Claims:1. A composition comprising:
a) asparagus extract; and
b) red pearl onion extract,
wherein the asparagus extract to the red pearl onion extract weight ratio in said composition is in the range of 5:1 - 4:5.
2. The composition as claimed in claim 1, wherein the asparagus extract is having a weight percentage in the range of 0.003% to 0.01%, and wherein the red pearl onion extract is having a weight percentage in the range of 0.001% to 0.006%.
3. The composition as claimed in claim 1, wherein the asparagus extract to the red pearl onion extract w/w ratio said composition is in the range of 4:1 – 1:1.
4. The composition as claimed in claim 1, wherein the asparagus extract to the red pearl onion extract w/w ratio is 4:1.
5. The composition as claimed in claim 1, wherein the asparagus extract to the red pearl onion extract w/w ratio is 3:2.
6. The composition as claimed in claim 1, wherein the asparagus extract to the red pearl onion extract w/w ratio is 1:1.
7. The composition as claimed in any of the claims 1-6, further comprising at least one carrier, at least one diluent, at least one excipient, and combinations thereof.
8. A method for preparing a composition as claimed in claim 1, said method comprising:
a) obtaining asparagus extract;
b) obtaining red pearl onion extract; and
c) contacting the asparagus extract, and the red pearl onion extract to obtain said composition.
9. A method for preparing a composition as claimed in any of the claims 1-7, said method comprising:
a) obtaining asparagus extract;
b) obtaining red pearl onion extract;
c) obtaining at least one carrier;
d) obtaining at least one diluent;
e) obtaining at least one excipient; and
f) contacting the asparagus extract, the red pearl onion extract, the at least one carrier, the at least one diluent, and the at least one excipient to obtain said composition.
10. The composition as claimed in any of the claims 1-7, wherein said composition exhibits anti-inflammatory effect.
11. The composition as claimed in claim 10, wherein said composition exhibits anti-inflammatory effect by inhibiting production of Interleukin 8 (IL-8).
, Description:[002] Inflammation is an integral part of immune system which plays an important role in defending the human body against pathogens such as bacteria, virus, and other parasites. Inflammation assists in healing process and limits the damage to a certain area which further speeds up the curing process, and it gets rid of damaged cells from the body. The process of inflammation employs certain cellular messengers such as NF-?B, certain cytokines, and chemokines for carrying out the desired effects on the body. NF-?B is a master regulator of inflammation and is highly activated at sites of inflammation in various diseases. Cytokines such as IL-1ß, IL-6, IL-8, and TNF-a are pro-inflammatory cytokines that are involved in the upregulation of inflammatory reactions, whereas there are also certain anti-inflammatory cytokines like IL-1ra, IL-4, IL-10, IL-11, and IL-13 which inhibit the inflammatory reactions.
[003] Inflammatory response also has harmful effects, majority of which is attributed to the lysosomal enzymes which are secreted by the inflammatory cells. The extended or unwanted inflammatory response causes destruction or swelling of normal tissues. There are certain diseases like atherosclerosis which are caused due to chronic inflammation, and prolonged inflammation may lead to stroke progression, and thrombosis. Prolonged inflammatory responses are therefore unfavorable, and a balance is required between the upregulation of inflammatory response and its timely inhibition for a perfect immune response. Such a timely response involves a precise interplay of pro-inflammatory and anti-inflammatory molecules for executing an immune response.
[004] Chronic inflammatory diseases can be treated by inhibiting the activity of some highly active pro-inflammatory molecules or by targeting an inflammatory response pathway. Non-steroidal anti-inflammatory drugs (NSAIDS) are one of the commonly prescribed drugs for treating inflammation and NSAIDS exert their effects by inhibiting synthesis of prostaglandins. Though it is commonly prescribed, it exerts certain amount of side effects. Using NSAIDS mainly increase the risk of gastrointestinal and cardiovascular complications (Sostres, Carlos, et al. “Best practice & research Clinical gastroenterology 24.2 (2010): 121-132.). Also, other synthetic drugs being used as inhibitors of inflammation are associated with a certain amount of risks to the health. Therefore, there is a pressing need of some alternatives that has reduced risks of side complications and at the same time are effective anti-inflammatory substances.
[005] US4393076 discloses an anti-inflammatory analgesic gel composition comprising ketoprofen and/or flurbiprofen, a glycol, and other necessary excipients.
[006] WO2006084334 discloses a composition comprising an effective amount of an analgesically and/or anti-inflammatory active fraction separated from a mixture of plasma and/or serum and at least one metal, metal ion or metal salt thereof.
[007] However, considering the established side-effects of drugs, the use of an efficacious herbal formulation to tackle inflammation is of utmost importance.
SUMMARY OF THE INVENTION
[008] In an aspect of present invention, there is provided a composition comprising: (a) asparagus extract; and (b) red pearl onion extract, wherein the asparagus extract to the red pearl onion extract weight ratio in said composition is in the range of 5:1 - 4:5.
[009] In an aspect of present invention, there is provided a method for preparing a composition comprising: (a) asparagus extract; and (b) red pearl onion extract, wherein the asparagus extract to the red pearl onion extract weight ratio in said composition is in the range of 5:1 - 4:5, said method comprising: (i) obtaining the asparagus extract; (ii) obtaining the red pearl onion extract; and (iii) contacting the asparagus extract, and the red pearl onion extract to obtain said composition.
[0010] In an aspect of present invention, there is provided a method for preparing a composition comprising: (a) asparagus extract; (b) red pearl onion extract; (c) at least one carrier; (d) at least one diluent; and (e) at least one excipient, wherein the asparagus extract to the red pearl onion extract weight ratio in said composition is in the range of 5:1 - 4:5, said method comprising: (i) obtaining the asparagus extract; (ii) obtaining the red pearl onion extract; (iii) obtaining the at least one carrier; (iv) obtaining the at least one diluent; (v) obtaining the at least one excipient, and (vi) contacting the asparagus extract, the red pearl onion extract, the at least one carrier, the at least one diluent, and the at least one excipient to obtain said composition.
[0011] These and other features, aspects, and advantages of the present subject matter will be better understood with reference to the following description and appended claims. This summary is provided to introduce a selection of concepts in a simplified form. This summary is not intended to identify key features or essential features of the claimed subject matter, nor is it intended to be used to limit the scope of the claimed subject matter.
BRIEF DESCRIPTION OF ACCOMPANYING DRAWINGS
[001] The following drawings form a part of the present specification and are included to further illustrate aspects of the present disclosure. The disclosure may be better understood by reference to the drawings in combination with the detailed description of the specific embodiments presented herein.
[002] Figure 1 depicts a GCMS chromatogram for Asparagus extract, in accordance with an embodiment of the present disclosure.
[003] Figure 2 depicts a GCMS chromatogram for Red pearl onion extract, in accordance with an embodiment of the present disclosure.
[004] Figure 3 depicts the DPPH value of chloroform fraction and methanol/water fraction of asparagus extract and red pearl onion extract, in accordance with an embodiment of the present disclosure.
[005] Figure 4 depicts the cell viability in presence of asparagus extract, in accordance with an embodiment of the present disclosure.
[006] Figure 5 depicts the cell viability in presence of red pearl onion extract, in accordance with an embodiment of the present disclosure.
[007] Figure 6 depicts the dose response study of asparagus extract in inhibiting IL-8 production, in accordance with an embodiment of the present disclosure.
[008] Figure 7 depicts the dose response study of red pearl onion extract in inhibiting IL-8 production, in accordance with an embodiment of the present disclosure.
[009] Figure 8 depicts the dose response study of asparagus extract in inhibiting IL-8 production, in accordance with an embodiment of the present disclosure.
[0010] Figure 9 depicts the dose response study of red pearl onion extract in inhibiting NF?B phosphorylation, in accordance with an embodiment of the present disclosure.
[0011] Figure 10 depicts the effect of various combinations of asparagus extract and red pearl onion extract in inhibiting NF?B phosphorylation, in accordance with an embodiment of the present disclosure.
[0012] Figure 11 depicts the effect of various combinations of asparagus extract and red pearl onion extract in inhibiting NF?B phosphorylation, in accordance with an embodiment of the present disclosure.

DETAILED DESCRIPTION OF THE INVENTION
[0012] Those skilled in the art will be aware that the present disclosure is subject to variations and modifications other than those specifically described. It is to be understood that the present disclosure includes all such variations and modifications. The disclosure also includes all such steps, features, compositions, and compounds referred to or indicated in this specification, individually or collectively, and any and all combinations of any or more of such steps or features.
Definitions
[0013] For convenience, before further description of the present disclosure, certain terms employed in the specification, and examples are delineated here. These definitions should be read in the light of the remainder of the disclosure and understood as by a person of skill in the art. The terms used herein have the meanings recognized and known to those of skill in the art, however, for convenience and completeness, particular terms and their meanings are set forth below.
[0014] The articles “a”, “an” and “the” are used to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article.
[0015] The terms “comprise” and “comprising” are used in the inclusive, open sense, meaning that additional elements may be included. It is not intended to be construed as “consists of only”.
[0016] Throughout this specification, unless the context requires otherwise the word “comprise”, and variations such as “comprises” and “comprising”, will be understood to imply the inclusion of a stated element or step or group of element or steps but not the exclusion of any other element or step or group of element or steps.
[0017] The term “including” is used to mean “including but not limited to”. “Including” and “including but not limited to” are used interchangeably.
[0018] Carriers are substances that serve as mechanisms to improve the delivery and the effectiveness of drugs.
[0019] A diluent (also referred to as filler, dilutant, or thinner) is a diluting agent.
[0020] An excipient is an inactive substance that serve as the vehicle or medium for a drug or other active substance. Excipients include colouring agents, humectants, preservatives, emollients, and combinations thereof.
