 238000000605 extraction Methods 0.000 claims description 16
- 150000002500 ions Chemical class 0.000 claims description 10
- 230000014759 maintenance of location Effects 0.000 claims description 9
- 239000000243 solution Substances 0.000 claims description 9
- 238000012937 correction Methods 0.000 claims description 7
- 238000001035 drying Methods 0.000 claims description 7
- RKEBXTALJSALNU-LDCXZXNSSA-N 3-[(3R,21S,22S)-16-ethenyl-11-ethyl-4-hydroxy-3-methoxycarbonyl-12,17,21,26-tetramethyl-7,23,24,25-tetrazahexacyclo[18.2.1.15,8.110,13.115,18.02,6]hexacosa-1,4,6,8(26),9,11,13(25),14,16,18(24),19-undecaen-22-yl]propanoic acid Chemical compound CCC1=C(C2=NC1=CC3=C(C4=C([C@@H](C(=C5[C@H]([C@@H](C(=CC6=NC(=C2)C(=C6C)C=C)N5)C)CCC(=O)O)C4=N3)C(=O)OC)O)C)C RKEBXTALJSALNU-LDCXZXNSSA-N 0.000 claims description 6
- USFZMSVCRYTOJT-UHFFFAOYSA-N Ammonium acetate Chemical compound N.CC(O)=O USFZMSVCRYTOJT-UHFFFAOYSA-N 0.000 claims description 6
- 239000005695 Ammonium acetate Substances 0.000 claims description 6
- JUDGRMABQJKRPW-XIADSQHASA-N CCC1=C(/C=C2\N=C(/C(\CC3=O)=C(/[C@@H](CCC(O)=O)[C@@H]4C)\N/C\4=C\C(C(C)=C4C=C)=N/C\4=C4)C3=C\2C)NC/4=C1C Chemical compound CCC1=C(/C=C2\N=C(/C(\CC3=O)=C(/[C@@H](CCC(O)=O)[C@@H]4C)\N/C\4=C\C(C(C)=C4C=C)=N/C\4=C4)C3=C\2C)NC/4=C1C JUDGRMABQJKRPW-XIADSQHASA-N 0.000 claims description 6
- 229940043376 ammonium acetate Drugs 0.000 claims description 6
- 235000019257 ammonium acetate Nutrition 0.000 claims description 6
- ANWUQYTXRXCEMZ-NYABAGMLSA-L chlorophyllide a Chemical compound C1([C@H](C2=O)C(=O)OC)=C(N3[Mg]N45)C2=C(C)\C3=C\C(=N2)C(CC)=C(C)\C2=C\C4=C(C=C)C(C)=C5\C=C/2[C@@H](C)[C@H](CCC(O)=O)C1=N\2 ANWUQYTXRXCEMZ-NYABAGMLSA-L 0.000 claims description 6
- 238000005516 engineering process Methods 0.000 claims description 6
- 229930002868 chlorophyll a Natural products 0.000 claims description 5
- 238000010828 elution Methods 0.000 claims description 5
- ZQGOYEJYAYJFTL-BTMCAZCFSA-N pheophytin b Chemical compound N1C(C=C2[C@H]([C@H](CCC(=O)OC\C=C(/C)CCC[C@H](C)CCC[C@H](C)CCCC(C)C)C(=N2)C2=C3NC(=C4)C(C)=C3C(=O)[C@@H]2C(=O)OC)C)=C(C)C(C=C)=C1C=C1C(C=O)=C(CC)C4=N1 ZQGOYEJYAYJFTL-BTMCAZCFSA-N 0.000 claims description 5
- 238000011002 quantification Methods 0.000 claims description 5
- 238000004445 quantitative analysis Methods 0.000 claims description 5
- FDKRLXBXYZKWRZ-UWJYYQICSA-N 3-[(21S,22S)-16-ethenyl-11-ethyl-4-hydroxy-12,17,21,26-tetramethyl-7,23,24,25-tetrazahexacyclo[18.2.1.15,8.110,13.115,18.02,6]hexacosa-1,4,6,8(26),9,11,13(25),14,16,18(24),19-undecaen-22-yl]propanoic acid Chemical compound CCC1=C(C2=NC1=CC3=C(C4=C(CC(=C5[C@H]([C@@H](C(=CC6=NC(=C2)C(=C6C)C=C)N5)C)CCC(=O)O)C4=N3)O)C)C FDKRLXBXYZKWRZ-UWJYYQICSA-N 0.000 claims description 4
- KZTYPOGXRFTJBN-MMLDQJOMSA-N 3-[(3R,12E,21S,22S)-16-ethenyl-11-ethyl-12-(hydroxymethylidene)-3-methoxycarbonyl-17,21,26-trimethyl-4-oxo-7,23,24,25-tetrazahexacyclo[18.2.1.15,8.110,13.115,18.02,6]hexacosa-1,5(26),6,8,10,13(25),14,16,18(24),19-decaen-22-yl]propanoic acid Chemical compound CCC\1=C2C=C3C(=C4C(=O)[C@@H](C(=C5[C@H]([C@@H](C(=CC6=NC(=CC(=N2)/C1=C/O)C(=C6C)C=C)N5)C)CCC(=O)O)C4=N3)C(=O)OC)C KZTYPOGXRFTJBN-MMLDQJOMSA-N 0.000 claims description 4
- ATNHDLDRLWWWCB-DVXFRRMCSA-M chlorophyll a' Chemical compound CCC(C(C=C(C(C)=C1C([C@H]2C(OC)=O)=O)[N-]C1=C2C([C@@H](CCC(OC/C=C(\C)/CCC[C@H](C)CCC[C@H](C)CCCC(C)C)=O)[C@@H]1C)=NC1=C1)=N2)=C(C)C2=CC2=C(C=C)C(C)=C1[N-]2.[Mg+2] ATNHDLDRLWWWCB-DVXFRRMCSA-M 0.000 claims description 4
- 229930002869 chlorophyll b Natural products 0.000 claims description 4
- NSMUHPMZFPKNMZ-VBYMZDBQSA-M chlorophyll b Chemical compound C1([C@@H](C(=O)OC)C(=O)C2=C3C)=C2N2C3=CC(C(CC)=C3C=O)=[N+]4C3=CC3=C(C=C)C(C)=C5N3[Mg-2]42[N+]2=C1[C@@H](CCC(=O)OC\C=C(/C)CCC[C@H](C)CCC[C@H](C)CCCC(C)C)[C@H](C)C2=C5 NSMUHPMZFPKNMZ-VBYMZDBQSA-M 0.000 claims description 4
- QPDWBRHRBKXUNS-IEEIVXFASA-L chlorophyllide b Chemical compound C1([C@H](C2=O)C(=O)OC)=C(N3[Mg]N45)C2=C(C)\C3=C\C(=N2)C(CC)=C(C=O)\C2=C\C4=C(C=C)C(C)=C5\C=C/2[C@@H](C)[C@H](CCC(O)=O)C1=N\2 QPDWBRHRBKXUNS-IEEIVXFASA-L 0.000 claims description 4
- CQIKWXUXPNUNDV-AXRVZGOCSA-N pheophytin a Chemical compound N1C(C=C2[C@H]([C@H](CCC(=O)OC\C=C(/C)CCC[C@H](C)CCC[C@H](C)CCCC(C)C)C(=N2)C2=C3NC(=C4)C(C)=C3C(=O)[C@@H]2C(=O)OC)C)=C(C)C(C=C)=C1C=C1C(C)=C(CC)C4=N1 CQIKWXUXPNUNDV-AXRVZGOCSA-N 0.000 claims description 4
- RYCNUMLMNKHWPZ-SNVBAGLBSA-N 1-acetyl-sn-glycero-3-phosphocholine Chemical compound CC(=O)OC[C@@H](O)COP([O-])(=O)OCC[N+](C)(C)C RYCNUMLMNKHWPZ-SNVBAGLBSA-N 0.000 claims description 3
- 108700032487 GAP-43-3 Proteins 0.000 claims description 3
- 238000004364 calculation method Methods 0.000 claims description 3
- 238000000227 grinding Methods 0.000 claims description 3
- 238000002347 injection Methods 0.000 claims description 3
- 239000007924 injection Substances 0.000 claims description 3
