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Abstract

Moringa oleifera, often referred to as the "Miracle Tree," is known for its vast range of medicinal properties, including anti-inflammatory activity. This study aimed to design and develop an anti-inflammatory cream using medicated oil derived from Moringa oleifera leaves. Phytochemical screening of the aqueous extract of M. oleifera leaves revealed the presence of flavonoids, tannins, saponins, alkaloids, glycosides, and volatile oils. In vitro anti-inflammatory activity was assessed using the protein denaturation inhibition assay, demonstrating a significant inhibition rate of 49.25% at 800 µg/ml, comparable to standard Ibuprofen at the same concentration. The results suggest that the formulated herbal cream possesses anti-inflammatory properties, making it a potential natural remedy for inflammation-related skin conditions

Keywords

Moringa oleifera, medicated oil, protein denaturation inhibition assay, herbal cream.

Introduction

Moringa oleifera L. is the most widely cultivated of the 13 species of the family Moringaceae that is native to the sub-Himalayan tracts of India, Pakistan, Bangladesh, Afghanistan, and Nepal, which is now indigenous to many regions of Africa, Southeast Asia, and South [1]. A plethora of studies that have been published in the decade describe the popularity of moringa as a “Miracle tree’’. It is a well-known plant for nutritional as well as potential antioxidant [2], anticancer, anti-inflammatory, antidiabetic, and antimicrobial agent, and possesses numerous medicinal qualities[3]. The leaves have been established as a rich source of phenolics and glucosinolates [4].[5], carotenoids [6], isothiocyanates [7] and various protein, vitamins, amino acids, and phenolics [8].  Cream containing leaf extract is a natural effective ingredient for improving hydration, which can be used in moisturizing cosmetic formulations and also to complement the treatment of dry skin.[9],[10],[11].


Table 1: Plant profile of Moringa oleifera

Kingdom

Plantae

Division

Magnoliophyta

Class

Magnoliopsida

Subclass

Dilleniidae

Order

Brassicales

Family

Moringaceae

Genus

Moringa

Species

oleifera

 


Table 2: Reported chemical constituents of Moringa oleifera leaves[12][13][14]

 

Name

Structure

Kaempferol

Quercetin

Rutin

Lutein

?-carotene

Niazirin

Niazirinin

Ellagic acid

Caffeic acid

4-(?-Lrhamnopyranosyloxy) benzyl glucosinolate

Niaziminin


MATERIALS AND METHODS

Collection and Authentication of Moringa oleifera leaves

The Moringa oleifera leaves were collected from Peringottukara, Thrissur, Kerala. The collected herbal leaves were authenticated by Dr. C T Anitha, Head of Department, Department of Botany, Sree Narayana College, Nattika, Thrissur, Kerala

Preparation of medicated oil from Moringa oleifera leaves [15]


The medicated oil was prepared from Moringa oleifera leaves using the formula given below.

Rx

Moringa leaves     -        25g

Coconut oil           -        100ml

Water                    -         400ml


  • Weigh the required amount of crushed moringa leaves and the coconut oil as an oil base.
  • Mix base oil with the water in an accurate ratio and add Kalka (paste of herb used, moringa leaves) in it.
  • Heat the mixture on low flame till only oil remains. Water and the moisture content get evaporated.
  • Do not forget to stir the mixture throughout the process to prevent charring.
  • When the oil is ready, leave it for several hours, so that the temperature falls naturally.
  • After that, filter the oil and discard the residues. Preserve the oil in a suitable tight container and evaluated its colour, odor, and texture

Preparation of herbal cream from prepared medicated oil[16]

The cream was prepared using the formula given below.


Rx

Moringa oil           -       8g

White beeswax     -        3.33g

Borax                    -        0.18g

Vitamin E             -        1 capsule

Water                    -        6.5ml


Method of preparation

  • The required quantity of prepared Moringa oil and beeswax was taken in a China dish and heated in a water bath up to 75°C.
  • In another China dish take borax and water, heated up to 75°C.
  • Mix both the solution slowly with continuous stirring till a cream-like consistency is obtained.
  • Add vitamin E capsule into the cream.
  • Pack it in a suitable tight container

Evaluation of the formulated cream

Organoleptic properties

The prepared cream was evaluated for its colour, odour, and texture by manually[17].

Physicochemical analysis of cream

The physicochemical analysis of cream includes pH, viscosity, homogeneity, appearance, softness, washability, and irritancy[18].