[0021] Asparagus is an herbaceous perennial plant known for its rich nutrient content. The metabolites of the plant have been beneficial for various purposes. Previous studies have identified many phytochemicals from asparagus which includes steroidal saponins (shatavarin 1-IV), sitosterol, 4, 6-dihydryxy-2-O (-2-hydroxy isobutyl) benzaldehyde, undecanyl cetanoate, oligospirostanoside (immunoside), essential fatty acids-Gamma linoleinic acids, diosgenin, quercetin 3-glucourbnides, furan compound-racemofuran, vitamins A, B1, B2, C, E, Mg, P, Ca, Fe, folic acid, essential oils, asparagine, arginine, tyrosine, flavonoids (kaempferol, quercetin, and rutin), resin, tannin, sarsasapogenin, new isoflavone (8-methoxy-5,6,4’-trihydroxyisoflavone-7-O-ß-D-glucopyranoside) polycyclic alkaloid (asparagamine A), 9,10-dihydrophenanthrene racemosol, kaempferol, oligofurostanosides and spirostanosides (Negi et al., 2010. Pharmacogn Rev. 4(8):215-20; Shashi et al., 2013. Asian Pac J Trop Dis. 3(3): 242–251).
[0022] Red pearl onion are also studied for the presence of different phytochemicals including organosulfur compounds (cepaenes and thiosulfinates) and pigments (anthocyanins: cyanidin 3-O-(3"-O-beta-glucopyranosyl-6"-O-malonyl-beta-glucopyranoside)-4'-O-beta-glucopyranoside, cyanidin 7-O-(3"-O-beta-glucopyranosyl-6"-O-malonyl-beta-glucopyranoside)-4'-O-beta-glucopyranoside, cyanidin 3,4'-di-O-beta-glucopyranoside, cyanidin 4'-O-beta-glucoside, peonidin 3-O-(6"-O-malonyl-beta-glucopyranoside)-5-O-beta-glucopyranoside and peonidin 3-O-(6"-O-malonyl-beta-glucopyranoside), flavonoids (quercetin and Kaempferol) Quercetin 3,7,4'-O-ß-triglucopyranoside, taxifolin-4'-O-ß-glucopyranoside (Torgils et al., 1998 Phytochemistry. 47:2, 281-285; Dorsch and Wagner., 1991. Int Arch Allergy Appl Immunol. 94: 262-265; Goldman et al., 1996. Thrombosis and Haemostasis. 450-452; Dorant 1994. Cancer Research 54: 6148-6153 and Fossen et al., 2003. Phytochemistry. 64(8):1367-74).
[0023] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the disclosure, the preferred methods, and materials are now described. All publications mentioned herein are incorporated herein by reference.
[0024] The present disclosure is not to be limited in scope by the specific embodiments described herein, which are intended for the purposes of exemplification only. Functionally-equivalent products, compositions, and methods are clearly within the scope of the disclosure, as described herein.
[0025] The extended use of synthetic drugs for treating chronic inflammatory diseases are accompanied by many ill-effects some of which are increased risk of cardiac disorder, renal failure, gastrointestinal disorder, and risk of allergic reactions. More preferred alternatives are being investigated and naturally occurring sources are a promising alternative because of the fact of being relatively safer to use and less side effects associated with them. Plants produce many metabolites during their metabolism for various functions. For example, primary metabolites are made during various stages of plant growth. On the other hand, secondary metabolites, are made for defense against biotic or abiotic stresses. Plant secondary metabolites belonging to the terpenoids, alkaloids and flavonoids are currently used as drug or a dietary supplement to cure or prevent various diseases. The present invention discloses an anti-inflammatory composition made up of a combination of asparagus extract and red pearl onion extract. The composition disclosed in the invention display a synergistic anti-inflammatory effect by inhibiting phosphorylation of NF-?B and by inhibiting production of IL-8 (interleukin). It is suggested that the secondary metabolites present in the extract play an active role in the anti-inflammatory response.
[0026] The present document discloses a composition comprising asparagus extract and red pearl onion extract, wherein the asparagus extract and the red pearl onion extract w/w ratio is in the range of 5:1 to 4:5. Also disclosed are processes for preparing the composition. The composition displays a synergistic inhibition of NF-?B and IL-8 as compared to individual additive effect of the asparagus extract and the red pearl onion extract.
[0027] In an embodiment of the present disclosure, there is provided a composition comprising: (a) asparagus extract; and (b) red pearl onion extract, wherein the asparagus extract to the red pearl onion extract weight ratio in said composition is in the range of 5:1 – 4:5.
[0028] In an embodiment of the present disclosure, there is provided a composition comprising: (a) asparagus extract; and (b) red pearl onion extract, wherein the asparagus extract to the red pearl onion extract weight ratio in said composition is in the range of 5:1 – 4:5, and wherein the asparagus extract is having a weight percentage in the range of 0.003% to 0.01%, and the red pearl onion extract is having a weight percentage in the range of 0.001% to 0.006%.
[0029] In an embodiment of the present disclosure, there is provided a composition comprising: (a) asparagus extract; and (b) red pearl onion extract, wherein the asparagus extract to the red pearl onion extract weight ratio in said composition is in the range of 5:1 – 4:5, and wherein the asparagus extract is having a weight percentage in the range of 0.005% to 0.009%, and the red pearl onion extract is having a weight percentage in the range of 0.002% to 0.005%.
[0030] In an embodiment of the present disclosure, there is provided a composition comprising: (a) asparagus extract; and (b) red pearl onion extract, wherein the asparagus extract to the red pearl onion extract weight ratio in said composition is in the range of 4:1 – 1:1.
[0031] In an embodiment of the present disclosure, there is provided a composition comprising: (a) asparagus extract; and (b) red pearl onion extract, wherein the asparagus extract to the red pearl onion extract weight ratio in said composition is in the range of 4:1 – 1:1, and wherein the asparagus extract is having a weight percentage in the range of 0.003% to 0.01%, and the red pearl onion extract is having a weight percentage in the range of 0.001% to 0.006%.
[0032] In an embodiment of the present disclosure, there is provided a composition comprising: (a) asparagus extract; and (b) red pearl onion extract, wherein the asparagus extract to the red pearl onion extract weight ratio in said composition is in the range of 4:1 – 1:1, and wherein the asparagus extract is having a weight percentage in the range of 0.005% to 0.009%, and the red pearl onion extract is having a weight percentage in the range of 0.002% to 0.005%.
[0033] In an embodiment of the present disclosure, there is provided a composition comprising: (a) asparagus extract; and (b) red pearl onion extract, wherein the asparagus extract to the red pearl onion extract weight ratio in said composition is 4:1.
[0034] In an embodiment of the present disclosure, there is provided a composition comprising: (a) asparagus extract; and (b) red pearl onion extract, wherein the asparagus extract to the red pearl onion extract weight ratio in said composition is 4:1, and wherein the asparagus extract is having a weight percentage in the range of 0.003% to 0.01%, and the red pearl onion extract is having a weight percentage in the range of 0.001% to 0.006%.
[0035] In an embodiment of the present disclosure, there is provided a composition comprising: (a) asparagus extract; and (b) red pearl onion extract, wherein the asparagus extract to the red pearl onion extract weight ratio in said composition is 4:1, and wherein the asparagus extract is having a weight percentage in the range of 0.005% to 0.009%, and the red pearl onion extract is having a weight percentage in the range of 0.002% to 0.005%.
[0036] In an embodiment of the present disclosure, there is provided a composition comprising: (a) asparagus extract; and (b) red pearl onion extract, wherein the asparagus extract to the red pearl onion extract weight ratio in said composition is 3:2.
[0037] In an embodiment of the present disclosure, there is provided a composition comprising: (a) asparagus extract; and (b) red pearl onion extract, wherein the asparagus extract to the red pearl onion extract weight ratio in said composition is 3:2, and wherein the asparagus extract is having a weight percentage in the range of 0.003% to 0.01%, and the red pearl onion extract is having a weight percentage in the range of 0.001% to 0.006%.
[0038] In an embodiment of the present disclosure, there is provided a composition comprising: (a) asparagus extract; and (b) red pearl onion extract, wherein the asparagus extract to the red pearl onion extract weight ratio in said composition is 3:2, and wherein the asparagus extract is having a weight percentage in the range of 0.005% to 0.009%, and the red pearl onion extract is having a weight percentage in the range of 0.002% to 0.005%.
[0039] In an embodiment of the present disclosure, there is provided a composition comprising: (a) asparagus extract; and (b) red pearl onion extract, wherein the asparagus extract to the red pearl onion extract weight ratio in said composition is 1:1.
[0040] In an embodiment of the present disclosure, there is provided a composition comprising: (a) asparagus extract; and (b) red pearl onion extract, wherein the asparagus extract to the red pearl onion extract weight ratio in said composition is 1:1, and wherein the asparagus extract is having a weight percentage in the range of 0.003% to 0.01%, and the red pearl onion extract is having a weight percentage in the range of 0.001% to 0.006%.
[0041] In an embodiment of the present disclosure, there is provided a composition comprising: (a) asparagus extract; and (b) red pearl onion extract, wherein the asparagus extract to the red pearl onion extract weight ratio in said composition is 1:1, and wherein the asparagus extract is having a weight percentage in the range of 0.005% to 0.009%, and the red pearl onion extract is having a weight percentage in the range of 0.002% to 0.005%.