- 238000001819 mass spectrum Methods 0.000 claims description 3
- 238000002156 mixing Methods 0.000 claims description 3
- WTJKGGKOPKCXLL-RRHRGVEJSA-N phosphatidylcholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCCC=CCCCCCCCC WTJKGGKOPKCXLL-RRHRGVEJSA-N 0.000 claims description 3
- 239000000843 powder Substances 0.000 claims description 3
- VLMUKTSQARJLRE-WLQLOANESA-N pyropheophytin b Chemical compound N1C(C=C2C(=C(CC)C(C=C3C(=C4C(=O)CC5=C4N3)C)=N2)C=O)=C(C=C)C(C)=C1C=C1C(C)C(CCC(=O)OC/C=C(C)/CC(C)CC(C)CCCC(C)CCCCC)C5=N1 VLMUKTSQARJLRE-WLQLOANESA-N 0.000 claims description 3
- 238000007873 sieving Methods 0.000 claims description 3
- 238000005507 spraying Methods 0.000 claims description 3
- 238000004704 ultra performance liquid chromatography Methods 0.000 claims description 3
- 229910021642 ultra pure water Inorganic materials 0.000 claims description 3
- 239000012498 ultrapure water Substances 0.000 claims description 3
- 238000005303 weighing Methods 0.000 claims description 3
- 238000002546 full scan Methods 0.000 claims description 2
- -1 pyropheophytin a Chemical compound 0.000 claims description 2
- 238000006731 degradation reaction Methods 0.000 abstract description 14
- 230000015556 catabolic process Effects 0.000 abstract description 13
- 238000001514 detection method Methods 0.000 abstract description 9
- 230000006652 catabolic pathway Effects 0.000 abstract description 8
- 108090000790 Enzymes Proteins 0.000 abstract description 6
- 102000004190 Enzymes Human genes 0.000 abstract description 6
- 230000001404 mediated effect Effects 0.000 abstract description 4
- 230000009897 systematic effect Effects 0.000 abstract description 3
- 241000196324 Embryophyta Species 0.000 abstract description 2
- 235000013305 food Nutrition 0.000 abstract description 2
- 244000269722 Thea sinensis Species 0.000 description 73
- 235000013616 tea Nutrition 0.000 description 44
- 235000009569 green tea Nutrition 0.000 description 29
- 239000000049 pigment Substances 0.000 description 5
- 238000012545 processing Methods 0.000 description 5
- 239000000126 substance Substances 0.000 description 5
- 238000006243 chemical reaction Methods 0.000 description 4
- 238000000926 separation method Methods 0.000 description 4
- 230000000694 effects Effects 0.000 description 3
- 239000002932 luster Substances 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 238000010415 tidying Methods 0.000 description 3
- 230000009471 action Effects 0.000 description 2
- 108010025790 chlorophyllase Proteins 0.000 description 2
- 239000001752 chlorophylls and chlorophyllins Substances 0.000 description 2
- 238000006114 decarboxylation reaction Methods 0.000 description 2
- 230000002255 enzymatic effect Effects 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 238000006317 isomerization reaction Methods 0.000 description 2
- 238000004811 liquid chromatography Methods 0.000 description 2
- 238000007069 methylation reaction Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 238000010239 partial least squares discriminant analysis Methods 0.000 description 2
- 238000003068 pathway analysis Methods 0.000 description 2
- 238000005096 rolling process Methods 0.000 description 2
- 235000014347 soups Nutrition 0.000 description 2
- 238000002798 spectrophotometry method Methods 0.000 description 2
- 230000007480 spreading Effects 0.000 description 2
- 238000003892 spreading Methods 0.000 description 2
- 238000004809 thin layer chromatography Methods 0.000 description 2
- 230000009466 transformation Effects 0.000 description 2
- IPMYMEWFZKHGAX-UHFFFAOYSA-N Isotheaflavin Natural products OC1CC2=C(O)C=C(O)C=C2OC1C(C1=C2)=CC(O)=C(O)C1=C(O)C(=O)C=C2C1C(O)CC2=C(O)C=C(O)C=C2O1 IPMYMEWFZKHGAX-UHFFFAOYSA-N 0.000 description 1
- 240000007817 Olea europaea Species 0.000 description 1
- UXRMWRBWCAGDQB-UHFFFAOYSA-N Theaflavin Natural products C1=CC(C2C(CC3=C(O)C=C(O)C=C3O2)O)=C(O)C(=O)C2=C1C(C1OC3=CC(O)=CC(O)=C3CC1O)=CC(O)=C2O UXRMWRBWCAGDQB-UHFFFAOYSA-N 0.000 description 1
- 238000010306 acid treatment Methods 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 125000002057 carboxymethyl group Chemical group [H]OC(=O)C([H])([H])[*] 0.000 description 1
- 238000009960 carding Methods 0.000 description 1
- 235000021466 carotenoid Nutrition 0.000 description 1
- 150000001747 carotenoids Chemical class 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 238000004040 coloring Methods 0.000 description 1
- 238000010835 comparative analysis Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 230000009849 deactivation Effects 0.000 description 1
- 230000007515 enzymatic degradation Effects 0.000 description 1
- 229930003935 flavonoid Natural products 0.000 description 1
- 235000017173 flavonoids Nutrition 0.000 description 1
- 150000002215 flavonoids Chemical class 0.000 description 1
