CN113295802A - Liquid beverage containing sodium hyaluronate and method for detecting content of sodium hyaluronate in liquid beverage - Google Patents
Liquid beverage containing sodium hyaluronate and method for detecting content of sodium hyaluronate in liquid beverage Download PDFInfo
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- CN113295802A CN113295802A CN202110783571.7A CN202110783571A CN113295802A CN 113295802 A CN113295802 A CN 113295802A CN 202110783571 A CN202110783571 A CN 202110783571A CN 113295802 A CN113295802 A CN 113295802A
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- sodium hyaluronate
- solution
- vitamin
- mobile phase
- liquid beverage
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- 229920002385 Sodium hyaluronate Polymers 0.000 title claims abstract description 131
- 229940010747 sodium hyaluronate Drugs 0.000 title claims abstract description 131
- YWIVKILSMZOHHF-QJZPQSOGSA-N sodium;(2s,3s,4s,5r,6r)-6-[(2s,3r,4r,5s,6r)-3-acetamido-2-[(2s,3s,4r,5r,6r)-6-[(2r,3r,4r,5s,6r)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2- Chemical compound [Na+].CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 YWIVKILSMZOHHF-QJZPQSOGSA-N 0.000 title claims abstract description 131
- 239000007788 liquid Substances 0.000 title claims abstract description 65
- 235000013361 beverage Nutrition 0.000 title claims abstract description 49
- 238000000034 method Methods 0.000 title claims abstract description 45
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims abstract description 54
- DLRVVLDZNNYCBX-UHFFFAOYSA-N Polydextrose Polymers OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(O)O1 DLRVVLDZNNYCBX-UHFFFAOYSA-N 0.000 claims abstract description 34
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 claims abstract description 32
- GVJHHUAWPYXKBD-UHFFFAOYSA-N (±)-α-Tocopherol Chemical compound OC1=C(C)C(C)=C2OC(CCCC(C)CCCC(C)CCCC(C)C)(C)CCC2=C1C GVJHHUAWPYXKBD-UHFFFAOYSA-N 0.000 claims abstract description 28
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 21
- 235000021552 granulated sugar Nutrition 0.000 claims abstract description 19
- 239000008213 purified water Substances 0.000 claims abstract description 19
- 239000001509 sodium citrate Substances 0.000 claims abstract description 19
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 claims abstract description 19
- CHHHXKFHOYLYRE-UHFFFAOYSA-M 2,4-Hexadienoic acid, potassium salt (1:1), (2E,4E)- Chemical compound [K+].CC=CC=CC([O-])=O CHHHXKFHOYLYRE-UHFFFAOYSA-M 0.000 claims abstract description 18
- DFPAKSUCGFBDDF-ZQBYOMGUSA-N [14c]-nicotinamide Chemical compound N[14C](=O)C1=CC=CN=C1 DFPAKSUCGFBDDF-ZQBYOMGUSA-N 0.000 claims abstract description 18
- 229940069338 potassium sorbate Drugs 0.000 claims abstract description 18
- 235000010241 potassium sorbate Nutrition 0.000 claims abstract description 18
- 239000004302 potassium sorbate Substances 0.000 claims abstract description 18
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 claims abstract description 17
- 229920001100 Polydextrose Polymers 0.000 claims abstract description 17
- 235000013856 polydextrose Nutrition 0.000 claims abstract description 17
- 239000001259 polydextrose Substances 0.000 claims abstract description 17
- 229940035035 polydextrose Drugs 0.000 claims abstract description 17
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 claims abstract description 16
- 239000004376 Sucralose Substances 0.000 claims abstract description 16
- 229930003268 Vitamin C Natural products 0.000 claims abstract description 16
- 235000019408 sucralose Nutrition 0.000 claims abstract description 16
- BAQAVOSOZGMPRM-QBMZZYIRSA-N sucralose Chemical compound O[C@@H]1[C@@H](O)[C@@H](Cl)[C@@H](CO)O[C@@H]1O[C@@]1(CCl)[C@@H](O)[C@H](O)[C@@H](CCl)O1 BAQAVOSOZGMPRM-QBMZZYIRSA-N 0.000 claims abstract description 16
- 239000011718 vitamin C Substances 0.000 claims abstract description 16
- 235000019154 vitamin C Nutrition 0.000 claims abstract description 16
- 229930003427 Vitamin E Natural products 0.000 claims abstract description 14
- WIGCFUFOHFEKBI-UHFFFAOYSA-N gamma-tocopherol Natural products CC(C)CCCC(C)CCCC(C)CCCC1CCC2C(C)C(O)C(C)C(C)C2O1 WIGCFUFOHFEKBI-UHFFFAOYSA-N 0.000 claims abstract description 14
- 235000019165 vitamin E Nutrition 0.000 claims abstract description 14
- 239000011709 vitamin E Substances 0.000 claims abstract description 14
- 229940046009 vitamin E Drugs 0.000 claims abstract description 14
- 235000015165 citric acid Nutrition 0.000 claims abstract description 10
- 239000000686 essence Substances 0.000 claims abstract description 10
- 235000011083 sodium citrates Nutrition 0.000 claims abstract description 10
- PJVXUVWGSCCGHT-ZPYZYFCMSA-N (2r,3s,4r,5r)-2,3,4,5,6-pentahydroxyhexanal;(3s,4r,5r)-1,3,4,5,6-pentahydroxyhexan-2-one Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C=O.OC[C@@H](O)[C@@H](O)[C@H](O)C(=O)CO PJVXUVWGSCCGHT-ZPYZYFCMSA-N 0.000 claims abstract description 8
- 239000006188 syrup Substances 0.000 claims abstract description 8
- 235000020357 syrup Nutrition 0.000 claims abstract description 8
- 239000000243 solution Substances 0.000 claims description 84
- 239000013558 reference substance Substances 0.000 claims description 36
- 238000001514 detection method Methods 0.000 claims description 27
- 239000012088 reference solution Substances 0.000 claims description 21
- 239000012085 test solution Substances 0.000 claims description 21
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 claims description 19
- 239000008351 acetate buffer Substances 0.000 claims description 19
- 239000008055 phosphate buffer solution Substances 0.000 claims description 19
- 229910052938 sodium sulfate Inorganic materials 0.000 claims description 19
- 235000011152 sodium sulphate Nutrition 0.000 claims description 19
- 238000002156 mixing Methods 0.000 claims description 18
- 238000002347 injection Methods 0.000 claims description 15
