CA3235325A1 - Methods for preparing white heirloom tomato-based compositions for skin lightening via enhanced glutathione, and associated compositions thereof - Google Patents
Methods for preparing white heirloom tomato-based compositions for skin lightening via enhanced glutathione, and associated compositions thereof Download PDFInfo
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- CA3235325A1 CA3235325A1 CA3235325A CA3235325A CA3235325A1 CA 3235325 A1 CA3235325 A1 CA 3235325A1 CA 3235325 A CA3235325 A CA 3235325A CA 3235325 A CA3235325 A CA 3235325A CA 3235325 A1 CA3235325 A1 CA 3235325A1
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- 235000007688 Lycopersicon esculentum Nutrition 0.000 title claims abstract description 57
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 title claims abstract description 33
- 238000000034 method Methods 0.000 title claims abstract description 30
- 239000000203 mixture Substances 0.000 title claims abstract description 26
- 229960003180 glutathione Drugs 0.000 title claims abstract description 15
- 108010024636 Glutathione Proteins 0.000 title claims abstract description 10
- 206010040829 Skin discolouration Diseases 0.000 title claims abstract description 4
- 240000003768 Solanum lycopersicum Species 0.000 title claims description 50
- 235000013399 edible fruits Nutrition 0.000 claims abstract description 36
- 239000000843 powder Substances 0.000 claims abstract description 33
- 239000011550 stock solution Substances 0.000 claims abstract description 14
- 239000012153 distilled water Substances 0.000 claims abstract description 4
- 238000004108 freeze drying Methods 0.000 claims abstract description 4
- 238000005070 sampling Methods 0.000 claims abstract description 4
- 238000005406 washing Methods 0.000 claims abstract description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 4
- 238000004519 manufacturing process Methods 0.000 claims abstract 2
- 210000004027 cell Anatomy 0.000 claims description 33
- 108010063907 Glutathione Reductase Proteins 0.000 claims description 9
- 102100036442 Glutathione reductase, mitochondrial Human genes 0.000 claims description 9
- YVLPJIGOMTXXLP-UHFFFAOYSA-N 15-cis-phytoene Chemical compound CC(C)=CCCC(C)=CCCC(C)=CCCC(C)=CC=CC=C(C)CCC=C(C)CCC=C(C)CCC=C(C)C YVLPJIGOMTXXLP-UHFFFAOYSA-N 0.000 claims description 8
- 230000000694 effects Effects 0.000 claims description 8
- YVLPJIGOMTXXLP-UUKUAVTLSA-N 15,15'-cis-Phytoene Natural products C(=C\C=C/C=C(\CC/C=C(\CC/C=C(\CC/C=C(\C)/C)/C)/C)/C)(\CC/C=C(\CC/C=C(\CC/C=C(\C)/C)/C)/C)/C YVLPJIGOMTXXLP-UUKUAVTLSA-N 0.000 claims description 4
- YVLPJIGOMTXXLP-BAHRDPFUSA-N 15Z-phytoene Natural products CC(=CCCC(=CCCC(=CCCC(=CC=C/C=C(C)/CCC=C(/C)CCC=C(/C)CCC=C(C)C)C)C)C)C YVLPJIGOMTXXLP-BAHRDPFUSA-N 0.000 claims description 4
- OVSVTCFNLSGAMM-KGBODLQUSA-N cis-phytofluene Natural products CC(=CCCC(=CCCC(=CCCC(=CC=C/C=C(C)/C=C/C=C(C)/CCC=C(/C)CCC=C(C)C)C)C)C)C OVSVTCFNLSGAMM-KGBODLQUSA-N 0.000 claims description 4
- 238000012258 culturing Methods 0.000 claims description 4
- 239000012895 dilution Substances 0.000 claims description 4
- 238000010790 dilution Methods 0.000 claims description 4
- 210000002950 fibroblast Anatomy 0.000 claims description 4
- 239000012909 foetal bovine serum Substances 0.000 claims description 4
- 235000011765 phytoene Nutrition 0.000 claims description 4
- OVSVTCFNLSGAMM-UZFNGAIXSA-N phytofluene Chemical compound CC(C)=CCCC(C)=CCCC(C)=CCCC(C)=CC=C\C=C(/C)\C=C\C=C(C)CCC=C(C)CCC=C(C)C OVSVTCFNLSGAMM-UZFNGAIXSA-N 0.000 claims description 4
- 235000002677 phytofluene Nutrition 0.000 claims description 4
- ZYSFBWMZMDHGOJ-SGKBLAECSA-N phytofluene Natural products CC(=CCCC(=CCCC(=CCCC(=CC=C/C=C(C)/CCC=C(/C)C=CC=C(/C)CCC=C(C)C)C)C)C)C ZYSFBWMZMDHGOJ-SGKBLAECSA-N 0.000 claims description 4
- ZIUDAKDLOLDEGU-UHFFFAOYSA-N trans-Phytofluen Natural products CC(C)=CCCC(C)CCCC(C)CC=CC(C)=CC=CC=C(C)C=CCC(C)CCCC(C)CCC=C(C)C ZIUDAKDLOLDEGU-UHFFFAOYSA-N 0.000 claims description 4