- 239000003205 fragrance Substances 0.000 description 1
- 235000021384 green leafy vegetables Nutrition 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 238000001558 permutation test Methods 0.000 description 1
- 125000001189 phytyl group Chemical group [H]C([*])([H])/C([H])=C(C([H])([H])[H])/C([H])([H])C([H])([H])C([H])([H])[C@@](C([H])([H])[H])([H])C([H])([H])C([H])([H])C([H])([H])[C@@](C([H])([H])[H])([H])C([H])([H])C([H])([H])C([H])([H])C(C([H])([H])[H])([H])C([H])([H])[H] 0.000 description 1
- 150000004032 porphyrins Chemical group 0.000 description 1
- 238000002203 pretreatment Methods 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000007127 saponification reaction Methods 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 230000001953 sensory effect Effects 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 235000014620 theaflavin Nutrition 0.000 description 1
- IPMYMEWFZKHGAX-ZKSIBHASSA-N theaflavin Chemical compound C1=C2C([C@H]3OC4=CC(O)=CC(O)=C4C[C@H]3O)=CC(O)=C(O)C2=C(O)C(=O)C=C1[C@@H]1[C@H](O)CC2=C(O)C=C(O)C=C2O1 IPMYMEWFZKHGAX-ZKSIBHASSA-N 0.000 description 1
- 229940026509 theaflavin Drugs 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 238000000844 transformation Methods 0.000 description 1
- 238000011282 treatment Methods 0.000 description 1
- 230000016776 visual perception Effects 0.000 description 1
- 238000003260 vortexing Methods 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/96—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation using ion-exchange
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N2030/022—Column chromatography characterised by the kind of separation mechanism
- G01N2030/027—Liquid chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
- G01N2030/065—Preparation using different phases to separate parts of sample
Landscapes
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
Abstract
An analysis method for chlorophyll and degradation product components thereof in tea leaves belongs to the field of analysis and detection of food or plant quality components. Which comprises the following steps: extracting chlorophyll and degradation products thereof from tea to be detected; LC-MS analysis of chlorophyll and degradation products thereof in the tea to be detected; and (4) relatively quantitatively analyzing chlorophyll and degradation products thereof in the tea to be detected. The invention has the following advantages: (1) the sample pretreatment is simple and quick, and chlorophyll with different solubilities and degradation product components (water solubility and fat solubility) thereof can be extracted simultaneously; (2) the analysis speed is high, and the LC-MS analysis time of each sample only needs 20 minutes; (3) the detection coverage of chlorophyll and degradation product components thereof in the tea is high, and the comprehensive coverage of 19 components in the chlorophyll degradation pathway in total on the enzyme-mediated degradation and non-enzyme thermal degradation pathways can be realized. In a word, the method is simple and rapid, and the analysis is more systematic and comprehensive.
Description
Technical Field
The invention belongs to the field of food or plant quality component analysis and detection, and particularly relates to an analysis method for chlorophyll and degradation product components thereof in tea.
Background
The color and luster of the tea is one of the important factors of the quality attributes of the color, the fragrance, the taste and the shape of the tea, is the most direct visual perception of consumers, and the beautiful and smooth color and luster is the important basis for the selection of the consumers. The color of the tea comprises three aspects of dry tea, liquor color and leaf bottom. Taking green tea as an example, green tea is popular with consumers because of its quality characteristics of "three greens", including green tea soup, dry green tea, green leaf base. The tea color generation substance comprises chlorophyll and its degradation derivative, carotenoid, flavonoid, tea pigment (theaflavin, tea red, and theabrownin), etc.; they can be classified into water-soluble pigments and fat-soluble pigments due to their different solubilities. The color substance of the tea soup is mainly water-soluble pigment, while the main color substance of the dry tea and the tea bottom is fat-soluble pigment. Among them, chlorophyll is an important coloring substance. In the tea processing process, chlorophyll can generate a series of transformations (such as isomerization, demagging, grafting removal and the like) under the triggering of chlorophyllase mediated or non-enzyme mediated thermal reaction and other conditions, and various products with different colors can be derived by degradation.
