CN116908352A - Detection and identification method for loquat honey - Google Patents
Detection and identification method for loquat honey Download PDFInfo
- Publication number
- CN116908352A CN116908352A CN202310916981.3A CN202310916981A CN116908352A CN 116908352 A CN116908352 A CN 116908352A CN 202310916981 A CN202310916981 A CN 202310916981A CN 116908352 A CN116908352 A CN 116908352A
- Authority
- CN
- China
- Prior art keywords
- acid
- mobile phase
- coumaroyl
- tri
- honey
- 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
- 235000012907 honey Nutrition 0.000 title claims abstract description 127
- 241001092070 Eriobotrya Species 0.000 title claims abstract description 78
- 235000009008 Eriobotrya japonica Nutrition 0.000 title claims abstract description 77
- 238000000034 method Methods 0.000 title claims abstract description 70
- 238000001514 detection method Methods 0.000 title claims abstract description 22
- ZEYHEAKUIGZSGI-UHFFFAOYSA-N 4-methoxybenzoic acid Chemical compound COC1=CC=C(C(O)=O)C=C1 ZEYHEAKUIGZSGI-UHFFFAOYSA-N 0.000 claims abstract description 60
- PZIRUHCJZBGLDY-UHFFFAOYSA-N Caffeoylquinic acid Natural products CC(CCC(=O)C(C)C1C(=O)CC2C3CC(O)C4CC(O)CCC4(C)C3CCC12C)C(=O)O PZIRUHCJZBGLDY-UHFFFAOYSA-N 0.000 claims abstract description 48
- 238000004128 high performance liquid chromatography Methods 0.000 claims abstract description 41
- FVQOMEDMFUMIMO-UHFFFAOYSA-N Hyperosid Natural products OC1C(O)C(O)C(CO)OC1OC1C(=O)C2=C(O)C=C(O)C=C2OC1C1=CC=C(O)C(O)=C1 FVQOMEDMFUMIMO-UHFFFAOYSA-N 0.000 claims abstract description 32
- RAGZUCNPTLULOL-KQJPBSFVSA-N 3-Feruloylquinic acid Chemical compound C1=C(O)C(OC)=CC(\C=C\C(=O)O[C@H]2[C@H]([C@H](O)C[C@@](O)(C2)C(O)=O)O)=C1 RAGZUCNPTLULOL-KQJPBSFVSA-N 0.000 claims abstract description 25
- CWVRJTMFETXNAD-FWCWNIRPSA-N 3-O-Caffeoylquinic acid Natural products O[C@H]1[C@@H](O)C[C@@](O)(C(O)=O)C[C@H]1OC(=O)\C=C\C1=CC=C(O)C(O)=C1 CWVRJTMFETXNAD-FWCWNIRPSA-N 0.000 claims abstract description 25
- RAGZUCNPTLULOL-MDRZDLKXSA-N 5-O-feruloylquinic acid Natural products O[C@@]1(C[C@H]([C@@H]([C@@H](C1)O)O)OC(C=CC1=CC(=C(C=C1)O)OC)=O)C(=O)O RAGZUCNPTLULOL-MDRZDLKXSA-N 0.000 claims abstract description 25
- RAGZUCNPTLULOL-JSHWQEIDSA-N 5-O-feruoylquinic acid Natural products O[C@@]1(C[C@H]([C@@H]([C@@H](C1)OC(C=CC1=CC(=C(C=C1)O)OC)=O)O)O)C(=O)O RAGZUCNPTLULOL-JSHWQEIDSA-N 0.000 claims abstract description 25
- OVSQVDMCBVZWGM-SJWGPRHPSA-N Hyperin Natural products O[C@H]1[C@H](O)[C@@H](O)[C@@H](CO)O[C@H]1OC1=C(C=2C=C(O)C(O)=CC=2)OC2=CC(O)=CC(O)=C2C1=O OVSQVDMCBVZWGM-SJWGPRHPSA-N 0.000 claims abstract description 25
- CWVRJTMFETXNAD-KLZCAUPSSA-N Neochlorogenin-saeure Natural products O[C@H]1C[C@@](O)(C[C@@H](OC(=O)C=Cc2ccc(O)c(O)c2)[C@@H]1O)C(=O)O CWVRJTMFETXNAD-KLZCAUPSSA-N 0.000 claims abstract description 25
- CWVRJTMFETXNAD-JUHZACGLSA-N chlorogenic acid Chemical compound O[C@@H]1[C@H](O)C[C@@](O)(C(O)=O)C[C@H]1OC(=O)\C=C\C1=CC=C(O)C(O)=C1 CWVRJTMFETXNAD-JUHZACGLSA-N 0.000 claims abstract description 25
- 229940074393 chlorogenic acid Drugs 0.000 claims abstract description 25
- FFQSDFBBSXGVKF-KHSQJDLVSA-N chlorogenic acid Natural products O[C@@H]1C[C@](O)(C[C@@H](CC(=O)C=Cc2ccc(O)c(O)c2)[C@@H]1O)C(=O)O FFQSDFBBSXGVKF-KHSQJDLVSA-N 0.000 claims abstract description 25
- 235000001368 chlorogenic acid Nutrition 0.000 claims abstract description 25
- BMRSEYFENKXDIS-KLZCAUPSSA-N cis-3-O-p-coumaroylquinic acid Natural products O[C@H]1C[C@@](O)(C[C@@H](OC(=O)C=Cc2ccc(O)cc2)[C@@H]1O)C(=O)O BMRSEYFENKXDIS-KLZCAUPSSA-N 0.000 claims abstract description 25
- BBFYUPYFXSSMNV-UHFFFAOYSA-N quercetin-7-o-galactoside Natural products OC1C(O)C(O)C(CO)OC1OC1=CC(O)=C2C(=O)C(O)=C(C=3C=C(O)C(O)=CC=3)OC2=C1 BBFYUPYFXSSMNV-UHFFFAOYSA-N 0.000 claims abstract description 25
- RAGZUCNPTLULOL-UHFFFAOYSA-N trans-3-feruloylquinic acid Natural products C1=C(O)C(OC)=CC(C=CC(=O)OC2C(C(O)CC(O)(C2)C(O)=O)O)=C1 RAGZUCNPTLULOL-UHFFFAOYSA-N 0.000 claims abstract description 25
- BMRSEYFENKXDIS-QHAYPTCMSA-N 3-O-Coumaroylquinic acid Chemical compound O[C@H]1[C@H](O)C[C@](O)(C(O)=O)C[C@H]1OC(=O)\C=C\C1=CC=C(O)C=C1 BMRSEYFENKXDIS-QHAYPTCMSA-N 0.000 claims abstract description 24
- BMRSEYFENKXDIS-UNIGVISCSA-N 5-p-coumaroylquinic acid Natural products O[C@H]1[C@H](O)C[C@](O)(C(O)=O)C[C@H]1OC(=O)C=CC1=CC=C(O)C=C1 BMRSEYFENKXDIS-UNIGVISCSA-N 0.000 claims abstract description 24
- OVSQVDMCBVZWGM-DTGCRPNFSA-N quercetin 3-O-beta-D-galactopyranoside Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1OC1=C(C=2C=C(O)C(O)=CC=2)OC2=CC(O)=CC(O)=C2C1=O OVSQVDMCBVZWGM-DTGCRPNFSA-N 0.000 claims abstract description 24
- CWVRJTMFETXNAD-BMNNCGMMSA-N (1s,3r,4s,5r)-3-[(e)-3-(3,4-dihydroxyphenyl)prop-2-enoyl]oxy-1,4,5-trihydroxycyclohexane-1-carboxylic acid Chemical compound O[C@H]1[C@H](O)C[C@@](O)(C(O)=O)C[C@H]1OC(=O)\C=C\C1=CC=C(O)C(O)=C1 CWVRJTMFETXNAD-BMNNCGMMSA-N 0.000 claims abstract description 23
- KRZBCHWVBQOTNZ-DLDRDHNVSA-N isochlorogenic acid Natural products O[C@@H]1[C@H](C[C@@](O)(C[C@H]1OC(=O)C=Cc2ccc(O)c(O)c2)C(=O)O)OC(=O)C=Cc3ccc(O)c(O)c3 KRZBCHWVBQOTNZ-DLDRDHNVSA-N 0.000 claims abstract description 23
- 238000001228 spectrum Methods 0.000 claims abstract description 15
- 238000004445 quantitative analysis Methods 0.000 claims abstract description 8
- QJTYCCFDQWFJHU-UHFFFAOYSA-N Quercetin-5-O-beta-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1=CC(O)=CC2=C1C(=O)C(O)=C(C=1C=C(O)C(O)=CC=1)O2 QJTYCCFDQWFJHU-UHFFFAOYSA-N 0.000 claims abstract description 7
- GXMWXESSGGEWEM-UHFFFAOYSA-N isoquercitrin Natural products OCC(O)C1OC(OC2C(Oc3cc(O)cc(O)c3C2=O)c4ccc(O)c(O)c4)C(O)C1O GXMWXESSGGEWEM-UHFFFAOYSA-N 0.000 claims abstract description 7
- SZDMSNWMQAMVTJ-UHFFFAOYSA-N quercetin-3-O-glucoside Natural products OC1OC(COC2=C(C(=O)c3cc(O)cc(O)c3O2)c4ccc(O)c(O)c4)C(O)C(O)C1O SZDMSNWMQAMVTJ-UHFFFAOYSA-N 0.000 claims abstract description 7
- FZKBNCDAGYDHTP-UHFFFAOYSA-N quercetin-3-O-glycoside Natural products OC1C(O)C(O)C(O)OC1COC1=C(C=2C=C(O)C(O)=CC=2)OC2=CC(O)=CC(O)=C2C1=O FZKBNCDAGYDHTP-UHFFFAOYSA-N 0.000 claims abstract description 7
- OVSQVDMCBVZWGM-QSOFNFLRSA-N quercetin 3-O-beta-D-glucopyranoside Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=C(C=2C=C(O)C(O)=CC=2)OC2=CC(O)=CC(O)=C2C1=O OVSQVDMCBVZWGM-QSOFNFLRSA-N 0.000 claims abstract 3
- 239000012071 phase Substances 0.000 claims description 87
- 150000001875 compounds Chemical class 0.000 claims description 77
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 57