[0042] In an embodiment of the present disclosure, there is provided a composition comprising: (a) asparagus extract; (b) red pearl onion extract; and (c) at least one carrier, at least one diluent, at least one excipient, and combinations thereof, wherein the asparagus extract to the red pearl onion extract weight ratio in said composition is in the range of 5:1 - 4:5, and wherein, the at least one carrier, at least one diluent, and at least one excipient is selected from the group consisting of lactose-anhydrous, spray dried lactose, hydrolyzed starch, gelatine, polyethylene glycol, glycerol, lactates, polysorbates, sorbitan esters, simethicone, saccharin, aspartame, peppermint, lemon oils, butterscotch, methyl paraben, propyl paraben, sorbitol, carob bean gum, mannitol, gum tragacanth, guar gum, benzoic acid, sodium benzoate, garlic and oil of garlic, oil of rue, propyl gallate, gum ghatti, gum arabic, sterculia gum, clove bud extract, clove bud oil, clove bud oleoresin, clove leaf oil, clove stem oil, cholic acid, desoxycholic acid, glycocholic acid, ox bile extract, taurocholic acid, sorbose (packaging), sodium thiosulfate, mustard oil , glycyrrhiza, ammoniated glycyrrhizin, licorice, caprylic acid, stannous chloride, ammonium bicarbonate, ammonium carbonate, ammonium chloride, ammonium hydroxide, ammonium phosphate dibasic, ammonium phosphate monobasic, ammonium sulfate, calcium iodate, potassium iodate, potassium iodide, aconitic acid, calcium carbonate, potassium bicarbonate, sodium bicarbonate, sodium carbonate, sodium sesquicarbonate, glycerin and glycerides, dextran, dextrins, corn, dextrins, calcium acetate, calcium chloride, calcium gluconate, calcium phytate, calcium hydroxide, calcium oxide, succinic acid, butylated hydroxytoluene (BHT), ammonium phosphate dibasic calcium hexametaphosphate, calcium phosphate dibasic, calcium phosphate monobasic, calcium phosphate tribasic, calcium pyrophosphate, phosphoric acid, potassium phosphate dibasic, potassium phosphate monobasic, potassium phosphate tribasic, potassium polymetaphosphate, potassium pyrophosphate, potassium tripolyphosphate, sodium acid pyrophosphate, sodium hexametaphosphate, sodium metaphosphate, sodium phosphate dibasic, sodium phosphate monobasic, sodium phosphate tribasic, sodium pyrophosphate, tetrabasic, sodium tetrametaphosphate, sodium tetraphosphate, sodium trimetaphosphate, sodium tripolyphosphate, sulfuric acid, alpha-tocopherol acetate, tocopherols, choline bitartrate, choline chloride, aluminum ammonium sulfate, aluminum hydroxide, aluminum oleate, aluminum palmitate, aluminum potassium sulfate, aluminum sodium sulfate, aluminum sulfate, sodium aluminate, sodium aluminum phosphate, acidic, sodium aluminum phosphate, basic, sodium phosphoaluminate, beeswax, japan wax, carnauba wax, corn sugar, corn syrup, invert sugar, inositol, calcium stearate, hydrogenated tallow, stearic acid, tallow, malic acid, L-Malic acid, calcium sorbate, potassium sorbate, sodium sorbate, sorbic acid, sulfamic acid, sodium hydrosulfite zinc hydrosulfite, tall oil, fish oil, hydrogenated, sucrose, agar-agar, ammonium alginate, calcium alginate, potassium alginate, propylene glycol alginate, sodium alginate, propylene glycol, propylene glycol monostearate, brown algae, red algae, calcium glycerophosphate, manganese glycerophosphate, magnesium glycerophosphate, potassium glycerophosphate, potassium hydroxide, sodium hydroxide, potassium metabisulfite, sodium bisulfite, sodium metabisulfite, sodium sulphite, sulfur dioxide, magnesium phosphate, dibasic, magnesium carbonate, magnesium chloride, magnesium hydroxide, magnesium oxide, magnesium stearate, magnesium sulfate, magnesium phosphate, tribasic, adipic acid, hydrogenated soybean oil, ethyl formate, formic acid, sodium formate, carrageenan, nutmeg and mace, zinc acetate, zinc carbonate, zinc chloride, zinc oxide, zinc sulfate, caramel, lard, lard oil, papain, gum guaiac, coconut oil, linoleic acid, oleic acid, peanut oil, calcium hypophosphite, manganous hypophosphite, potassium hypophosphite, sodium hypophosphite, pectin, amidated, pectin, high ester, pectin, low acid, pectinates, pectinic acid, carboxymethyl cellulose, cellulose acetate, ethyl cellulose, hydroxypropylmethyl cellulose, methylcellulose, sodium carboxymethyl cellulose, rennet, tannic acid, acetic acid, sodium acetate, sodium diacetate, pyridoxine, pyridoxine hydrochloride, sodium oleate, sodium palmitate, ethyl acrylate, monomeric, methyl acrylate, monomeric, ethyl acrylate, polymeric, methyl acrylate, polymeric, bentonite, clay, corn silk, ammonium citrate, calcium citrate, citric acid, isopropyl citrate, potassium citrate, sodium citrate, stearyl citrate, triethyl citrate, biotin, enzymatically hydrolyzed casein, acid hydrolyzed proteins, enzymatically hydrolyzed protein, soy sauces, yeast autolyzates, caffeine, L-glutamic acid, L-glutamic acid hydrochloride, monoammonium L-glutamate, monopotassium L-glutamate, monosodium L-glutamate, calcium lactate, L(+)-calcium lactate, D(-)-lactic acid, lactic acid, L(+)-lactic acid, butylated hydroxyanisole (BHA), D- or DL- calcium pantothenate, D-pantothenyl alcohol, D- or DL- sodium pantothenate, urea, thiamine hydrochloride, thiamine mononitrate, magnesium gluconate, potassium gluconate, sodium gluconate, zinc gluconate, vitamin B12, vitamin D2, vitamin D3, potassium chloride, sodium chloride, soy protein isolate, hydrochloric acid, copper (cupric) gluconate, copper (cupric) sulfate, cuprous iodide, calcium caseinate, casein, sodium caseinate, aluminum calcium silicate, calcium silicate, diatomaceous earth, magnesium silicate, perlite, potassium silicate, silica aerogel, silicon dioxides, sodium aluminosilicate, sodium calcium aluminosilicate, sodium silicate, basic magnesium silicate, tricalcium silicate, L(+)-potassium acid tartrate, L(+)-sodium tartrate, L(+)-tartaric acid, manganous chloride, manganous citrate, manganous gluconate, manganous oxide, manganous sulfate, lecithin, lecithin, hydrogen peroxide bleached, riboflavin, riboflavin-5'-phosphate, calcium propionate, dilauryl thiodipropionate, propionic acid, sodium propionate, thiodipropionic acid, hydrogen peroxide, carbon dioxide, nickel, niacin, niacinamide, carotene, l-ascorbic acid, ascorbyl palmitate, calcium l-ascorbate, erythorbic acid, sodium erythorbate, sodium l-ascorbate, acetylated distarch adipate, acetylated distarch glycerol, acetylated distarch phosphate, acetylated distarch oxypropanol, acid modified starch, arrowroot starch, bleached starch, ,cornstarch, distarch glycerol, distarch oxypropanol, distarch phosphate, high amylose cornstarch, hydroxypropyl distarch glycerol, hydroxypropyl distarch phosphate, hydroxypropyl starch, hydroxypropyl starch, oxidized, milo starch, monostarch phosphate, potato starch, pregelatinized starch, rice starch, sodium hydroxide gelatinized starch, starch acetate, starch aluminum octenyl succinate, starch sodium succinate, starch sodium octenyl succinate, succinyl distarch glycerol, tapioca starch, waxy maize starch, wheat starch, phosphated distarch phosphate, starch, sodium hypochlorite oxidized, vitamin A, vitamin A acetate, vitamin A palmitate, diacetyl, starter distillate, carbonyl iron, carbonyl iron, electrolytic iron, ferric ammonium citrate, ferric chloride, ferric citrate, ferric oxide, ferric phosphate, ferric pyrophosphate, ferric sodium pyrophosphate, ferric sulfate, ferrous ascorbate, ferrous carbonate, ferrous citrate, ferrous fumarate, ferrous gluconate, ferrous lactate, ferrous sulfate, iron caprylate, iron linoleate, iron naphthenate, iron oxides, iron peptonate, iron polyvinylpyrrolidone, iron tallate, iron, elemental, reduced iron, reduced iron, sodium ferric EDTA, sodium ferricitropyrophosphate, dietary iron, ferric oxide, ammonium phosphate monobasic, potassium carbonate, calcium glycerophosphate, cellulose, cellulose, microcrystalline, and combinations thereof.
[0043] In an embodiment of the present disclosure, there is provided a composition comprising: (a) asparagus extract; (b) red pearl onion extract; and (c) at least one carrier, at least one diluent, at least one excipient, and combinations thereof, wherein the asparagus extract to the red pearl onion extract weight ratio in said composition is in the range of 5:1 - 4:5, and wherein the asparagus extract is having a weight percentage in the range of 0.003% to 0.01%, and the red pearl onion extract is having a weight percentage in the range of 0.001% to 0.006%.
[0044] In an embodiment of the present disclosure, there is provided a composition comprising: (a) asparagus extract; (b) red pearl onion extract; and (c) at least one carrier, at least one diluent, at least one excipient, and combinations thereof, wherein the asparagus extract to the red pearl onion extract weight ratio in said composition is in the range of 5:1 – 4:5, and wherein the asparagus extract is having a weight percentage in the range of 0.005% to 0.009%, and the red pearl onion extract is having a weight percentage in the range of 0.002% to 0.005%.
[0045] In an embodiment of the present disclosure, there is provided a composition comprising: (a) asparagus extract; (b) red pearl onion extract; and (c) at least one carrier, at least one diluent, at least one excipient, and combinations thereof, wherein the asparagus extract to the red pearl onion extract weight ratio in said composition is in the range of 4:1 - 1:1.
[0046] In an embodiment of the present disclosure, there is provided a composition comprising: (a) asparagus extract; (b) red pearl onion extract; and (c) at least one carrier, at least one diluent, at least one excipient, and combinations thereof, wherein the asparagus extract to the red pearl onion extract weight ratio in said composition is in the range of 4:1 - 1:1, and wherein the asparagus extract is having a weight percentage in the range of 0.003% to 0.01%, and the red pearl onion extract is having a weight percentage in the range of 0.001% to 0.006%.
[0047] In an embodiment of the present disclosure, there is provided a composition comprising: (a) asparagus extract; (b) red pearl onion extract; and (c) at least one carrier, at least one diluent, at least one excipient, and combinations thereof, wherein the asparagus extract to the red pearl onion extract weight ratio in said composition is 4:1.