The prior literature reports that common methods for measuring chlorophyll in tea leaves include spectrophotometry, thin-layer chromatography and liquid chromatography. Spectrophotometry involves complicated pre-treatments such as acid treatment and saponification treatment, and only can roughly measure chlorophyll components, and has low sensitivity. Thin layer chromatography takes a long time and the separation effect between components is poor. Liquid chromatography detection, while improving separation between different components, also requires complex and time-consuming pretreatment. The detection coverage of the method for chlorophyll and degradation components thereof needs to be improved, the simultaneous extraction and determination of a plurality of derivative products (water solubility and fat solubility) on a chlorophyll degradation pathway are difficult to realize, and epimers with close chemical structures and properties are difficult to separate. The limitations of the above technology are difficult to satisfy the requirements for detailed characterization of chlorophyll components in tea leaves and systematic study of their transformation processes.
Based on this, the applicant has designed a composition based on methyl tert-butyl ether (MTBE)/methanol (MeOH)/water (H)2O) system two-phase extraction and UHPLC-q-active technology, and has the advantages of simplicity, rapidness, high detection coverage, good specificity and good repeatability.
Disclosure of Invention
Aiming at the problems in the prior art, the invention aims to provide an analysis method for components of chlorophyll and degradation products thereof in tea. The invention adopts methyl tert-butyl ether (MTBE)/methanol (MeOH)/water (H)2O) and combining a two-phase mixing strategy to extract; the UHPLC-q-active high-resolution liquid chromatography-mass spectrometry technology is adopted for analysis, so that the separation and detection (including 6 pairs of epimers) of 19 kinds of chlorophyll and degradation and derivative products thereof in the tea can be realized within 20 minutes; and further realizing rapid relative quantitative analysis through corresponding internal standard correction. The method has the characteristics of simplicity, rapidness, high detection coverage, good specificity and good repeatability, and can provide effective technical support for researches on chlorophyll composition analysis of different tea leaves, dynamic change analysis of chlorophyll and degradation product components thereof in the tea leaf processing process, influence of different tea leaf processing technologies on chlorophyll degradation and the like.
The invention is realized by adopting the following technical scheme:
the method for analyzing the components of chlorophyll and degradation products thereof in the tea is characterized by comprising the following steps of:
1) extraction of chlorophyll and degradation product thereof in tea to be detected
Adopting a two-phase extraction technology based on methyl tert-butyl ether/methanol/water, combining a two-phase mixing strategy, extracting, and introducing an internal standard compound in the extraction process;
2) LC-MS analysis of chlorophyll and degradation products thereof in tea to be detected
Based on UHPLC-q-active high-resolution liquid chromatography-mass spectrometry technology, performing high-resolution full-scan analysis by adopting optimized chromatographic gradient elution conditions;
3) relative quantitative analysis of chlorophyll and degradation products thereof in tea to be detected
And (3) extracting the high-resolution extraction ion current chromatograms of the target compound and the internal standard compound to obtain the intensity of the high-resolution extraction ion current chromatograms, and correcting the target compound and the corresponding internal standard compound to perform relative quantitative analysis on the target compound.
The method for analyzing the components of chlorophyll and degradation products thereof in the tea is characterized in that the step 1) specifically comprises the following steps:
firstly, grinding a tea sample to be detected, and then sieving the ground tea sample with a 30-mesh sieve for later use;
weighing about 20.0mg +/-0.1 mg of tea powder into a centrifuge tube, adding 300 mu L of methanol solution containing an internal standard compound, and performing vortex operation for 30 s; the internal standard compound is lysophosphatidylcholine LPC19:0, and the concentration is 2.00 mu g/mL; phosphatidylcholine PC38:0, concentration 1.33 mug/mL;
③ adding 1mL of methyl tert-butyl ether, and whirling for 30 min;
fourthly, 300 mu L of ultrapure water is added, and vortex is carried out for 30 s;
standing for 10min, and centrifuging at 10000rpm for 10min to layer;
sixthly, taking 600 mu L of the upper layer and 250 mu L of the lower layer, transferring the upper layer and the lower layer into a new centrifugal tube, and centrifuging at low temperature, concentrating and drying for later use.
The method for analyzing the components of chlorophyll and degradation products thereof in the tea is characterized in that the step 2) specifically comprises the following steps:
before sample injection analysis, redissolving a sample subjected to low-temperature centrifugal concentration and drying, wherein the redissolution is acetonitrile/isopropanol/water solution, and the volume ratio of the acetonitrile/isopropanol/water is 65/30/5;
the UHPLC-q-active analysis conditions are as follows:
the instrument comprises the following steps: UHPLC-ESI-q-Orbitrap;
chromatographic conditions are as follows: the chromatographic column is a UPLC T3 chromatographic column, and the mobile phase A is acetonitrile/water (6/4 v/v) and contains 10mM ammonium acetate; the mobile phase B is isopropanol/acetonitrile 9/1v/v, containing 10mM ammonium acetate; the flow rate was 0.26mL/min and the gradient elution procedure was: 0-2.0min, 32% B; 2.0-4.0min, 32% B-60% B; 4.0-13.0min, 60% B-97% B; 13.0-17.0min, 97% B; 17.0-17.1min, 97% B-32% B; 17.1-20.0min, 32% B;
mass spectrum conditions: and performing high-resolution full scanning in an ESI positive ion mode, wherein the m/z range is 200-1200, the spraying voltage, the capillary temperature and the gas heater temperature are respectively 3.2kV, 320 ℃ and 450 ℃, and the flow rates of the sheath gas and the auxiliary gas are respectively 40arb and 10 arb.
