This document provides a synopsis for an M.Pharmacy thesis on formulating and evaluating a herbal gel as a multifunctional anti-aging product. The proposed work involves developing a hydrogel incorporating red wine and honey using carbopol as a base. The gel will undergo pharmaceutical, chemical, pharmacological, and microbial evaluations as well as stability studies over 2 months. The evaluations will assess properties such as viscosity, swelling, spreadability, diffusion, and anti-oxidant and anti-inflammatory activities. Excision wound healing, burn wound healing, and anti-microbial effects will also be examined using animal and microbiological models. The aim is to formulate an effective, natural anti-aging product at a reasonable price point

1. FORMULATION AND EVALUATION OF HERBAL GEL :
AMULTIFUNCTIONAL ANTIAGEING PRODUCT
SYNOPSIS FOR M.PHARMACY THESIS
Submitted By:TarunThukral
Course:M.Pharmacy(PHARMACEUTICS)
Supervisor:Anshu Gupta

2. FORMULATION AND EVALUATION OF HERBAL GEL :
AMULTIFUNCTIONAL ANTIAGEING PRODUCT
INTRODUCTION:Ageing is the phase of gradual decline of body efficiency and metabolic
activities after reaching a maturity stage[1]. Skin ageing is the result of continual deterioration
process because of damage of cellular DNA and protein[2] which is linked to the conjunction of
intrinsic factors [chronological ageing] and extrinsic factors [fundamentally photo ageing [26]
[27]. In elderly people, cells are less likely to proliferate, they have shorter life spans, are less
responsive to cytokines and epithelialization of skin tissue is longer[3].
Sequential skin ageing is universal and predictable process characterized by physiological
alteration in skin function. In the ageing process keratinocytes are unable to form a functional
stratum cornium and rate of formation from neutral lipids slows down, resulting in dry pale skin
with wrinkle [28] [29]. In contrast photo ageing is caused by over exposure to UV rays from
sunlight. It is characterized by, pale and shallow skin displaying wrinkles as well as deep furrows
caused by the disorganization of epidermal and dermal components associated with elastosis and
heliodermatitis[2]. This ageing is enhanced due to free radicles that are generated in the body
through various metabolic pathways due to oxidation of fatty food and constant exposure to
radiations [1]. This oxidation damages cell membranes and other structures including cellular
proteins, lipids and DNA. When oxygen is metabolized, it creates free radicals which steal
electrons from other molecules causing damage[28] [30]. The body can cope with some free
radicals and needs them to function effectively however on overload of these radicals leads to
number of health problems including those related to skin too[4].
REVIEW OF LITERATURE:In India, medicines based on herbal origin have been the
basis of treatment and cure for various diseases.Moreover, Indian folk medicine comprises
numerous prescriptions for therapeutic purposes such as healing of wounds, inflammation, skin
infections,leprosy, diarrhea, scabies, venereal disease, ulcers, snake bite, etc. More than 80% of
the world’s population still depends upon traditional medicines for various skin diseases. Herbal
medicines in wound management involve disinfection, debridement and providing a moist
environment to encourage the establishment of the suitable environment for natural healing
process [42, 43]. A large number of plants are used by folklore traditions in India for treatment
of cuts, wounds and burns.Ayurveda remains one of the most ancient and yet alive tradition
practiced widely in India, Sri Lanka and other countries that have a sound philosophical and
experimental basis.Atharvaveda[1200 BC],CharakSamhita and SushrutSamhita[1000-500 BC]
are main classes that give detailed description over 700 herbs.Ayurveda and traditional and
Chinese medicinal system share many common approaches and have long history of practice.
Ayurveda has several formulations for management of ageing and related condition. Ayurveda
literature describes more than 200 herbs, minerals, fats for skin care [3]. We selected honey and
red wine for incorporating in a topical hydrogel base which are used individually to counteract
the main ageing symptoms which involve skin tightening, wound healing, epithelialization,
cellulite removal, anti oxidant, anti inflammatoryetc [26,27,28,29,30,33,34,35].