- 239000007924 injection Substances 0.000 claims description 15
- 229940088594 vitamin Drugs 0.000 claims description 15
- 229930003231 vitamin Natural products 0.000 claims description 15
- 235000013343 vitamin Nutrition 0.000 claims description 15
- 239000011782 vitamin Substances 0.000 claims description 15
- 150000003722 vitamin derivatives Chemical class 0.000 claims description 13
- 229930003779 Vitamin B12 Natural products 0.000 claims description 9
- FDJOLVPMNUYSCM-WZHZPDAFSA-L cobalt(3+);[(2r,3s,4r,5s)-5-(5,6-dimethylbenzimidazol-1-yl)-4-hydroxy-2-(hydroxymethyl)oxolan-3-yl] [(2r)-1-[3-[(1r,2r,3r,4z,7s,9z,12s,13s,14z,17s,18s,19r)-2,13,18-tris(2-amino-2-oxoethyl)-7,12,17-tris(3-amino-3-oxopropyl)-3,5,8,8,13,15,18,19-octamethyl-2 Chemical compound [Co+3].N#[C-].N([C@@H]([C@]1(C)[N-]\C([C@H]([C@@]1(CC(N)=O)C)CCC(N)=O)=C(\C)/C1=N/C([C@H]([C@@]1(CC(N)=O)C)CCC(N)=O)=C\C1=N\C([C@H](C1(C)C)CCC(N)=O)=C/1C)[C@@H]2CC(N)=O)=C\1[C@]2(C)CCC(=O)NC[C@@H](C)OP([O-])(=O)O[C@H]1[C@@H](O)[C@@H](N2C3=CC(C)=C(C)C=C3N=C2)O[C@@H]1CO FDJOLVPMNUYSCM-WZHZPDAFSA-L 0.000 claims description 9
- 238000001914 filtration Methods 0.000 claims description 9
- 238000007689 inspection Methods 0.000 claims description 9
- 238000004811 liquid chromatography Methods 0.000 claims description 9
- 239000011259 mixed solution Substances 0.000 claims description 9
- 238000002360 preparation method Methods 0.000 claims description 9
- 238000003756 stirring Methods 0.000 claims description 9
- 235000019163 vitamin B12 Nutrition 0.000 claims description 9
- 239000011715 vitamin B12 Substances 0.000 claims description 9
- 238000005303 weighing Methods 0.000 claims description 9
- 235000021433 fructose syrup Nutrition 0.000 claims description 3
- 229930003270 Vitamin B Natural products 0.000 abstract description 10
- 235000019156 vitamin B Nutrition 0.000 abstract description 10
- 239000011720 vitamin B Substances 0.000 abstract description 10
- 239000012071 phase Substances 0.000 description 66
- KIUKXJAPPMFGSW-DNGZLQJQSA-N (2S,3S,4S,5R,6R)-6-[(2S,3R,4R,5S,6R)-3-Acetamido-2-[(2S,3S,4R,5R,6R)-6-[(2R,3R,4R,5S,6R)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2-carboxylic acid Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 KIUKXJAPPMFGSW-DNGZLQJQSA-N 0.000 description 17
- 229920002674 hyaluronan Polymers 0.000 description 17
- 229960003160 hyaluronic acid Drugs 0.000 description 17
- 210000003491 skin Anatomy 0.000 description 13
- 238000010812 external standard method Methods 0.000 description 8
- 238000007865 diluting Methods 0.000 description 7
- 235000019534 high fructose corn syrup Nutrition 0.000 description 7
- 238000004128 high performance liquid chromatography Methods 0.000 description 7
- 238000010828 elution Methods 0.000 description 6
- 239000000203 mixture Substances 0.000 description 6
- UJOBWOGCFQCDNV-UHFFFAOYSA-N 9H-carbazole Chemical compound C1=CC=C2C3=CC=CC=C3NC2=C1 UJOBWOGCFQCDNV-UHFFFAOYSA-N 0.000 description 4
- 230000032683 aging Effects 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 102000008186 Collagen Human genes 0.000 description 3
- 108010035532 Collagen Proteins 0.000 description 3
- 229920001436 collagen Polymers 0.000 description 3
- 239000002537 cosmetic Substances 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 235000013402 health food Nutrition 0.000 description 3
- 238000010829 isocratic elution Methods 0.000 description 3
- 230000003796 beauty Effects 0.000 description 2
- 150000001720 carbohydrates Chemical class 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 210000002615 epidermis Anatomy 0.000 description 2
- 210000001508 eye Anatomy 0.000 description 2
- 238000000855 fermentation Methods 0.000 description 2
- 230000004151 fermentation Effects 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- WCDDVEOXEIYWFB-VXORFPGASA-N (2s,3s,4r,5r,6r)-3-[(2s,3r,5s,6r)-3-acetamido-5-hydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-4,5,6-trihydroxyoxane-2-carboxylic acid Chemical compound CC(=O)N[C@@H]1C[C@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](C(O)=O)O[C@@H](O)[C@H](O)[C@H]1O WCDDVEOXEIYWFB-VXORFPGASA-N 0.000 description 1
- 206010003210 Arteriosclerosis Diseases 0.000 description 1
- 208000024806 Brain atrophy Diseases 0.000 description 1
- LZZYPRNAOMGNLH-UHFFFAOYSA-M Cetrimonium bromide Chemical compound [Br-].CCCCCCCCCCCCCCCC[N+](C)(C)C LZZYPRNAOMGNLH-UHFFFAOYSA-M 0.000 description 1
- 206010016807 Fluid retention Diseases 0.000 description 1
- 230000005526 G1 to G0 transition Effects 0.000 description 1
- 229920002683 Glycosaminoglycan Polymers 0.000 description 1
- 206010053615 Thermal burn Diseases 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 208000011775 arteriosclerosis disease Diseases 0.000 description 1
- 206010003246 arthritis Diseases 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008033 biological extinction Effects 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 238000004737 colorimetric analysis Methods 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 210000004207 dermis Anatomy 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 230000002500 effect on skin Effects 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- 210000001339 epidermal cell Anatomy 0.000 description 1
- 230000029142 excretion Effects 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 238000005227 gel permeation chromatography Methods 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 230000035876 healing Effects 0.000 description 1
- 238000011141 high resolution liquid chromatography Methods 0.000 description 1
- 229940014041 hyaluronate Drugs 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000001802 infusion Methods 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- 230000031700 light absorption Effects 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 230000001050 lubricating effect Effects 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 238000004452 microanalysis Methods 0.000 description 1