The method for analyzing chlorophyll and degradation product components thereof in tea is characterized in that the UHPLC-q-Exactive analysis method in the step 2) can detect 19 chlorophyll and degradation product components thereof, namely chlorophyll a, chlorophyll a ', chlorophyll b', pheophytin a ', hydroxypheophytin a', pheophytin b, pyropheophytin a, pyropheophytin b, pheophorbide a ', pheophorbide b, pyropheophorbide a, pyropheophorbide b, chlorophyllide a' and chlorophyllide b, and the molecular formulas, retention times, ionization modes and mass-to-charge ratios of the 19 compounds are shown in Table 1.
TABLE 119 molecular formulae, retention times, ionization patterns and Mass-to-Charge ratios of chlorophyll and its degradation product Components
The method for analyzing the components of chlorophyll and degradation products thereof in the tea is characterized in that the step 3) specifically comprises the following steps:
performing high-resolution extraction ion current chromatogram (EIC) analysis on the target compound by using Xcalibur software based on retention time and mass-to-charge ratio in the table 1, and performing EIC analysis on the internal standard compound to obtain peak intensity;
normalizing the target compound to a corresponding internal standard compound for correction to obtain a relative quantitative result, wherein the internal standard compound corresponding to the target compound chlorophyll a, chlorophyll a ', chlorophyll b', pheophytin a ', hydroxyphenylpheophytin a', pheophytin b, pyropheophytin a and pyropheophytin b is PC38:0, the internal standard compound corresponding to the target compound pheophorbide a, pheophorbide a ', pheophorbide b, pyropheophorbide a, pyropheophorbide b, chlorophyllide a' and chlorophyllide b is LPC19:0, and the calculation method for relative quantification after correction is as follows: target compound relative intensity-target compound peak area/corresponding internal standard compound peak area.
Compared with the prior art, the method has the advantages that,
the invention has the following advantages: (1) the sample pretreatment is simple and quick, and chlorophyll with different solubilities and degradation product components (water solubility and fat solubility) thereof can be extracted simultaneously; (2) the analysis speed is high, and the LC-MS analysis time of each sample only needs 20 minutes; (3) the detection coverage of chlorophyll and degradation product components thereof in the tea is high, and the comprehensive coverage of 19 components in the chlorophyll degradation pathway in total on the enzyme-mediated degradation and non-enzyme thermal degradation pathways can be realized. In a word, the method is simple and rapid, and the analysis is more systematic and comprehensive.
Drawings
Fig. 1 shows the chlorophyll degradation pathway during green tea processing.
FIG. 2 is a graph of the score of the orthogonal partial least squares discriminant analysis (OPLS-DA) (A), a graph of the 20-time replacement test (B) and a graph of the factor load (C) of chlorophyll and degradation product components thereof in green tea in five different full-fire modes.
Detailed Description
The invention is further explained below with reference to examples and figures. This example is intended to be illustrative only and is not intended to limit the scope of the invention, which is to be construed as broadly as the invention is entitled to any variations or modifications made in accordance with the teachings of the invention.
Example 1
1) Extraction of chlorophyll and degradation product thereof in tea to be detected
Firstly, grinding a tea sample to be detected, and then sieving the ground tea sample with a 30-mesh sieve for later use;
② weighing about 20.0mg +/-0.1 mg tea powder into a centrifuge tube, adding 300 mu L methanol solution containing the internal standard compound, and whirling for 30 s. The internal standard compound is lysophosphatidylcholine LPC19:0, and the concentration is 2.00 mu g/mL; phosphatidylcholine PC38:0, concentration 1.33 μ g/mL;
adding 1mLMTBE, and vortexing for 30 min;
fourthly, 300 mu L of ultrapure water is added, and vortex is carried out for 30 s;
standing for 10min, and centrifuging at high speed (10000rpm 10min) to layer;
sixthly, taking 600 mu L of the upper layer and 250 mu L of the lower layer, transferring the upper layer and the lower layer into a new centrifugal tube, and centrifuging at low temperature, concentrating and drying for later use.
2) LC-MS analysis of chlorophyll and degradation products thereof in tea to be detected
Before sample injection analysis, redissolving a sample concentrated and dried by low-temperature centrifugation, wherein the redissolution is acetonitrile/isopropanol/water (65/30/5) (v/v/v);
the established analytical method of the UHPLC-q-active-based chlorophyll and degradation products thereof is as follows:
the instrument comprises the following steps: UHPLC-ESI-q-Orbitrap (thermo Fisher);
chromatographic conditions are as follows: the column was a UPLC T3 column (2.1 × 100mM, 1.8 μm, Waters) and mobile phase a was acetonitrile/water 6/4(v/v) containing 10mM ammonium acetate; mobile phase B was isopropanol/acetonitrile 9/1(v/v) with 10mM ammonium acetate at a flow rate of 0.26mL/min, and the gradient elution procedure was: 0-2.0min, 32% B; 2.0-4.0min, 32% B-60% B; 4.0-13.0min, 60% B-97% B; 13.0-17.0min, 97% B; 17.0-17.1min, 97% B-32% B; 17.1-20.0min, 32% B;
mass spectrum conditions: and performing high-resolution full scanning in an ESI positive ion mode, wherein the m/z range is 200-1200, the spraying voltage, the capillary temperature and the gas heater temperature are respectively 3.2kV, 320 ℃ and 450 ℃, and the flow rates of the sheath gas and the auxiliary gas are respectively 40arb and 10 arb.
Under the analysis conditions, the molecular formula, retention time, ionization mode and mass-to-charge ratio of 19 chlorophyll and degradation product components thereof can be detected by the method disclosed by the invention are shown in table 1.
3) And (4) relatively quantitatively analyzing chlorophyll and degradation products thereof in the tea to be detected.
Using Xcalibur software (version 2.2, Thermo Fisher, usa), based on retention time and mass-to-charge ratio in table 1, high resolution extraction ion current chromatogram (EIC) analysis was performed on the target compound, and similarly, EIC analysis was also performed on the internal standard compound to obtain its peak intensity;
normalizing the target compounds to corresponding internal standard compounds for correction to obtain relative quantification results, wherein the internal standard compounds corresponding to the target compounds 1-11 in the table 1 are PC38:0, the internal standard compounds corresponding to the target compounds 12-19 are LPC19:0, and the calculation method for performing relative quantification after correction comprises the following steps: target compound relative intensity-target compound peak area/corresponding internal standard compound peak area.