Preparation and Phytochemical analysis of Moringa leaves extract

Preparation of aqueous extract of leaves

50g of the powdered leaves were weighed and poured into a 250ml conical flask in which 200ml of distilled water was added. The mixture was kept for 12 hours with constant agitation at 30 minute intervals. The extract was filtered using Whatman No.1 filter paper. The filtrate was concentrated in a Rotary vacuum evaporator. The semi-solid extract obtained was stored in a refrigerator for further use[18].

Phytochemical analysis of the extract[19]

  1. Test for flavonoids

Shinoda Test:  To the extract, add 5ml 95% ethanol, a few drops of conc. Hydrochloric acid and 0.5 g magnesium turnings. Orange, pink, red to purple colour indicate the presence of flavonoids.

  1. Test for Tannins

Add few drops of aqueous basic lead acetate solution to 1ml of aqueous solution of crude extract using a test tube. Reddish brown bulky precipitate indicates the presence of tannins.

  1. Test for saponins

1g of extract was taken in a test tube containing 5ml of sodium bicarbonate and 5ml of water and the contents were shaken vigorously. The formation of a stable froth suggests the presence of saponins.

  1. Test for Alkaloids

Take 5ml solution of extract and then add 2ml of Wagner’s reagent to it. The appearance of brown precipitate indicates the presence of alkaloids.

  1. Test for Glycosides

To 5ml extract add dilute sulphuric acid and boil it for 15 minutes. Cool and neutralize with 10% NaOH, then 5ml of Fehling’s solution A and B was added. A brick-red precipitate of reducing sugar indicates the presence of glycosides.

  1. Test for Volatile oils

2ml of extract solution was shaken with 0.1ml of sodium hydroxide and a small quantity of dilute HCl. A white precipitate was formed with volatile oils.

In vitro Anti-Inflammatory activity

Inhibition of Protein Denaturation Assay

Protein denaturation results loose of biological properties of protein molecules. Protein denaturation has been correlated with the formation of inflammatory disorders like rheumatoid arthritis, diabetes and cancer. Therefore, ability of substance to prevent the protein denaturation may also help to prevent the inflammatory disorders[20].  The 0.2 ml of egg albumin was added to vials containing 2.5ml of 0.1M Phosphate Buffer Saline (PBS) pH 6.4. 2 ml of sample was added to vials (in triplicate) to achieve the final concentration of 200, 400, 800 µg/ml. PBS (2.5 ml) and egg albumin were added to control vials. Then, the mixture was incubated at 37°C ± 2°C in an incubator for 15 min and then heated at 70°C for 5 min. After cooling, their absorbance was measured at 660 nm using the vehicle as blank. The percentage inhibition of protein denaturation of the test should be compared with negative control[21].

% Inhibition of denaturation = (1-D/C) X100.

Where D is the absorbance of test sample and C is the absorbance of the negative control (without the test sample or reference drug).

RESULTS AND DISCUSSION

Preliminary analysis of Moringa Taila


Table 3: Preliminary analysis of Moringa  Taila

Colour

Greenish colour

Odour

Characteristic odour

Touch

Greasy and soft

Consistency

Smooth and homogeneous

Texture

Smooth liquid, opaque


Preliminary analysis of cream

Table 4: Preliminary analysis of cream

Colour

Light green colour

Odour

Characteristic odour

Touch

Greasy and soft

Consistency

Smooth, homogeneous

Texture

Smooth creamy thick, non-gritty

 


Table 5: Physicochemical analysis of cream

 

pH

5.6-7.0

Viscosity

2.6-3.7 cps

Homogeneity

The formulation produced uniform distribution of extract in cream. This was confirmed by visual appearance and by touch.

Appearance

Formulation was kept for 3 months, it was found that there was no change in colour of cream.

Softness

Slipperiness, and amount of residue left after the application of the fixed amount of cream were found.

Washability

The cream applied on the skin were easily removed by washing with tap water and showed good washability.

Irritancy

The cream showed no redness, inflammation or irritation during irritancy studies. These formulations are safe to use for skin.

 


Phytochemical analysis of Moringa leaves extract

 

Table 6: Phytochemical analysis of Moringa leaves extract

 

Flavonoids

Present

Tannins

Present

Saponins

Present

Alkaloids

Present

Glycosides

Present

Volatile oil

Present


Inhibition of protein denaturation assay

The effect of aqueous extract of M. oleifera leaves in protein denaturation with egg albumin is given in Table 7. The results obtained indicated that the formulation showed significant anti-inflammatory activity by reducing the protein denaturation.  The highest inhibition rate was observed at the concentration of 800 µg/ml (Table 7). The observed inhibition rate was comparable with standard Ibuprofen at 800 µg/ml which was 88.88%[22].