- 230000014759 maintenance of location Effects 0.000 claims description 38
- 239000000243 solution Substances 0.000 claims description 31
- 239000000126 substance Substances 0.000 claims description 28
- 239000003480 eluent Substances 0.000 claims description 18
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 claims description 18
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 15
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 15
- 239000006228 supernatant Substances 0.000 claims description 12
- 239000000523 sample Substances 0.000 claims description 11
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 claims description 9
- 239000008367 deionised water Substances 0.000 claims description 9
- 229910021641 deionized water Inorganic materials 0.000 claims description 9
- 235000019253 formic acid Nutrition 0.000 claims description 9
- 238000011068 loading method Methods 0.000 claims description 8
- 238000002414 normal-phase solid-phase extraction Methods 0.000 claims description 7
- 239000012488 sample solution Substances 0.000 claims description 6
- 229910021642 ultra pure water Inorganic materials 0.000 claims description 6
- 239000012498 ultrapure water Substances 0.000 claims description 6
- 229910001868 water Inorganic materials 0.000 claims description 6
- ZCHPKWUIAASXPV-UHFFFAOYSA-N acetic acid;methanol Chemical group OC.CC(O)=O ZCHPKWUIAASXPV-UHFFFAOYSA-N 0.000 claims description 5
- 238000001914 filtration Methods 0.000 claims description 5
- 239000004480 active ingredient Substances 0.000 claims description 4
- 230000008859 change Effects 0.000 claims description 4
- 238000002347 injection Methods 0.000 claims description 4
- 239000007924 injection Substances 0.000 claims description 4
- 230000001105 regulatory effect Effects 0.000 claims description 4
- 238000005303 weighing Methods 0.000 claims description 4
- 230000003213 activating effect Effects 0.000 claims description 3
- 239000007864 aqueous solution Substances 0.000 claims description 3
- 238000001035 drying Methods 0.000 claims description 3
- 238000010828 elution Methods 0.000 claims description 3
- 239000012528 membrane Substances 0.000 claims description 3
- 238000002156 mixing Methods 0.000 claims description 3
- 239000000047 product Substances 0.000 claims description 3
- 238000000605 extraction Methods 0.000 claims description 2
- 239000011148 porous material Substances 0.000 claims description 2
- 239000012086 standard solution Substances 0.000 claims description 2
- -1 trans-coumaroyl spermidine Chemical compound 0.000 claims description 2
- ATHGHQPFGPMSJY-UHFFFAOYSA-N Spermidine Natural products NCCCCNCCCN ATHGHQPFGPMSJY-UHFFFAOYSA-N 0.000 claims 1
- 239000002253 acid Substances 0.000 claims 1
- 238000003556 assay Methods 0.000 claims 1
- 238000004587 chromatography analysis Methods 0.000 claims 1
- 229940063673 spermidine Drugs 0.000 claims 1
- 238000013441 quality evaluation Methods 0.000 abstract description 2
- 230000035945 sensitivity Effects 0.000 abstract description 2
- 150000002500 ions Chemical class 0.000 description 38
- 238000010521 absorption reaction Methods 0.000 description 18
- 238000004458 analytical method Methods 0.000 description 18
- 238000001819 mass spectrum Methods 0.000 description 17
- 230000008569 process Effects 0.000 description 12
- 238000004811 liquid chromatography Methods 0.000 description 11
- 241000196324 Embryophyta Species 0.000 description 9
- 238000013467 fragmentation Methods 0.000 description 4
- 238000006062 fragmentation reaction Methods 0.000 description 4
- OPJZLUXFQFQYAI-GNPVFZCLSA-N quercetin 3-O-beta-D-glucofuranoside Chemical compound O[C@@H]1[C@@H](O)[C@@H]([C@H](O)CO)O[C@H]1OC1=C(C=2C=C(O)C(O)=CC=2)OC2=CC(O)=CC(O)=C2C1=O OPJZLUXFQFQYAI-GNPVFZCLSA-N 0.000 description 4
- 239000002994 raw material Substances 0.000 description 4
- 238000012795 verification Methods 0.000 description 4
- 244000020518 Carthamus tinctorius Species 0.000 description 3
- 235000003255 Carthamus tinctorius Nutrition 0.000 description 3
- 240000008620 Fagopyrum esculentum Species 0.000 description 3
- 235000009419 Fagopyrum esculentum Nutrition 0.000 description 3
- 240000005323 Hoya carnosa Species 0.000 description 3
- 241000207925 Leonurus Species 0.000 description 3
- 235000010654 Melissa officinalis Nutrition 0.000 description 3
- 239000013543 active substance Substances 0.000 description 3
- 238000004949 mass spectrometry Methods 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 210000000051 wattle Anatomy 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 235000000604 Chrysanthemum parthenium Nutrition 0.000 description 2
- 235000000802 Leonurus cardiaca ssp. villosus Nutrition 0.000 description 2
- 235000017784 Mespilus germanica Nutrition 0.000 description 2
- 244000182216 Mimusops elengi Species 0.000 description 2
- 235000000560 Mimusops elengi Nutrition 0.000 description 2
- 240000003152 Rhus chinensis Species 0.000 description 2
- 235000014220 Rhus chinensis Nutrition 0.000 description 2
- 235000007837 Vangueria infausta Nutrition 0.000 description 2
- RDOXTESZEPMUJZ-UHFFFAOYSA-N anisole Chemical compound COC1=CC=CC=C1 RDOXTESZEPMUJZ-UHFFFAOYSA-N 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 238000011156 evaluation Methods 0.000 description 2
- 239000007789 gas Substances 0.000 description 2
- NQYPTLKGQJDGTI-FCVRJVSHSA-N hyperoside Natural products OC[C@H]1O[C@@H](OC2=C(Oc3cc(O)cc(O)c3[C@H]2O)c4ccc(O)c(O)c4)[C@H](O)[C@@H](O)[C@H]1O NQYPTLKGQJDGTI-FCVRJVSHSA-N 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 150000002772 monosaccharides Chemical class 0.000 description 2
- 230000000050 nutritive effect Effects 0.000 description 2
- AAWZDTNXLSGCEK-LNVDRNJUSA-N (3r,5r)-1,3,4,5-tetrahydroxycyclohexane-1-carboxylic acid Chemical compound O[C@@H]1CC(O)(C(O)=O)C[C@@H](O)C1O AAWZDTNXLSGCEK-LNVDRNJUSA-N 0.000 description 1
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 1
- AAWZDTNXLSGCEK-UHFFFAOYSA-N Cordycepinsaeure Natural products OC1CC(O)(C(O)=O)CC(O)C1O AAWZDTNXLSGCEK-UHFFFAOYSA-N 0.000 description 1
- 206010011224 Cough Diseases 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 229930091371 Fructose Natural products 0.000 description 1
- 239000005715 Fructose Substances 0.000 description 1
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 241000257303 Hymenoptera Species 0.000 description 1
