EVALUATION OF ANTIOXIDANT AND ANTI-INFLAMMATORY ACTIVITIES OF SOME NUTRITIONAL SUPPLEMENTS

EVALUATION OF ANTIOXIDANT AND ANTI-INFLAMMATORY ACTIVITIES OF SOME NUTRITIONAL SUPPLEMENTS

P. Swetha^*, Harish C. Rodda and T. Sandhya

Department of Pharmacognosy and Phytochemistry, Pathfinder Instituent of Pharmacy Education and Research, Opposite Warangal Airport, Mamnoor, Warangal-506166, Telangana, India.

ABSTRACT: The nutritional supplements are the natural food supplements, and other related products used to boost the nutritional content of the diet. These nutritional supplements have been of interest to research because of their antioxidant property by consumption of food and drinks. The present study deals with a comparative evaluation of antioxidant and anti-inflammatory effects of marketed nutritional supplements using cytoprotection, lipidperoxdiation and membrane stabilization activities on RBC under in-vitro condition. The marketed nutritional supplements exhibited cytoprotection or prevented hemolysis in erythrocytes and provided better protection against H₂O₂ induced oxidation and lipid peroxidation and exhibited good membrane stabilization when compared to standard. The Women Horlicks showed significant protection against H₂O₂ induced cytotoxicity in erythrocytes when compared to other supplements, the nutritional supplement Boost provided better protection against lipidperoxidation induced by H₂O₂ in erythrocytes. The nutritional supplement Maltova showed better membrane stabilization activity when compared to standard and other nutritional supplement.

Alternative uses of drugs, Antioxidant activity, Cytoprotection activity, Membrane stabilization activity, Add-on therapy

INTRODUCTION: Nutrition can be defined as all processes used by an adult    or child to take in food and to digest, absorb, transport, utilize, and excrete food substances, components or substances  found in food are called as nutrition ¹. Nutritional supplements include vitamins, minerals, herbs, meal supplements, sports nutrition products, natural food supplements, and other related products used to boost the nutritional improve the content of the diet. Nutritional supplements are used for many purposes.

They can be added to the diet to boost overall health and energy; to provide immune system support and reduce the risks of illness and age-related conditions; to improve performance in athletic and mental activities; and to support the healing process during illness and disease ². However, most of these products are treated as food and not regulated as drugs. Nutritional supplement drinks are available for different purposes and different age groups children's, women’s, sport persons, diseased persons, pregnant ladies, feeding mothers ².

In present study antioxidant, cytoprotection and membrane stabilization activities of different nutritional supplements was studied on RBC.

MATERIALS AND METHODS: Human red blood cell, Saline phosphate buffer containing 0.44 mol/l  H₂O₂, TCA containing    0.1 M sodium Meta aresenite, thiobarbituric acid (TBA), malondialdehyde (MDA), 1,1,3,3-tetra ethoxypropane (TEP), ascorbic acid, hydrogen peroxide, hypotonic saline, isotonic saline, diclofenac sodium, sodium hydroxides.

Nutritional supplements:

Boost, Horlicks, Women Horlicks, Junior Horlicks and Kissan Nutria Gro (Glaxo Smith Kine, Punjab), Complan Memory, Complan Nutri Gro, Complan Kesar Badam (Heinz India Pvt Limited, Mumbai) Pediasure (Abbott Singapore Private Limited, Singapore), Bournvita, Bournvita 5 Star (Cabdury India Limited, India), Protinex Vanilla (Wock Hardt Limited, Mumbai).

Methodology:

Sample preparation:

700mg of nutritional supplements was transferred into a beaker and 5ml of distilled water was added and mixed well ⁴.