Table 7: Percentage inhibition rate of Protein denaturation of aqueous extract of Moringa oleifera

Concentration         Rate of inhibition (%)

     (µg/ml)

200                                  17.91±3.28

400                                  31.34±1.20

800                                  49.25±0.41


Results are shown as mean±SEM. SEM: Standard error of mean

       
            Graph showing the result of Inhibition of protein denaturation assay.png
       

Concentration (µg/ml)

Figure 3: Graph showing the result of Inhibition of protein denaturation assay

CONCLUSION

The present study aims at developing a herbal cream using medicated oil of Moringa oleifera leaves. The physicochemical evaluation of the herbal cream was performed. Phytochemical screening of the aqueous extract revealed the presence of flavonoids, tannins, saponins, alkaloids, glycosides and volatile oil. The in vitro antinflammatory evaluation of the aqueous extract was performed by ‘Inhibition of albumin denaturation assay’. . The anti-inflammatory activity of the aqueous extract of  Moringa leaves was found to be 49.25% at the concentration of 800 µg/ml.  Hence it was concluded that the medicated oil and the formulated herbal cream possess Anti-Inflammatory activities. Further studies can be directed inorder to find out the phytochemical in the plant responsible for the anti-inflammatory action and to develop formulations using it.

REFERENCES

  1. Fahey JW. Moringa oleifera: A review of the medicinal potential. InI International Symposium on Moringa 1158 2015 Nov 15 (pp. 209-224).
  2. Bharali R, Tabassum J, Azad MR. Chemomodulatory effect of Moringa oleifera, Lam, on hepatic carcinogen metabolising enzymes, antioxidant parameters and skin papillomagenesis in mice. Asian Pacific Journal of Cancer Prevention. 2003 Apr 24;4(2):131-40.
  3. Mahmood KT, Mugal T, Haq IU. Moringa oleifera: a natural gift-A review. Journal of Pharmaceutical Sciences and Research. 2010 Nov 1;2(11):775.
  4. Sreelatha S, Padma PR. Antioxidant activity and total phenolic content of Moringa oleifera leaves in two stages of maturity. Plant foods for human nutrition. 2009 Dec;64:303-11.
  5. Amaglo NK, Bennett RN, Curto RB, Rosa EA, Turco VL, Giuffrida A, Curto AL, Crea F, Timpo GM. Profiling selected phytochemicals and nutrients in different tissues of the multipurpose tree Moringa oleifera L., grown in Ghana. Food Chemistry. 2010 Oct 15;122(4):1047-54.
  6. Saini RK, Harish Prashanth KV, Shetty NP, Giridhar P. Elicitors, SA and MJ enhance carotenoids and tocopherol biosynthesis and expression of antioxidant related genes in Moringa oleifera Lam. leaves. Acta physiologiae plantarum. 2014 Oct;36:2695-704.
  7. Waterman C, Cheng DM, Rojas-Silva P, Poulev A, Dreifus J, Lila MA, Raskin I. Stable, water extractable isothiocyanates from Moringa oleifera leaves attenuate inflammation in vitro. Phytochemistry. 2014 Jul 1;103:114-22.