- 244000241838 Lycium barbarum Species 0.000 description 1
- 235000015459 Lycium barbarum Nutrition 0.000 description 1
- 241000219071 Malvaceae Species 0.000 description 1
- AAWZDTNXLSGCEK-ZHQZDSKASA-N Quinic acid Natural products O[C@H]1CC(O)(C(O)=O)C[C@H](O)C1O AAWZDTNXLSGCEK-ZHQZDSKASA-N 0.000 description 1
- 235000004789 Rosa xanthina Nutrition 0.000 description 1
- 241000220222 Rosaceae Species 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 241000793823 Tilia amurensis Species 0.000 description 1
- 240000007313 Tilia cordata Species 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 235000011114 ammonium hydroxide Nutrition 0.000 description 1
- 238000000889 atomisation Methods 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 238000005842 biochemical reaction Methods 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 150000001793 charged compounds Chemical class 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 235000009508 confectionery Nutrition 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000010812 external standard method Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 239000003205 fragrance Substances 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 239000001307 helium Substances 0.000 description 1
- 229910052734 helium Inorganic materials 0.000 description 1
- SWQJXJOGLNCZEY-UHFFFAOYSA-N helium atom Chemical compound [He] SWQJXJOGLNCZEY-UHFFFAOYSA-N 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 238000010813 internal standard method Methods 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
- UZKWTJUDCOPSNM-UHFFFAOYSA-N methoxybenzene Substances CCCCOC=C UZKWTJUDCOPSNM-UHFFFAOYSA-N 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000004451 qualitative analysis Methods 0.000 description 1
- 230000002040 relaxant effect Effects 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 238000004088 simulation Methods 0.000 description 1
- 238000011895 specific detection Methods 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 238000012916 structural analysis Methods 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 238000001196 time-of-flight mass spectrum Methods 0.000 description 1
- 230000017260 vegetative to reproductive phase transition of meristem Effects 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
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
- 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
- G01N30/02—Column chromatography
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/34—Control of physical parameters of the fluid carrier of fluid composition, e.g. gradient
-
- 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
- G01N30/86—Signal analysis
- G01N30/8675—Evaluation, i.e. decoding of the signal into analytical information
- G01N30/8686—Fingerprinting, e.g. without prior knowledge of the sample components
-
- 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
- G01N2030/042—Standards
- G01N2030/047—Standards external
-
- 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
- G01N2030/062—Preparation extracting sample from raw material
-
- 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
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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)
- Engineering & Computer Science (AREA)
- Library & Information Science (AREA)
- Treatment Of Liquids With Adsorbents In General (AREA)
Abstract
The invention relates to a detection and identification method of loquat honey, which adopts a high performance liquid chromatography combined with a fingerprint spectrum method and a high performance liquid chromatography quantitative analysis method to detect and identify the loquat honey, wherein the high performance liquid chromatography combined with the fingerprint spectrum indicates that the loquat honey contains anisoic acid, chlorogenic acid, isochlorogenic acid, 3-O-p-coumaroyl quinic acid, 5-O-feruloyl quinic acid, hyperin (namely quercetin-3-O-glucoside), cis-tri-p-coumaroyl spermidine, cis, trans-tri-p-coumaroyl spermidine, cis, trans, cis-tri-p-coumaroyl spermidine and trans, trans-tri-p-coumaroyl spermidine simultaneously, and the honey to be detected can be judged to be the loquat honey. The invention provides a new method for constructing the loquat honey authenticity and quality evaluation system, has high specificity and good detection sensitivity, and has good economic value and social significance.
Description
Technical Field
The invention relates to the technical field of food detection, in particular to a detection and identification method for loquat honey.
Background
Loquat is a evergreen arbor plant of Eriobotrya genus of Malidae of Rosaceae, and is native to Sichuan and Hubei areas of China, and is distributed in the south of Yangtze river basin, and is mostly cultivated in hilly and low mountain areas and plain areas, the optimal honey secretion temperature is 15-22 ℃, the honey can be produced in one flowering phase by 10-20 jin, and the honey can be produced in each group by more than 30 jin during high yield. The loquat honey is light amber in color, easy to crystallize, white in shape, unique in fragrance, fine in honey quality and sweet in taste. The loquat honey has higher nutritive value, the main components are glucose and fructose, and the loquat honey also contains a small amount of active substances such as sucrose, protein, vitamins, organic acid, minerals, enzymes and the like, the most representative nutritive substances are monosaccharide and active substances, the monosaccharide can supplement a large amount of energy for a human body, and the active substances are catalysts for various biochemical reactions in the human body. As with other honey, loquat honey also has the functions of relaxing bowel, moistening lung, relieving cough, detoxifying, treating sore, relieving pain, tonifying middle-jiao and Qi, and harmonizing various medicines.
Therefore, the loquat honey is driven by economic benefits, has adulteration phenomenon, seriously disturbs the normal order of the honey market, and restricts the development of the special economy of the loquat honey industry. Therefore, establishing a set of methods for identifying the authenticity of loquat honey is a technical problem to be solved.
Disclosure of Invention
Aiming at the problems in the prior art, the invention aims to provide a detection and identification method for loquat honey.