[0048] In an embodiment of the present disclosure, there is provided a composition comprising: (a) asparagus extract; (b) red pearl onion extract; and (c) at least one carrier, at least one diluent, at least one excipient, and combinations thereof, wherein the asparagus extract to the red pearl onion extract weight ratio in said composition is 4:1, and wherein the asparagus extract is having a weight percentage in the range of 0.003% to 0.01%, and the red pearl onion extract is having a weight percentage in the range of 0.001% to 0.006%.
[0049] In an embodiment of the present disclosure, there is provided a composition comprising: (a) asparagus extract; (b) red pearl onion extract; and (c) at least one carrier, at least one diluent, at least one excipient, and combinations thereof, wherein the asparagus extract to the red pearl onion extract weight ratio in said composition is 4:1, and wherein the asparagus extract is having a weight percentage in the range of 0.005% to 0.009%, and the red pearl onion extract is having a weight percentage in the range of 0.002% to 0.005%.
[0050] In an embodiment of the present disclosure, there is provided a composition comprising: (a) asparagus extract; (b) red pearl onion extract; and (c) at least one carrier, at least one diluent, at least one excipient, and combinations thereof, wherein the asparagus extract to the red pearl onion extract weight ratio in said composition is 3:2.
[0051] In an embodiment of the present disclosure, there is provided a composition comprising: (a) asparagus extract; (b) red pearl onion extract; and (c) at least one carrier, at least one diluent, at least one excipient, and combinations thereof, wherein the asparagus extract to the red pearl onion extract weight ratio in said composition is 3:2, and wherein the asparagus extract is having a weight percentage in the range of 0.003% to 0.01%, and the red pearl onion extract is having a weight percentage in the range of 0.001% to 0.006%.
[0052] In an embodiment of the present disclosure, there is provided a composition comprising: (a) asparagus extract; (b) red pearl onion extract; and (c) at least one carrier, at least one diluent, at least one excipient, and combinations thereof, wherein the asparagus extract to the red pearl onion extract weight ratio in said composition is 3:2, and wherein the asparagus extract is having a weight percentage in the range of 0.005% to 0.009%, and the red pearl onion extract is having a weight percentage in the range of 0.002% to 0.005%.
[0053] In an embodiment of the present disclosure, there is provided a composition comprising: (a) asparagus extract; (b) red pearl onion extract; and (c) at least one carrier, at least one diluent, at least one excipient, and combinations thereof, wherein the asparagus extract to the red pearl onion extract weight ratio in said composition is 1:1.
[0054] In an embodiment of the present disclosure, there is provided a composition comprising: (a) asparagus extract; (b) red pearl onion extract; and (c) at least one carrier, at least one diluent, at least one excipient, and combinations thereof, wherein the asparagus extract to the red pearl onion extract weight ratio in said composition is 1:1, and wherein the asparagus extract is having a weight percentage in the range of 0.003% to 0.01%, and the red pearl onion extract is having a weight percentage in the range of 0.001% to 0.006%.
[0055] In an embodiment of the present disclosure, there is provided a composition comprising: (a) asparagus extract; (b) red pearl onion extract; and (c) at least one carrier, at least one diluent, at least one excipient, and combinations thereof, wherein the asparagus extract to the red pearl onion extract weight ratio in said composition is 1:1, and wherein the asparagus extract is having a weight percentage in the range of 0.005% to 0.009%, and the red pearl onion extract is having a weight percentage in the range of 0.002% to 0.005%.
[0056] In an embodiment of the present disclosure, there is provided a method for preparing a composition comprising: (a) asparagus extract; and (b) red pearl onion extract, said method comprising: (i) obtaining the asparagus extract; (ii) obtaining the red pearl onion extract; and (iii) contacting the asparagus extract, and the red pearl onion extract to obtain said composition, wherein the asparagus extract to the red pearl onion extract weight ratio in said composition is in the range of 5:1 - 4:5.
[0057] In an embodiment of the present disclosure, there is provided a method for preparing a composition comprising: (a) asparagus extract; and (b) red pearl onion extract, said method comprising: (i) obtaining the asparagus extract; (ii) obtaining the red pearl onion extract; and (iii) contacting the asparagus extract, and the red pearl onion extract to obtain said composition, wherein the asparagus extract to the red pearl onion extract weight ratio in said composition is in the range of 4:1 - 1:1.
[0058] In an embodiment of the present disclosure, there is provided a method for preparing a composition comprising: (a) asparagus extract; and (b) red pearl onion extract, said method comprising: (i) obtaining the asparagus extract; (ii) obtaining the red pearl onion extract; and (iii) contacting the asparagus extract, and the red pearl onion extract to obtain said composition, wherein the asparagus extract to the red pearl onion extract weight ratio in said composition is 4:1.
[0059] In an embodiment of the present disclosure, there is provided a method for preparing a composition comprising: (a) asparagus extract; and (b) red pearl onion extract, said method comprising: (i) obtaining the asparagus extract; (ii) obtaining the red pearl onion extract; and (iii) contacting the asparagus extract, and the red pearl onion extract to obtain said composition, wherein the asparagus extract to the red pearl onion extract weight ratio in said composition is 3:2.
[0060] In an embodiment of the present disclosure, there is provided a method for preparing a composition comprising: (a) asparagus extract; and (b) red pearl onion extract, said method comprising: (i) obtaining the asparagus extract; (ii) obtaining the red pearl onion extract; and (iii) contacting the asparagus extract, and the red pearl onion extract to obtain said composition, wherein the asparagus extract to the red pearl onion extract weight ratio in said composition is 1:1.
[0061] In an embodiment of the present disclosure, there is provided a method for preparing a composition comprising: (a) asparagus extract; (b) red pearl onion extract; (c) at least one carrier; (d) at least one diluent; and (e) at least one excipient, said method comprising: (i) obtaining the asparagus extract; (ii) obtaining the red pearl onion extract; (iii) obtaining the at least one carrier; (iv) obtaining the at least one diluent; (v) obtaining the at least one excipient, and (vi) contacting the asparagus extract, the red pearl onion extract, the at least one carrier, the at least one diluent, and the at least one excipient to obtain said composition, wherein the asparagus extract to the red pearl onion extract weight ratio in said composition is in the range of 5:1 - 4:5.
[0062] In an embodiment of the present disclosure, there is provided a method for preparing a composition comprising: (a) asparagus extract; and (b) red pearl onion extract, said method comprising: (i) obtaining the asparagus extract; (ii) obtaining the red pearl onion extract; and (iii) contacting the asparagus extract, and the red pearl onion extract to obtain said composition, wherein the asparagus extract to the red pearl onion extract weight ratio in said composition is in the range of 4:1 - 1:1.
[0063] In an embodiment of the present disclosure, there is provided a method for preparing a composition comprising: (a) asparagus extract; (b) red pearl onion extract; (c) at least one carrier; (d) at least one diluent; and (e) at least one excipient, said method comprising: (i) obtaining the asparagus extract; (ii) obtaining the red pearl onion extract; (iii) obtaining the at least one carrier; (iv) obtaining the at least one diluent; (v) obtaining the at least one excipient, and (vi) contacting the asparagus extract, the red pearl onion extract, the at least one carrier, the at least one diluent, and the at least one excipient to obtain said composition, wherein the asparagus extract to the red pearl onion extract weight ratio in said composition is 4:1.
[0064] In an embodiment of the present disclosure, there is provided a method for preparing a composition comprising: (a) asparagus extract; (b) red pearl onion extract; (c) at least one carrier; (d) at least one diluent; and (e) at least one excipient, said method comprising: (i) obtaining the asparagus extract; (ii) obtaining the red pearl onion extract; (iii) obtaining the at least one carrier; (iv) obtaining the at least one diluent; (v) obtaining the at least one excipient, and (vi) contacting the asparagus extract, the red pearl onion extract, the at least one carrier, the at least one diluent, and the at least one excipient to obtain said composition, wherein the asparagus extract to the red pearl onion extract weight ratio in said composition is 3:2.
[0065] In an embodiment of the present disclosure, there is provided a method for preparing a composition comprising: (a) asparagus extract; (b) red pearl onion extract; (c) at least one carrier; (d) at least one diluent; and (e) at least one excipient, said method comprising: (i) obtaining the asparagus extract; (ii) obtaining the red pearl onion extract; (iii) obtaining the at least one carrier; (iv) obtaining the at least one diluent; (v) obtaining the at least one excipient, and (vi) contacting the asparagus extract, the red pearl onion extract, the at least one carrier, the at least one diluent, and the at least one excipient to obtain said composition, wherein the asparagus extract to the red pearl onion extract weight ratio in said composition is 1:1.
[0066] In an embodiment of the present disclosure, there is provided a method for preparing a composition comprising: (i) asparagus extract; and (ii) red pearl onion extract, said method comprising: (a) obtaining a solution A of the asparagus extract; (b) obtaining a solution B of the red pearl onion extract; and (c) contacting solution A and solution B to obtain said composition, wherein the asparagus extract to the red pearl onion extract w/w ratio in said composition is in the range of 5:1-4:5.
[0067] In an embodiment of the present disclosure, there is provided a method for preparing a composition comprising: (i) asparagus extract; and (ii) red pearl onion extract, said method comprising: (a) obtaining a solution A of the asparagus extract; (b) obtaining a solution B of the red pearl onion extract; and (c) contacting solution A and solution B to obtain said composition, wherein the asparagus extract is having a weight percentage in the range of 0.003% to 0.01%, and wherein the red pearl onion extract is having a weight percentage in the range of 0.001%-0.006%, wherein the asparagus extract to the red pearl onion extract w/w ratio in said composition is in the range of 5:1-4:5.
[0068] In an embodiment of the present disclosure, there is provided a method for preparing a composition comprising: (i) asparagus extract; and (ii) red pearl onion extract, said method comprising: (a) obtaining a solution A of the asparagus extract; (b) obtaining a solution B of the red pearl onion extract; and (c) contacting solution A and solution B to obtain said composition, wherein the asparagus extract is having a weight percentage in the range of 0.003% to 0.01%, and wherein the red pearl onion extract is having a weight percentage in the range of 0.001%-0.006%, wherein the asparagus extract to the red pearl onion extract w/w ratio in said composition is in the range of 4:1-1:1.