  8. Anwar F, Latif S, Ashraf M, Gilani AH. Moringa oleifera: a food plant with multiple medicinal uses. Phytotherapy Research: An International Journal Devoted to Pharmacological and Toxicological Evaluation of Natural Product Derivatives. 2007 Jan;21(1):17-25.
  9. Sahu T, Patel T, Sahu S, Gidwani B. Skin cream as topical drug delivery system: a review. Journal of Pharmaceutical and Biological Sciences. 2016 Sep 1;4(5):149.
  10. Ali A, Akhtar N, Mumtaz AM, Khan MS, Iqbal FM, Zaidi SS. In vivo skin irritation potential of a cream containing Moringa oleifera leaf extract. Afr J Pharm Pharmacol. 2013 Feb 15;7(6):289-93.
  11. Gyawali R, Bhattarai D, Bartaula N, Bastola R, Bhatta S, Suwal S, Paudel PN. Formulation and Evaluation of Cream Using Moringa oleifera L. Leaf Extract. Journal of Institute of Science and Technology. 2022 Dec 29;27(2):61-6.
  12. Pandey A, Pandey RD, Tripathi P, Gupta PP, Haider J, Bhatt S, Singh AV. Moringa Oleifera Lam.(Sahijan)-A Plant with a Plethora of Diverse Therapeutic Benefits: An Updated Retrospection. Pandeyet al. Medicinal Aromatic Plants 2012 [Internet]. 2012
  13. Ragasa CY, Medecilo MP, Shen CC. Chemical constituents of Moringa oleifera Lam. leaves. delta. 2015;1:1a.
  14. Leone A, Spada A, Battezzati A, Schiraldi A, Aristil J, Bertoli S. Cultivation, genetic, ethnopharmacology, phytochemistry and pharmacology of Moringa oleifera leaves: An overview. International journal of molecular sciences. 2015 Jun 5;16(6):12791-835.
  15. Panda P, Das B, Sahu DS, Meher SK, Bhuyna GC, Das BK, Rao MM. Taila Kalpana (Medicated Oil) in Ayurveda. Research Journal of Pharmacology and Pharmacodynamics. 2016;8(1):39-41.
  16. Sharma AN, Banyal MA, Gupta JY, Joshi SW. Formulation and evaluation of herbal cold cream. IJARIIE. 2019;9(3):2578-87.
  17. Badwaik CB, Lade UB, Agarwal T, Barsagade P, Nandgave M, Gaddamwar N. Formulation and Evaluation of Herbal Face Cream. International Journal of Pharmaceutical Research and Applications. 2022 Jan;7(1):955-60.
  18. K. R. Khandelwal, “Practical Pharmacognosy,” Ninth Edition, Nirali prakashan, Delhi, 2002, pp. 157-160
  19. Shaikh JR, Patil M. Qualitative tests for preliminary phytochemical screening: An overview. International Journal of Chemical Studies. 2020 Mar 1;8(2):603-8.
  20. Bailey-Shaw YA, Williams LA, Green CE, Rodney S, Smith AM. In-vitro evaluation of the anti-inflammatory potential of selected jamaican plant extracts using the bovine serum albumin protein denaturation assay. International Journal of Pharmaceutical Sciences Review and Research. 2017;47(1):145-53.
  21. Mirke NB, Shelke PS, Malavdkar PR, Jagtap PN. In vitro protein denaturation inhibition assay of Eucalyptus globulus and Glycine max for potential anti-inflammatory activity. Innovations in Pharmaceuticals and Pharmacotherapy. 2020;8(2):28.
  22. Saleem U, Zaib S, Khalid S, Anwar F, Akhtar MF, Ahmad B. Chemical characterization, docking studies, anti-arthritic activity and acute oral toxicity of Convolvulus arvensis L. leaves. Asian Pacific Journal of Tropical Biomedicine. 2020 Oct 1;10(10):442-51.