Honey is an ancient remedy for the treatment of infected wounds, which has recently been
'rediscovered' by the medical profession, particularly where conventional modern therapeutic

3. agents are failing. There are now many published reports describing the effectiveness of honey in
rapidly clearing infection from wounds, with no adverse effects to slow the healing process,
there is also some evidence to suggest that honey may actively promote healing[35] [36] [37]
[38] [39] [40] [41] [42] [43] [44].
In nature, resveratrol functions as part of a plant’s defensive arsenal. Resveratrol is an
antimicrobial substance produced by plants in response to stress, infection, or strong UV
radiation [24] [25] [26]. In recent years, resveratrol in particular has become the subject of
intense interest due to its powerful antioxidant and anti-ageing properties. The antioxidant ability
of resveratrol is known to be both potent and efficient. In fact, it has been shown to be greater
than that of vitamins E and C. One study demonstrated that resveratrol was 95% efficient at
preventing lipid peroxidation, compared to 65% for vitamin E and 37% for vitamin C [23] [24]
[25] [31] [32] [33] [34].
NEED OF THE WORK:With the changing trends people have turned too much beauty
conscious but with age senescence of beauty and health is unavoidable to everyone so a wide
range of cosmetic products are available in market to keep their beauty up to date. Despite great
demand, many anti-ageing products and treatments are synthetic or semi synthetic in nature and
have not been proven to give lasting or major positive effects. One study found that the best
performing creams reduced wrinkles by less than 10% over 12 weeks, which is not noticeable to
the human eye. Another study found that cheap moisturizers were as effective as high-priced
anti-wrinkle creams. So with an idea to bring an effective approach on reasonable prices and
safer levels we decided to formulate a polyherbal preparation.
PLAN OF WORK :steps to be taken are as followed
MATERIAL REQUIRED :
S.NO INGREDIENTS SOURCE REFERENCE
1 Red Wine To Be Purchased 8 , 9
2 Honey To Be Purchased 10 , 11
3 Carbopol 934P To Be Purchased 7
4 Triethanolamine To Be Purchased 7
5 Glycerol To Be Purchased 7
6 Potassium Ferricyanide To Be Purchased 4
7 Ferric Chloride To Be Purchased 4
8 Hydrogen Peroxide To Be Purchased 4
9 Trichloroacetic Acid To Be Purchased 4
10 Dansyl chloride/DihydroxyAcetone (DHA) To Be Purchased 1
11 Pantobarbitone / Diethyl Ether To Be Purchased 3
12 Agar Media Broth To Be Purchased 12,13
13 Propionibacterium acnes To Be Purchased 12,13
14 Staphylococcus epidermidis To Be Purchased 12,13
15 Brookfield Viscometer To Be Purchased 2
16 SpreadibilityTesting Instrument To Be Purchased 2
17 Franz Diffusion Cell To Be Purchased 2
18 Albino Female Rats (15) To Be Purchased 3
19 UV Spectrophotometer To Be Purchased 2
20 pH Meter To Be Purchased 2
21 Stability Chamber To Be Purchased 2
22 Microbial Incubator To Be Purchased 2

4. MATERIAL METHOD: The branded red wine[8][9]and honey[10][11] to be procured and
formulations were developed using carbopol and other excepients obtained from renowned
suppliers. The associate of these active ingredients has been selected according to specific profile
of single substance. The development will took into account not only the efficacy of active
ingredients to counteract the causes of skin ageing but also the pleasantness of the final
formulation in order to achieve the best possible compliance [5].
EXPERIMENTALCRITERIA:
A)PREPARATION OF HYDROGEL:Carbopol 934P NF 1gm and measured quantity of honey
and wine is to be dispersed in 80ml of distilled water and mixed by stirring continuously in a
magnetic stirrer at 800rpm for 1 h. Glycerol 5ml is to be added to the mixture under continuous
stirring. The mixture is to be neutralized by drop-wise addition of 50% triethanolamine (w/w).