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- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 150000002772 monosaccharides Chemical class 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 239000012466 permeate Substances 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
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- 150000003254 radicals Chemical class 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
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- 230000008439 repair process Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000002000 scavenging effect Effects 0.000 description 1
- 230000037380 skin damage Effects 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 230000000475 sunscreen effect Effects 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 238000000825 ultraviolet detection Methods 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
- 230000037303 wrinkles Effects 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
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/38—Other non-alcoholic beverages
-
- 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
- G01N30/32—Control of physical parameters of the fluid carrier of pressure or speed
-
- 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
- G01N30/50—Conditioning of the sorbent material or stationary liquid
- G01N30/52—Physical parameters
- G01N30/54—Temperature
-
- 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
- G01N30/62—Detectors specially adapted therefor
- G01N30/74—Optical detectors
-
- 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
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- 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
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/32—Control of physical parameters of the fluid carrier of pressure or speed
- G01N2030/324—Control of physical parameters of the fluid carrier of pressure or speed speed, flow rate
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- 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)
- Nutrition Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Non-Alcoholic Beverages (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
The invention discloses a liquid beverage containing sodium hyaluronate and a method for detecting the content of sodium hyaluronate in the liquid beverage, wherein the components of the liquid beverage comprise sodium hyaluronate, white granulated sugar, fructose-glucose syrup, polydextrose, citric acid, sodium citrate, sucralose, vitamin C, vitamin B, vitamin E, edible essence, nicotinamide, potassium sorbate, EDTA and purified water.
Description
Technical Field
The invention relates to the technical field of detection of sodium hyaluronate content, in particular to a liquid beverage containing sodium hyaluronate and a method for detecting the content of sodium hyaluronate in the liquid beverage.
Background
Sodium Hyaluronate (HA) is also called Hyaluronic acid, and is the most important substance for keeping moisture in human tissues, Hyaluronic acid is natural biological molecules inherent to skin, and Hyaluronic acid with small molecular weight can permeate into the epidermal layer of the skin to promote the supply of nutrition and the excretion of waste, so that the skin is prevented from aging, and the effects of beautifying and nourishing the skin are achieved.
Hyaluronic acid can promote the regeneration of the skin at the injured part by promoting the proliferation and differentiation of epidermal cells and scavenging free radicals. The hyaluronic acid can simultaneously reduce the transmission of ultraviolet rays and repair skin damage caused by a small amount of transmitted ultraviolet rays, and has a sunscreen effect. When the skin suffers from mild burns and scalds, the surface of the skin is coated with water containing hyaluronic acid and cosmetics, so that the pain can be relieved, and the healing of the skin at the injured part can be accelerated.
The edible hyaluronic acid beauty health care product increases the content of endogenous hyaluronic acid from dermis to epidermis, activates cells and can play a role in whole body beauty from inside to outside. The dermal matrix is mainly composed of mucopolysaccharides such as collagen and hyaluronic acid as the main components of the skin, and aging of the skin is caused by changes in collagen due to the decrease in hyaluronic acid having a water-retaining effect. Therefore, hyaluronic acid must be supplemented to maintain the function of collagen. The hyaluronic acid health food can be taken orally, can improve the water retention of the skin, improve the skin, enable the skin to be rich in elasticity and reduce wrinkles, plays an important physiological role in lubricating and retaining water in tissues such as articular cavities, blood vessels, eyes, brain and the like, supplements hyaluronic acid in vivo, can reduce the occurrence of arthritis, arteriosclerosis, eye aging and brain atrophy, prevents aging and maintains the health of aged people.
At present, the commonly used detection methods of hyaluronic acid include an HPLC method, a colorimetric method, a CTAB turbidimetric method and a carbazole chromogenic method. For example, patent document 1 discloses a method for quantitatively detecting the content of hyaluronic acid in a hyaluronic acid fermentation broth by carbazole coloration, but it is necessary to remove interfering factors such as residual heterosugars and pigments in the fermentation broth before detection. Patent document 2 discloses a method for measuring the content of hyaluronic acid by an enzymatic method in combination with high performance liquid chromatography. However, the method needs to carry out enzymolysis on the sample before detection, if the enzymolysis is incomplete, the detection effect is influenced, the hydrolysis time is long, and the operation process is complex.
