BIOLOGIA (PAKISTAN) 2012, 58 (1&2), 161-166

PK ISSN 0006 - 3096

Determination of skin irritancy by essential oils from some

members of Family Rutaceae of Pakistan
FERHAT MEHMOOD & ZAHEER UD DIN KHAN
Department of Botany GC University, Lahore, Pakistan.
ABSTRACT
In this study essential oils from some members of family Rutaceae were tested
for their possible potential to cause dermatitis on human skin using Patch test. Essential
oils from root of Boenninghausinia albiflora showed maximum positive results regarding
Irritant Contact Dermatitis (ICD) i.e. 71.42% while essential oils from Zanthoxylum
armatum leaves exhibited minimum i.e. 21.42%. The essential oils from other plant
species showed weak positive while some even negative results. Gender based results
indicated variations, as females of 12-16 years age group 25.57%, 20-22 years age group
31.5% and above 40 years 46.6% positive results. Among males, 12-16 years age group
39.84%, 20 years age group 33.2% and above 40 years 37.7% positive results. The
results can help the manufacturers of sundry products using essential oils, for human use.
Key words: Essential oils; Rutaceae; Patch test; Boenninghausinia albiflora; Zanthoxylum
armatum

INTRODUCTION
Skin reactions due to plants are common, as plants are ubiquitous and
exhibit a number of physical and chemical properties that may result in harmful
effects. These reactions are of two types, i.e. Irritant Contact Dermatitis (ICD)
and Allergic Contact Dermatitis (ACD). In ICD localized rash or irritation of the
skin occurs (Lovell, 1993; Gunjan et al., 2009), while ACD represents a delayedtype hypersensitivity reaction (Type IV hypersensitivity). The ICD comprises 80%
of all contact dermatitis reactions. Severe acute toxic reactions with necrosis may
be seen following exposure to potent alkaloids from plant sap (Lovell, 1997;
Morris et al., 2002). In type IV reactions, an allergen easily penetrates the
stratum corneum and covalently binds to keratinocytes in the stratum spinosum
below and elicits an immune response that is remembered on subsequent
allergen exposures (Yunginger, 2003; Yiannias, 2004; Cohen & Heidary, 2004).
Irritation and its severity depend on the functional integrity of the individual's skin,
the potency and amount of irritant to which one is exposed, and the duration of
irritant exposure. According to Cavani & Luca (2010), chemical irritants are
responsible for chemical ICD (CICD). Diterpene esters are among the most
irritant of plant-derived chemicals (Hughes et al., 2002; Asilian & Faghihi, 2004).
The skin and eyes are particularly susceptible to ICD (Raison et al., 2003;
Cappiello & Shadow, 2005).
Family Rutaceae has genera, many of which produce edible fruits and
essential oils are obtained from many species, which are used for medicinal
purposes, aroma-therapy, perfume industry, flavor and fragrance (Sawamura,
2011). Apart from the useful effects some of essential oils can cause irritation or
provoke an allergic reaction (Heskel et al., 1983).
Correspondence; ferhatmehmood786@yahoo.com

162

F. MEHMOOD & Z. KHAN

BIOLOGIA (PAKISTAN)

Essential oils have wide and varied applications in scenting and

flavouring, finished products, which contribute directly to our health, happiness
and well-being. At the same time the essential oils may cause dermatitis. Some
essential oils are photosensitizers (Sams, 1941). Oil of lemon peel could cause
irritation. Peelers of citrus fruits suffer from dermatitis and paronchya (Opdyke,
1974).
The present study has been planned for the first time in Pakistan,
keeping in view, the ever increasing use of essential oils in medicinal, food
articles, aroma-therapy and especially cosmetics (which might come under use
of any age group) and the possibility of essential oils to cause Contact
Dermatitis. This study has been carried out with hope that its results would be
helpful not only for people at large regarding health aspects but also to the
commercial entrepreneurs from economic point of view as well.
MATERIALS AND METHODS
The essential oils of the following plants and their parts of members of
family Rutaceae were considered for their Irritant Contact Dermatitis (ICD)
effects:
Table 1: List of plants and their parts used
Botanical Name
Aegle marmelos (Linn.) Correa
Boenninghausenia albiflora (Hook.)
Reichb.ExHeynh.
Citrus reticulata (Blanco) cv.honey
Murraya koenigii (Linn.) Spreng

Local name
Bel
Pissu mar buti,
Honey
Curry

patta

Murraya paniculata (Linn) Jack cv.

China
Murraya paniculata (Linn) Jack cv. Desi.
Skimmia laureola (DC.) Zucc. ex Walp

Marwa

China

Zanthoxylum aromatum DC

Timber

Marwa desi
Nair

Part/s used
Rind and leaves
Stem,
root
and
leaves
Rind, leaves
Leaves
Leaves
Leaves
Leaves, stem and
root
Leaves, seeds

The plants were selected for the present study on the basis of their
ethno-botanical uses in Pakistan. The plants were collected from their natural
habitat (Abbotabad, Kooza Gali, Murree Hills and Nathia Gali) identified and
authenticated by a working taxonomist in Dr. Sultan Herbarium, GC University,
Lahore. The respective plant parts were separated and subjected to hydrodistillation for about four hours. The essential oils thus obtained were dried over
anhydrous sodium sulphate and stored in dark colored glass bottle at
o
temperature of about 4 C.
GC-MS Analysis of Essential Oils
GC-MS analyses were performed using DB-5MS column in triplicate with
a blank run after every analysis on a Shimadzu GCMS-QP2010A system, and EI