In order to achieve the object, the technical scheme of the invention is as follows:
in a first aspect of the invention, a detection and identification method for loquat honey is provided, which adopts a high performance liquid chromatography combined with fingerprint method and a high performance liquid chromatography quantitative analysis method to detect and identify the loquat honey,
wherein, the high performance liquid chromatography combines with fingerprint spectrum to identify that the loquat honey contains anisoic acid, which has the structure shown in formula (I):
further, the retention time of the anisic acid is 51.94min;
in the high performance liquid chromatography, the mobile phase A is acetic acid aqueous solution with the concentration of 0.18-0.22 v/v%, and the mobile phase B is acetic acid methanol solution with the concentration of 0.18-0.22 v/v%; still more preferably, in the high performance liquid chromatography, the mobile phase A is an aqueous acetic acid solution with a concentration of 0.2v/v%, and the mobile phase B is a methanol acetic acid solution with a concentration of 0.2 v/v%;
further, in the high performance liquid chromatography, the gradient elution procedure is:
0-11 min, the mobile phase B is increased from 5v/v% to 11v/v%; 11-14 min, the mobile phase B is increased from 11v/v% to 14v/v%; the flow phase B rises from 14v/v% to 15v/v% for 14-17 min; 17-24 min, the mobile phase B is increased from 15v/v% to 16v/v%; the mobile phase B rises from 16% to 17% after 24-28 min; 28-30 min, the mobile phase B rises from 17v/v% to 22v/v%; 30-38 min, the mobile phase B is increased from 22v/v% to 25v/v%; the flow phase B rises from 25v/v% to 30v/v% for 38-41 min; 41-46 min, the mobile phase B is increased from 30v/v% to 33v/v%; 46-55 min, the mobile phase B is kept unchanged at 33%; 55-60 min, the mobile phase B is increased from 33v/v% to 34v/v%; 60-70 min, the mobile phase B is increased from 34v/v% to 36v/v%; 70-80 min, and the mobile phase B rises from 36% to 40%; 80-90 min, the mobile phase B is increased from 40v/v% to 45v/v%; 90-100 min, the mobile phase B is increased from 45v/v% to 52v/v%; 100-110 min, the mobile phase B is increased from 52v/v% to 57v/v%; 110-120 min, the mobile phase B rises from 57% to 65%; 120-130 min, the mobile phase B rises from 65v/v% to 70v/v%; 130-135 min, the mobile phase B is increased from 70v/v% to 80v/v%; mobile phase a changes with the volume change of mobile phase B, and the sum of the volumes of the mobile phase a and the mobile phase B is 100%; mobile phase a changes with the volume change of mobile phase B, and the sum of the volumes of the mobile phase a and the mobile phase B is 100%;
further, the high performance liquid chromatography includes the following chromatographic conditions:
the chromatographic column is Phenomenex Gemini C 18 The chromatographic column has the flow rate of 0.6-0.8 mL/min, the sample injection amount of 10-30 mu L, the column temperature of 34-36 ℃ and the detection wavelength of 280nm; further preferably, the sample injection amount is 20 mu L, the flow rate is 0.7mL/min, and the column temperature is 35 ℃;
further, the high performance liquid chromatography further includes a pretreatment step:
s1, mixing honey to be detected with a proper amount of water, fully dissolving the honey to obtain honey water solution, regulating the pH value of the honey water solution to be 6.5-7, centrifuging the honey water solution at a rotating speed of 8800-9200rpm for 18-22 min, and taking supernatant for later use;
s2: extracting the supernatant obtained in S1 by using a Strata-X-A solid phase extraction column, activating the solid phase extraction column by using methanol, balancing the extraction column by using ultrapure water, loading the supernatant, eluting by using deionized water, eluting by using a methanol solution containing 9-11 v/v% formic acid, extracting active ingredients in the supernatant to obtain an eluent, drying the obtained eluent, re-dissolving by using a proper amount of methanol, and filtering to obtain the product, wherein the dosage of the methanol, the ultrapure water, the deionized water and the methanol solution containing 9-11 v/v% formic acid is 2-4 times of the column volume;
further, in S1, the pH value is regulated to 6.8, the centrifugal rotating speed is 9000rpm, and the centrifugal time is 20min;
further, in S2, the pore diameter of the filtering membrane is 0.22 μm;
further, in S2, the eluent contains 10v/v% formic acid in methanol;
in S2, the dosage of the methanol, the ultrapure water, the deionized water and the methanol solution containing 9-11 v/v percent of formic acid is 3 times of the column volume;
further, the high performance liquid chromatography is combined with fingerprint to identify chlorogenic acid, isochlorogenic acid, 3-O-p-coumaroyl quinic acid, 5-O-feruloyl quinic acid, hyperoside (namely quercetin-3-O-glucoside), cis-tri-p-coumaroyl spermidine, cis, trans, cis-tri-p-coumaroyl spermidine, trans-tri-p-coumaroyl spermidine;
further, the retention time t of chlorogenic acid R Retention time t of isochlorogenic acid of 22.21min R Retention time t of 3-O-p-coumaroyl quinic acid of 30.17min R Retention time t of 5-O-feruloylquinic acid of 34.82min R A retention time t of hyperin (i.e. quercetin-3-O-glucoside) of 40.53min R 59.59min, cis-tri-p-coumaroyl spermidine retention time t R A retention time t of 87.06min for cis, trans-tri-p-coumaroyl spermidine R Retention time t of 96.22min, cis, trans, cis-tri-p-coumaroyl spermidine R Retention time t of 102.07min, trans-tri-p-coumaroyl spermidine R 107.36min;
further, the quantitative analysis method of high performance liquid chromatography comprises the following steps:
(1) Chromatographic conditions: combining with the high performance liquid chromatography and fingerprint method;
(2) Preparing a loquat honey standard substance solution: weighing appropriate amounts of chlorogenic acid, isochlorogenic acid, 3-O-p-coumaroyl quinic acid, 5-O-feruloyl quinic acid, anisic acid, hyperin and tri-p-coumaroyl spermidine to prepare standard substance solutions with concentrations of 0.22mg/ml,0.8mg/ml,0.64mg/ml,0.32mg/ml,0.6mg/ml,0.56mg/ml and 3.6mg/ml respectively, and drawing loquat honey standard curves as shown in the following table 1: correcting according to the sample loading amount x and the peak area y and the gradient sample loading amount of 5-10 uL by adopting a 5-point method; preferably, the concentration of chlorogenic acid, isochlorogenic acid, 3-O-p-coumaroyl quinic acid, 5-O-feruloyl quinic acid, anisic acid, hyperin and tri-p-coumaroyl spermidine standard solution is 0.22mg/ml,0.8mg/ml,0.64mg/ml,0.32mg/ml,0.6mg/ml,0.56mg/ml and 3.6mg/ml respectively;
TABLE 1 information about standards
(3) Content determination method: injecting the sample solution according to the method in the step (1), and injecting the sample solution according to the corresponding t R Obtaining the content of the compound in honey by substituting the area of chromatographic peaks into a corresponding standard curve, wherein the content of chlorogenic acid is not lower than 0.12-1.22 mg/kg, the content of iso-chlorogenic acid is not lower than 0.11-0.94mg/kg, the content of 3-O-p-coumaroyl quinic acid is 0.28-1.72 mg/kg, the content of 5-O-feruloylquinic acid is not remarkably lower than 0.14-0.58 mg/kg, the content of anisic acid is not lower than 3.30-9.64mg/kg, the content of hyperin is not lower than 0.14-2.15 mg/kg, the content of cis, cis-tri-p-coumaroyl spermidine is not lower than 0.50-3.07 mg/kg, the content of cis, trans-tri-p-coumaroyl spermidine is not lower than 0.68-2.45 mg/kg, the content of trans-tri-p-coumaroyl spermidine is not lower than 0.74-2.85 mg/kg, and the content of trans-tri-p-coumaroyl spermidine is not remarkably lower than 0.37 mg/kg;
in a second aspect the invention provides the use of anisic acid in the identification or detection of loquat honey.
Advantageous effects
1. The invention adopts a solid phase extraction method to extract and separate the active ingredients of the loquat honey, and adopts an HPLC combined fingerprint spectrum method and an HPLC quantitative analysis method to establish a qualitative analysis and a quantitative analysis method of plant compounds in the loquat honey.
2. The analysis method identifies that the loquat honey contains representative compounds which are chlorogenic acid, isochlorogenic acid, 3-O-p-coumaroyl quinic acid, 5-O-feruloyl quinic acid, hyperin (namely quercetin-3-O-glucoside), cis-tri-p-coumaroyl spermidine, cis, trans-tri-p-coumaroyl spermidine, cis, trans, cis-tri-p-coumaroyl spermidine, trans-tri-p-coumaroyl spermidine and the obtained loquat honey plant compound HPLC (high performance liquid chromatography) map. The spectrum is imported into a traditional Chinese medicine chromatographic fingerprint similarity evaluation system, and the standard loquat honey fingerprint with representative significance is generated through preliminary simulation.
By comparing with HPLC patterns of four large amounts of single nectar such as locust honey, linden honey, wattle honey and rape honey and analyzing and comparing with plant compounds such as collected other masses of honey such as safflower honey, chinese gall honey, buckwheat honey, motherwort honey, matrimony vine honey and the like, chlorogenic acid, isochlorogenic acid, 3-O-p-coumaroyl quinic acid, 5-O-feruloyl quinic acid, anisic acid, hyperin and cis, cis-tri-p-coumaroyl spermide, cis, trans-tri-p-coumaroyl spermide, cis, trans-tri-p-coumaroyl spermide and the like are not detected in other single nectar except loquat honey. The invention proves that the 10 components can be used as characteristic plant compound markers of loquat honey, and relates to a detection and identification method of loquat honey, which adopts a high performance liquid chromatography combined with a fingerprint method and a high performance liquid chromatography quantitative analysis method to detect and identify the loquat honey, wherein the high performance liquid chromatography combined with the fingerprint method can identify that the loquat honey contains anisoic acid, chlorogenic acid, isochlorogenic acid, 3-O-p-coumaryl quinic acid, 5-O-feruloyl quinic acid, hyperoside (namely quercetin-3-O-glucoside), cis-tri-p-coumaroyl spermide, cis, trans-tri-p-coumaroyl spermide, cis, trans-tri-p-coumaroyl spermide and trans, trans-tri-p-coumaroyl spermide simultaneously, and then the honey to be detected can be determined to be the loquat honey.