Reference

  1. Fahey JW. Moringa oleifera: A review of the medicinal potential. InI International Symposium on Moringa 1158 2015 Nov 15 (pp. 209-224).
  2. Bharali R, Tabassum J, Azad MR. Chemomodulatory effect of Moringa oleifera, Lam, on hepatic carcinogen metabolising enzymes, antioxidant parameters and skin papillomagenesis in mice. Asian Pacific Journal of Cancer Prevention. 2003 Apr 24;4(2):131-40.
  3. Mahmood KT, Mugal T, Haq IU. Moringa oleifera: a natural gift-A review. Journal of Pharmaceutical Sciences and Research. 2010 Nov 1;2(11):775.
  4. Sreelatha S, Padma PR. Antioxidant activity and total phenolic content of Moringa oleifera leaves in two stages of maturity. Plant foods for human nutrition. 2009 Dec;64:303-11.
  5. Amaglo NK, Bennett RN, Curto RB, Rosa EA, Turco VL, Giuffrida A, Curto AL, Crea F, Timpo GM. Profiling selected phytochemicals and nutrients in different tissues of the multipurpose tree Moringa oleifera L., grown in Ghana. Food Chemistry. 2010 Oct 15;122(4):1047-54.
  6. Saini RK, Harish Prashanth KV, Shetty NP, Giridhar P. Elicitors, SA and MJ enhance carotenoids and tocopherol biosynthesis and expression of antioxidant related genes in Moringa oleifera Lam. leaves. Acta physiologiae plantarum. 2014 Oct;36:2695-704.
  7. Waterman C, Cheng DM, Rojas-Silva P, Poulev A, Dreifus J, Lila MA, Raskin I. Stable, water extractable isothiocyanates from Moringa oleifera leaves attenuate inflammation in vitro. Phytochemistry. 2014 Jul 1;103:114-22.
  8. Anwar F, Latif S, Ashraf M, Gilani AH. Moringa oleifera: a food plant with multiple medicinal uses. Phytotherapy Research: An International Journal Devoted to Pharmacological and Toxicological Evaluation of Natural Product Derivatives. 2007 Jan;21(1):17-25.
  9. Sahu T, Patel T, Sahu S, Gidwani B. Skin cream as topical drug delivery system: a review. Journal of Pharmaceutical and Biological Sciences. 2016 Sep 1;4(5):149.
  10. Ali A, Akhtar N, Mumtaz AM, Khan MS, Iqbal FM, Zaidi SS. In vivo skin irritation potential of a cream containing Moringa oleifera leaf extract. Afr J Pharm Pharmacol. 2013 Feb 15;7(6):289-93.
  11. Gyawali R, Bhattarai D, Bartaula N, Bastola R, Bhatta S, Suwal S, Paudel PN. Formulation and Evaluation of Cream Using Moringa oleifera L. Leaf Extract. Journal of Institute of Science and Technology. 2022 Dec 29;27(2):61-6.
  12. Pandey A, Pandey RD, Tripathi P, Gupta PP, Haider J, Bhatt S, Singh AV. Moringa Oleifera Lam.(Sahijan)-A Plant with a Plethora of Diverse Therapeutic Benefits: An Updated Retrospection. Pandeyet al. Medicinal Aromatic Plants 2012 [Internet]. 2012
  13. Ragasa CY, Medecilo MP, Shen CC. Chemical constituents of Moringa oleifera Lam. leaves. delta. 2015;1:1a.
  14. Leone A, Spada A, Battezzati A, Schiraldi A, Aristil J, Bertoli S. Cultivation, genetic, ethnopharmacology, phytochemistry and pharmacology of Moringa oleifera leaves: An overview. International journal of molecular sciences. 2015 Jun 5;16(6):12791-835.
  15. Panda P, Das B, Sahu DS, Meher SK, Bhuyna GC, Das BK, Rao MM. Taila Kalpana (Medicated Oil) in Ayurveda. Research Journal of Pharmacology and Pharmacodynamics. 2016;8(1):39-41.
  16. Sharma AN, Banyal MA, Gupta JY, Joshi SW. Formulation and evaluation of herbal cold cream. IJARIIE. 2019;9(3):2578-87.
  17. Badwaik CB, Lade UB, Agarwal T, Barsagade P, Nandgave M, Gaddamwar N. Formulation and Evaluation of Herbal Face Cream. International Journal of Pharmaceutical Research and Applications. 2022 Jan;7(1):955-60.
  18. K. R. Khandelwal, “Practical Pharmacognosy,” Ninth Edition, Nirali prakashan, Delhi, 2002, pp. 157-160
  19. Shaikh JR, Patil M. Qualitative tests for preliminary phytochemical screening: An overview. International Journal of Chemical Studies. 2020 Mar 1;8(2):603-8.
  20. Bailey-Shaw YA, Williams LA, Green CE, Rodney S, Smith AM. In-vitro evaluation of the anti-inflammatory potential of selected jamaican plant extracts using the bovine serum albumin protein denaturation assay. International Journal of Pharmaceutical Sciences Review and Research. 2017;47(1):145-53.
  21. Mirke NB, Shelke PS, Malavdkar PR, Jagtap PN. In vitro protein denaturation inhibition assay of Eucalyptus globulus and Glycine max for potential anti-inflammatory activity. Innovations in Pharmaceuticals and Pharmacotherapy. 2020;8(2):28.
  22. Saleem U, Zaib S, Khalid S, Anwar F, Akhtar MF, Ahmad B. Chemical characterization, docking studies, anti-arthritic activity and acute oral toxicity of Convolvulus arvensis L. leaves. Asian Pacific Journal of Tropical Biomedicine. 2020 Oct 1;10(10):442-51.

Photo
Prasanth B.
Corresponding author

St James’ College of Pharmaceutical Sciences, Chalakudy, Kerala- 680307.

Photo
Theertha S. V.
Co-author

St James’ College of Pharmaceutical Sciences, Chalakudy, Kerala- 680307.

Photo
Krishnakumar K.
Co-author

St James’ College of Pharmaceutical Sciences, Chalakudy, Kerala- 680307.

Photo
Neghla E. N.
Co-author

St James’ College of Pharmaceutical Sciences, Chalakudy, Kerala- 680307.

Photo
Riyamol V. L.
Co-author

St James’ College of Pharmaceutical Sciences, Chalakudy, Kerala- 680307.

Photo
Sahla Shaimol
Co-author

St James’ College of Pharmaceutical Sciences, Chalakudy, Kerala- 680307.

Prasanth B.*, Theertha S. V., Krishnakumar K., Neghla E. N, Riyamol V. L., Sahla Shaimol, Design and Development of Anti-Inflammatory Cream from Prepared Medicated Oil of Moringa Oleifera, Int. J. of Pharm. Sci., 2025, Vol 3, Issue 1, 1956-1963. https://doi.org/10.5281/zenodo.14723218

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