Literature "content of sodium hyaluronate by gel chromatography" on the basis of "Kongyun, Koelreuterin, Guozhei", a method for measuring sodium hyaluronate by liquid chromatography is provided, but this method employs a differential detector, and is only applicable to isocratic elution, but not to gradient elution. The complex composition of the mixture is not favorable for fast elution and separation of components.
The document "research on measuring sodium hyaluronate content by high performance liquid chromatography" provides a liquid phase detection method for an ultraviolet detector, such as chenyuanjuan, chenyianWen, and Qiaoli apple, but an isocratic elution method is also adopted.
Prior art documents patent documents
Patent document 1: CN 108362686A;
patent document 2: CN 109298112A.
Disclosure of Invention
The invention provides a liquid beverage containing sodium hyaluronate and a method for detecting the content of sodium hyaluronate in the liquid beverage, aiming at solving the technical problems in the prior art, aiming at carrying out gradient elution on a gel column for analyzing molecular weight by matching with a mobile phase containing multiple components, rapidly and accurately separating sodium hyaluronate from other components, and having the advantages of high column efficiency, good durability and simple and convenient operation process.
In order to achieve the purpose, the invention provides the following technical scheme: a liquid beverage containing sodium hyaluronate comprises, by weight, 0.02-0.3% of sodium hyaluronate, 0-10% of white granulated sugar, 0-5% of fructose-glucose syrup, 0-5% of polydextrose, 0.01-2.5% of citric acid, 0.01-2.5% of sodium citrate, 0-0.5% of sucralose, 0-0.5% of vitamin C0, 0-0.5% of vitamin B60, 120-0.5% of vitamin B, 0-0.5% of vitamin E0, 0-0.5% of edible essence, 0-0.5% of nicotinamide, 0-0.5% of potassium sorbate, 0-0.5% of EDTA 0-0.5% of purified water and 85-99%.
The method for detecting the content of the sodium hyaluronate in the liquid beverage comprises the following steps:
(1) preparing a mobile phase: preparing a sodium sulfate solution, a phosphate buffer solution and an acetate buffer solution, and filtering and ultrasonically treating the sodium sulfate solution, the phosphate buffer solution and the acetate buffer solution;
(2) preparation of a reference solution: precisely weighing 25mg of sodium hyaluronate reference substance, placing the sodium hyaluronate reference substance in a 25ml volumetric flask, adding the mobile phase obtained in the step (1), shaking to dissolve the sodium hyaluronate reference substance, adding a mobile phase solution to dilute the sodium hyaluronate reference substance to a scale, and shaking uniformly;
(3) precisely measuring 4ml of the sodium hyaluronate solution dissolved in the step (2), placing the sodium hyaluronate solution in a 50ml volumetric flask, adding the mobile phase solution to dilute the solution to a scale, and shaking up;
(4) preparing a test solution: mixing sodium hyaluronate, white granulated sugar, fructose syrup, polydextrose, citric acid, sodium citrate, sucralose, vitamin C, vitamin B12, vitamin E, edible essence, nicotinamide, potassium sorbate and EDTA according to a formula ratio, adding purified water to stir and dissolve after uniformly mixing to obtain a liquid beverage mixed solution, precisely measuring a proper amount, adding a mobile phase to dissolve and dilute to a scale, and shaking uniformly;
(5) setting a liquid chromatograph inspection method, including a mobile phase gradient program, a column temperature, a detection wavelength and a sample injection amount;
(6) and detecting the reference solution and the test solution after the base line is balanced, recording a chromatogram, and calculating the content of the sodium hyaluronate by adopting an external standard method according to the peak area.
Preferably, in step (1), the flow rate of the mobile phase is 0.5 to 1.0 ml/min.
Preferably, in the step (5), the column temperature of the column used in the liquid chromatography is 20 to 60 ℃.
Preferably, in the step (5), the detection wavelength in the liquid chromatography is 195-250 nm.
Preferably, in the step (5), the sample amount in the liquid chromatography is 10 to 20. mu.L.
Preferably, in step (6), the reference solution and the test solution after the base line is balanced are detected by using an ultraviolet detector.
Compared with the prior art, the invention has the beneficial effects that:
the invention relates to a liquid beverage containing sodium hyaluronate and a method for detecting the content of sodium hyaluronate in the liquid beverage, which can quickly and accurately separate sodium hyaluronate from other components by matching a gel column for analyzing molecular weight with a plurality of component mobile phases for gradient elution, and has the advantages of high column efficiency, good durability and simple and convenient operation process. The ultraviolet detector is matched, so that the sensitivity is high, the noise is low, the linear range is wide, the selectivity is good, and the ultraviolet detector is not sensitive to the environmental temperature, the composition change of a mobile phase and the flow rate fluctuation, so that the ultraviolet detector can be used for isocratic elution and gradient elution. The ultraviolet detector is insensitive to flow rate and temperature, and can be used for preparing chromatogram. Due to the high sensitivity, even those with low light absorption and low extinction coefficient can be analyzed in microanalysis with a UV detector. The detection method can be used for determining the content of the sodium hyaluronate in the multi-component mixture of health food, medicines, cosmetics and the like, is simple to operate, does not need pretreatment such as enzymolysis and the like, is not influenced by components of saccharides and vitamins, and is suitable for detecting the content of the sodium hyaluronate in complex components.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Examples
The invention provides a liquid beverage containing sodium hyaluronate and a technical scheme of a method for detecting the content of the sodium hyaluronate in the liquid beverage: a liquid beverage containing sodium hyaluronate comprises, by weight, 0.02-0.3% of sodium hyaluronate, 0-10% of white granulated sugar, 0-5% of fructose-glucose syrup, 0-5% of polydextrose, 0.01-2.5% of citric acid, 0.01-2.5% of sodium citrate, 0-0.5% of sucralose, 0-0.5% of vitamin C0, 0-0.5% of vitamin B60, 120-0.5% of vitamin B, 0-0.5% of vitamin E0, 0-0.5% of edible essence, 0-0.5% of nicotinamide, 0-0.5% of potassium sorbate, 0-0.5% of EDTA 0-0.5% of purified water and 85-99%.