The invention provides a new method for constructing the loquat honey authenticity and quality evaluation system, has high specificity and good detection sensitivity, and has good economic value and social significance.
Drawings
FIG. 1 is a primary mass spectrum of anisic acid, wherein FIG. 1A is a primary mass spectrum of anisic acid in a positive ion mode; FIG. 1B is a first-order mass spectrum of anisic acid in negative ion mode.
FIG. 2 is a secondary mass spectrum of anisic acid;
FIG. 3 shows the mass fragmentation pathway of anisic acid.
FIG. 4 is a chromatogram of loquat honey of example 1 of the present invention.
Fig. 5 is an HPLC overlay of the honey of ten different sources of loquat in example 2 of the present invention. In the figure, R represents a standard fingerprint spectrum generated based on 5 loquat raw materials of different production places, and S1-S15 represent 5 loquat raw materials of different production places.
FIG. 6 is a graph showing the verification of the mixing of compounds with a molecular weight of 152.
FIG. 7 is a diagram showing the verification of loquat honey and anisole standard.
Detailed Description
Preferred embodiments of the present invention will be described in detail below with reference to examples. It is to be understood that the following examples are given for illustrative purposes only and are not intended to limit the scope of the present invention. Various modifications and alterations of this invention may be made by those skilled in the art without departing from the spirit and scope of this invention.
The experimental methods used in the following examples are conventional methods unless otherwise specified. Materials, reagents and the like used in the examples described below are commercially available unless otherwise specified.
Example 1: separation, analysis and content determination method of loquat honey components and establishment of fingerprint
1) The analysis method of the loquat honey comprises the following steps:
s1, preparation of loquat honey solution: accurately weighing 20.0g of loquat honey in a beaker, adding 80mL of deionized water, stirring with a glass rod until the honey is fully dissolved, adding 5v/v% ammonia water solution to adjust the pH value to about 6.8, centrifuging at 9000rpm for 20min, and taking supernatant for later use;
s2, extracting and enriching the loquat honey plant compound: extracting the supernatant obtained in the step S1 by using a Strata-X-A solid phase extraction column, treating the Strata-X-A solid phase extraction column by using 3mL of methanol as an activating agent and 3mL of deionized water as a balancing agent, sampling the supernatant, eluting with 3mL of deionized water, eluting with 3mL of methanol solution containing 10v/v% formic acid, extracting active ingredients in the supernatant to obtain an eluent, drying the obtained eluent, re-dissolving with 2.0mL of methanol, and filtering with a 0.22 mu m filter membrane to obtain an extract for later use;
s3, separating and detecting the loquat honey plant compound: and (3) separating and detecting the extracting solution obtained in the step S2 by using high performance liquid chromatography, wherein the detection conditions comprise:
chromatographic column: phenomenex Gemini C 18 A chromatographic column;
mobile phase: the mobile phase A is acetic acid aqueous solution with the concentration of 0.2v/v%, and the mobile phase B is acetic acid methanol solution with the concentration of 0.2 v/v%;
the gradient elution procedure was: 0-11 min, the mobile phase B is increased from 5v/v% to 11v/v%; 11-14 min, the mobile phase B is increased from 11v/v% to 14v/v%; the flow phase B rises from 14v/v% to 15v/v% for 14-17 min; 17-24 min, the mobile phase B is increased from 15v/v% to 16v/v%; the mobile phase B rises from 16% to 17% after 24-28 min; 28-30 min, the mobile phase B rises from 17v/v% to 22v/v%; 30-38 min, the mobile phase B is increased from 22v/v% to 25v/v%; the flow phase B rises from 25v/v% to 30v/v% for 38-41 min; 41-46 min, the mobile phase B is increased from 30v/v% to 33v/v%; 46-55 min, the mobile phase B is kept unchanged at 33%; 55-60 min, the mobile phase B is increased from 33v/v% to 34v/v%; 60-70 min, the mobile phase B is increased from 34v/v% to 36v/v%; 70-80 min, and the mobile phase B rises from 36% to 40%; 80-90 min, the mobile phase B is increased from 40v/v% to 45v/v%; 90-100 min, the mobile phase B is increased from 45v/v% to 52v/v%; 100-110 min, the mobile phase B is increased from 52v/v% to 57v/v%; 110-120 min, the mobile phase B rises from 57% to 65%; 120-130 min, the mobile phase B rises from 65v/v% to 70v/v%; 130-135 min, the mobile phase B is increased from 70v/v% to 80v/v%; mobile phase a changes with the volume change of mobile phase B, and the sum of the volumes of the mobile phase a and the mobile phase B is 100%;
flow rate: the flow rate is 0.7mL/min, the sample injection amount is 20 mu L, the column temperature is 35 ℃, and the detection wavelength is 280nm;
2) The content determination method comprises the following steps:
(1) Chromatographic conditions: combining with the high performance liquid chromatography and fingerprint method;
(2) Preparing a loquat honey standard substance solution: weighing a proper amount of chlorogenic acid, isochlorogenic acid, 3-O-p-coumaroyl quinic acid, 5-O-feruloyl quinic acid, anisic acid, hyperin and tri-p-coumaroyl spermidine to prepare standard substance solutions, wherein the concentrations of the standard substance solutions of chlorogenic acid, isochlorogenic acid, 3-O-p-coumaroyl quinic acid, 5-O-feruloyl quinic acid, anisic acid, hyperin and tri-p-coumaroyl spermidine are respectively 0.22mg/ml,0.8mg/ml,0.64mg/ml,0.32mg/ml,0.6mg/ml,0.56mg/ml and 3.6mg/ml, and drawing a loquat honey standard curve as shown in the following table 1: correcting according to the sample loading amount x and the peak area y and the gradient sample loading amount of 5-10 uL by adopting a 5-point method; preferably, the method comprises the steps of,
TABLE 1 information about standards
(3) Content determination method: injecting the sample solution according to the method in the step (1), and injecting the sample solution according to the corresponding t R Obtaining the content of the compound in honey by substituting the area of chromatographic peaks into a corresponding standard curve, wherein the content of chlorogenic acid is not lower than 0.12-1.22 mg/kg, the content of iso-chlorogenic acid is not lower than 0.11-0.94mg/kg, the content of 3-O-p-coumaroyl quinic acid is 0.28-1.72 mg/kg, the content of 5-O-feruloylquinic acid is not remarkably lower than 0.14-0.58 mg/kg, the content of anisic acid is not lower than 3.30-9.64mg/kg, the content of hyperin is not lower than 0.14-2.15 mg/kg, the content of cis, cis-tri-p-coumaroyl spermidine is not lower than 0.50-3.07 mg/kg, the content of cis, trans-tri-p-coumaroyl spermidine is not lower than 0.68-2.45 mg/kg, the content of trans-tri-p-coumaroyl spermidine is not lower than 0.74-2.85 mg/kg, and the content of trans-tri-p-coumaroyl spermidine is not remarkably lower than 0.37 mg/kg;
3) The structure identification method comprises the following steps: detection, component separation were performed according to the above conditions, and a chromatogram was recorded, see fig. 4. The eluate at each peak time was collected and identified. The method for detecting the tandem diode array detector (HPLC-PDA) by the liquid chromatography and detecting the tandem quadrupole/time-of-flight mass spectrum (HPLC-QTOF) by the liquid chromatography is specifically adopted to carry out category identification on the plant compounds obtained by effective separation and enrichment.
The specific identification method is described in detail as follows:
(1) Carrying out structural identification on plant compound components in the loquat honey after detection under the liquid chromatography conditions:
liquid phase mass spectrometry conditions: the ion source is ESI source (electrospray), the ion source spray voltage is 4kV, the outlet voltage is 130V, the heating temperature is 350 ℃, and nitrogen (N) 2 ) The flow rate is 11L/min, the collision gas is helium, the flow rate of the atomization gas is 80kPa, the pressure of the atomizer is 40psi, and the mass scanning range m/z=100-900 Da.