Mixing is to be continued until a transparent gel is formed [6][10][15].
B)PHARMACEUTICAL EVALUATION :
a)Solubilty: Normally the hydrogel content of a given material is estimated by measuring
its insoluble part in dried sample after immersion in deionised water for 16 h or 48 h at
room temperature. The sample is to be prepared at a dilute concentration (typically ~ 1%)
to ensure that hydrogel material is fully dispersed in water [22].
b) Viscosity:Viscosity of gel is to be determined by Brookfield viscometer at room
temperature[2] [20].
c) Swelling Property:The swelling is measured according to the Japanese Industrial
Standard (JIS). The dry gel is to be immersed in deionized water for 16 h at room
temperature. After swelling, the hydrogel is to be filtered using a stainless-steel net of
100-mesh and weighed [22].
d)Colour: The appraisal colour of the obtained product is to be noted [2].
e)SpreadibiltyTesting: Spreadability is expressed in terms of timein seconds taken by two
slides to slip off from the formulation, placed between, under the application of a certain
load. Lesser the time taken for the separation ofthe two, better the spreadability[2] [20].
f)Texture: The texture of the obtained product is to be felt by application[2].
g) Diffusion Profile: Diffusion of the gel is to be noted by Agar plate diffusion method
and Franz Diffusion Cell[1].
h) ExtrudabilityIndex:Extrudability is defined as the weight ingrams required for
extruding 0.5cm long ribbon offormulation in 10 secs which is to be determined using
collapsible tubes [13].
i) Drug Content:The drug content of the gelformulations is to be determined by
dissolving an accuratelyweighed quantity 1gm of gel in 100ml of solvent( a mixtureof
ethanol and phosphate buffer pH 6.8 (60:40) for formulations of coriander oil). The
solutions are to be kept forshaking for 4hrs and then kept for 6hrs for completedissolution
of the formulations. Then the solutions are to befiltered through 0.45mm membrane

5. filters and properdilutions should be made and solutions should be subjected to
theSpectrophotometric analysis [13].
j) Homogenecity:All developed gels are to be tested for homogeneity by visual inspection
after the gels have been set in the container. They are to be tested for their appearance
and presence of any aggregates [20].
k) pH Evaluation: is to be measured using pH meter[2].
C)CHEMICAL EVALUATION:
a) Determination of reducing power: 2.5 ml of solution of different formulations are to be
mixed with 2.5ml PO4 buffer solution (pH 6.6) and 2.5 ml potassium ferricyanide
solution (1 %w/v). Mixtures are to be placed in water bath/Incubate at 500C for 20 min.
After incubation resulting solution are to be cooled & mixed with 2.5 ml 10% Trichloro
acetic acid to each test tube. The mixtures are to be centrifuged at 650 rpm for 10 min.2.5
ml upper solution layer are to be mixed with 5 ml of deionised water & 0.5 ml ferric
chloride (1%w/v). Absorbanceis to be measured at 700 nm [4].
b)Hydrogen peroxide scavenging activity:Hydrogen peroxide 2mm/L solution are to be
prepared with standard (PO4 buffer pH-7.4).Dilutions of prepared gels are to be prepared
in distilled water.1ml of solution of themare to be added to 0.6 ml hydrogen peroxide
solution. After 10 min Absorbance is to be measured at 230 nm against blank solution
containing PO4 Buffer without hydrogen peroxide [4].
D)PHARMACOLOGICAL EVALUATION:
a)Excision wound model: The animals are to be taken and weighed and anaesthetized and
shaved dorsally. The excision is to be made and than the application of the gel is to be
done over the wound which should be checked regularly with time and noted [3] [31]
[32] [33] [40] [42] [44].
b)Anti inflammatory activity: is to be observed by application over rats by albumin
induced paw edema method [7].
c) Acid induced wound healing: The acid induced wound is to be created and initial
wound area is to be measured and all the gels are to be applied topically and observed [3].
d) Burn wound model: The burn wound is to be created and initial wound area is to be
measured and all the gels will be applied topically and observed [7].
e)Epithelialization: Time taken for full epithelialization of the wounds is to be measured
by recording the days [7] [24] [25].