The method for detecting the content of the sodium hyaluronate in the liquid beverage comprises the following steps:
(1) preparing a mobile phase: preparing a sodium sulfate solution, a phosphate buffer solution and an acetate buffer solution, and filtering and ultrasonically treating the sodium sulfate solution, the phosphate buffer solution and the acetate buffer solution;
(2) preparation of a reference solution: precisely weighing 25mg of sodium hyaluronate reference substance, placing the sodium hyaluronate reference substance in a 25ml volumetric flask, adding the mobile phase obtained in the step (1), shaking to dissolve the sodium hyaluronate reference substance, adding a mobile phase solution to dilute the sodium hyaluronate reference substance to a scale, and shaking uniformly;
(3) precisely measuring 4ml of the sodium hyaluronate solution dissolved in the step (2), placing the sodium hyaluronate solution in a 50ml volumetric flask, adding the mobile phase solution to dilute the solution to a scale, and shaking up;
(4) preparing a test solution: mixing sodium hyaluronate, white granulated sugar, high fructose corn syrup, polydextrose, citric acid, sodium citrate, sucralose, vitamin C, vitamin B12, vitamin E, edible essence, nicotinamide, potassium sorbate and EDTA according to a formula ratio, adding purified water after uniformly mixing, and stirring and dissolving to obtain a liquid beverage mixed solution. Precisely measuring a proper amount, adding a mobile phase for dissolving and diluting to a scale, and shaking up;
(5) setting a liquid chromatograph inspection method, including a mobile phase gradient program, a column temperature, a detection wavelength and a sample injection amount;
(6) and detecting the reference solution and the test solution after the base line is balanced, recording a chromatogram, and calculating the content of the sodium hyaluronate by adopting an external standard method according to the peak area.
Wherein, in the step (1), the flow rate of the mobile phase is 0.5-1.0 ml/min.
Wherein, in the step (5), the column temperature of the chromatographic column used in the liquid chromatography is 20-60 ℃.
Wherein, in the step (5), the detection wavelength in the liquid chromatography is 195-250 nm.
Wherein, in the step (5), the sample amount in the liquid chromatogram is 10-20 μ L.
In the step (6), the reference solution and the test solution after the base line is balanced are detected by adopting an ultraviolet detector.
Specifically, the mobile phase: 0.001-0.10mol/L sodium sulfate solution, or 0.001-0.10mol/L phosphate buffer solution, or 0.001-0.10mol/L acetate buffer solution, and passing through the chromatographic column at different time stages and different proportions.
Specifically, the conditions of the high performance liquid chromatography are as follows: a gel column, specifically a chromatographic column, is used: TSK G2000SWXL or TSK G4000 SWXL.
Specifically, in the multi-component mixture, the molecular weight of other components is greatly different from that of sodium hyaluronate, and the components can be vitamins, monosaccharides, small molecular polysaccharides and the like, and each component can be separated according to the molecular weight by using a gel column, and the content of the sodium hyaluronate can be detected by using an ultraviolet detection method after separation.
Specifically, the high performance liquid chromatography is also called "high performance liquid chromatography", "high resolution liquid chromatography", "modern column chromatography", and the like. High performance liquid chromatography is an important branch of chromatography, liquid is used as a mobile phase, a high-pressure infusion system is adopted, mobile phases such as single solvents with different polarities or mixed solvents, buffer solutions and the like with different proportions are pumped into a chromatographic column filled with a stationary phase, and after components in the column are separated, the mobile phases enter a detector for detection, so that analysis of a sample is realized.
The mobile phase gradient program was as follows:
example one
Example two
EXAMPLE III
Example four
Example five
The optimal proportion of the mobile phase can be selected according to different beverage formulas.
Example 1
A liquid beverage containing sodium hyaluronate comprises, by weight, 0.03% of sodium hyaluronate, 2.5% of white granulated sugar, 1% of fructose-glucose syrup, 1.4% of polydextrose, 0.014% of citric acid, 0.015% of sodium citrate, 0.01% of sucralose, 0.005% of vitamin C, vitamin B60.003%, vitamin B120.002%, 0.005% of vitamin E, 0.006% of edible essence, 0.005% of nicotinamide, 0.005% of EDTA and 95% of purified water.
The method for detecting the content of the sodium hyaluronate in the liquid beverage comprises the following steps:
(1) preparing a mobile phase: preparing a sodium sulfate solution, a phosphate buffer solution and an acetate buffer solution, and filtering and ultrasonically treating the sodium sulfate solution, the phosphate buffer solution and the acetate buffer solution, wherein the flow rate of the mobile phase is 0.7 ml/min;
(2) preparation of a reference solution: precisely weighing 25mg of sodium hyaluronate reference substance, placing the sodium hyaluronate reference substance in a 25ml volumetric flask, adding the mobile phase obtained in the step (1), shaking to dissolve the sodium hyaluronate reference substance, adding a mobile phase solution to dilute the sodium hyaluronate reference substance to a scale, and shaking uniformly;
(3) precisely measuring 4ml of the sodium hyaluronate solution dissolved in the step (2), placing the sodium hyaluronate solution in a 50ml volumetric flask, adding the mobile phase solution to dilute the solution to a scale, and shaking up;
(4) preparing a test solution: mixing sodium hyaluronate, white granulated sugar, high fructose corn syrup, polydextrose, citric acid, sodium citrate, sucralose, vitamin C, vitamin B12, vitamin E, edible essence, nicotinamide, potassium sorbate and EDTA according to a formula ratio, adding purified water after uniformly mixing, and stirring and dissolving to obtain a liquid beverage mixed solution. Precisely measuring a proper amount, adding a mobile phase for dissolving and diluting to a scale, and shaking up;
(5) setting a liquid chromatograph inspection method, which comprises a mobile phase gradient program, a column temperature, a detection wavelength and a sample injection amount, wherein the column temperature of a chromatographic column used in the liquid chromatograph is 22 ℃, the detection wavelength is 200nm, and the sample injection amount is 12 mu L;
(6) and detecting the reference solution and the test solution after the base line is balanced, recording a chromatogram, and calculating the content of the sodium hyaluronate by adopting an external standard method according to the peak area.