(2) The structural identification analysis process and the structural identification analysis result of the plant compounds in the loquat honey:
a. the structure analysis process of the compound in the eluent with the retention time of 22.21min is as follows:
the compound has m/z 355.2485[ M+H ] in the positive ion state] + Ion peak of (C), M/z377.2443[ M+Na ]] + In the negative ion state, m/z 353.0824[ M-H ]]Ion peak, m/z 707.1622[2M-H] - The molecular weight of the compound is determined to be 354 based on mass spectrum information in the known positive and negative ion modes. The compound has a maximum ultraviolet absorption peak at a wavelength of 326 nm. And combining mass spectrum data, retention time of chlorogenic acid standard substance spectrum and maximum ultraviolet absorption wavelength, and simultaneously verifying according to chlorogenic acid standard substance liquid chromatography to finally determine that the compound is chlorogenic acid.
b. The structure analysis process of the compound in the eluent with the retention time of 30.17min is as follows:
the compound has m/z 377.2441[ M+Na ] in positive ion state] + In the negative ion state, m/z 353.0833[ M-H ]] - Therefore, the molecular weight of the compound was determined to be 354, which is isochlorogenic acid. The compound has a maximum ultraviolet absorption peak at a wavelength of 324 nm.
c. The structure analysis process of the compound in the eluent with the retention time of 34.82min is as follows:
the compound has ion peak m/z 339.2427[ M+H ] under positive ion state] + Having an ion peak m/z 337.0723[ M-H ] in the negative ion state] - Thus initiallyThe molecular weight of this compound was determined to be 338, which had a maximum ultraviolet absorbance peak at 311 wavelengths. And combining mass spectrum data, retention time of a 3-O-p-coumaroyl quinic acid standard substance spectrum and maximum ultraviolet absorption wavelength, and simultaneously verifying according to a 3-O-p-coumaroyl quinic acid standard substance liquid chromatography to finally determine that the compound is 3-O-p-coumaroyl quinic acid.
d. The structure analysis process of the compound in the eluent with the retention time of 40.53min is as follows:
the compound has m/z 391.2678[ M+Na ] in the positive ion state] + Ion peaks of (C), M/z369.2728[ M+H ]] + Has an ion peak of m/z 367.0772[ M-H ] in the negative ion state] - And therefore the molecular weight of the compound was determined to be 368, the compound having a maximum ultraviolet absorption peak at a wavelength of 324 nm. And combining mass spectrum data, retention time of a 5-O-feruloyl quinic acid standard substance spectrum and maximum ultraviolet absorption wavelength, and simultaneously verifying according to a liquid chromatography of the 5-O-feruloyl quinic acid standard substance to finally determine that the compound is 5-O-feruloyl quinic acid.
e. The structural analysis process of the compound in the eluent with retention time of 51.94min is as follows, the specific primary mass spectrum is shown in figure 1, and the secondary mass spectrum is shown in figure 2:
the compound has a molecular weight of 153.0521M/z [ M+H ] in positive ion mode] + And 175.0341M/z [ M+Na] + Molecular ion peak of 151.0410M/z [ M-H ] in negative ion mode] - Molecular ion peak, judge the molecular weight of this compound to be 152 according to the first-order mass spectrum information, this compound has the maximum ultraviolet absorption peak at 256nm wavelength. Combining the secondary mass spectrum result and the maximum ultraviolet absorption wavelength of the compound to determine the molecular formula as C 8 H 8 O 3 The structure of the compound is shown as a formula (I):
its chemical name is 4-Methoxybenzoic acid, and the compound is finally determined to be anisic acid.
The secondary mass spectrometry analysis process is as follows: H2O is removed from m/z 153 to m/z 135; the compound contains a carboxyl group as indicated by a difference of 44 from m/z 153 to m/z 109; the fragmentation fraction with m/z 77 indicates that the compound contains a benzene ring. The structure of the compound was subjected to a speculative analysis and the mass fragmentation path of the compound is shown in figure 3. By the principle of secondary mass spectrometry fragmentation we have speculated on the structure of this compound by 5 compounds of the speculated structure: the standard substances of the compound, namely the anisic acid, are verified by an internal standard method, namely the standard substances of the compound, namely the anisic acid, are shown in figure 7, the maximum ultraviolet absorption wavelength and the peak time of the compound are consistent with those of the anisic acid, and the compound is determined to be the anisic acid (p-methoxybenzoic acid).
f. The structure analysis process of the compound in the eluent with the retention time of 59.59min is as follows: the compound has m/z 487.3102[ M+Na ] in positive ion state] + Ion peaks of (C), M/z465.3148[ M+H ]] + Ion peaks of (2); in the negative ion state, m/z 463.0645[ M-H ]] - And therefore the molecular weight of the compound was determined to be 464. The compound has a maximum ultraviolet absorption peak at 354nm wavelength. And combining mass spectrum data, the retention time of the hyperin standard substance spectrum and the maximum ultraviolet absorption wavelength, and simultaneously verifying according to the hyperin standard substance liquid chromatography to finally determine the hyperin compound.
g. The structure analysis process of the compound in the eluent with the retention time of 87.06min is as follows: the compound has m/z 606.5580[ M+Na ] in positive ion state] + Ion peaks of (C), M/z584.5628[ M+H ]] + In the negative ion state, M/z582.2670[ M-H ]]Ion peak, thus initially determining the molecular weight of the compound as 584. The compound has a maximum ultraviolet absorption peak at 269nm wavelength. And combining mass spectrum data, retention time and maximum ultraviolet absorption wavelength of a cis, cis-tri-p-coumaroyl spermidine standard substance map, and finally determining that the compound is cis, cis-tri-p-coumaroyl spermidine according to the liquid chromatography verification of the cis, cis-tri-p-coumaroyl spermidine standard substance.
h. The structure analysis process of the compound in the eluent with the retention time of 96.22min is as follows: the compound has m/z in the positive ion state606.5571[M+Na] + Ion peaks of (C), M/z584.5618[ M+H ]] + In the negative ion state, M/z582.2494[ M-H ]] - Thus, the molecular weight of the compound was initially determined to be 584. The maximum wavelength of ultraviolet absorption of the compound is 279. And combining mass spectrum data, retention time and maximum ultraviolet absorption wavelength of a standard substance spectrum of cis, trans-tri-p-coumaroyl spermidine, and finally determining that the compound is cis, trans-tri-p-coumaroyl spermidine according to liquid chromatography verification of the standard substance of cis, trans-tri-p-coumaroyl spermidine.
i. The structure analysis process of the compound in the eluent with the retention time of 102.07min is as follows:
the compound has m/z 606.5575[ M+Na ] in positive ion state] + Ion peaks of (C), M/z584.5618[ M+H ]] + In the negative ion state, m/z 582.2356[ M-H ]] - Thus, the molecular weight of the compound was initially determined to be 584. The maximum wavelength of ultraviolet absorption of the compound is 290. And combining mass spectrum data, namely, the retention time and the maximum ultraviolet absorption wavelength of a spectrum of the standard substance of the trans, cis, trans-tri-p-coumaroyl spermidine, and simultaneously verifying according to the liquid chromatography of the standard substance of the trans, cis, trans-tri-p-coumaroyl spermidine to finally determine that the compound is the trans, cis, trans-tri-p-coumaroyl spermidine.
j. The structure analysis process of the compound in the eluent with the retention time of 107.36min is as follows:
the compound has m/z 606.5570[ M+Na ] in positive ion state] + Ion peaks of (C), M/z584.5619[ M+H ]] + In the negative ion state, there is m/z 582.2307[ M-H ]] - Thus, the molecular weight of the compound was initially determined to be 584. And combining the retention time and the maximum ultraviolet absorption wavelength of the spectrum of the standard substance of the anti-tri-p-coumaroyl spermidine with mass spectrum data, and simultaneously verifying according to the liquid chromatography of the standard substance of the anti-tri-p-coumaroyl spermidine to finally determine that the compound is the anti-tri-p-coumaroyl spermidine.
Example 2
The loquat honey of ten bees was detected by the high performance liquid chromatography of example 1, and the components of the loquat honey detected were quantitatively analyzed by an external standard method.
Specific honey source information is as follows table 1:
table 1 loquat raw materials honey
The obtained chromatographic superposition patterns of the loquat honey of five different producing areas are shown in figure 5.
