ABSTRACT Dandruff is the most common problem that affects many people leading to an embarrassing condition. It results from three main factors namely: Malassezia fungi, sebaceous secretion and individual sensitivity. The liphophillic yeast Malassezia furfur is the most common dandruff causing agent in India. In this study dandruff flakes were isolated from patient and cultured in appropriate media like Potato dextrose agar and Sabouraud&apos;s dextrose agar. The antidandruff activity of three selected medicinal plants-Piper cubeba, Cissus quadrangularis and Bauhinia vahlii was studied by well diffusion and broth dilution assay. On screening, methanol extract of Piper cubeba showed good activity and further on broth dilution assay gave a Minimum inhibitory concentration (MIC) at 100µg/ml and IC50 at 800µg/ml. On partial purification through TLC and bio-autography an R f value of 0.705 was obtained where the active compound had activity against dandruff

Arumugam P

Babu Hariram

Nirmala Nithya

Sekar Rohini M

Shanmathi S

Sindhu S

CiteSeerX

          ISSN: 0975-8585 
 
July-September      2013           RJPBCS              Volume 4 Issue 3    Page No.392 
Research Journal of Pharmaceutical, Biological and Chemical 
Sciences 
Exploring the Anti-dandruff Potential of Selected Medicinal Plants 
        
Nirmala Nithya R, Shanmathi S, Rohini M, Sekar Babu Hariram, Arumugam P 
and Sindhu S 
 
Department of Biotechnology, Veltech Hightech, Dr. Rangarajan, Dr. Sakunthala, Engineering College, Avadi, 
Chennai.62 
 
 
 
 
ABSTRACT 
 
           Dandruff is the most common problem that affects many people leading to an embarrassing condition. It 
results from three main factors namely: Malassezia fungi, sebaceous secretion and individual sensitivity. The 
liphophillic yeast Malassezia furfur is the most common dandruff causing agent in India. In this study dandruff 
flakes were isolated from patient and cultured in appropriate media like Potato dextrose agar and Sabouraud’s 
dextrose agar. The antidandruff activity of three selected medicinal plants-Piper cubeba, Cissus quadrangularis and 
Bauhinia vahlii was studied by well diffusion and broth dilution assay. On screening, methanol extract of Piper 
cubeba showed good activity and further on broth dilution assay gave a Minimum inhibitory concentration (MIC) 
at 100µg/ml and IC50 at 800µg/ml. On partial purification through TLC and bio-autography an Rf value of 0.705 was 
obtained where the active compound had activity against dandruff. 
Keywords: Dandruff; Malassezia furfur; Potato dextrose agar; MIC; IC50. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
*Corresponding author 
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July-September      2013           RJPBCS              Volume 4 Issue 3    Page No.393 
INTRODUCTION 
 
Dandruff is a common scalp disorder defined as a slight-to-moderate scaling of the scalp 
with varying degrees of irritation or erythema. The characteristic flaking and scaling of the scalp 
suggest impairment in the desquamation process. The etiology of dandruff is multi-factorial, 
influenced by Malassezia, sebum production and individual susceptibility.The commensal yeast 
Malassezia is a strong contributory factor to dandruff formation. The lipophilic yeast 
Malassezia utilizes sebum lipids as a nutrient source, and sebum production is hypothesized to 
be required to support growth of Malassezia. It has been widely theorized that this increase in 
sebum production and Malassezia proliferation triggers the development of dandruff. The 
exact mechanism of dandruff formation is now believed to be the result of the formation of 
enzymes called lipases. The Malassezia fungus uses these enzymes to break down sebum to 
oleic acid. The oleic acid then penetrates the top layer of skin and causes increased skin cell 
turnover in susceptible people. This in turn causes dandruff flakes and sometimes leads to 
itching and redness.  
 
The current strategy for treating dandruff with antidandruff products and most are 
chemicals too crude for daily use.Several fungistatic compounds have been shown to improve 
dandruff condition. Besides the chemical substances, there is a wide range of herbal ingredients 
like pepper extract, basil extract, neem extract, rosemary oil, basil oil, clove oil, coleus oil, tea 
tree oil which have been documented to have good antidandruff activity.Herbal medicine, now 
a days are gaining importance for treating many diseases due to their significant effect and 
lesser side effects as compared to allopathic medicines.In view of the above said facts, the 
present study was designed to investigate and determine the antidandruff activity of three 
different medicinal plants – Piper cubeba (Valmilagu, Kavab cini ), Bauhinia Vahlii (Mandarai, 
Malu), Cissus quadrangularis (Pirandai, Hadjod). 
 
MATERIALS AND METHODS 
 
The fresh leaves of Bauhinia vahlii, stem of cissus quadrangularis and seeds of Piper 
cubeba were collected from local sources and then they were grind into powder and stored in 
room temperature. Finely grinded plant material was extracted with hexane, ethyl acetate and 
methanol in the ratio of 1:10 in conical flask in shaking condition for overnight. The extract was 
filtered through the Whatmann No. 1 filter paper in a separate container. The process was 
repeated three times and the same plant material was used with fresh solvent. The solvent was 
removed by placing the extracts in distillation unit and maintained in respective temperature. 
The extracted residues were weighed and re-dissolved in different solvents to yield 10mg/ml 
solutions for further analysis. 
 