E)MICROBIAL EVALUATION:The cylinder-plate method depends upon diffusion of the
antibiotic from a vertical cylinder through a solidified agar layer in a Petri dish or plate to an
extent such that growth of the added micro-organism is prevented entirely in a zone around the
cylinder containing a solution of the antibiotic [12] [13] [35] [38] [42] [43] [44].
F) STABILITY STUDIES: To assess the drugand formulation stability, stability studiesshould
be done according to ICH guidelines.The stability studies is to be carried out asper ICH

6. guidelines. The gel is to be filled inbottle and kept in humidity chambermaintained at 30 ± 2 °C/
65 ± 5 % RHand 40 ± 2 °C / 75 ± 5 % RH for twomonths. At the end of studies, samplesare tobe
analyzed for the physical propertiesand viscosity [2].
TIME MANAGEMENT :
S.No EXPERIMENT PROPOSED
TIME
MONTHS
1 Preparation And Formulation Of
Hydrogel
1 month
September
2 Pharmaceutical Evaluation
3 Chemical Evaluation 2 months October &
November4 Pharmacological Evaluation
5 Microbiological Evaluation 1 month December
6 Stability Studies
REFERENCES :
[1]VidyaSabale, Harish Kunjwani, PrafullaSabale, Formulation and in vitro evaluation of the
topical anti ageing preparation of the fruit of Benincasahispida, J Ayurveda Integr Med.
2011;2[3]:124-128.
[2]Sujit S Nair, Molly Mathew, Sreena k, Formulation and Evaluation of Herbal Cream
containing Curcuma longa, International Journal Of Pharmaceutical and Chemical Sciences;
ISSN:2277-5005.
[3]Hema Sharma Datta, Shankar Kumar Mitra, BhushanPatwardhan, Wound Healing Activity of
Topical Application Forms Based on Ayurveda,Evid Based Compliment Alternet
Med.2011:2011:134318.
[4]PradnyaOnkar, JitendraBangar, RevanKarodi, Evaluation of anti oxidant activity of traditional
formulation Giloysatva and hydroalcoholic extract of the Curuligoorchioidesgaertn; Journal of
Applied Pharmaceutical Science 02[06];2012:209-213.
[5] Adele Sparavigna, Giancarlo Gugulielmini, Stefano Togni Et al, Evaluation of Anti cellulite
efficacy, A topical cosmetic treatment for cellulite blemishes- A multifunctional formulation : J.
Cosmet. Sci ,62, 305-316[2011].
[6]Imokawa G., Recent advances in characterizing biological mechanisms underlying UV-
induced wrinkles: a pivotal role of fibroblast-derived elastase.
[7] SB. Ramane, VN. Syed , KR. Biyani , Evaluation of Wound Healing Activity of Polyherbal
Gel – A Novel Herbal Formulation: International Journal of Research in Pharmaceutical and
Biomedical Sciences: ISSN: 2229-3701.
[8] Resveratrol Ver. 1.0TK: For Anti-ageing,Anti-oxidation, Neuron Protection & Metabolic
Syndrome : ORYZA OIL & FAT CHEMICAL CO., LTD.
[9] Resveratrol; Natural Standard Professional Monograph, Copyright © 2013
[www.naturalstandard.com].

7. [10] The Wound Healing and Management Node Group,The use of medical-grade honey in
wound care: The Joanna Briggs Institute.
[11]Mwipatayi BP, Angel D, Norrish J, The use of honey in chronic leg ulcers: a literature
review : Primary Intention 2004; 12[3]: 107-108, 110-112.
[12]H.A.Sawarkar,S.S.Khadabadi, D.M.Mankar, Development and Biological Evaluation of
HerbalAnti-Acne Gel :International Journal of PharmTech Research :Vol.2, No.3, pp 2028-2031,
2010.