The content of chlorogenic acid in fifteen honey samples is not obviously lower than 0.12-1.22 mg/kg, and the average content is 0.53+/-0.09 mg/kg; the content of the iso-chlorogenic acid is not remarkably lower than 0.11-0.94mg/kg, and the average content is 0.42+/-0.07 mg/kg; 3-O-p-coumaroyl quinic acid average content 0.87mg + -0.09/kg; the average content of 5-O-feruloyl quinic acid is 0.32+/-0.04 mg/kg; the average content of the anisic acid is 5.20+/-0.43 mg/kg; the average content of hyperin is 0.85+/-0.15 mg/kg, and the average content of cis, cis-tri-p-coumaroyl spermidine is 1.94+/-0.21 mg/kg; the average content of cis, trans-tri-p-coumaroyl spermidine is 1.22+/-0.14 mg/kg; the average content of trans, cis, trans-tri-p-coumaroyl spermidine is 1.75+/-0.22 mg/kg; the average content of trans, trans-tri-p-coumaroyl spermidine is 1.04+/-0.21 mg/kg.
Example 3
The high performance liquid chromatography of example 1 was used to perform component analysis (specific raw material honey source information is shown in table 3) on locust honey, tilia amurensis honey, wattle honey, rape honey, safflower honey, gallnut honey, buckwheat honey, leonurus honey and medlar honey by high performance liquid chromatography, and the above honey was compared with the HPLC profile analysis of loquat honey of example 1. The results show that the compounds chlorogenic acid, isochlorogenic acid, 3-O-p-coumaroyl quinic acid, 5-O-feruloyl quinic acid, anisoic acid, hyperin, cis-tri-p-coumaroyl spermidine, cis, trans-tri-p-coumaroyl spermidine, cis, trans, cis-tri-p-coumaroyl spermidine, trans-tri-p-coumaroyl spermidine are not detected simultaneously in other single nectar except loquat honey. The specific detection results are shown in Table 4.
TABLE 3 other raw Honey Source information Table
/>
/>
Example 4
Twelve commercial (different brands from different sales platforms) loquat honey samples were tested using the hplc method of example 1. The quality of the loquat honey of seven brands was better by HPLC chromatogram comparison analysis, the compounds were detected, the specific average contents were respectively 0.69mg/kg, 0.33mg/kg, 0.67mg/kg, 0.35mg/kg, 5.54mg/kg, 0.43mg/kg, 9.87mg/kg, 0.88mg/kg, 0.81mg/kg, 0.60mg/kg, and the other five brands of loquat honey were not detected at the same time, and the products were judged to be not loquat honey.
In summary, by comparing the high performance liquid chromatography of the loquat honey and the rape honey, the locust honey, the wattle honey and the basswood honey and analyzing and comparing the plant compound components of other collected small-people honey such as the motherwort honey, the Chinese gall honey, the safflower honey, the buckwheat honey, the medlar honey and the like, the compound chlorogenic acid, the isochlorogenic acid, the 3-O-p-coumaroyl quinic acid, the 5-O-feruloyl quinic acid, the anisic acid, the hyperin and cis, cis-tri-p-coumaroyl spermide, cis, trans-tri-p-coumaroyl spermide and trans, trans-tri-p-coumaroyl spermide are not detected in other single nectar except the loquat honey. Thus, chlorogenic acid, isochlorogenic acid, 3-O-p-coumaroyl quinic acid, 5-O-feruloyl quinic acid, anisoic acid, hyperin and cis, cis-tri-p-coumaroyl spermidine, cis, trans-tri-p-coumaroyl spermidine, cis, trans, cis-tri-p-coumaroyl spermidine, trans-tri-p-coumaroyl spermidine can be used as characteristic plant compound markers of loquat honey, and are applied to construction of an evaluation system of authenticity and quality of loquat honey.
While the invention has been described in detail in the foregoing general description and with reference to specific embodiments thereof, it will be apparent to one skilled in the art that modifications and improvements can be made thereto. Accordingly, such modifications or improvements may be made without departing from the spirit of the invention and are intended to be within the scope of the invention as claimed.
Claims (10)
1. A detection and identification method for loquat honey is characterized in that the method adopts a high performance liquid chromatography combined with fingerprint spectrum method and a high performance liquid chromatography quantitative analysis method to detect and identify the loquat honey,
wherein, the high performance liquid chromatography combines with fingerprint spectrum to identify that the loquat honey contains anisoic acid, which has the structure shown in formula (I):
further, the retention time of the anisic acid was 51.94in.
2. The detection and identification method of loquat honey according to claim 1, wherein in the high performance liquid chromatography, a mobile phase A is an acetic acid aqueous solution with the concentration of 0.18-0.22 v/v%, and a mobile phase B is an acetic acid methanol solution with the concentration of 0.18-0.22 v/v%; still more preferably, in the high performance liquid chromatography, the mobile phase A is an aqueous acetic acid solution having a concentration of 0.2v/v%, and the mobile phase B is a methanol acetic acid solution having a concentration of 0.2 v/v%.
3. The method for detecting and identifying loquat honey according to claim 1, wherein the gradient elution procedure in the high performance liquid chromatography is as follows: 0-11 min, the mobile phase B is increased from 5v/v% to 11v/v%; 11-14 min, the mobile phase B is increased from 11v/v% to 14v/v%; the flow phase B rises from 14v/v% to 15v/v% for 14-17 min; 17-24 min, the mobile phase B is increased from 15v/v% to 16v/v%; the mobile phase B rises from 16% to 17% after 24-28 min; 28-30 min, the mobile phase B rises from 17v/v% to 22v/v%; 30-38 min, the mobile phase B is increased from 22v/v% to 25v/v%; the flow phase B rises from 25v/v% to 30v/v% for 38-41 min; 41-46 min, the mobile phase B is increased from 30v/v% to 33v/v%; 46-55 min, the mobile phase B is kept unchanged at 33%; 55-60 min, the mobile phase B is increased from 33v/v% to 34v/v%; 60-70 min, the mobile phase B is increased from 34v/v% to 36v/v%; 70-80 min, and the mobile phase B rises from 36% to 40%;
80-90 min, the mobile phase B is increased from 40v/v% to 45v/v%; 90-100 min, the mobile phase B is increased from 45v/v% to 52v/v%; 100-110 min, the mobile phase B is increased from 52v/v% to 57v/v%; 110-120 min, the mobile phase B rises from 57% to 65%; 120-130 min, the mobile phase B rises from 65v/v% to 70v/v%; 130-135 min, the mobile phase B is increased from 70v/v% to 80v/v%; mobile phase a changes with the volume change of mobile phase B, and the sum of the volumes of the mobile phase a and the mobile phase B is 100%; mobile phase a varied with the volume of mobile phase B, and the sum of the volumes was 100%.
4. The method for detecting and identifying loquat honey according to claim 1, 3 or 4, wherein the high performance liquid chromatography comprises the following chromatographic conditions:
the chromatographic column is Phenomenex Gemini C 18 Chromatographic column with flow rate of 0.6-0.8 mL/min, sample feeding amount of 10-30 muL and column temperature of 34 deg.cThe detection wavelength is 280nm at 36 ℃; further preferably, the sample injection amount is 20. Mu.L, the flow rate is 0.7mL/min, and the column temperature is 35 ℃.
5. The method for detecting and identifying loquat honey according to claim 1, further comprising a pretreatment step of:
s1: mixing honey to be detected with a proper amount of water, fully dissolving the honey to obtain aqueous honey solution, regulating the pH value of the aqueous honey solution to be 6.5-7, centrifuging the aqueous honey solution at a rotating speed of 8800-9200rpm for 18-22 min, and taking supernatant for later use; preferably, the pH value is regulated to 6.8, the centrifugal rotating speed is 9000rpm, and the centrifugal time is 20min;
s2: extracting the supernatant obtained in S1 by using a Strata-X-A solid phase extraction column, activating the solid phase extraction column by using methanol, balancing the extraction column by using ultrapure water, loading the supernatant, eluting by using deionized water, eluting by using a methanol solution containing 9-11 v/v% formic acid, extracting active ingredients in the supernatant to obtain an eluent, drying the obtained eluent, re-dissolving by using a proper amount of methanol, and filtering to obtain the product, wherein the dosage of the methanol, the ultrapure water, the deionized water and the methanol solution containing 9-11 v/v% formic acid is 2-4 times of the column volume;
further, in S2, the pore diameter of the filtering membrane is 0.22 μm;
further, in S2, the eluent contains 10v/v% formic acid in methanol;
in S2, the amount of methanol, ultrapure water, deionized water and a methanol solution containing 9-11 v/v% formic acid is 3 times the column volume.