- 230000035899 viability Effects 0.000 claims description 4
- 238000003556 assay Methods 0.000 claims description 3
- 230000005779 cell damage Effects 0.000 claims description 3
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 claims description 2
- 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 claims description 2
- 231100000002 MTT assay Toxicity 0.000 claims description 2
- 238000000134 MTT assay Methods 0.000 claims description 2
- 239000008103 glucose Substances 0.000 claims description 2
- 230000001404 mediated effect Effects 0.000 claims description 2
- 239000002609 medium Substances 0.000 claims description 2
- 241000227653 Lycopersicon Species 0.000 abstract 7
- 210000003491 skin Anatomy 0.000 description 15
- 230000002087 whitening effect Effects 0.000 description 11
- 102000003855 L-lactate dehydrogenase Human genes 0.000 description 6
- 108700023483 L-lactate dehydrogenases Proteins 0.000 description 6
- 230000003712 anti-aging effect Effects 0.000 description 5
- QIGBRXMKCJKVMJ-UHFFFAOYSA-N Hydroquinone Chemical compound OC1=CC=C(O)C=C1 QIGBRXMKCJKVMJ-UHFFFAOYSA-N 0.000 description 4
- 235000021466 carotenoid Nutrition 0.000 description 4
- 150000001747 carotenoids Chemical class 0.000 description 4
- 231100000135 cytotoxicity Toxicity 0.000 description 4
- 230000003013 cytotoxicity Effects 0.000 description 4
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 3
- 238000002835 absorbance Methods 0.000 description 3
- 239000002537 cosmetic Substances 0.000 description 3
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 2
- XUMBMVFBXHLACL-UHFFFAOYSA-N Melanin Chemical compound O=C1C(=O)C(C2=CNC3=C(C(C(=O)C4=C32)=O)C)=C2C4=CNC2=C1C XUMBMVFBXHLACL-UHFFFAOYSA-N 0.000 description 2
- 238000004061 bleaching Methods 0.000 description 2
- 238000010195 expression analysis Methods 0.000 description 2
- 229930003935 flavonoid Natural products 0.000 description 2
- 150000002215 flavonoids Chemical class 0.000 description 2
- 235000017173 flavonoids Nutrition 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 230000003647 oxidation Effects 0.000 description 2
- 238000007254 oxidation reaction Methods 0.000 description 2
- 150000008442 polyphenolic compounds Chemical class 0.000 description 2
- 150000005207 1,3-dihydroxybenzenes Chemical class 0.000 description 1
- WVXRAFOPTSTNLL-NKWVEPMBSA-N 2',3'-dideoxyadenosine Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@H]1CC[C@@H](CO)O1 WVXRAFOPTSTNLL-NKWVEPMBSA-N 0.000 description 1
- YQHMWTPYORBCMF-ZZXKWVIFSA-N 2',4,4',6'-tetrahydroxychalcone Chemical compound C1=CC(O)=CC=C1\C=C\C(=O)C1=C(O)C=C(O)C=C1O YQHMWTPYORBCMF-ZZXKWVIFSA-N 0.000 description 1
- JMGZEFIQIZZSBH-UHFFFAOYSA-N Bioquercetin Natural products CC1OC(OCC(O)C2OC(OC3=C(Oc4cc(O)cc(O)c4C3=O)c5ccc(O)c(O)c5)C(O)C2O)C(O)C(O)C1O JMGZEFIQIZZSBH-UHFFFAOYSA-N 0.000 description 1
- 235000005979 Citrus limon Nutrition 0.000 description 1
- 244000131522 Citrus pyriformis Species 0.000 description 1
- 240000008067 Cucumis sativus Species 0.000 description 1
- 235000010799 Cucumis sativus var sativus Nutrition 0.000 description 1
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 description 1
- 201000004624 Dermatitis Diseases 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 231100000416 LDH assay Toxicity 0.000 description 1
- 240000003394 Malpighia glabra Species 0.000 description 1
- 235000014837 Malpighia glabra Nutrition 0.000 description 1
- 241000220225 Malus Species 0.000 description 1
- YQHMWTPYORBCMF-UHFFFAOYSA-N Naringenin chalcone Natural products C1=CC(O)=CC=C1C=CC(=O)C1=C(O)C=C(O)C=C1O YQHMWTPYORBCMF-UHFFFAOYSA-N 0.000 description 1
- 241000220317 Rosa Species 0.000 description 1
- 206010040880 Skin irritation Diseases 0.000 description 1