[0069] In an embodiment of the present disclosure, there is provided a method for preparing a composition comprising: (i) asparagus extract; and (ii) red pearl onion extract, said method comprising: (a) obtaining a solution A of the asparagus extract; (b) obtaining a solution B of the red pearl onion extract; and (c) contacting solution A and solution B to obtain said composition, wherein the asparagus extract is having a weight percentage in the range of 0.003% to 0.01%, and wherein the red pearl onion extract is having a weight percentage in the range of 0.001%-0.006%, wherein the asparagus extract to the red pearl onion extract w/w ratio in said composition is 4:1.
[0070] In an embodiment of the present disclosure, there is provided a method for preparing a composition comprising: (i) asparagus extract; and (ii) red pearl onion extract, said method comprising: (a) obtaining a solution A of the asparagus extract; (b) obtaining a solution B of the red pearl onion extract; and (c) contacting solution A and solution B to obtain said composition, wherein the asparagus extract is having a weight percentage in the range of 0.003% to 0.01%, and wherein the red pearl onion extract is having a weight percentage in the range of 0.001%-0.006%, wherein the asparagus extract to the red pearl onion extract w/w ratio in said composition is 3:2.
[0071] In an embodiment of the present disclosure, there is provided a method for preparing a composition comprising: (i) asparagus extract; and (ii) red pearl onion extract, said method comprising: (a) obtaining a solution A of the asparagus extract; (b) obtaining a solution B of the red pearl onion extract; and (c) contacting solution A and solution B to obtain said composition, wherein the asparagus extract is having a weight percentage in the range of 0.003% to 0.01%, and wherein the red pearl onion extract is having a weight percentage in the range of 0.001%-0.006%, wherein the asparagus extract to the red pearl onion extract w/w ratio in said composition is 1:1.
[0072] In an embodiment of the present disclosure, there is provided a method as described herein, wherein the asparagus extract and the red pearl onion extract is an aqueous/methanolic extract. In another embodiment, the asparagus extract and the red pearl onion extract is a chloroform extract. It is described in the example that the aqueous/methanolic extract of the asparagus extract and the red pearl onion extract exhibits a higher anti-inflammatory effect as compared to the chloroform extract.
[0073] In an embodiment of the present disclosure, there is provided a composition for use in preparing formulations, said composition comprising: (a) asparagus extract; and (b) red pearl onion extract, wherein the asparagus extract to the red pearl onion extract weight ratio in said composition is in the range of 5:1 - 4:5.
[0074] In an embodiment of the present disclosure, there is provided a composition for use in preparing formulations, said composition comprising: (a) asparagus extract; and (b) red pearl onion extract, wherein the asparagus extract is having a weight percentage in the range of 0.003% to 0.01%, and wherein the red pearl onion extract is having a weight percentage in the range of 0.001%-0.006%, and wherein the asparagus extract to the red pearl onion extract w/w ratio in said composition is in the range of 5:1-4:5.
[0075] In an embodiment of the present disclosure, there is provided a composition for use in preparing formulations, said composition comprising: (a) asparagus extract; and (b) red pearl onion extract, wherein the asparagus extract to the red pearl onion extract weight ratio in said composition is in the range of 4:1 - 1:1.
[0076] In an embodiment of the present disclosure, there is provided a composition for use in preparing formulations, said composition comprising: (a) asparagus extract; and (b) red pearl onion extract, wherein the asparagus extract is having a weight percentage in the range of 0.003% to 0.01%, and wherein the red pearl onion extract is having a weight percentage in the range of 0.001%-0.006%, and wherein the asparagus extract to the red pearl onion extract w/w ratio in said composition is in the range of 4:1-1:1.
[0077] In an embodiment of the present disclosure, there is provided a composition for use in preparing formulations, said composition comprising: (a) asparagus extract; and (b) red pearl onion extract, wherein the asparagus extract to the red pearl onion extract weight ratio in said composition is 4:1.
[0078] In an embodiment of the present disclosure, there is provided a composition for use in preparing formulations, said composition comprising: (a) asparagus extract; and (b) red pearl onion extract, wherein the asparagus extract is having a weight percentage in the range of 0.003% to 0.01%, and wherein the red pearl onion extract is having a weight percentage in the range of 0.001%-0.006%, and wherein the asparagus extract to the red pearl onion extract w/w ratio in said composition is 4:1.
[0079] In an embodiment of the present disclosure, there is provided a composition for use in preparing formulations, said composition comprising: (a) asparagus extract; and (b) red pearl onion extract, wherein the asparagus extract to the red pearl onion extract weight ratio in said composition is 3:2.
[0080] In an embodiment of the present disclosure, there is provided a composition for use in preparing formulations, said composition comprising: (a) asparagus extract; and (b) red pearl onion extract, wherein the asparagus extract is having a weight percentage in the range of 0.003% to 0.01%, and wherein the red pearl onion extract is having a weight percentage in the range of 0.001%-0.006%, and wherein the asparagus extract to the red pearl onion extract w/w ratio in said composition is 3:2.
[0081] In an embodiment of the present disclosure, there is provided a composition for use in preparing formulations, said composition comprising: (a) asparagus extract; and (b) red pearl onion extract, wherein the asparagus extract to the red pearl onion extract weight ratio in said composition is 1:1.
[0082] In an embodiment of the present disclosure, there is provided a composition for use in preparing formulations, said composition comprising: (a) asparagus extract; and (b) red pearl onion extract, wherein the asparagus extract is having a weight percentage in the range of 0.003% to 0.01%, and wherein the red pearl onion extract is having a weight percentage in the range of 0.001%-0.006%, and wherein the asparagus extract to the red pearl onion extract w/w ratio in said composition is 1:1.
[0083] In an embodiment of the present disclosure, there is provided a composition as described herein, wherein said composition exhibits anti-inflammatory effect.
[0084] In an embodiment of the present disclosure, there is provided a composition as described herein, wherein said composition exhibits anti-inflammatory effect by inhibiting production of Interleukin 8 (IL-8).
[0085] In an embodiment of the present disclosure, there is provided a composition as described herein, wherein the asparagus extract comprises 4H-pyran-4-one, 2,3-dihydro-3,5-dihydroxy-6-methyl (C6H8O4) in a weight percentage in the range of 1% to 30% with respect to the extract. In another embodiment, the asparagus extract comprises 4H-pyran-4-one, 2,3-dihydro-3,5-dihydroxy-6-methyl (C6H8O4) in a weight percentage in the range of 5% to 30% with respect to the extract.
[0086] In an embodiment of the present disclosure, there is provided a composition as described herein, wherein the asparagus extract comprises 1,2-Dithiolane-3-carboxylic in a weight percentage in the range of 1% to 20% with respect to the extract.
[0087] In an embodiment of the present disclosure, there is provided a composition as described herein, wherein the asparagus extract comprises 4H-pyran-4-one, 2,3-dihydro-3,5-dihydroxy-6-methyl (C6H8O4) in a weight percentage in the range of 1% to 30% with respect to the extract, and 1,2-Dithiolane-3-carboxylic in a weight percentage in the range of 1% to 20% with respect to the extract.
[0088] In an embodiment of the present disclosure, there is provided a composition as described herein, wherein the red pearl onion extract comprises 5-hydroxymethylfurfural in a weight percentage in the range of 1% to 60% with respect to the extract.
[0089] In an embodiment of the present disclosure, there is provided a composition as described herein, wherein the red pearl onion extract comprises furfural in a weight percentage in the range of 1% to 20% with respect to the extract.
[0090] In an embodiment of the present disclosure, there is provided a composition as described herein, wherein said composition can be in the form of an aerosol.
[0091] In an embodiment of the present disclosure, there is provided a composition as described herein, wherein said composition can be in the form of a lotion.
[0092] In an embodiment of the present disclosure, there is provided a composition as described herein, wherein said composition can be in form of an oil.
[0093] In an embodiment of the present disclosure, there is provided a composition as described herein, wherein said composition can be in the form of a serum.
EXAMPLES
[0094] The disclosure will now be illustrated with working examples, which is intended to illustrate the working of disclosure and not intended to be taken restrictively or to imply any limitations on the scope of the present disclosure. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood to one of ordinary skill in the art to which this disclosure belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice of the disclosed methods and compositions, the exemplary methods, devices and materials are described herein. It is to be understood that this disclosure is not limited to particular methods, and experimental conditions described, as such methods and conditions may apply.
[0095] Although the subject matter has been described in considerable detail with reference to certain examples and implementations thereof, other implementations are possible.
[0096] The examples describe the fingerprint analysis of asparagus extract and red pearl onion extract followed by the compositions comprising different w/w ratios of both the extracts. The anti-inflammatory effects of the compositions have been shown in terms of their ability to inhibit IL-8 production and to inhibit phosphorylation of NF?B, thus highlighting their ability to inhibit inflammatory response.
Example 1
Fingerprint analysis of asparagus extract and red pearl onion extract
Asparagus extract
[0097] The asparagus extract and red pearl onion extract were subjected to gas chromatography/mass spectrometry analysis (GCMS). The main purpose was to characterize the presence of secondary metabolites in the extract. Figure 1 depicts a chromatogram of the asparagus extract and Table 1 depicts a concise form of report of the peak profile. One of the secondary metabolite - 4H-pyran-4-one, 2,3-dihydro-3,5-dihydroxy-6-methyl (C6H8O4) is present in a weight percentage in the range of 1% to 30% with respect to the asparagus extract. One other secondary metabolite to be present in higher quantities is 1,2-Dithiolane-3-carboxylic present in a weight percentage in the range of 1% to 20%. It can be appreciated that the asparagus extract has a high concentration of secondary metabolite 4H-pyran-4-one, 2,3-dihydro-3,5-dihydroxy-6-methyl (C6H8O4) representing a peak area of 28.57%. The asparagus extract disclosed in the present disclosure was found to have a higher percentage of C6H8O4 as compared to a report published previously (Vembarasi et al., 2013 International Journal of Natural Products Research. 2(1): 17-19) which was found to have only 1.95% of C6H8O4. The presence of some of the other active ingredients in the extract are enlisted in Table 1.