Example 2
The liquid beverage comprises, by weight, 0.024% of sodium hyaluronate, 2.4% of white granulated sugar, 1% of fructose-glucose syrup, 1.5% of polydextrose, 0.015% of citric acid, 0.015% of sodium citrate, 0.01% of sucralose, 0.01% of vitamin C, 0. 60.005% of vitamin B, 120.005% of vitamin E, 0.006% of edible essence, 0.005% of potassium sorbate and 95% of purified water.
The method for detecting the content of the sodium hyaluronate in the liquid beverage comprises the following steps:
(1) preparing a mobile phase: preparing a sodium sulfate solution, a phosphate buffer solution and an acetate buffer solution, and filtering and ultrasonically treating the sodium sulfate solution, the phosphate buffer solution and the acetate buffer solution, wherein the flow rate of the mobile phase is 0.8 ml/min;
(2) preparation of a reference solution: precisely weighing 25mg of sodium hyaluronate reference substance, placing the sodium hyaluronate reference substance in a 25ml volumetric flask, adding the mobile phase obtained in the step (1), shaking to dissolve the sodium hyaluronate reference substance, adding a mobile phase solution to dilute the sodium hyaluronate reference substance to a scale, and shaking uniformly;
(3) precisely measuring 4ml of the sodium hyaluronate solution dissolved in the step (2), placing the sodium hyaluronate solution in a 50ml volumetric flask, adding the mobile phase solution to dilute the solution to a scale, and shaking up;
(4) preparing a test solution: mixing sodium hyaluronate, white granulated sugar, high fructose corn syrup, polydextrose, citric acid, sodium citrate, sucralose, vitamin C, vitamin B12, vitamin E, edible essence, nicotinamide, potassium sorbate and EDTA according to a formula ratio, adding purified water after uniformly mixing, and stirring and dissolving to obtain a liquid beverage mixed solution. Precisely measuring a proper amount, adding a mobile phase for dissolving and diluting to a scale, and shaking up;
(5) setting a liquid chromatograph inspection method which comprises a mobile phase gradient program, a column temperature, a detection wavelength and a sample injection amount, wherein the column temperature of a chromatographic column used in the liquid chromatograph is 55 ℃, the detection wavelength is 205nm, and the sample injection amount is 18 mu L;
(6) and detecting the reference solution and the test solution after the base line is balanced, recording a chromatogram, and calculating the content of the sodium hyaluronate by adopting an external standard method according to the peak area.
Example 3
A liquid beverage containing sodium hyaluronate comprises, by weight, 0.025% of sodium hyaluronate, 3.5% of white granulated sugar, 1.4% of fructose syrup, 0.017% of citric acid, 0.018% of sodium citrate, 0.01% of sucralose, 0.01% of vitamin C, 60.001% of vitamin B, 120.002% of vitamin B, 0.005% of edible essence, 0.005% of nicotinamide, 0.005% of potassium sorbate, 0.002% of EDTA and 95% of purified water.
The method for detecting the content of the sodium hyaluronate in the liquid beverage comprises the following steps:
(1) preparing a mobile phase: preparing a sodium sulfate solution, a phosphate buffer solution and an acetate buffer solution, and filtering and ultrasonically treating the sodium sulfate solution, the phosphate buffer solution and the acetate buffer solution, wherein the flow rate of the mobile phase is 0.8 ml/min;
(2) preparation of a reference solution: precisely weighing 25mg of sodium hyaluronate reference substance, placing the sodium hyaluronate reference substance in a 25ml volumetric flask, adding the mobile phase obtained in the step (1), shaking to dissolve the sodium hyaluronate reference substance, adding a mobile phase solution to dilute the sodium hyaluronate reference substance to a scale, and shaking uniformly;
(3) precisely measuring 4ml of the sodium hyaluronate solution dissolved in the step (2), placing the sodium hyaluronate solution in a 50ml volumetric flask, adding the mobile phase solution to dilute the solution to a scale, and shaking up;
(4) preparing a test solution: mixing sodium hyaluronate, white granulated sugar, high fructose corn syrup, polydextrose, citric acid, sodium citrate, sucralose, vitamin C, vitamin B12, vitamin E, edible essence, nicotinamide, potassium sorbate and EDTA according to a formula ratio, adding purified water after uniformly mixing, and stirring and dissolving to obtain a liquid beverage mixed solution. Precisely measuring a proper amount, adding a mobile phase for dissolving and diluting to a scale, and shaking up;
(5) setting a liquid chromatograph inspection method which comprises a mobile phase gradient program, a column temperature, a detection wavelength and a sample injection amount, wherein the column temperature of a chromatographic column used in the liquid chromatograph is 45 ℃, the detection wavelength is 203nm, and the sample injection amount is 11 mu L;
(6) and detecting the reference solution and the test solution after the base line is balanced, recording a chromatogram, and calculating the content of the sodium hyaluronate by adopting an external standard method according to the peak area.