- 229930003268 Vitamin C Natural products 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 208000037887 cell injury Diseases 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000001086 cytosolic effect Effects 0.000 description 1
- 230000008030 elimination Effects 0.000 description 1
- 238000003379 elimination reaction Methods 0.000 description 1
- IVTMALDHFAHOGL-UHFFFAOYSA-N eriodictyol 7-O-rutinoside Natural products OC1C(O)C(O)C(C)OC1OCC1C(O)C(O)C(O)C(OC=2C=C3C(C(C(O)=C(O3)C=3C=C(O)C(O)=CC=3)=O)=C(O)C=2)O1 IVTMALDHFAHOGL-UHFFFAOYSA-N 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 238000002843 lactate dehydrogenase assay Methods 0.000 description 1
- 210000002752 melanocyte Anatomy 0.000 description 1
- 201000001441 melanoma Diseases 0.000 description 1
- 150000002978 peroxides Chemical class 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 235000013824 polyphenols Nutrition 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- FDRQPMVGJOQVTL-UHFFFAOYSA-N quercetin rutinoside Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC=2C(C3=C(O)C=C(O)C=C3OC=2C=2C=C(O)C(O)=CC=2)=O)O1 FDRQPMVGJOQVTL-UHFFFAOYSA-N 0.000 description 1
- IKGXIBQEEMLURG-BKUODXTLSA-N rutin Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](C)O[C@@H]1OC[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](OC=2C(C3=C(O)C=C(O)C=C3OC=2C=2C=C(O)C(O)=CC=2)=O)O1 IKGXIBQEEMLURG-BKUODXTLSA-N 0.000 description 1
- ALABRVAAKCSLSC-UHFFFAOYSA-N rutin Natural products CC1OC(OCC2OC(O)C(O)C(O)C2O)C(O)C(O)C1OC3=C(Oc4cc(O)cc(O)c4C3=O)c5ccc(O)c(O)c5 ALABRVAAKCSLSC-UHFFFAOYSA-N 0.000 description 1
- 235000005493 rutin Nutrition 0.000 description 1
- 229960004555 rutoside Drugs 0.000 description 1
- 230000036556 skin irritation Effects 0.000 description 1
- 231100000475 skin irritation Toxicity 0.000 description 1
- 230000036555 skin type Effects 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 235000019154 vitamin C Nutrition 0.000 description 1
- 239000011718 vitamin C Substances 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9783—Angiosperms [Magnoliophyta]
- A61K8/9789—Magnoliopsida [dicotyledons]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/84—Products or compounds obtained by lyophilisation, freeze-drying
Abstract
A method for preparing white heirloom tomato-based composition for skin lightening via enhance glutathione production, the method including the steps of, sampling and decontaminating a tomato fruit sample via vigorous washing; freeze-drying and homogenising the tomato fruit sample to produce tomato fruit powder; preparing tomato fruit powder stock solution using the tomato fruit powder; and, dissolving and mixing the tomato fruit powder stock solution (100mg/ml) in distilled water.
Description
METHODS FOR PREPARING WHITE HEIRLOOM TOMATO-BASED
COMPOSITIONS FOR SKIN LIGHTENING VIA ENHANCED
GLUTATHIONE, AND ASSOCIATED COMPOSITIONS THEREOF
FIELD OF THE INVENTION
Embodiments of the present invention generally relate to compositions for tomato-based compositions, and, more particularly, to methods for preparing white heirloom tomato-based compositions for anti-ageing purposes, and associated compositions thereof.
BACKGROUND OF THE INVENTION
As it is well known that for the whitening of the skin to a lighter skin tone or for the elimination of dark spots from the skin, whitening compositions are useful.
Further, these whitening or skin bleaching compositions are effective as these inhibits the formation of melanin (Skin colour is determined by this pigment).
However, such skin bleaching or whitening compositions contain harsh chemical, such as peroxides, glutathione, formaldehyde and the like, that has severe effects on the skin.
As disclosed in conventional prior art disclosure, the use of hydroquinone suppresses the melanocyte activity and helps in whitening of the skin.
However, such less stringent composition has other disadvantage i.e., the hydroquinone is oxidised by air and light. Further, the oxidised product causes skin irritation and inflammation.