Table 1: GCMS report of asparagus extract
Peak# R. Time Area Area% Height Name Base m/z
1 3.782
97272 0.34 41457 2-Propenoic acid, 2-methyl- 86.00
2 4.369
166285 0.57 60250 Pyrazine, methyl- 94.00

3 4.559 2
173502 0.60 71376 -Furancarboxylic acid, 4-methylpentyl ester 96.00
4 5.019
437981 1.51 115410 2-Furanmethanol 98.00
5 5.252
326679 1.13 100657 2-Propenamide 71.05
6 5.624
98360 0.34 31265 4-Cyclopentene-1,3-dione 96.00
7 6.083
82647 0.29 32142 Methional 104.00
8 6.219 189548 0.65 81440 Cyclopropane, 2-(1,1-dimethyl-2-pentenyl)-1,1-dimethyl-
95.00
9 6.261
324878 1.12 123576 Butanoic acid, 4-hydroxy-
86.00
10 6.325
128305 0.44 47718 2(3H)-Furanone 55.05
11 6.526

147571 0.51 47119 2-Ethylbutyl propionate
57.05
12 6.866
88320 0.30 27518 1-Butanamine, 2-methyl-N-(2-methylbutylidene)-
98.10
13 6.960
59457 0.21 26242 1H-Azepine, hexahydro-3,3,5-trimethyl- 56.05
14 7.355
244424 0.84 48385 2-Furanmethanol, 5-methyl- 95.05
15 7.529
210955 0.73 82883 2-Furancarboxaldehyde, 5-methyl-
110.00
16 7.928
420196 1.45 178801 2,4-Dihydroxy-2,5-dimethyl-3(2H)-furan-3-one
101.00
17 8.022
917360 3.16 122298 Silane, [3-(2,3-epoxypropoxy)propyl]ethoxydimethyl- 103.00
18 8.389 157256 0.54 49716 Cyclohexane, 1,4-dimethyl-, trans-
55.00
19 8.582
473882 1.63 121735 Cyclohexane, bromo- 83.05
20 9.477
53935 0.19 25693 1-Butanamine, 2-methyl-N-(2-methylbutylidene)-
98.10
21 9.722
685737 2.37 218942 N-carbobenzyloxy-glycyl-proline 91.05
22 9.885
54117 0.19 26148 2-Thiazolamine, 4,5-dihydro- 102.05
23 10.236
818409 2.82 187926 2,5-Dimethyl-4-hydroxy-3(2H)-furanone 128.05
24 10.464
81741 0.28 27644 2-Pyrrolidinone
85.05
25 10.670
47636 0.16 16703 Erythro-dl-O-ethylthreonine 120.00
26 10.816
518537 1.79 106075 Thymine
126.00
27 10.919 262792 0.91 71719 2,4,5-Trihydroxypyrimidine 128.00
28 10.995 82978 0.29 31819 Thymidine, 3',5'-bis(trifluoroacetate)
126.00
29 11.249 801113 2.76 134098 1-Oxetan-2-one, 4-methyl-3-methylene-
83.05
30 11.399
149614 0.52 30909 1,2-Cyclobutanedicarboxylic acid, trans- 126.00
31 12.167
208986 0.72 66063 3-Hexanone, 2,4-dimethyl-
57.05
32 12.464
268688 0.93 83871 Ethanamine, N-ethyl-N-nitroso-
102.05
33 12.694
8281966 28.57 2054965 4H-Pyran-4-one, 2,3-dihydro-3,5-dihydroxy-6-methyl-
144.00
34 13.194 442421 1.53 146852 3-[N'-(3H-Indol-3-ylmethylene)-hydrazino]-5-methyl-[1,2,4]triazol 142.00
35 13.336
1957835 6.75 300623 3-[N'-(3H-Indol-3-ylmethylene)-hydrazino]-5-methyl-[1,2,4]triazol- 142.05
36 13.560
492345 1.70 115627 5-Methoxypyrrolidin-2-one 84.05
37 13.660
349936 1.21 48520 (S)-5-Hydroxymethyl-2[5H]-furanone 84.00
38 13.797
306261 1.06 94014 4H-Pyran-4-one, 3,5-dihydroxy-2-methyl- 142.00
39 14.889
1246607 4.30 195670 2-Oxepanone, 7-methyl-
55.00
40 15.116
505630 1.74 83207 5-Hydroxymethylfurfural
97.00
41 16.702
776177 2.68 101552 beta.-Alanine, N-acryloyl-, isobutyl ester 55.00
42 17.355
294879 1.02 58121 N-(2-Hydroxyethyl)-2-pyrrolidone 98.05
43 19.595 520935 1.80 110171 3-[N'-(3H-Indol-3-ylmethylene)-hydrazino]-5-methyl-[1,2,4]triazol- 142.05
44 20.490
155356 0.54 24219 2,3,4,4-Tetramethyl-pentane-1,3-diol 101.00
45 20.696
3378903 11.66 749761 1,2-Dithiolane-3-carboxylic acid 149.95
46 24.390
116756 0.40 34310 Diethyl Phthalate 149.00
47 24.470 165786 0.57 52345 Ethyl N-(o-anisyl)formimidate
179.05
48 28.950
71799 0.25 27464 Cyclopentanone, 2-(3-dimethylaminoprop-2-enylideno)- 165.00
49 29.188
82952 0.29 25140 Pyrrolo[1,2-a]pyrazine-1,4-dione, hexahydro-3-(2-methylpropyl)- 154.05
50 29.406
102799 0.35 27352 4(1H)-Pteridinone, 2-amino-6-methyl- 177.05
51 31.491
129128 0.45 45415 Pyrrolo[1,2-a]pyrazine-1,4-dione, hexahydro-3-(2-methylpropyl)- 154.05
52 32.116 97186 0.34 38083 Pentadecanoic acid 73.05
53 35.384 305960 1.06 48290 8-Methoxy-1,3,4,5-tetrahydro-2H-1-benzazepin-2-one
136.00
54 40.699
131574 0.45 47256 (2,3-Diphenylcyclopropyl)methyl phenyl sulfoxide, trans- 91.05
55 42.286
99139 0.34 33919 (2,3-Diphenylcyclopropyl)methyl phenyl sulfoxide, trans- 91.05
56 42.533 198863 0.69 61498 (2,3-Diphenylcyclopropyl)methyl phenyl sulfoxide, trans- 91.05
28990354 100.00 6961972

Red pearl onion extract
[0098] The red pearl onion extract was subjected to GCMS analysis for identification of major actives present in the extract. Figure 2 depicts a chromatogram of the red pearl onion extract and Table 2 depicts a concise report of the peak profile. One secondary metabolite - 5-hydroxymethylfurfural is present in a weight percentage in the range of 1% to 60% with respect to the extract, one other secondary metabolite – furfural is present in a weight percentage in the range of 1% to 20% with respect to the extract. Referring to Table 2, it can be appreciated that 5-hydroxymethylfurfural (C6H6O3) is a major active present in the extract representing around 55% of the peak area. Table 2 enlists the presence of other actives in the red pearl onion extract.