Example 4
The liquid beverage comprises, by weight, 0.02% of sodium hyaluronate, 3.9% of white granulated sugar, 1% of polydextrose, 0.04% of sodium citrate, 0.004% of vitamin C, 60.005% of vitamin B, 0.008% of vitamin E, 0.006% of edible essence, 0.006% of nicotinamide, 0.004% of potassium sorbate, 0.007% of EDTA and 95% of purified water.
The method for detecting the content of the sodium hyaluronate in the liquid beverage comprises the following steps:
(1) preparing a mobile phase: preparing a sodium sulfate solution, a phosphate buffer solution and an acetate buffer solution, and filtering and ultrasonically treating the sodium sulfate solution, the phosphate buffer solution and the acetate buffer solution, wherein the flow rate of the mobile phase is 0.9 ml/min;
(2) preparation of a reference solution: precisely weighing 25mg of sodium hyaluronate reference substance, placing the sodium hyaluronate reference substance in a 25ml volumetric flask, adding the mobile phase obtained in the step (1), shaking to dissolve the sodium hyaluronate reference substance, adding a mobile phase solution to dilute the sodium hyaluronate reference substance to a scale, and shaking uniformly;
(3) precisely measuring 4ml of the sodium hyaluronate solution dissolved in the step (2), placing the sodium hyaluronate solution in a 50ml volumetric flask, adding the mobile phase solution to dilute the solution to a scale, and shaking up;
(4) preparing a test solution: mixing sodium hyaluronate, white granulated sugar, high fructose corn syrup, polydextrose, citric acid, sodium citrate, sucralose, vitamin C, vitamin B12, vitamin E, edible essence, nicotinamide, potassium sorbate and EDTA according to a formula ratio, adding purified water after uniformly mixing, and stirring and dissolving to obtain a liquid beverage mixed solution. Precisely measuring a proper amount, adding a mobile phase for dissolving and diluting to a scale, and shaking up;
(5) setting a liquid chromatograph inspection method which comprises a mobile phase gradient program, a column temperature, a detection wavelength and a sample injection amount, wherein the column temperature of a chromatographic column used in the liquid chromatograph is 55 ℃, the detection wavelength is 202nm, and the sample injection amount is 18 mu L;
(6) and detecting the reference solution and the test solution after the base line is balanced, recording a chromatogram, and calculating the content of the sodium hyaluronate by adopting an external standard method according to the peak area.
Example 5
A liquid beverage containing sodium hyaluronate comprises, by weight, 0.026% of sodium hyaluronate, 2.4% of white granulated sugar, 1.5% of fructose-glucose syrup, 1% of polydextrose, 0.016% of citric acid, 0.011% of sodium citrate, 0.009% of vitamin C, 0. 60.008% of vitamin B, 120.005% of vitamin B, 0.008% of edible essence, 0.007% of nicotinamide, 0.001% of potassium sorbate, 0.009% of EDTA and 95% of purified water.
The method for detecting the content of the sodium hyaluronate in the liquid beverage comprises the following steps:
(1) preparing a mobile phase: preparing a sodium sulfate solution, a phosphate buffer solution and an acetate buffer solution, and filtering and ultrasonically treating the sodium sulfate solution, the phosphate buffer solution and the acetate buffer solution, wherein the flow rate of the mobile phase is 0.8 ml/min;
(2) preparation of a reference solution: precisely weighing 25mg of sodium hyaluronate reference substance, placing the sodium hyaluronate reference substance in a 25ml volumetric flask, adding the mobile phase obtained in the step (1), shaking to dissolve the sodium hyaluronate reference substance, adding a mobile phase solution to dilute the sodium hyaluronate reference substance to a scale, and shaking uniformly;
(3) precisely measuring 4ml of the sodium hyaluronate solution dissolved in the step (2), placing the sodium hyaluronate solution in a 50ml volumetric flask, adding the mobile phase solution to dilute the solution to a scale, and shaking up;
(4) preparing a test solution: mixing sodium hyaluronate, white granulated sugar, high fructose corn syrup, polydextrose, citric acid, sodium citrate, sucralose, vitamin C, vitamin B12, vitamin E, edible essence, nicotinamide, potassium sorbate and EDTA according to a formula ratio, adding purified water after uniformly mixing, and stirring and dissolving to obtain a liquid beverage mixed solution. Precisely measuring a proper amount, adding a mobile phase for dissolving and diluting to a scale, and shaking up;
(5) setting a liquid chromatograph inspection method, which comprises a mobile phase gradient program, a column temperature, a detection wavelength and a sample injection amount, wherein the column temperature of a chromatographic column used in the liquid chromatograph is 24 ℃, the detection wavelength is 208nm, and the sample injection amount is 15 mu L;
(6) and detecting the reference solution and the test solution after the base line is balanced, recording a chromatogram, and calculating the content of the sodium hyaluronate by adopting an external standard method according to the peak area.
Example 6
A liquid beverage containing sodium hyaluronate comprises, by weight, 0.03% of sodium hyaluronate, 2.9% of white granulated sugar, 2% of fructose-glucose syrup, 0.016% of citric acid, 0.012% of sodium citrate, 0.005% of vitamin C, 120.007% of vitamin B, 0.006% of vitamin E, 0.009% of edible essence, 0.008% of nicotinamide, 0.005% of potassium sorbate, 0.002% of EDTA and 95% of purified water.