Furthermore, there has been disclosed in the prior art disclosure about the dermatological compositions that use paucilamellar liposomal carriers that help in treating Melanoma caused by exposure to sunlight. However, such compositions are not effective in whitening the skin to a lighter skin tone.
Additionally, some conventional prior art disclosure discloses the use of naturally derived materials in dermatological compositions, such as but not limited to, lemons, oranges, cucumber, rose fruit and the like. However, such naturally derived materials limit their usefulness based on age, skin type, colour and vary greatly in different subjects.
As disclosed in various prior art disclosures, the use of cosmetics composition containing carotenoids with the extract of plant of the genus malus, containing polyphenols helps in skin whitening. However, these compositions containing carotenoids are typically highly coloured and are sensitive to light and oxidation that considerably limits their usefulness.
Furthermore, prior art disclosures also disclose the use of polyphenolic compounds, mostly resorcinols having skin whitening properties. However, these compounds being susceptible to air and UV oxidation leads to colour instability that causes loss of skin tightening efficacy. Additionally, as per related prior art disclosure, the extract obtained from fruits of acerola containing Vitamin C, are incorporated into cosmetics to provide a whitening effect.
Therefore, the prior art disclosures briefly discuss about the compositions that are useful for whitening of the skin. However, the element of usage to increase natural glutathione level is yet to be introduced in the market.
Thus, there is a recognized need for, and it would be highly beneficial to have alternative, effective natural cocktail of flavonoid and carotenoid preparations having minimal adverse side effects, which are effective for whitening skin and which are suitable for the cosmetic industry. Accordingly, there remains a need
COMPOSITIONS FOR SKIN LIGHTENING VIA ENHANCED
GLUTATHIONE, AND ASSOCIATED COMPOSITIONS THEREOF
FIELD OF THE INVENTION
Embodiments of the present invention generally relate to compositions for tomato-based compositions, and, more particularly, to methods for preparing white heirloom tomato-based compositions for anti-ageing purposes, and associated compositions thereof.
BACKGROUND OF THE INVENTION
As it is well known that for the whitening of the skin to a lighter skin tone or for the elimination of dark spots from the skin, whitening compositions are useful.
Further, these whitening or skin bleaching compositions are effective as these inhibits the formation of melanin (Skin colour is determined by this pigment).
However, such skin bleaching or whitening compositions contain harsh chemical, such as peroxides, glutathione, formaldehyde and the like, that has severe effects on the skin.
As disclosed in conventional prior art disclosure, the use of hydroquinone suppresses the melanocyte activity and helps in whitening of the skin.
However, such less stringent composition has other disadvantage i.e., the hydroquinone is oxidised by air and light. Further, the oxidised product causes skin irritation and inflammation.
Furthermore, there has been disclosed in the prior art disclosure about the dermatological compositions that use paucilamellar liposomal carriers that help in treating Melanoma caused by exposure to sunlight. However, such compositions are not effective in whitening the skin to a lighter skin tone.
Additionally, some conventional prior art disclosure discloses the use of naturally derived materials in dermatological compositions, such as but not limited to, lemons, oranges, cucumber, rose fruit and the like. However, such naturally derived materials limit their usefulness based on age, skin type, colour and vary greatly in different subjects.
As disclosed in various prior art disclosures, the use of cosmetics composition containing carotenoids with the extract of plant of the genus malus, containing polyphenols helps in skin whitening. However, these compositions containing carotenoids are typically highly coloured and are sensitive to light and oxidation that considerably limits their usefulness.
Furthermore, prior art disclosures also disclose the use of polyphenolic compounds, mostly resorcinols having skin whitening properties. However, these compounds being susceptible to air and UV oxidation leads to colour instability that causes loss of skin tightening efficacy. Additionally, as per related prior art disclosure, the extract obtained from fruits of acerola containing Vitamin C, are incorporated into cosmetics to provide a whitening effect.
Therefore, the prior art disclosures briefly discuss about the compositions that are useful for whitening of the skin. However, the element of usage to increase natural glutathione level is yet to be introduced in the market.
Thus, there is a recognized need for, and it would be highly beneficial to have alternative, effective natural cocktail of flavonoid and carotenoid preparations having minimal adverse side effects, which are effective for whitening skin and which are suitable for the cosmetic industry. Accordingly, there remains a need
2 in the art for innovative, novel, efficient methods for preparing white heirloom tomato-based compositions for anti-ageing purposes, and associated compositions thereof.
SUMMARY OF THE INVENTION
The embodiments of the present disclosure have several features, no single one of which is solely responsible for their desirable attributes. Without limiting the scope of the present embodiments as expressed by the claims that follow, their more prominent features will now be discussed briefly. After considering this discussion, and particularly after reading the section entitled "Detailed Description", one will understand how the features of the present embodiments provide advantages.