 Dandruff causing agent samples were collected by scraping the lesions of patients and 
stored in sterile containers in refrigerator until use. Different media formulations (potato 
dextrose agar, Sabouraud’s dextrose agar) were supplemented with olive oil and inoculated 
with the sample. The plates were incubated at 37°C for 3- 5 days .The culture was stained with 
          ISSN: 0975-8585 
 
July-September      2013           RJPBCS              Volume 4 Issue 3    Page No.394 
lactophenol cotton blue stain and examined under the high power objective of microscope and 
the characters were recorded. 
 
Various concentrations (250-1000µg/ml) of the extracts were prepared in DMSO from 
the resultant extract to determine its antidandruff activity. Control experiments were 
performed by using DMSO with identical concentration used to test the extract. Isolates from 
dandruff were inoculated by swabbing on the surface of gelled media plates. Wells of 6 mm in 
diameter were performed in the PDA media, and each well was filled with 50 µl of certain 
concentration of extract. The plates were kept in laminar air flow for 30 minutes for proper 
diffusion of the extract and thereafter incubated at 37ºC for 3-5days. The radius for the zone of 
inhibition was in millimeters and recorded against the corresponding concentration.  
 
Broth Dilution assays are standard method used to compare the inhibition efficiency of 
the antimicrobial agents. 5ml of the potato dextrose broth, 0.1ml of the 24 h growing culture 
(M. furfur) and the different concentration (100µg, 200µg…1000µg) of the crude extract 
dissolved in Dimethyl sulphoxide. The tubes were incubated at 37ºC for 24 h. The optical 
densities were measured spectrometically at 600 nm (Cos et al., 2006). The percentage of 
viable cells was calculated by using the following formula. 
 
% of inhibition =               Control O.D - Test O.D X 100 
                       Control O.D 
 
O.D = Optical density 
 
The organism was biochemically analysed by assays such as Catalase test, Nitrate 
reduction test and Urea hydrolysis test using standard procedures. 
 
RESULTS 
 
The extract of three plants Piper cubeba, Bauhinia vahlii, Cissus quadrangularis are 
obtained by direct solvent extraction method. In this method, different solvents were used 
namely hexane, ethyl acetate and methanol .The flakes from different patients were collected 
and cultured in potato dextrose broth and sub-cultured in Sabouraud Dextrose Agar (SDA)and  
Potato Dextrose agar(PDA)  by swabbing and streaking method.The significant growth was 
observed in  PDA. Well diffusion was done to determine the anti-fungal activity of three 
selected plants Piper cubeba, Bauhinia vahlii, Cissus quadrangularis with different solvents at 
different concentrations (250, 500, 750, 1000) µg/ml. The activity was found to be effective in 
the methanol extract of Piper cubeba and it was taken for further assay. Table 1   Broth dilution 
assay was performed to know the minimum inhibitory concentration of Piper cubeba. The MIC 
was 100µg/mL and IC 50 was found to be 800µg/mL [.Table 2 and Graph 1] 
                              
  
 
 
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July-September      2013           RJPBCS              Volume 4 Issue 3    Page No.395 
Table 1: Well Diffusion 
      
 
Table 2: Broth Dilution Assay 
 
 
    
          
 
 
 
 
 
 
 
 
 
 
 
Figure 2: Broth Dilution Assay 
 
DISCUSSION 
 
 Currently available treatments for dandruff include therapeutic use of Zinc Pyrithione, 
salicylic acid, Imidazole derivatives, Glycolic acid, Steroids, Sulfur and Tar derivatives. However, 
these tools have limitations in the application, or due to poor clinical efficacy or due to poor 
response to their use. Hence the herbal drugs are prove to be eco-friendly, alternative for 
synthetic drugs. In order to avoid the harmful effects of synthetic drugs for controlling dandruff, 
it is suggested to choose herbal medicines for its control. In the present study among the three 
 
S .No 
 
Conc. 
(µg/mL) 
Zone of inhibition (mm) 
Piper cubeba Bauhinia vahlii Cissus quadrangularis 
Methanol Ethyl 
acetate 
Hexane Methanol Ethyl 
acetate 
Hexane Methanol Ethyl 
acetate 
Hexane 
1 250 - - - - - - - - - 
2 500 12 - - - - - - - - 
3 750 16 - - 13 - - 13 - - 
4 1000 18 - - 16 - - 15 - - 
S .No Concentration(µg/ml) % Inhibition 
1 100 32.45 
2 200 35.81 
3 300 37.62 
4 400 40.47 
5 500 43.26 
6 600 45.45 
7 700 48.99 
8 800 51.65 
9 900 55.33 
10 1000 60.93 
          ISSN: 0975-8585 
 
July-September      2013           RJPBCS              Volume 4 Issue 3    Page No.396 
medicinal plants, methanol extract of Piper cubeba shows significant antidandruff activity 
against Malassezia furfur giving a zone of inhibition of 18mm at 1000µg/ml and an IC50 value at 
800µg/ml. Thus, methanol extract of Piper cubeba can be suggested as effective and safe 
therapeutic agent against dandruff. Hence such type of study will certainly bring out scope for 
further research to identify the potent bioactive compound having remarkable therapeutic 
value against dandruff. 
 
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Exploring the Anti-dandruff Potential of Selected Medicinal Plants

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