The method for detecting the content of the sodium hyaluronate in the liquid beverage comprises the following steps:
(1) preparing a mobile phase: preparing a sodium sulfate solution, a phosphate buffer solution and an acetate buffer solution, and filtering and ultrasonically treating the sodium sulfate solution, the phosphate buffer solution and the acetate buffer solution, wherein the flow rate of the mobile phase is 0.6 ml/min;
(2) preparation of a reference solution: precisely weighing 25mg of sodium hyaluronate reference substance, placing the sodium hyaluronate reference substance in a 25ml volumetric flask, adding the mobile phase obtained in the step (1), shaking to dissolve the sodium hyaluronate reference substance, adding a mobile phase solution to dilute the sodium hyaluronate reference substance to a scale, and shaking uniformly;
(3) precisely measuring 4ml of the sodium hyaluronate solution dissolved in the step (2), placing the sodium hyaluronate solution in a 50ml volumetric flask, adding the mobile phase solution to dilute the solution to a scale, and shaking up;
(4) preparing a test solution: mixing sodium hyaluronate, white granulated sugar, high fructose corn syrup, polydextrose, citric acid, sodium citrate, sucralose, vitamin C, vitamin B12, vitamin E, edible essence, nicotinamide, potassium sorbate and EDTA according to a formula ratio, adding purified water after uniformly mixing, and stirring and dissolving to obtain a liquid beverage mixed solution. Precisely measuring a proper amount, adding a mobile phase for dissolving and diluting to a scale, and shaking up;
(5) setting a liquid chromatograph inspection method, which comprises a mobile phase gradient program, a column temperature, a detection wavelength and a sample injection amount, wherein the column temperature of a chromatographic column used in the liquid chromatograph is 24 ℃, the detection wavelength is 208nm, and the sample injection amount is 12 mu L;
(6) and detecting the reference solution and the test solution after the base line is balanced, recording a chromatogram, and calculating the content of the sodium hyaluronate by adopting an external standard method according to the peak area.
The gel column for analyzing molecular weight is matched with a multi-component mobile phase for gradient elution, so that the sodium hyaluronate can be quickly and accurately separated from other components, the column efficiency is high, the durability is good, the operation process is simple and convenient, the detection method can be used for determining the content of the sodium hyaluronate in a multi-component mixture of health food, medicines, cosmetics and the like, the operation is simple, pretreatment such as enzymolysis is not needed, the influence of saccharides and vitamin components is avoided, and the method is suitable for detecting the content of the sodium hyaluronate in complex components.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (7)
1. The liquid beverage containing sodium hyaluronate is characterized by comprising, by weight, 0.02-0.3% of sodium hyaluronate, 0-10% of white granulated sugar, 0-5% of fructose-glucose syrup, 0-5% of polydextrose, 0.01-2.5% of citric acid, 0.01-2.5% of sodium citrate, 0-0.5% of sucralose, 0-0.5% of vitamin C0, 0-0.5% of vitamin B60, 0-0.5% of vitamin B120, 0-0.5% of vitamin E0, 0-0.5% of edible essence, 0-0.5% of nicotinamide, 0-0.5% of potassium sorbate, 0-0.5% of EDTA0 and 85-99% of purified water.
2. The liquid beverage and the method for detecting the content of the sodium hyaluronate in the liquid beverage according to claim 1, which is characterized by comprising the following steps:
(1) preparing a mobile phase: preparing a sodium sulfate solution, a phosphate buffer solution and an acetate buffer solution, and filtering and ultrasonically treating the sodium sulfate solution, the phosphate buffer solution and the acetate buffer solution;
(2) preparation of a reference solution: precisely weighing 25mg of sodium hyaluronate reference substance, placing the sodium hyaluronate reference substance in a 25ml volumetric flask, adding the mobile phase obtained in the step (1), shaking to dissolve the sodium hyaluronate reference substance, adding a mobile phase solution to dilute the sodium hyaluronate reference substance to a scale, and shaking uniformly;
(3) precisely measuring 4ml of the sodium hyaluronate solution dissolved in the step (2), placing the sodium hyaluronate solution in a 50ml volumetric flask, adding the mobile phase solution to dilute the solution to a scale, and shaking up;
(4) preparing a test solution: mixing sodium hyaluronate, white granulated sugar, fructose syrup, polydextrose, citric acid, sodium citrate, sucralose, vitamin C, vitamin B12, vitamin E, edible essence, nicotinamide, potassium sorbate and EDTA according to a formula ratio, adding purified water to stir and dissolve after uniformly mixing to obtain a liquid beverage mixed solution, precisely measuring a proper amount, adding a mobile phase to dissolve and dilute to a scale, and shaking uniformly;
(5) setting a liquid chromatograph inspection method, including a mobile phase gradient program, a column temperature, a detection wavelength and a sample injection amount;
(6) and detecting the reference solution and the test solution after the base line is balanced, recording a chromatogram, and calculating the content of the sodium hyaluronate according to the peak area.
3. The method for detecting the content of the sodium hyaluronate in the liquid beverage as claimed in claim 2, wherein: in step (1), the flow rate of the mobile phase is 0.5-1.0 ml/min.
4. The method for detecting the content of the sodium hyaluronate in the liquid beverage as claimed in claim 2, wherein: in the step (5), the column temperature of the column used in the liquid chromatography is 20 to 60 ℃.
5. The method for detecting the content of the sodium hyaluronate in the liquid beverage as claimed in claim 2, wherein: in the step (5), the detection wavelength in the liquid chromatography is 195-250 nm.
6. The method for detecting the content of the sodium hyaluronate in the liquid beverage as claimed in claim 2, wherein: in the step (5), the sample amount in the liquid chromatography is 10-20 μ L.
7. The method for detecting the content of the sodium hyaluronate in the liquid beverage as claimed in claim 2, wherein: in step (6), the reference solution and the test solution after the base line is balanced are detected by using an ultraviolet detector.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114081114A (en) * | 2021-11-12 | 2022-02-25 | 华中农业大学 | Sodium hyaluronate beverage with function of regulating intestinal flora and preparation method thereof |
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