Table 2: GCMS report of red pearl onion extract
Peak# R. Time Area Area% Height Name Base m/z
1 4.389
238680 0.11 81376 Homoserine 74.05
2 4.577
9847649 4.42 1372132 Furfural 96.00
3 5.018
4033971 1.81 632563 2-Furanmethanol 98.05

4 5.616 3311786 1.49 348469 1,5-Pentanedithiol
102.00
5 6.069 1161828 0.52 291591 Thiophene, 2,5-dimethyl-
111.00
6 6.558 368672 0.17 84044 Glutamine, N-methyl-
98.00
7 6.911 288606 0.13 122667 1,3-Dithiane
120.00
8 7.354 422641 0.19 124267 2-Furanmethanol, 5-methyl-
95.05
9 7.541 2170524 0.97 445309 2-Furancarboxaldehyde, 5-methyl-
110.00
10 7.692 613116 0.28 185234 Dimethyl trisulfide
125.95
11 7.932 2380821 1.07 602533 2,4-Dihydroxy-2,5-dimethyl-3(2H)-furan-3-one
101.00
12 8.393 1185838 0.53 151586 Triethylenediamine
55.00
13 9.728 767006 0.34 296560 Benzeneacetaldehyde
91.05
14 9.891 385594 0.17 130201 2-Methyl-4-(methylthio)-1-phenyl-but-3-en-1-ol
102.00
15 10.326 2497050 1.12 493403 2,5-Dimethyl-4-hydroxy-3(2H)-furanone
128.05
16 10.693 424751 0.19 130869 2,5-Furandicarboxaldehyde
124.00
17 10.915 6468840 2.90 834705 2,2-Dimethylpropyl ethylphosphonofluoridate
126.00
18 12.294 2067952 0.93 237925 1-Butanol, 3-methyl-, formate
55.05
19 12.550 1903432 0.85 466547 Ethanamine, N-ethyl-N-nitroso-
102.00
20 12.770 2125895 9.54 3547077 4H-Pyran-4-one, 2,3-dihydro-3,5-dihydroxy-6-methyl-
144.00
21 13.086 152504 0.07 67323 1-Propene, 1-(methylthio)-, (Z)-
88.00
22 13.225 636483 0.29 136558 2,3,4-Trimethyllevoglucosan
88.00
23 13.322 160020 0.07 63681 1-Methyl-1-(3-methylbutyl)oxy-1-silacyclobutane
142.00
24 13.778 399776 0.18 123855 L-Threonine, ethyl ester
74.05
25 13.874 1631299 0.73 453100 4H-Pyran-4-one, 3,5-dihydroxy-2-methyl-
142.00
26 14.137 578900 0.26 144696 (S)-5-Hydroxymethyl-2[5H]-furanone
84.00
27 14.992 1549392 0.70 377044 5-Acetoxymethyl-2-furaldehyde
126.00
28 15.487 124508037 55.88 8620721 5-Hydroxymethylfurfural
97.00
29 15.829 2887532 1.30 273217 Butanoic acid, 3-oxo-, hexyl ester
103.05
30 16.116 3654837 1.64 323721 Ethyl 1-methylcyclopropanecarboxylate
100.05
31 16.375 1193919 0.54 125404 3-Isopropylidene-5-methyl-hex-4-en-2-one
137.00
32 17.006 2799732 1.26 171472 3-Acetoxypentadecane
55.05
33 17.232 1796302 0.81 217662 5-Acetoxymethyl-2-furaldehyde
126.00
34 17.630 2207872 0.99 357594 2H-Pyran-2,4-dione, tetrahydro-3,5,5-trimethyl-6-propyl-
126.00
35 18.088 379270 0.17 100726 2,3-O-Benzylidene-.beta.-d-ribose
114.00
36 18.335 1083606 0.49 159332 D-Gluconic acid, .gamma.-lactone
114.05
37 18.426 1053734 0.47 149069 Adipic acid, cis-hex-3-enyl ethyl ester
111.05
38 18.706 542393 0.24 132435 2,3,5,6-Tetrafluorophenol
166.00
39 19.606 518526 0.23 97513 3-[N'-(3H-Indol-3-ylmethylene)-hydrazino]-5-methyl-[1,2,4]triazol 142.05
40 20.765 785137 0.35 216654 4-tert-Butylcyclohexyl methyl ethylphosphonate 125.00
41 23.832 264386 0.12 58306 Phenol, 4-methoxy-3-(methoxymethyl)- 135.05
42 24.490 337616 0.15 115153 Ethyl N-(o-anisyl)formimidate 179.05
43 24.605 1168115 0.52 364547 Isophoronediamine
68.05
44 25.264 224269 0.10 59418 1,7-Dimethylxanthine
68.05
45 26.419 1476688 0.66 188344 Hydrazinecarboxamide, 2-(2-methylcyclohexylidene)- 169.05
46 26.813 727777 0.33 98122 1,4-Dioxaspiro[4,5]decane, (E)-2,3-dimethyl-7-t-butyl- 169.00
47 27.045 1603380 0.72 195878 2,4-Hexadienoic acid, tert-butyldimethylsilyl ester, (E,E)- 169.05
48 29.670 1005256 0.45 133373 Pyrimidine-2,4(1H,3H)-dione, 6-hydroxy-5-methyliminomethyl- 169.05
49 30.432 416716 0.19 81514 2,3,3-Trimethyl-2-(4-methylpentanoyl)-cyclopentanone 111.05
50 30.787 297939 0.13 61968 Succinic acid, dodec-9-yn-1-yl nonyl ester 101.00
51 32.131 1145856 0.51 189287 3-(1,3,5-Cycloheptatrien-7-yl)-2,4-pentanedione 91.05

52 32.404 718256 0.32 88458 Borinic acid, diethyl-, 5-hexynyl ester 81.00
53 32.703 1384677 0.62 206200 2-Cyclohexene-1-carboxylic acid, 1-methyl-4-oxo-, ethyl ester 182.05
54 33.689 312472 0.14 69004 Perimidine, 2-ethyl-
195.05
55 37.703 200837 0.09 54603 Diethylphenylphosphine 109.00
56 37.930 411048 0.18 59640 10-Methyl-8-tetradecen-1-ol acetate 111.05
57 38.579 786740 0.35 87025 Phenyl palmitate
239.00
222799015 100.00 25001675

Example 2
Preparation of asparagus extract and red pearl onion extract and its effect on DPPH radical scavenging activity
[0099] The asparagus and red pearl onion used for preparing the respective extracts were cultivated in Karnataka, India and obtained from local vegetable vendors. The Metabolite extraction from asparagus and red pearl onion were performed using different solvent systems. A mixture of 40% methanol, 40% chloroform and 20% water (v/v/v) was used as a solvent system, and the ratio was 1 g of grinded asparagus or red pearl onion to 10 mL of solvent. Samples were vortexed for 1 min and maintained in an orbital shaker for 15 min at room temperature. Samples were centrifuged at 1000g for 10 min for phase separation: the lower chloroform fraction and the upper aqueous/methanol fraction. Both fractions were further centrifuged for 5 min at 10,000g to remove debris and evaporated at 37° C. Subsequently, these were dissolved with DMSO and stored at -20º C until further analysis.
[00100] The chloroform fraction and the aqueous/methanol fraction of asparagus extract and red pearl onion extract were independently tested for radical scavenging activity (DPPH value) for assessing their anti-inflammation ability.
DPPH radical scavenging activity
[00101] DPPH (2,2-diphenyl-1-picrylhydrazyl) is a well-known radical and it also functions as a radical scavenger and is used as an indicator to detect the presence of radical scavenging activity of a compound. DPPH radical scavenging activity was determined according to the method of Blois, 1958. Nature, 26:1199-200 and Chang et al., 2006. Tajen J, 28:17-36. Briefly, 1 ml of 0.1 mM DPPH radical solution was mixed with 3 ml of different concentrations of different extracts of asparagus and red pearl onion. The mixture was then thoroughly vortexed and left in dark for 30 min at 40°C. For the baseline control, 3 ml of methanol was used. The absorbance was measured at 517 nm using an ELISA reader (Thermo Fisher, Germany) and the DPPH value in terms of percentage was calculated.
[00102] The chloroform fraction and aqueous/methanol fraction of asparagus and red pearl onion extract were tested for DPPH radical scavenging activity. Figure 3 depicts the radical scavenging activity of all the extracts, it can be appreciated that the methanol fraction of asparagus extract and red pearl onion extract is showing enhanced radical scavenging activity or anti-oxidant activity as compared to the chloroform fraction. Since, the aqueous/methanol fraction is displaying anti-oxidant effects and it is known that damage from free radicals is one of the causes of inflammation, therefore, the aqueous/methanol fraction of asparagus extract and red pearl onion extract was further tested for their anti-inflammation activity. The aqueous/methanol extract of the asparagus extract or the red pearl onion extract were subjected to fingerprint analysis to identify the actives, the results are depicted in Example 1.
Example 3
Determining cytotoxicity of asparagus extract and red pearl onion extract
[00103] The methanol fraction of asparagus extract and red pearl onion extract were independently tested for cytotoxicity on human PBMC (peripheral bone marrow cells).
Cytotoxicity assay - Human PBMC were treated with various concentrations of secondary metabolites containing extracts from asparagus and red onion. The cytotoxicity was determined by reduction of Thiazolyl Blue Tetrazolium Blue (MTT, Sigma) to formazan. Cells were seeded in 96 well culture plates and cultured for 24h. Cells were treated with various concentrations of the extracts and incubated for 24 hours. After 24 hours, the cells were analyzed for cytotoxicity by MTT assay. MTT (2 mg/ml in PBS, 100 µl) was added to each well. Cells were incubated at 37°C for 30 min and DMSO (100 µl) was added to dissolve the formazan crystals. The absorbance was measured at 560 nm using an ELISA reader (Thermo Fisher, Germany).
[00104] The concentrations of both the extracts ranging from 10µg/ml to 1000µg/ml were independently tested for their cytotoxicity on human PBMCs. Figure 4 depicts the cytotoxicity of the asparagus extract, it can be appreciated that no distinct cytotoxicity is observed till 500µg/ml but cytotoxic effects of the extract is observed above 500µg/ml.
[00105] Figure 5 depicts the cytotoxicity of the red pearl onion extract, it can be appreciated that distinct cytotoxicity is observed above 250µg/ml of the extract. Several commercial drugs suffer from poor bio-tolerance, the cell-viability studies indicate the high bio-tolerance of these natural extracts.
Example 4
Dose response study of asparagus extract and red pearl onion extract in inhibiting IL-8 production
[00106] IL-8 is a well-known pro-inflammatory cytokine, thus, the anti-inflammatory ability of asparagus extract and red pearl onion extract was tested on the inhibition of IL-8 production.
IL-8 cytokine estimation - Human PBMC were treated with secondary metabolites for 24 hours. To quantify IL-8 production, culture supernatants were collected and the levels of secreted cytokines were determined using an ELISA kit (Krishgen BioSystems, India).
[00107] Figure 6 depicts the effect of different concentrations (20µg/ml, 40µg/ml, 50µg/ml, 60µg/ml, and 80µg/ml) of asparagus extract on IL-8 production inhibition. A well-known anti-inflammatory molecule – curcumin was used as a positive control in the present example. It can be appreciated that the inhibition of IL-8 production varies from 11.8% (20µg/ml of asparagus extract) to a maximum of 43.1% in case of 80µg/ml of asparagus extract. The effect of red pearl onion extract was further tested.
[00108] Figure 7 depicts the effect of different concentrations (20µg/ml, 40µg/ml, 50µg/ml, 60µg/ml, and 80µg/ml) of red pearl onion extract on IL-8 production inhibition. The inhibitory effect varies from 13.9% for 20µg/ml red pearl onion extract to 41.6% for 80µg/ml red pearl onion extract.
[00109] The inhibition displayed by curcumin is the highest (68.5%) in both the cases. The effects of various combinations of asparagus extract and red pearl onion extract was further tested on the ability to inhibit IL-8 production.
Example 5
Effect of combinations of asparagus extract and red pearl onion extract in inhibiting IL-8 production
[00110] It was prudent to investigate the anti-inflammatory effects of various combinations of asparagus extract and red pearl onion extract on inhibition of IL-8 production. Table 3 enlists the various combinations those were tested for anti-inflammatory effects.
Table 3: Combinations of asparagus extract and red pearl onion extract
Ratios Asparagus Red pearl onion
Asp: Po (80:20) – 4:1 (w/w) (80 µg/ml) (20 µg/ml)
Asp: Po (60:40) – 3:2 (w/w) (60 µg/ml) (40 µg/ml)
Asp: Po (50:50) – 1:1 (w/w) (50 µg/ml) (50 µg/ml)
Asp: Po (40:60) – 2:3 (w/w) (40 µg/ml) (60 µg/ml)
Asp: Po (20:80) – 1:4 (w/w) (20 µg/ml) (80 µg/ml)

[00111] Five different combinations of asparagus extract and red pearl onion extract was used to test the anti-inflammatory effects. The composition comprising asparagus extract and red pearl onion extract in a w/w ratio of 4:1, 3:2, 1:1. 2:3, and 1:4 were used in the experiment.