[13]AditiVats1,Pranav Sharma, FORMULATION AND EVALUATION OF TOPICAL ANTI
ACNEFORMULATION OF CORIANDER OIL,International Journal of Pharmacy and
Pharmaceutical Science Research ,ISSN: 2249-0337.
[14]Nikunj B Gurjar, Bhavesh S Barot, Pragna K Shelat, Development and Evaluation of
Cinnamon and Aloevera containing Anti Acne Gel : International Journal of Pharmacy and
Pharmaceutical Science Research, ISSN : 2231 -3656.
[15]ArunRasheed, ShaikNeelufarShama, JyothiMulanjananiyil Joy, Formulation and
evaluation of herbal anti-acne moisturizer, International Journal of Pharmacy and
Pharmaceutical Science Research, ISSN : 2312 -3165
[16] SB. Ramane, VN. Syed and KR. Biyani, Evaluation of Wound Healing Activity of
Polyherbal Gel – A Novel Herbal Formulation, International Journal of Research in
Pharmaceutical and Biomedical Sciences ISSN: 2229-3701.
[17] Thomas C. Tsai, BS and Basil M. Hantash ,Cosmeceutical Agents: A
Comprehensive Review of the Literature , Program for Regenerative Medicine, Stanford
University School of Medicine, Stanford, CA.
[18]TahiraFarooqui, Ph.D., Honey: An Anti-Aging Remedy to Keep you Healthy in a Natural
Way,The Ohio State University.
[19]Fabbrocini G, Staibano S, De Rosa G,Resveratrol-containing gel for the treatment of acne
vulgaris: a single-blind, vehicle-controlled, pilot study:Am J ClinDermatol. 2011 Apr
1;12(2):133-41. doi: 10.2165/11530630-000000000-00000.
[20]SumitDwivedi,Shailesh Gupta, FORMULATION AND EVALUATION OF HERBAL GEL
CONTAINING SESBANIA GRANDIFLORA (L.) POIR. LEAF EXTRACT,ActaChim. Pharm.
Indica: 2(1), 2012, 54-59, ISSN 2277-288X.
[21]Todd R. Hoare a, Daniel S. Kohane,Hydrogels in drug delivery: Progress and
challenges,Science Direct ,Polymer 49 (2008) 1993e2007.
[22]Syed K. H. Gulrez, Saphwan Al-Assaf, Glyn O Phillips, Hydrogels: Methods of Preparation,
Characterization and Applications ,Glyn O Phillips Hydrocolloids Research Centre Glyndwr
University, Wrexham United Kingdom.
[23]Botden IP, Oeseburg H, Durik M, Et al, Red wine extract protects against oxidative-stress-
induced endothelial senescence, ClinSci (Lond). 2012 Oct;123(8):499-507. doi: 10.1042/CS2011.

8. [24] Park SJ, Ahmad F, Philp A, Baar K, Et al, Resveratrol ameliorates aging-related metabolic
phenotypes by inhibiting cAMPphosphodiesterases, Cell. 2012 Feb 3;148(3):421-33. doi:
10.1016/j.cell.2012.01.017.
[25]Panaro MA, Carofiglio V, Acquafredda A, Et al, Anti-inflammatory effects of resveratrol
occur via inhibition of lipopolysaccharide-induced NF-κB activation in Caco-2 and SW480
human colon cancer cells, Br J Nutr. 2012 Nov 14;108(9):1623-32. doi:
10.1017/S0007114511007227. Epub 2012 Jan 17.
[26] N kengne A, Roure R, Rossi AB, The skin aging index: a new approach for documenting
anti-aging products or procedures, Skin Res Technol. 2013 Aug;19(3):291-8. doi:
10.1111/srt.12040. Epub 2013 Apr 11.
[27]Ganceviciene R, Liakou AI, Theodoridis A, Et al, Skin anti-aging strategies,
Dermatoendocrinol. 2012 Jul 1;4(3):308-19. doi: 10.4161/derm.22804.