In one embodiment, a method for preparing white heirloom tomato-based composition for anti-ageing purposes, the method including the steps of, sampling and decontaminating a tomato fruit sample via vigorous washing;
freeze-drying and homogenising the tomato fruit sample to produce tomato fruit powder; preparing tomato fruit powder stock solution using the tomato fruit powder; and, dissolving and mixing the tomato fruit powder stock solution (100mg/m1) in distilled water.
DETAILED DESCRIPTION
Various embodiments of the present invention are aimed at providing novel and inventive solutions focused on methods for preparing white heirloom tomato-based compositions for anti-ageing purposes, and associated compositions thereof.
In accordance with an embodiment of the present invention, a method for preparing white heirloom tomato-based composition for anti-ageing purposes,
SUMMARY OF THE INVENTION
The embodiments of the present disclosure have several features, no single one of which is solely responsible for their desirable attributes. Without limiting the scope of the present embodiments as expressed by the claims that follow, their more prominent features will now be discussed briefly. After considering this discussion, and particularly after reading the section entitled "Detailed Description", one will understand how the features of the present embodiments provide advantages.
In one embodiment, a method for preparing white heirloom tomato-based composition for anti-ageing purposes, the method including the steps of, sampling and decontaminating a tomato fruit sample via vigorous washing;
freeze-drying and homogenising the tomato fruit sample to produce tomato fruit powder; preparing tomato fruit powder stock solution using the tomato fruit powder; and, dissolving and mixing the tomato fruit powder stock solution (100mg/m1) in distilled water.
DETAILED DESCRIPTION
Various embodiments of the present invention are aimed at providing novel and inventive solutions focused on methods for preparing white heirloom tomato-based compositions for anti-ageing purposes, and associated compositions thereof.
In accordance with an embodiment of the present invention, a method for preparing white heirloom tomato-based composition for anti-ageing purposes,
3 the method including the steps of, sampling and decontaminating a tomato fruit sample via vigorous washing; freeze-drying and homogenising the tomato fruit sample to produce tomato fruit powder; preparing tomato fruit powder stock solution using the tomato fruit powder; and, dissolving and mixing the tomato fruit powder stock solution (100mg/m1) in distilled water.
In accordance with an embodiment of the present invention, the method further includes the step of passing the tomato fruit powder stock solution through syringe filter for preparing different dilutions in plain medium (DMEM). In use, the tomato fruit powder stock solution is prepared by using dilutions of 10Oug/ml, 200ug/ml, 500ug/ml, 1000ug/m1 and 2000ug/ml.
In accordance with an embodiment of the present invention, the method further includes steps of, obtaining a NIH 3T3 (mouse fibroblast) cell line from a cell tissue culture; and, culturing the cell line in Dulbecco's Modified Eagle's Medium High Glucose (DMEM-HG) with 10% foetal bovine serum (FBS). In use, the method further includes the steps of, treating the NIH 3T3 (mouse fibroblast) cell lines with tomato fruit powder; culturing the cell lines onto 96 well plates;
treating the cell lines with 6mm H202 for approx. 4 hours to create ROS
mediated cellular damage; and, treating injured cells with the concentrations (10Oug/ml, 200ug/ml, 500ug/ml, 1000ug/m1 and 2000ug/m1) of the tomato fruit powder stock solution for a period of 24 hours.
In accordance with an embodiment of the present invention, the method further includes the step of, calculating viability of the NIH 3T3 cells via MTT
assay, evaluating the Glutathione reductase and reduced Gluthathione (GSH) levels in the NIH 3T3 cells, and, calculating the LDH activity of the NIH 3T3 cells via
In accordance with an embodiment of the present invention, the method further includes the step of passing the tomato fruit powder stock solution through syringe filter for preparing different dilutions in plain medium (DMEM). In use, the tomato fruit powder stock solution is prepared by using dilutions of 10Oug/ml, 200ug/ml, 500ug/ml, 1000ug/m1 and 2000ug/ml.
In accordance with an embodiment of the present invention, the method further includes steps of, obtaining a NIH 3T3 (mouse fibroblast) cell line from a cell tissue culture; and, culturing the cell line in Dulbecco's Modified Eagle's Medium High Glucose (DMEM-HG) with 10% foetal bovine serum (FBS). In use, the method further includes the steps of, treating the NIH 3T3 (mouse fibroblast) cell lines with tomato fruit powder; culturing the cell lines onto 96 well plates;
treating the cell lines with 6mm H202 for approx. 4 hours to create ROS
mediated cellular damage; and, treating injured cells with the concentrations (10Oug/ml, 200ug/ml, 500ug/ml, 1000ug/m1 and 2000ug/m1) of the tomato fruit powder stock solution for a period of 24 hours.