Example 2: chlorophyll degradation pathway analysis in green tea
(1) A green tea sample was collected and analyzed by the method of example 1. Each tea sample was taken 3 portions separately and tested as technical replicates.
(2) Analysis of chlorophyll degradation pathway in green tea: the results show that 19 chlorophylls and their degradation products involved in the present invention can be detected in green tea samples, covering enzymatic, non-enzymatic degradation products of different nature. Pathway analysis (fig. 1) shows that the following reactions are involved in the green tea processing: carrying out isomerization: chlorophyll a andb are converted into the respective isomers a 'and b'. Enzymatic phytyl removal reaction: hydrolysis reaction is carried out under the action of chlorophyllase to generate water-soluble chlorophyllide. ③ demagging degradation reaction: promoting Mg on porphyrin ring under acidic or heating condition2+Loss of chlorophyll converts chlorophyll to pheophytin. The chlorophyllide is easy to generate a demagging reaction under the same conditions and is degraded into the pheophorbide. Decarboxylation methylation reaction: under the action of long-term heat treatment, the carboxymethyl is further removed to produce pyropheophytin or pyropheophorbide.
Example 3: comparative analysis of chlorophyll and its degradation products in green tea under different full fire modes
(1) Preparing green tea: naturally spreading (indoor temperature is 25-28 ℃, relative humidity is 70-80%, and water content is reduced to about 70%) → roller type enzyme deactivation (leaf throwing amount is 150kg/h, 260 ℃ and 75s) → spreading for cooling (1h) → rolling (30kg leaf throwing amount, rotation speed is 50-52 r/min, and gentle rolling is 30min) → drying with a hair fire (110 ℃ and 20min, and water content is reduced to about 20%). And respectively drying with five different foot fire modes (far infrared foot fire, microwave foot fire, aroma raising machine foot fire, tea carding machine foot fire and hexagonal hot pot foot fire) to obtain green tea.
(2) The green tea prepared by the different ways of full fire was analyzed according to the method of example 1. Each tea sample was taken 3 portions separately and tested as technical replicates.
(3) Orthogonal partial least squares discriminant analysis (OPLS-DA) of green tea chlorophyll components under five different underfoot fire modes: and (3) carrying out OPLS-DA analysis by taking the contents of 19 different chlorophyll and degradation components thereof as X variables and five different full-fire modes as Y variables. The results showed significant separation of the tea samples obtained with different full fire regimes (R2X-0.998, R2Y-0.902, Q2-0.715) (fig. 2A). Wherein, the green tea prepared by microwave and far infrared foot fire is far away on the score chart, which indicates that the chlorophyll composition difference between the green tea and the score chart is more obvious. The heat transfer modes of the aroma raising machine foot fire, the strip tidying foot fire and the hexagonal hot pot foot fire are relatively close on a score chart, and the effects of the aroma raising machine foot fire, the strip tidying foot fire and the hexagonal hot pot foot fire on chlorophyll degradation are prompted to be closer. Model Y variables were cross-validated (20 permutation tests) (fig. 2B) with respective intercepts of R2 ═ 0.0,0.335 and Q2 ═ 0.0, -1.48, indicating that the model was not over-fitted. Load analysis of OPLS-DA (FIG. 2C) showed that the chlorophyll components that were more affected by microwave foot fire were Phe a, Phe b, Chl a, Phe a', Chl b, etc., while those that were more affected by far infrared foot fire were PyrPhe a, PyrPhe b, Phe b, etc.
(4) Comparison of the effects of different foot fire patterns on the changes in the components of chlorophyll and its degradation products in green tea:
one-factor anova was performed on 19 chlorophylls and their degradation components in green tea prepared in different full-fire modes (table 2). Significant differences (p <0.05) were found in the content of PyrPo a, Chl b, Ch1 b ', Chl a, Pheb, Chl a', HydPhe a, PyrPhe b, HydPhe a ', Phe a', PyrPhe a in the green tea prepared by different foot fire modes. Wherein except water-soluble pyropheophorbide (PyrPo a), the others are all fat-soluble. The contents of Chl b, Chl b ', Chl a, Phe b, Phe a and Phe a' are the highest in microwave foot fire and the lowest in far infrared foot fire, and the contents of the corresponding components are 1.6, 1.4, 2.4, 1.1, 1.3 and 1.3 times lower than that of microwave foot fire. These chlorophyll components are all green, olive green or dark green. Meanwhile, the content of the components of Pyrpo b, PyrPo a, PyrPhe b and PyrPhe a is the highest in the green tea prepared by far infrared full fire, and is respectively 10.8, 4.4, 4.3 and 3.8 times higher than that of the green tea prepared by microwave full fire, and the content of the corresponding components in the green tea prepared by microwave full fire is the lowest. The four chlorophyll degradation components are all generated by demagging degradation and decarboxylation methylation reactions and are all brown. The contents of chlorophyll and degradation components of the aroma raising machine full fire, the strip tidying full fire and the hexagonal hot pot full fire are all between the microwave full fire and the far infrared full fire. Based on the results, the retention of chlorophyll in tea leaves is higher due to microwave underfire, which is more beneficial to the color of green tea, and the damage degree of chlorophyll is higher due to far infrared underfire, which results in the color of green tea being deteriorated. The color and luster sensory analysis of the tea samples prepared by the five different foot fire modes also shows that the green tea prepared by the microwave foot fire is bright and smooth, while the green tea prepared by the far infrared foot fire is slightly dark, which is consistent with the result of the component analysis.