6. The method according to any one of claims 1 to 5, wherein the high performance liquid chromatography is combined with fingerprint chromatography to identify chlorogenic acid, isochlorogenic acid, 3-O-p-coumaroyl quinic acid, 5-O-feruloyl quinic acid, hyperin (i.e. quercetin-3-O-glucoside), cis-tri-p-coumaroyl spermidine, cis, trans-tri-p-coumaroyl spermidine, cis, trans, cis-tri-p-coumaroyl spermidine, and trans, trans-tri-p-coumaroyl spermidine.
7. The method for detecting and identifying loquat honey according to claim 6, wherein the retention time t of chlorogenic acid R Retention time t of isochlorogenic acid of 22.21min R Retention time t of 3-O-p-coumaroyl quinic acid of 30.17min R Retention time t of 5-O-feruloylquinic acid of 34.82min R A retention time t of hyperin (i.e. quercetin-3-O-glucoside) of 40.53min R 59.59min, cis-tri-p-coumaroyl spermidine retention time t R A retention time t of 87.06min for cis, trans-tri-p-coumaroyl spermidine R Retention time t of 96.22min, cis, trans, cis-tri-p-coumaroyl spermidine R Retention time t of 102.07min, trans-tri-p-coumaroyl spermidine R 107.36min.
8. The detection and identification method of loquat honey according to claim 1, wherein the quantitative analysis method of high performance liquid chromatography comprises the following steps:
(1) Chromatographic conditions: combining with the high performance liquid chromatography and fingerprint method;
(2) Preparing a loquat honey standard substance solution: weighing a proper amount of chlorogenic acid, isochlorogenic acid, 3-O-p-coumaroyl quinic acid, 5-O-feruloyl quinic acid, anisic acid, hyperin and tri-p-coumaroyl spermidine to prepare standard substance solutions, wherein the concentrations of the standard substance solutions are respectively 0.2-0.3 mg/ml, 0.7-0.9 mg/ml, 0.6-0.7 mg/ml, 0.3-0.4 mg/ml, 0.5-0.7 mg/ml, 0.5-0.6 mg/ml and 3-4 mg/ml, and drawing a loquat honey standard curve as shown in the following table 1: correcting according to the sample loading amount x and the peak area y and the gradient sample loading amount of 5-10 uL by adopting a 5-point method; preferably, the concentration of chlorogenic acid, isochlorogenic acid, 3-O-p-coumaroyl quinic acid, 5-O-feruloyl quinic acid, anisic acid, hyperin and tri-p-coumaroyl spermidine standard solution is 0.22mg/ml,0.8mg/ml,0.64mg/ml,0.32mg/ml,0.6mg/ml,0.56mg/ml and 3.6mg/ml respectively;
TABLE 1 information about standards
(3) Content determination method: injecting the sample solution according to the method in the step (1), and injecting the sample solution according to the corresponding t R Obtaining the chromatographic peak area and substituting the chromatographic peak area into a corresponding standard curve to obtain the content of the compound in honey, wherein the content of chlorogenic acid is not lower than 0.12-1.22 mg/kg, the content of iso-chlorogenic acid is not lower than 0.11-0.94mg/kg, the content of 3-O-p-coumaroyl quinic acid is 0.28-1.72 mg/kg, the content of 5-O-feruloylquinic acid is not significantly lower than 0.14-0.58 mg/kg, the content of anisic acid is not lower than 3.30-9.64mg/kg, the content of hyperin is not lower than 0.14-2.15 mg/kg, the content of cis, cis-tri-p-coumaroyl spermidine is not lower than 0.50-3.07 mg/kg, the content of cis, trans-tri-p-coumaroyl spermidine is not lower than 0.68-2.45 mg/kg, the content of trans, trans-tri-p-coumaroyl spermidine is not significantly lower than 0.74-2.85 mg/kg, and trans-coumaroyl spermidine is not lower than 0.37 mg/kg.
9. Use of anisonic acid in identification or detection of loquat honey, characterized in that identification or detection is performed by the method of loquat honey according to claim 1.
10. The use according to claim 9, wherein the assay identification method also simultaneously identifies chlorogenic acid, isochlorogenic acid, 3-O-p-coumaroyl quinic acid, 5-O-feruloyl quinic acid, anisoic acid, hyperin, cis-tri-p-coumaroyl spermidine, cis, trans-tri-p-coumaroyl spermidine, trans-tri-p-coumaroyl spermidine.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310916981.3A CN116908352A (en) | 2023-07-25 | 2023-07-25 | Detection and identification method for loquat honey |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310916981.3A CN116908352A (en) | 2023-07-25 | 2023-07-25 | Detection and identification method for loquat honey |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN116908352A true CN116908352A (en) | 2023-10-20 |
Family
ID=88352910
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310916981.3A Pending CN116908352A (en) | 2023-07-25 | 2023-07-25 | Detection and identification method for loquat honey |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116908352A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114088827A (en) * | 2021-10-29 | 2022-02-25 | 中国农业科学院蜜蜂研究所 | Application of shikimic acid and quinic acid as characteristic markers of linden honey |
-
2023
- 2023-07-25 CN CN202310916981.3A patent/CN116908352A/en active Pending
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114088827A (en) * | 2021-10-29 | 2022-02-25 | 中国农业科学院蜜蜂研究所 | Application of shikimic acid and quinic acid as characteristic markers of linden honey |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN116908352A (en) | Detection and identification method for loquat honey | |
| CN109125258B (en) | Preparation method of strong loquat syrup | |
| CN111249321B (en) | Method for simultaneously preparing black currant polyphenol and polysaccharide | |
| CN111398499B (en) | Application of 3-amino-2-naphthoic acid in identifying apis cerana honey and apis mellifera honey | |
| Martin et al. | Natural isotope fractionation in the discrimination of sugar origins | |
| CN114181176B (en) | Shishan kunmiki A and preparation method thereof, and identification method of bauhinia championii honey | |
| CN113736662A (en) | Yeast selenoprotein and preparation method and application thereof | |
| Takatsuki et al. | Liquid chromatographic determination of free and added niacin and niacinamide in beef and pork | |
| CN111398498B (en) | Application of indole-3-methyl acetate in identifying apis cerana honey and apis mellifera honey | |
| CN105717227A (en) | Judgment method for flavor quality of concentrated apple juice and application thereof | |
| CN107449837B (en) | Method for identifying honey and syrup by using non-sugar characteristic substances | |
| CN111380965A (en) | Method for detecting metabolite in microbial fermentation liquid | |
| CN104977383A (en) | Method for rapidly and quantitatively detecting microcystins in spirulina food | |
| CN114324650A (en) | Method for identifying bauhinia championii honey based on (Z) -EDTO characteristic marker | |
| CN111398497B (en) | Application of kynurenine in identifying Chinese bee honey and apis mellifera honey | |
| CN116930378A (en) | Detection and identification method for wild osmanthus honey | |
| CN117233292B (en) | Identification method of nine-dragon rattan honey | |
| CN115611840B (en) | Cold winter honey element and preparation method and application thereof | |
| CN106674184A (en) | Extraction method of anthocyanin | |
| CN112858554A (en) | Method for extracting anthocyanin substances from polluted membrane in membrane separation process | |
| CN112147249A (en) | UPC2-PDA-Q-Tof/MS detection method for 31 effective components in waxberry wine | |
| CN115385794B (en) | Method for separating and extracting chlorogenic acid in multi-juice by utilizing tamarind polysaccharide adsorption | |
| CN114910582B (en) | Method for rapidly identifying chemical components of bamboo leaf green wine based on UPLC-Q-TOF-MS technology | |
| CN117263790A (en) | Longan essence, preparation method thereof and identification method of longan honey | |
| Fracassetti et al. | Melatonin and its derivatives in red wine: contribution of fermenting microorganisms |
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 |