In accordance with an embodiment of the present invention, the method further includes the step of, calculating viability of the NIH 3T3 cells via MTT
assay, evaluating the Glutathione reductase and reduced Gluthathione (GSH) levels in the NIH 3T3 cells, and, calculating the LDH activity of the NIH 3T3 cells via
4 LDH released assay. In use, the heirloom white tomato fruit includes phytoene and Phytofluene.
In accordance with an embodiment of the present invention, Figure 1 illustrates 3T3 cell line, as the plot of cytotoxicity percentage (%) versus various concentrations of Tomesol heirloom white tomato (Tomesoral), wherein lower concentrations of tomato powder did not show significant cytotoxicity on the cell lines, whereas, the higher concentration like 1mg or 2mg showed some cytotoxicity. Subsequently, on the basis of cytotoxicities, IC50 value was calculated as 1.986mg/ml.
Also, the embodiment of the invention in Figure 1 (a) illustrates the Bar graph which further shows the relative values of absorbance after treating the 3T3 cells with MTT (3-(4, 5-dimethylthiazol-2-y1) - 2, 5-diphenyltetrazolium bromide) assay.
Furthermore, the embodiment of the invention as illustrated in Figure 1 (b) shows line graph that depicts the relative percentage of viabilities on different concentrations of tomato powder while box represents the IC50 value for tomato powder. Values were taken as mean SEM and * shows the significant difference between viabilities of treated groups and untreated controls (p<0.05).
In accordance with an embodiment of the present invention, as illustrated in Figure 2(a), the relative absorbance of post treated cells, measuring the reduced levels of cytoplasmic enzyme LDH (Lactate dehydrogenase) released (during cell injury or stress), in treatment groups of 3T3 cells, after performing LDH assay. Also, the embodiment of the invention as illustrated in Figure 2 (b) shows the Expression analysis of LDH release in post treated cells. Values
In accordance with an embodiment of the present invention, Figure 1 illustrates 3T3 cell line, as the plot of cytotoxicity percentage (%) versus various concentrations of Tomesol heirloom white tomato (Tomesoral), wherein lower concentrations of tomato powder did not show significant cytotoxicity on the cell lines, whereas, the higher concentration like 1mg or 2mg showed some cytotoxicity. Subsequently, on the basis of cytotoxicities, IC50 value was calculated as 1.986mg/ml.
Also, the embodiment of the invention in Figure 1 (a) illustrates the Bar graph which further shows the relative values of absorbance after treating the 3T3 cells with MTT (3-(4, 5-dimethylthiazol-2-y1) - 2, 5-diphenyltetrazolium bromide) assay.
Furthermore, the embodiment of the invention as illustrated in Figure 1 (b) shows line graph that depicts the relative percentage of viabilities on different concentrations of tomato powder while box represents the IC50 value for tomato powder. Values were taken as mean SEM and * shows the significant difference between viabilities of treated groups and untreated controls (p<0.05).
In accordance with an embodiment of the present invention, as illustrated in Figure 2(a), the relative absorbance of post treated cells, measuring the reduced levels of cytoplasmic enzyme LDH (Lactate dehydrogenase) released (during cell injury or stress), in treatment groups of 3T3 cells, after performing LDH assay. Also, the embodiment of the invention as illustrated in Figure 2 (b) shows the Expression analysis of LDH release in post treated cells. Values
5 were taken as mean SEM and *shows the significant difference between LDH
levels of treated groups and untreated controls (p<0.05).
In accordance with an embodiment of the present invention, Figure 3 (a) shows relative absorbance of post treated cells that further shows an increased Glutathione Reductase in Post Treated Groups; 3T3 cells 'intracellular glutathione reductase (GR) activity was measured (n = 5 replicates) before and after treatment with Tomesol heirloom white tomato (Tomesoral).
Subsequently, Glutathione reductase (GR) activity was significantly increased in post treated cells, wherein, the presence of white tomato powder in the media significantly boosted GR activity from 2.24 0.152 mU/mL prior to treatment to 5.07 0.323 mU/mL in the presence of Tomesol heirloom white tomato (Tomesoral) at a concentration of 500ug/ml. However, the addition of more tomato powder to a concentration of 1mg/m1 and 2mg/m1 will not boost the levels further. The peak value is observed in 500ug/m1 concentration.