Table 2 analysis of the content and difference of 19 chlorophyll and its degradation product components in green tea in five full-fire modes
Note: the data shown in the table are relative quantification of the content of chlorophyll components (mean ± standard deviation, n ═ 3), with the letters in the same row indicating significant differences (p < 0.05).
Finally, it should be noted that: the above embodiments are only used to illustrate the technical solution of the present invention, and not to limit the same; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some or all of the technical features may be equivalently replaced; and the modifications or the substitutions do not make the essence of the corresponding technical solutions depart from the scope of the technical solutions of the embodiments of the present invention.
Claims (5)
1. A method for analyzing components of chlorophyll and degradation products thereof in tea leaves is characterized by comprising the following steps:
1) extraction of chlorophyll and degradation product thereof in tea to be detected
Adopting a two-phase extraction technology based on methyl tert-butyl ether/methanol/water, combining a two-phase mixing strategy, extracting, and introducing an internal standard compound in the extraction process;
2) LC-MS analysis of chlorophyll and degradation products thereof in tea to be detected
Based on UHPLC-q-active high-resolution liquid chromatography-mass spectrometry technology, performing high-resolution full-scan analysis by adopting optimized chromatographic gradient elution conditions;
3) relative quantitative analysis of chlorophyll and degradation products thereof in tea to be detected
And (3) extracting the high-resolution extraction ion current chromatograms of the target compound and the internal standard compound to obtain the intensity of the high-resolution extraction ion current chromatograms, and correcting the target compound and the corresponding internal standard compound to perform relative quantitative analysis on the target compound.
2. The method for analyzing chlorophyll and degradation product components thereof in tea leaves according to claim 1, wherein the step 1) comprises the following steps:
firstly, grinding a tea sample to be detected, and then sieving the ground tea sample with a 30-mesh sieve for later use;
weighing about 20.0mg +/-0.1 mg of tea powder into a centrifuge tube, adding 300 mu L of methanol solution containing an internal standard compound, and performing vortex operation for 30 s; the internal standard compound is lysophosphatidylcholine LPC19:0, and the concentration is 2.00 mu g/mL; phosphatidylcholine PC38:0, concentration 1.33 mug/mL;
③ adding 1mL of methyl tert-butyl ether, and whirling for 30 min;
fourthly, 300 mu L of ultrapure water is added, and vortex is carried out for 30 s;
standing for 10min, and centrifuging at 10000rpm for 10min to layer;
sixthly, taking 600 mu L of the upper layer and 250 mu L of the lower layer, transferring the upper layer and the lower layer into a new centrifugal tube, and centrifuging at low temperature, concentrating and drying for later use.
3. The method for analyzing chlorophyll and degradation product components thereof in tea leaves according to claim 1, wherein the step 2) comprises the following steps:
before sample injection analysis, redissolving a sample subjected to low-temperature centrifugal concentration and drying, wherein the redissolution is acetonitrile/isopropanol/water solution, and the volume ratio of the acetonitrile/isopropanol/water is 65/30/5;
the UHPLC-q-active analysis conditions are as follows:
the instrument comprises the following steps: UHPLC-ESI-q-Orbitrap;
chromatographic conditions are as follows: the chromatographic column is a UPLC T3 chromatographic column, and the mobile phase A is acetonitrile/water (6/4 v/v) and contains 10mM ammonium acetate; the mobile phase B is isopropanol/acetonitrile 9/1v/v, containing 10mM ammonium acetate; the flow rate was 0.26mL/min and the gradient elution procedure was: 0-2.0min, 32% B; 2.0-4.0min, 32% B-60% B; 4.0-13.0min, 60% B-97% B; 13.0-17.0min, 97% B; 17.0-17.1min, 97% B-32% B; 17.1-20.0min, 32% B;
mass spectrum conditions: and performing high-resolution full scanning in an ESI positive ion mode, wherein the m/z range is 200-1200, the spraying voltage, the capillary temperature and the gas heater temperature are respectively 3.2kV, 320 ℃ and 450 ℃, and the flow rates of the sheath gas and the auxiliary gas are respectively 40arb and 10 arb.
4. The method for analyzing chlorophyll and its degradation product components in tea leaves according to claim 1 or 3, wherein the UHPLC-q-exact analysis method of step 2) is capable of detecting 19 chlorophyll and its degradation product components selected from the group consisting of chlorophyll a, chlorophyll a ', chlorophyll b ', pheophytin a ', pheophytin b, pyropheophytin a, pheophytin b, pheophorbide a ', pheophorbide b, pyropheophorbide a, pyropheophorbide b, chlorophyllide a ' and chlorophyllide b, and the molecular formula, retention time, ionization pattern and charge ratio of the 19 compounds are shown in Table 1.