[28] Wang Y, Wang M, Xiao XS, Et al, The anti-wrinkle efficacy of Argireline, J Cosmet Laser
Ther. 2013 Aug;15(4):237-41. doi: 10.3109/14764172.2013.769273. Epub 2013 Mar 6.
[29]Poljšak B, Dahmane RG, Godić A, Intrinsic skin aging: the role of oxidative stress,
ActaDermatovenerol Alp PanonicaAdriat. 2012;21(2):33-6.
[30]Felippi CC, Oliveira D, Ströher A, Safety and efficacy of antioxidants-loaded nanoparticles
for an anti-aging application, J Biomed Nanotechnol. 2012 Apr;8(2):316-21.
[31] Baxter RA, Anti-aging properties of resveratrol: review and report of a potent new
antioxidant skin care formulation, J CosmetDermatol. 2008 Mar;7(1):2-7. doi: 10.1111/j.1473-
2165.2008.00354.x.
[32]Orallo F, Trans-resveratrol: a magical elixir of eternal youth? Curr Med Chem.
2008;15(19):1887-98.
[33] Chan MM, Antimicrobial effect of resveratrol on dermatophytes and bacterial pathogens of
the skin, BiochemPharmacol. 2002 Jan 15;63(2):99-104.
[34]Bråkenhielm E, Cao R, Cao Y, Suppression of angiogenesis, tumor growth, and wound
healing by resveratrol, a natural compound in red wine and grapes, FASEB J. 2001
Aug;15(10):1798-800.
[35] Al-Waili N, Al-Ghamdi A, Ansari MJ, Et al, Synergistic effects of honey and propolis
toward drug multi-resistant Staphylococcus aureus, Escherichia coli and Candida albicans isolates
in single and polymicrobial cultures, Int J Med Sci. 2012;9(9):793-800. doi: 10.7150/ijms.4722.
Epub 2012 Oct 26.
[36]Alzahrani HA, Alsabehi R, Boukraâ L, Antibacterial and antioxidant potency of floral honeys
from different botanical and geographical origins, Molecules. 2012 Sep 4;17(9):10540-9. doi:
10.3390/molecules170910540.
[37]Kwakman PH, Zaat SA, Antibacterial components of honey, IUBMB Life. 2012
Jan;64(1):48-55. doi: 10.1002/iub.578. Epub 2011 Nov 17.
[38] Al-Waili NS, Salom K, Butler G, Honey and microbial infections: a review supporting the
use of honey for microbial control, J Med Food. 2011 Oct;14(10):1079-96. doi:
10.1089/jmf.2010.0161. Epub 2011 Aug 22.

9. [39]Deshpande SD, Kulkarni KS. In vitro effect of some Indian honeys on Staphylococcus aureus
from wounds, Indian J Exp Biol. 2010 Sep;48(9):931-5.
[40]Vandamme L, Heyneman A, Hoeksema H, Et al, Honey in modern wound care: A systematic
review, Burns. 2013 Jul 26.pii: S0305-4179(13)00197-6. doi: 10.1016/j.burns.2013.06.014.
[41]Vijaya KK, Nishteswar K, Wound healing activity of honey: A pilot study, Ayu. 2012
Jul;33(3):374-7. doi: 10.4103/0974-8520.108827.
[42] Tan MK, HasanAdli DS, Tumiran MA, Et al, The Efficacy of Gelam Honey Dressing
towards Excisional Wound Healing, Evid Based Complement Alternat Med. 2012;2012:805932.
doi: 10.1155/2012/805932. Epub 2012 Mar 28.
[43] Al-Waili N, Salom K, Al-Ghamdi AA, Honey for wound healing, ulcers, and burns; data
supporting its use in clinical practice, ScientificWorldJournal. 2011 Apr 5;11:766-87. doi:
10.1100/tsw.2011.78.
[44] Jull AB, Walker N, Deshpande S, Honey as a topical treatment for wounds, Cochrane
Database Syst Rev. 2013 Feb 28;2:CD005083. doi: 10.1002/14651858.CD005083.pub3.