FORMULATION OF ALCOHOL-BASED HAND RUBS AND NON ALCOHOL-BASED HAND SCRUBS USING Aloe vera AND SELECTED  ESSENTIAL OILS AND EVALUATION OF ANTIMICROBIAL EFFICACY

Hand hygiene is the primary measure proven to be effective in preventing healthcare 
associated infections (HAI). This study aimed to develop and evaluate alcohol-based hand 
rubs and non-alcohol-based hand scrubs using Aloe vera and selected essential oils (clove, 
cinnamon, castor, nutmeg). Four types of alcohol-based hand rubs (AF₁ – AF₄) and seven 
types of non-alcohol-based hand scrubs (NAF₁ – NAF₇) were formulated by incorporating 
A. vera, essential oils and 4% chlorhexidine gluconate into the base preparations. Further, 
the physical stability of formulations prepared was evaluated for three months at room 
temperature. Agar well diffusion method was performed against selected standard cultures 
of Gram-negative, Gram-positive bacteria and fungi [(Escherichia coli (ATCC 25922), 
Enterobacter cloacae (ATCC 23355), Acinetobacter baumannii (ATCC 19606), 
Pseudomonas aeruginosa (ATCC 27853), Klebsiella pneumoniae (ATCC 70603), 
Salmonella enterica Typhi (clinical isolate), Shigella sonnei (clinical isolate), Proteus 
mirabillis (ATCC 12453), Streptococcus pyogenes (ATCC 12384), Enterococcus faecium 
(ATCC 29212), Staphylococcus aureus (ATCC 25923) and Candida albicans (ATCC 
10231)] to evaluate in-vitro antimicrobial efficacy. The formulations were found to be 
homogenous, liquid, and milky white-pale yellow except NAF₇ reddish in colour with a 
pungent odour. The higher zones of inhibition of AF₁ were 20.00 ± 2.00, 20.30 ± 1.53, 
34.67 ± 1.15, 20.33 ± 0.58, and 22.67 ± 0.58 mm and of AF₃ were 22.30 ± 2.52, 20.00 ± 
1.00, 37.00 ± 1.73, 21.00 ± 1.00, and 21.33 ± 0.58 mm against A. baumannii, P. mirabillis, 
C. albicans, S. pyogenes, and S. aureus respectively. The higher zones of inhibition of 
NAF₁ were 20.00 ± 1.00, 23.67 ± 0.58, 25.33 ± 0.58, 23.00 ± 5.29, and 18.67 ± 1.53 mm 
and of NAF₃ were 23.33 ± 1.15, 20.33 ± 1.15, 34.67 ± 1.15, 21.00 ± 0.00 and 19.67 ± 1.53 
mm against A. baumannii, S. enterica Typhi, C. albicans, S. pyogenes and S. aureus 
respectively.  The higher zones of inhibition of NAF₇ were 25.00 ± 1.00, 34.33 ± 0.58, 
23.67 ± 0.58, 24.33 ± 0.58, and 27.33 ± 0.58 mm against S. enterica Typhi, C. albicans, S. 
pyogenes, E. faecium, and S. aureus respectively. Zero values were obtained for distilled 
water (N₁), glycerin (N₂), and 70% aqueous ethanol (P₂) for both conditions. The market 
product (P₁) showed inhibition zones of less than 12.33 ± 1.15 mm for NAF against E. 
cloacae, S. enterica Typhi, C. albicans, S. pyogenes, E. faecium, and S. aureus and less 
than 13.67 ± 0.58 mm for AF against S. sonnei, C. albicans, S. pyogenes, E. faecium, and 
S. aureus. The most active five formulations (AF₁, AF₃, NAF₁, NAF₃ and NAF₇) were 
subjected to phase II, in-vivo antimicrobial efficacy test, by using the ‘Fingertip method’ and it showed > 96% reductions of colony-forming units (CFUs). The results of self
administered questionnaire showed that the majority of participants (> 90%) had no 
hesitation, itching, irritation, rashes, or skin damage while > 85% was no skin-drying 
condition. More than 65% of participants responded with a moisturizing effect in tested 
formulations. In conclusion, the formulated hand rubs/scrubs are having promising 
antimicrobial activity against the tested pathogens and clinically proved the skin 
sensitivity, safety, and compatibility against human volunteers. Production cost per 100 
mL was calculated as Rs. 200.00 which suggest that not only formulations are efficient but 
also cost effective. 

Keywords - Alcohol-based hand rub, antimicrobial efficacy, essential oils, fungi, Gram
negative, Gram-positive, non-alcohol-based hand scrubs