Lipid peroxidation method:

An aliquot containing 1ml of RBC suspension in 9 ml saline phosphate buffer(pH 7.2) which contains 0.44 mol/L H₂0₂  was prepared. To it 1ml of test sample was added. About 2.5ml aliquot was withdrawn at zero hour and remaining aliquot (2.5ml) was incubated at 37⁰ for 2 hours. To all the tubes 1 ml of 28% trichloroacetic acid (TCA) was added and kept   aside.  The 3 ml of supernatant was taken and to it 1ml of 1% thiobarbituric acid (TBA) was added and boiled for 15mins. The tubes were then cooled and centrifuged for 15 min at 3000 rpm. Absorbance was read at 540 nm using Shimadzu spectrophotometer (UV-1800). All the determinations were performed in triplicate. The malondialdehyde (MDA) content was estimated ^{3, 4}.

Cytoprotection activity:

The samples were prepared as shown in Table 1.

TABLE 1: CYTOPROTECTION ACTIVITY

To the test, positive control, negative control, RBC suspension was added and incubated for 1 hr. Phosphate buffer and hydrogen peroxide were then added to the above solution and incubated for 1½ hr and the supernatant solutions were separated.

The hemoglobin released into the supernatant of the mixtures was measured at 540 nm using Shimadzu spectrophotometer (UV-1800). All the determinations were performed in triplicate ⁵.

Membrane stabilization: 

Test solution: A reaction mixture consisting of 2ml of hypotonic saline (0.25%w/v), 1ml of phosphate buffer (pH 7.4), 0.5ml of test sample, 0.5ml of 10%w/v human red blood cell in isotonic saline was prepared and finally the volume was made up to 4.5ml with hypotonic saline.

Blood control:

A control  mixture consisting of 2.5ml of hypotonic saline (0.25%w/v), 1ml of phosphate buffer (pH 7.4), and 0.5ml of 10%w/v human red blood cell in isotonic saline was prepared  and  finally the volume was made up to 4.5ml with hypotonic saline.

Standard solution:

A standard mixture containing 2ml of hypotonic saline (0.25%w/v), 1ml of phosphate buffer (pH 7.4), 0.5ml of diclofenac sodium, and 0.5ml of 10%w/v human red blood cell in isotonic saline was prepared  and finally the volume  was made up to 4.5ml with hypotonic saline.

Drug control:

A reaction mixture containing 2.5ml of hypotonic saline (0.25%w/v), 1ml of phosphate buffer (pH 7.4), and 0.5ml of diclofenac sodium solution was prepared and finally the volume was made up to 4.5ml with hypotonic saline ⁷. The mixtures were incubated at 56°C for 30 minutes. The tubes were cooled under running water for 20 minutes, and mixtures were centrifuged at 3000 rpm for 10min and the supernatant liquid was separated. The absorbance of supernatant was read at 560 nm using shimadzu spectrophotometer (UV-1800). All the determinations were performed in triplicate.^{6, 7}

RESULTS AND DISCUSSION:

Effect of different extracts on lipid peroxdiation:

Extent of lipid peroxidation is measured through malondialdehyde levels in the biological tissues and fluids, a pro-oxidant factor which determines the oxidative damage.

The standard graph of malondialdehye for determination of lipid peroxide levels is shown in Fig 1. The absorbance values and concentrations were found to be linear in the concentration range of 1-30 nM/ml, as confirmed by the correlation coefficient of 0.994. The absorbance (y) was proportional to the concentration of Ascorbic acid (x) following the regression equation y=0.0298+0.0378.

The results reveal that most of the nutritional supplements provided better protection against lipid peroxidation induced by H₂O₂ in erythrocytes. The nutritional supplements Women Horlicks, Proteinex V, Proteinex C, Kissan smart exhibited better results in lipid peroxidation induced by H₂O₂ in erythrocytes. (Table.2, Fig.1 and Fig.2)

TABLE 2:  EVALUATION OF LIPID PEROXIDATION ACTIVITY OF NUTRITIONAL SUPPLEMENTS

FIG.1. EVALUATION OF LIPIDPEROXIDATION ACTIVITY OF NUTRITIONAL SUPPLEMENTS AT 0-HOUR

Fig. 2: Evaluation of Lipid peroxidation activity of Nutritional supplements at 2-hour

Cytoprotection activity:

According to our results, the percentage of hemolysis in erythrocytes pretreated with the nutritional supplements was lower than the non pretreated erythrocytes suspension.  Nutritional supplements like Women Horlicks and Pediasure showed significant protective effect against H₂O₂ induced cytotoxicity in erythrocytes. (Fig.3 and Table 3)

FIG. 3: EFFECT AND PERCENTAGE OF DIFFERENT NUTRITIONAL SUPPLEMENTS ON CYTOTOXCITY

TABLE 3: CYTOPROTECTION AGAINST HYDROGEN PEROXIDE INDUCED ON HUMAN RBC

 Membrane stabilization method:

The percentage membrane stabilization of different nutritional supplements is shown in Table 4. The results indicate that the Maltova had highest in-vitro anti-inflammatory activity when compared to other products. (Fig.4 and Table.4)

FIG.4: PERCENTAGE OF MEMBRANE STABILIZATION OF DIFFERENT PRODUCT

TABLE 4: EFFECT OF DIFFERENT NUTRITIONAL SUPPLEMENTS ON MEMBRANE STABILIZATION

Summary: The results of the present investigation reveal that nutritional supplements like Horlicks and Bournvita exhibited the better antioxidant potential. Nutritional supplements like Protienx, Horlicks Junior, and Complan exhibited good protection against lipid peroxdiation induced by H₂O_2. Women Horlicks and Pediasure showed significant protection against H₂O₂ induced cytotoxicity in erythrocytes. Maltova had highest in-vitro anti-inflammatory activity when compared to other products.

CONCLUSION: The present study reveals that nutritional supplements were found to active against cytotoxicity, oxidation, and membrane destabilization. The comparative data obtained regarding the pharmacological activities of nutritional supplements in the present investigation can be used for further studies in other areas of interest and also for studies on healthy volunteers.

ACKNOWLEDGEMENTS: The authors wish to acknowledge the management of the pathfinder institute of pharmacy educational and research, for providing the necessary facilities.

REFERENCES:

1. Amin I, Tan SH: Antioxidant Activity of Selected Commercial Seaweeds. Indian Journal Nutrition, 2002; 8: 167-177.
2. Balch PA: Prescription for Nutritional Healing. East Rutherford, Nutritional Journal Penguin Group 2005; 3: 234-237.
3. Sudha K, Ashalatha VR, Suryanarayana R, Anjali R: Lipid peroxidation, hemolysis and antioxidant enzymes of erythrocytes in stroke. Indian Journal physiology and pharmacology. 2004; 48:199-205.
4. Sandesh P, Velu V and Singh RP: Antioxidant activities of tamarind (Tamarindus Indica) seed coat extracts using in vitro and in vivo models. Journal of Food Science and Technology. 2014; 51(9):1965-1973.

5. Kakansutra dhar, Tania binte wahed, and A.H.M Nazmul and Sonia binte wahed: Invitro antioxidant activities and cytotoxicity study of the methanolic extract of bark of Syzygium cymosum. Dharetal, IJPSR 2016; 7(3): 1021-1025.
6. Yerramsetty, Nagaharika, Valluri, kalyani, Shaik Rasheed, Ramadosskarthikeyan: Anti-inflammatory activity of leaves of Jatropha gossypifolia by hrbc membrane stabilization method. Journal of Acute Disease. 2013; 156-158.
7. Rajan Murugan, Thangaraj Parimelazhagan: Comparative evaluation of different extraction methods for antioxidant and anti-inflammatory properties from Osbeckia parvifolia – An in vitro approach. Journal of King Saud University – Science .2014; 26: 267–27.
   

   ---

   How to cite this article:

   Swetha P, Rodda HC and Sandhya T: Evaluation of antioxidant and anti-inflammatory activities of some nutritional supplements. Int J Pharm Sci Res 2016; 7(11): 4540-45.doi: 10.13040/IJPSR.0975-8232.7(11).4540-45.

   ---

   All © 2013 are reserved by International Journal of Pharmaceutical Sciences and Research. This Journal licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported License.

   ---