[00112] The assay to quantitate IL-8 levels was as described in Example 4. Figure 8 depicts the inhibition of IL-8 production using different combinations of both the extracts as described previously. The additive effects of the extracts (Figure 6 and Figure 7) were compared to the combinatorial inhibitory effect (Figure 8) of different weigh ratios, and the analysis has been depicted in Table 4:
Table 4: Effect of combinations of asparagus extract and red pearl onion extract in inhibiting IL-8 production
Individual Dosage Individual inhibitory effect Combination inhibitory effect
Asparagus Red Pearl Onion Asparagus
(Data from Figure 6) Red Pearl onion
(Data from Figure 7) Additive Synergy
(Data from Figure 8)
80 µg/ml 20 µg/ml 43.1% 13.9% 57% 62.9% (4:1)
60 µg/ml 40 µg/ml 36.3% 18.8% 55.1% 61.9%(3:2)
50 µg/ml 50 µg/ml 26.7% 21.3% 48% 57.2% (1:1)
40 µg/ml 60 µg/ml 24.3% 32.9% 57.2% 53.1%(2:3)
20 µg/ml 80 µg/ml 11.8% 41.6% 53.4% 44.7% (1:4)

[00113] It can be inferred from Figure 8 and Table 4 that the compositions comprising asparagus extract and red pearl onion extract in a w/w ratio of 4:1, 3:2 and 1:1 are showing a synergistic effect in inhibiting the production of IL-8. Table 4 clearly illustrates that the inhibitory effect exhibited by the compositions in a w/w ratio of 4:1, 3:2, and 1:1 is more than the additive effects of individual components therefore, exhibiting clear synergy. The composition comprising asparagus extract and red pearl onion extract in a w/w ratio of 4:1 exhibits 62.9% inhibition of IL-8 production, and the compositions with w/w ratio 3:2 and 1:1 exhibits 61.9% and 57.2% inhibition respectively. The mentioned combinations were found to perform comparably to curcumin in terms of their anti-inflammatory activity. Whereas, the compositions comprising asparagus extract and red pearl onion extract in a w/w ratio of 2:3, and 1:4 exhibits 53.1% and 44.7% inhibition of IL-8 production. As can be inferred from Table 4 and Figure 8 that these two compositions fail to display any synergistic effect. Therefore, considerable experimentation was required to show synergistic effect of a composition comprising asparagus extract and red pearl onion extract.

Example 6
Dose response study of Asparagus extract and red pearl onion extract in inhibiting NF?B phosphorylation
[00114] In the present example, the anti-inflammation ability of both the extracts by regulating the master regulator NF?B (nuclear factor ?B) was determined. The inhibition of NF?B phosphorylation is considered as an anti-inflammatory effect since NF?B is a pro-inflammatory molecule and its activity increases the downstream effects of inflammation.
NF?B p65 (RelA) ser536 assay – The assay is used to determine the phosphorylation of NF?B. Human PBMC were seeded into each well of 12-well plates until 80% confluent. NF?B p65 ser 536 phosphorylation was stimulated with (10 µg/ml) lipopolysaccharides (LPS). This treatment induces the inflammatory response within 90 minutes of stimulation. Simultaneously, the cells were treated with secondary metabolites. Detection of NF?B p65 ser 536 phosphorylation was done using sandwich ELISA (Cell Signalling Inc). Briefly, media was removed from the cells and the cells were rinsed with ice cold 0.1M phosphate buffer saline (1X). Cells were lysed with 0.5 mL ice cold 1X cell lysis buffer (20mM Tris-HCl (pH 7.5); 150mM NaCl; 1mM Na2EDTA; 1mM EGTA; 1% triton; 2.5mM sodium pyrophosphate; 1mM beta-glycerophosphate; 1mM Na3VO4; 1µg/mL leupeptin; 1mM PMSF (phenylmethylsulfonyl fluoride) (immediately before use) and incubated on ice for five minutes. The cells were subsequently scarped from the plate and transferred to a tube and lysed using probe sonication (sonication with one cycle of five seconds at 25% power, samples kept on ice during sonication to prevent excessive heating). The lysate was then spun at 14,000 RPM for 10 minutes at 4ºC and transferred to a new tube. Cell lysates when needed, were diluted with sample diluents (supplied with kit). The lysates were then transferred to 96 well plates and incubated overnight at 37ºC. Next, washing of the wells was done with 1X wash buffer (supplied with kit). To each well, 100µL of NF-?B p65 detection antibody (supplied with kit) was added and the plate was incubated at 37ºC for one-hour. The wells were then washed again and 100 µL of horseradish peroxidase (HRP) -linked secondary antibody was added to each well. The plate was then incubated for 30 minutes at 37°C. The wash step was repeated and 100 µL of 3, 3', 5, 5'-tetramethylbenzidine (TMB) buffer was added. Finally, the stop solution was added and absorbance read at 450 nm.
[00115] Figure 9 depicts the effect of varying concentrations of asparagus extract (20µg/ml, 40µg/ml, 50µg/ml, 60µg/ml, and 80µg/ml). A known herbal anti-inflammatory product – curcumin is considered as a positive control. It can be observed that the inhibition percentage varies from 15.4% in case of 20µg/ml asparagus extract to 20.6% in case of 80µg/ml of asparagus extract. It can be inferred that although, different concentrations are showing percentage inhibition of NF?B phosphorylation but it is lower than the effect displayed by curcumin (37.7%).
[00116] Figure 10 depicts the effect of varying concentrations of red pearl onion extract (20µg/ml, 40µg/ml, 50µg/ml, 60µg/ml, and 80µg/ml). It can be observed that the inhibition percentage varies from 14.2% in case of 20µg/ml red pearl onion extract to 24.8% in case of 80µg/ml of red pearl onion extract. As was observed with asparagus extract, the effect of NF?B phosphorylation inhibition with red pearl onion extract is lesser than that observed with curcumin (36.9%).
Example 7
Effect of combinations of asparagus extract and red pearl onion extract in inhibiting NF?B phosphorylation
[00117] The combinations of asparagus extract and red pearl onion extract as listed in Table 3 of Example 5 was used to test the effect on inhibiting NF?B phosphorylation. The composition comprising asparagus extract and red pearl onion extract in a w/w ratio of 4:1, 3:2, 1:1. 2:3, and 1:4 were used in the experiment.
[00118] The percentage of NF?B phosphorylation inhibition was measured as described previously in Example 6. Figure 11 depicts the effect of various combinations of asparagus extract and red pearl onion extract in inhibiting NF?B phosphorylation. As in previous examples, curcumin has been considered as a positive control. The additive effects of the extracts (Figure 9 and Figure 10) were compared to the combinatorial inhibitory effect (Figure 11) of different weight ratios, and the analysis has been depicted in Table 5:
Table 5: Effect of combinations of asparagus extract and red pearl onion extract in inhibiting NF?B phosphorylation
Individual Dosage Individual inhibitory effect Combination inhibitory effect
Asparagus Red Pearl Onion Asparagus
(Figure 9) Red Pearl onion
(Figure 10) Additive Synergy
(Figure 11)
80 µg/ml 20 µg/ml 20.6% 14.2% 34.8% 36.5% (4:1)
40 µg/ml 60 µg/ml 17.5% 22.8% 40.3% 26.1% (2:3)
20 µg/ml 80 µg/ml 15.4% 24.8% 40.2% 20.5% (1:4)

[00119] It can be appreciated from the Table 5 and Figure 11, that the composition comprising asparagus extract and red pearl onion extract in a w/w ratio of 4:1 is showing a synergistic inhibition effect on NF?B phosphorylation as compared to the other compositions. Further it can be observed that the composition showing a synergistic effect is displaying an enhanced level of inhibition which is even better than curcumin. Therefore, the composition comprising asparagus extract and red pearl onion extract in a w/w ratio of 4:1 has an enhanced high anti-inflammation ability and thus can be exploited in innumerable ways. It can be observed that the composition comprising asparagus extract and red pearl onion extract in a w/w ratio of 2:3 and 1:4 does not exhibit synergistic effect.
[00120] Overall, the present document discloses anti-inflammatory compounds comprising asparagus extract and red pearl onion extract in a w/w ratio of 4:1, 3:2, and 1:1. The compositions display synergistic anti-inflammatory effects in terms of inhibiting IL-8 production, and the composition pertaining to asparagus extract and red pearl onion extract in a w/w ratio of 4:1 has been shown to exert synergistic effect in inhibiting NF?B phosphorylation.
Advantages of the present disclosure
[00121] The present document discloses anti-inflammatory compounds that exhibits its effect by inhibiting the production of IL-8 (pro-inflammatory cytokine) and phosphorylation of NF?B (a well-established master regulator of inflammation). The compounds comprise asparagus extract and red pearl onion extract in different w/w ratios of 4:1, 3:2, and 1:1. The anti-inflammatory compounds comprise herbal components, hence exhibit high bio-tolerance. The extracts of asparagus and red pearl onion are relatively easier to prepare and exhibits distinct synergistic anti-inflammatory effects at disclosed ratios and the compounds can be used in preparing various anti-inflammatory formulations. The synergistic effects of the disclosed compositions (asparagus and red pearl onion extract in w/w ratio 4:1, 3:2 and 1:1) in terms of inhibiting IL-8 production is comparable to the anti-inflammatory effects of a well-studied compound – curcumin. The anti-inflammatory effects of the composition comprising asparagus extract and red pearl onion extract (in a w/w ratio of 4:1) in terms of inhibiting NF?B phosphorylation is higher than that of curcumin. Thus, the document discloses highly effective herbal compositions which exhibit enhanced anti-inflammatory effects.