TABLE 119 molecular formulae, retention times, ionization patterns and Mass-to-Charge ratios of chlorophyll and its degradation product Components
5. The method for analyzing chlorophyll and degradation product components thereof in tea leaves as claimed in claim 4, wherein said step 3) comprises the following steps:
performing high-resolution extraction ion current chromatogram (EIC) analysis on the target compound by using Xcalibur software based on retention time and mass-to-charge ratio in the table 1, and performing EIC analysis on the internal standard compound to obtain peak intensity;
normalizing the target compound to a corresponding internal standard compound for correction to obtain a relative quantitative result, wherein the internal standard compound corresponding to the target compound chlorophyll a, chlorophyll a ', chlorophyll b', pheophytin a ', hydroxyphenylpheophytin a', pheophytin b, pyropheophytin a and pyropheophytin b is PC38:0, the internal standard compound corresponding to the target compound pheophorbide a, pheophorbide a ', pheophorbide b, pyropheophorbide a, pyropheophorbide b, chlorophyllide a' and chlorophyllide b is LPC19:0, and the calculation method for relative quantification after correction is as follows: target compound relative intensity-target compound peak area/corresponding internal standard compound peak area.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110436988.6A CN113203806A (en) | 2021-04-22 | 2021-04-22 | Method for analyzing components of chlorophyll and degradation products thereof in tea |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110436988.6A CN113203806A (en) | 2021-04-22 | 2021-04-22 | Method for analyzing components of chlorophyll and degradation products thereof in tea |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113203806A true CN113203806A (en) | 2021-08-03 |
Family
ID=77028060
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110436988.6A Pending CN113203806A (en) | 2021-04-22 | 2021-04-22 | Method for analyzing components of chlorophyll and degradation products thereof in tea |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113203806A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115166095A (en) * | 2022-07-20 | 2022-10-11 | 江苏省食品药品监督检验研究院 | Method for detecting related substances of egg yolk lecithin |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20170360861A1 (en) * | 2014-12-12 | 2017-12-21 | Green Sky Labs, Inc. | Methods for extracting target compounds from cannabis |
| CN111323494A (en) * | 2018-12-14 | 2020-06-23 | 中国科学院大连化学物理研究所 | Rapid pretreatment method for extracting micromolecular metabolites in adherent cells |
| CN112435721A (en) * | 2020-11-03 | 2021-03-02 | 江南大学 | Method for constructing Longjing green tea quality discrimination model based on partial least squares |
-
2021
- 2021-04-22 CN CN202110436988.6A patent/CN113203806A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20170360861A1 (en) * | 2014-12-12 | 2017-12-21 | Green Sky Labs, Inc. | Methods for extracting target compounds from cannabis |
| CN111323494A (en) * | 2018-12-14 | 2020-06-23 | 中国科学院大连化学物理研究所 | Rapid pretreatment method for extracting micromolecular metabolites in adherent cells |
| CN112435721A (en) * | 2020-11-03 | 2021-03-02 | 江南大学 | Method for constructing Longjing green tea quality discrimination model based on partial least squares |
Non-Patent Citations (3)
| Title |
|---|
| LI J 等: "Comprehensive lipidome-wide profiling reveals dynamic changes of tea lipids during manufacturing process of black tea", 《JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY》 * |
| LI J 等: "Investigation on green tea lipids and their metabolic variations during manufacturing by nontargeted lipidomics" * |
| LI J 等: "Investigation on green tea lipids and their metabolic variations during manufacturing by nontargeted lipidomics", 《FOOD CHEMISTRY》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115166095A (en) * | 2022-07-20 | 2022-10-11 | 江苏省食品药品监督检验研究院 | Method for detecting related substances of egg yolk lecithin |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Wang et al. | Widely targeted metabolomic analysis reveals dynamic changes in non-volatile and volatile metabolites during green tea processing | |
| Liu et al. | Enhancing Robusta coffee aroma by modifying flavour precursors in the green coffee bean | |
| Xiong et al. | Effect of processing on the phenolic contents, antioxidant activity and volatile compounds of sorghum grain tea | |
| Arena et al. | Distribution of fatty acids and phytosterols as a criterion to discriminate geographic origin of pistachio seeds | |
| Sun et al. | Content variations in compositions and volatile component in jujube fruits during the blacking process | |
| Chen et al. | Non-targeted metabolomics analysis reveals dynamic changes of volatile and non-volatile metabolites during oolong tea manufacture | |
| Nemzer et al. | Betalainic and nutritional profiles of pigment-enriched red beet root (Beta vulgaris L.) dried extracts | |
| Zheng et al. | Screening the cultivar and processing factors based on the flavonoid profiles of dry teas using principal component analysis | |
| Procida et al. | Chemical composition and functional characterisation of commercial pumpkin seed oil | |
| Perrone et al. | Comprehensive analysis of major and minor chlorogenic acids and lactones in economically relevant Brazilian coffee cultivars | |
| Scalzo et al. | Anthocyanin composition of cauliflower (Brassica oleracea L. var. botrytis) and cabbage (B. oleracea L. var. capitata) and its stability in relation to thermal treatments | |
| Tran et al. | Assessing cocoa aroma quality by multiple analytical approaches | |
| Somporn et al. | Effects of roasting degree on radical scavenging activity, phenolics and volatile compounds of Arabica coffee beans (Coffea arabica L. cv. Catimor) | |
| Liu et al. | Multi-omics and enzyme activity analysis of flavour substances formation: Major metabolic pathways alteration during Congou black tea processing | |
| Teixido et al. | Liquid chromatography multi-stage mass spectrometry for the analysis of 5-hydroxymethylfurfural in foods | |
| Nenadis et al. | Applicability of PTR-MS in the quality control of saffron | |
| Zhang et al. | Untargeted metabolomics coupled with chemometrics analysis reveals potential non-volatile markers during oolong tea shaking | |
| Liu et al. | The effects of dynamic changes of malonyl ginsenosides on evaluation and quality control of Panax ginseng CA Meyer | |
| Yang et al. | Rapid profiling of volatile compounds in green teas using Micro-Chamber/Thermal Extractor combined with thermal desorption coupled to gas chromatography-mass spectrometry followed by multivariate statistical analysis | |
| Gras et al. | Determination of anthocyanins from black carrots by UHPLC-PDA after ultrasound-assisted extraction | |
| Canjura et al. | Separation of chlorophyll compounds and their polar derivatives by high-performance liquid chromatography | |
| Delpino-Rius et al. | A fast and reliable ultrahigh-performance liquid chromatography method to assess the fate of chlorophylls in teas and processed vegetable foodstuff | |
| Avellone et al. | Investigation on the influence of spray-drying technology on the quality of Sicilian Nero d’Avola wines | |
| Kallio et al. | Analysis of low erucic acid turnip rapeseed oil (Brassica campestris) by negative ion chemical ionization tandem mass spectrometry. A method giving information on the fatty acid composition in positions sn-2 and sn-1/3 of triacylglycerols | |
| Magi et al. | Characterization of cocoa liquors by GC–MS and LC–MS/MS: focus on alkylpyrazines and flavanols |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20210803 |
|
| RJ01 | Rejection of invention patent application after publication |