In accordance with an embodiment of the present invention, Figure 3(b) shows the expression analysis of reduced glutathione (GSH). Reduced glutathione (GSH) of H202 injured 3T3 cells was estimated (n = 5 replicates) pre and post-treatment with heirloom tomato powder. GSH was significantly increased in tomato powder treated cells. The presence of white tomato powder in the media significantly enhances GSH as untreated cells has value of 0.347 0.0152 while treated groups have GSH levels as: T.TP-10Oug/m1 (0.417 0.0426), T.TP-200ug/m1 (0.524 0.0426), T.TP-500ug/m1 (0.630 0.0323), T.TP-1mg/m1 (0.630 0.0104) and T.TP-2mg/m1 (0.629 0.0199) compared to stressed cells (0.201 0.0139). The optimum dose of tomato powder is 500ug/ml. The addition of more tomato powder to a concentration of 1mg/m1 and 2mg/m1 will not boost
levels of treated groups and untreated controls (p<0.05).
In accordance with an embodiment of the present invention, Figure 3 (a) shows relative absorbance of post treated cells that further shows an increased Glutathione Reductase in Post Treated Groups; 3T3 cells 'intracellular glutathione reductase (GR) activity was measured (n = 5 replicates) before and after treatment with Tomesol heirloom white tomato (Tomesoral).
Subsequently, Glutathione reductase (GR) activity was significantly increased in post treated cells, wherein, the presence of white tomato powder in the media significantly boosted GR activity from 2.24 0.152 mU/mL prior to treatment to 5.07 0.323 mU/mL in the presence of Tomesol heirloom white tomato (Tomesoral) at a concentration of 500ug/ml. However, the addition of more tomato powder to a concentration of 1mg/m1 and 2mg/m1 will not boost the levels further. The peak value is observed in 500ug/m1 concentration.
In accordance with an embodiment of the present invention, Figure 3(b) shows the expression analysis of reduced glutathione (GSH). Reduced glutathione (GSH) of H202 injured 3T3 cells was estimated (n = 5 replicates) pre and post-treatment with heirloom tomato powder. GSH was significantly increased in tomato powder treated cells. The presence of white tomato powder in the media significantly enhances GSH as untreated cells has value of 0.347 0.0152 while treated groups have GSH levels as: T.TP-10Oug/m1 (0.417 0.0426), T.TP-200ug/m1 (0.524 0.0426), T.TP-500ug/m1 (0.630 0.0323), T.TP-1mg/m1 (0.630 0.0104) and T.TP-2mg/m1 (0.629 0.0199) compared to stressed cells (0.201 0.0139). The optimum dose of tomato powder is 500ug/ml. The addition of more tomato powder to a concentration of 1mg/m1 and 2mg/m1 will not boost
6
7 the levels further. Further, Graph shows the reduced glutathione (GSH) levels in post treated cells. Values were taken as mean SEM and * shows the significant difference between GR levels of treated groups and untreated controls (p<0.05). Therefore, the present invention shows the use of Tomesol heirloom white tomato (Tomesoral) that is rich in carotenoids (phytoene and phytofluene) and flavonoids (chalconaringenin and rutin) unique natural cocktail which were specifically targeted to increase and forces glutathione level to increase excessively. In use, the heirloom white tomato fruit includes phytoene and Phytofluene.
The conditional language used herein, such as, among others, "can," "could,"
"might," "may," "e.g.," and the like, unless specifically stated otherwise, or otherwise understood within the context as used, is generally intended to convey that certain embodiments include, while other embodiments do not include, certain features, elements and/or steps. Thus, such conditional language is not generally intended to imply that features, elements and/or steps are in any way required for one or more embodiments or that one or more embodiments necessarily include logic for deciding, with or without author input or prompting, whether these features, elements and/or steps are included or are to be performed in any particular embodiment. The terms "comprising,"
"including," 'having," and the like are synonymous and are used inclusively, in an open-ended fashion, and do not exclude additional elements, features, acts, operations, and so forth. Also, the term "or" is used in its inclusive sense (and not in its exclusive sense) so that when used, for example, to connect a list of elements, the term "or" means one, some, or all of the elements in the list.
While there has been shown and described the preferred embodiment of the instant invention it is to be appreciated that the invention may be embodied otherwise than is herein specifically shown and described and that, within said embodiment, certain changes may be made in the form and arrangement of the parts without departing from the underlying ideas or principles of this invention as outlined in the Claims appended herewith. Therefore, the appended claims are to be construed to cover all equivalents falling within the true scope and spirit of the invention.
The conditional language used herein, such as, among others, "can," "could,"
"might," "may," "e.g.," and the like, unless specifically stated otherwise, or otherwise understood within the context as used, is generally intended to convey that certain embodiments include, while other embodiments do not include, certain features, elements and/or steps. Thus, such conditional language is not generally intended to imply that features, elements and/or steps are in any way required for one or more embodiments or that one or more embodiments necessarily include logic for deciding, with or without author input or prompting, whether these features, elements and/or steps are included or are to be performed in any particular embodiment. The terms "comprising,"
"including," 'having," and the like are synonymous and are used inclusively, in an open-ended fashion, and do not exclude additional elements, features, acts, operations, and so forth. Also, the term "or" is used in its inclusive sense (and not in its exclusive sense) so that when used, for example, to connect a list of elements, the term "or" means one, some, or all of the elements in the list.
While there has been shown and described the preferred embodiment of the instant invention it is to be appreciated that the invention may be embodied otherwise than is herein specifically shown and described and that, within said embodiment, certain changes may be made in the form and arrangement of the parts without departing from the underlying ideas or principles of this invention as outlined in the Claims appended herewith. Therefore, the appended claims are to be construed to cover all equivalents falling within the true scope and spirit of the invention.
8
Claims (9)
1. A method for preparing white heirloom tomato-based composition for skin lightening via enhance glutathione production, said method comprising the steps of:
sampling and decontaminating a tomato fruit sample via vigorous washing;
freeze-drying and homogenising said tomato fruit sample to produce tomato fruit powder;
preparing tomato fruit powder stock solution using said tomato fruit powder; and, dissolving and mixing said tomato fruit powder stock solution (100mg/ml) in distilled water.
sampling and decontaminating a tomato fruit sample via vigorous washing;
freeze-drying and homogenising said tomato fruit sample to produce tomato fruit powder;
preparing tomato fruit powder stock solution using said tomato fruit powder; and, dissolving and mixing said tomato fruit powder stock solution (100mg/ml) in distilled water.
2. The method as claimed in Claim 1, wherein said method further comprises the step of passing said tomato fruit powder stock solution through syringe filter for preparing different dilutions in plain medium (DMEM).
3. The method as claimed in claim 2, wherein said tomato fruit powder stock solution is prepared by using dilutions of 10Oug/ml, 200ugiml, 500ug/ml, 1000ug/ml and 2000ug/rnl.
4. The method as claimed in claim 1, wherein said rnethod further comprises steps of:
obtaining a NIH 3T3 (mouse fibroblast) cell line from a cell tissue culture;
and, culturing said cell line in Dulbecco's Modified Eagle's Medium High Glucose (DMEM-HG) with 10% foetal bovine serum (FBS).
obtaining a NIH 3T3 (mouse fibroblast) cell line from a cell tissue culture;
and, culturing said cell line in Dulbecco's Modified Eagle's Medium High Glucose (DMEM-HG) with 10% foetal bovine serum (FBS).
5. The method as claimed in claim 1, wherein said method further comprises the steps of:
culturing the said NIH 3T3 (mouse fibroblast) cell lines onto 96 well plates;
treating said cell lines with 6mm H202 for approx. 4 hours to create ROS
mediated cellular damage; and, treating injured cells with said concentrations of said tomato fruit powder stock solution for a period of 24 hours.
culturing the said NIH 3T3 (mouse fibroblast) cell lines onto 96 well plates;
treating said cell lines with 6mm H202 for approx. 4 hours to create ROS
mediated cellular damage; and, treating injured cells with said concentrations of said tomato fruit powder stock solution for a period of 24 hours.
6. The method as claimed in claim 5, wherein said method further comprises the step of calculating viability of said NIH 3T3 cells via MTT
assay.
assay.
7. The method as claimed in claim 1, wherein said method further comprises the step of evaluating the glutathione reductase and reduced Gluthathione (GSH) levels in said NIH 3T3 cells.
8. The method as claimed in claim 1, wherein said method further comprises the steps of calculating the LDH activity of said NIH 3T3 cells via LDH released assay.
9. The method as claimed in claim 1, wherein said heirloom white tomato fruit comprises phytoene and Phytofluene.
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|---|---|---|---|
| PCT/IB2021/053926 WO2022238727A1 (en) | 2021-05-08 | 2021-05-08 | Methods for preparing white heirloom tomato-based compositions for skin lightening via enhanced glutathione, and associated compositions thereof |
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| EP2844083B1 (en) * | 2012-04-23 | 2018-07-25 | University Of Oslo | Use of tomato extract as antihypertensive agent and process for making water soluble sugar free tomato extract |
| CN110192994A (en) * | 2019-06-25 | 2019-09-03 | 昆仑技术研究(广州)有限公司 | A kind of white tomato whitening piece and preparation method thereof |
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