VOL. 58 (1&2)

SKIN IRRITANCY BY ESSENTIAL OILS

163

mode (70eV) equipped with injector at 250 C. The data thus obtained was
processed using Shimadzu Lab Solution GCMS Post run Analysis software.
Comparing the mass fragmentation pattern of the reported data and NIST 147
and NIST 27 libraries, the components of essential oils were identified.
Patch Test
Patch test (Seidenari et al., 1990) was performed in collaboration with
Department of Dermatology, KEMC University, Lahore, after taking permission
(NOC) from Vice Chancellor, King Edward Medical College University, Lahore.
One hundred (100) normal volunteers, of either sex, ranging from 10-45 yrs of
age from different walks of life and professions e.g. Government servants,
Private job personals, students, family members, house maids, peons etc., were
enrolled after getting the consent form signed by them.
Test Procedure
It was made sure that skin of volunteers was clean, healthy and free of
ointments, lotions, powders, acne, dermatitis, scars, hair or any other condition
that might interfere the effect of samples of essential oils. Moreover it was
ensured that none of the volunteer was taking any sort of anti-allergic drug.
i.
About a drop of each sample (in petrolatum 5% w/w) was applied on
about 1 cm diameter dark colored food grade plastic disc.
ii.
The disc (with the applied sample side) was placed on the palm side
skin of forearm in vicinity of elbow and fixed with the help of
cellophane tape. At a time two to three samples were applied.
Sample number was written on the tape.
iii.
Samples were kept applied on the skin for 48 hours. Results were
taken after 15-20 minutes of removal of the discs, for better
observations.
iv.
An individual showing any positive result was given treatment as per
prescription of consultant dermatologist and Patch testing was done
again after complete subsidence of signs and symptoms and
withdrawal of the drugs.
Results were interpreted according to International Contact Dermatitis
Research Group Scale (Fregert, 1981 & Foussereau et al., 1982) given in Table
2. The Chi-Square test was applied to analyze the results statistically, using
STATA version 10, statistical software.
RESULTS AND DISCUSSION
GC-MS analyses revealed Limonene as the highest percentage
compound in essential oils from leaves and rind Aegle marmelos. This has
already been reported by Suvimol & Pranee (2008).
. Ketones dominated in essential oil from leaves of Zanthoxylum
armatum while Limonene, cymene and linalool dominated in EO from its seeds.
EO from leaves of Murraya koenigii had -Cymene and Linalool in agreement
with Rees et al.,(1989). Germacerene and Caryophylline were in high
concentration in leaves of M. paniculata cv. China and Nerodilol and Cubebene in leaves of M. paniculata cv. Desi. In Citrus reticulata, Blanco cv.
Honey leaf and rind essential oil showed Limonene in high percentage, which
was also reported by Al-Sheikh & Gad el-Rab (1996). The essential oils from

164

F. MEHMOOD & Z. KHAN

BIOLOGIA (PAKISTAN)

Boenninghausinia albiflora root, stem and leaves showed maximum number of

compounds, i.e. 38, 35 and 33 respectively. Apart from the variations in the
three oils, sesquiterpenes dominated in them.
Results obtained in patch test (Table 2) had been quite variable. Apart
from other factors such as, type, amount of allergen and exposure time, the
variation in positivity to the patch test could be probably due to the genetic
backgrounds of the individuals (Cohen & Heidary, 2004). Essential oils from
different parts of B. albiflora showed maximum positive results while essential
oils from M. koenigii and Z. armatum leaves exhibited minimum regarding Irritant
Contact Dermatitis (ICD). To be brief the intensity of ICD by different essential
oils was in order of B. albiflora root 71.42%, S. laureola root 66.66 %, B. albiflora
stem 64.28%, C. reticulata (honey) rind 59.52%, S. laureola stem 54.76%, B.
albiflora leaves 54.76%, M. koenigii leaves 28.57%, A. marmelos rind 38.09%,
and Z. armatum leaves 21.42%. Essential oils from leaves of S. laureola, M.
paniculata desi, M. paniculata china, C. reticulata (honey), A. marmelos and
from seeds of Z. armatum showed no reaction.
A significant difference was found between the ICD activities of all
essential oils used in this study after using Chi-Square test statistic, at P 0.05.
Table: 2 Results of Patch Tests in percentage
Names of Plant & part used
for EO
S. laureola leaves
S. laureola root
S. laureola stem
M. koenigii leaves
M. paniculata desi leaves
M. paniculata china leaves
C. reticulata (honey) rind
C. reticulata (honey) leaves
A. marmelos leaves
A. marmelos rind
B. albiflora leaves
B. albiflora stem
B. albiflora root
Z. armatum leaves
Z. armatum seeds

?
Nil
Nil
Nil
Nil
Nil
Nil
Nil
Nil
Nil
Nil
Nil
Nil
Nil
Nil
Nil

Nil
24
17
24
Nil
Nil
29
Nil
Nil
15
48
57
38
Nil
Nil

ICDRG Scale

Nil
Nil
100
Nil
Nil
33
Nil
Nil
45
Nil
Nil
71
Nil
Nil
100
Nil
Nil
100
Nil
Nil
40
Nil
Nil
100
Nil
Nil
100
Nil
Nil
64
Nil
Nil
45
Nil
Nil
36
Nil
Nil
29
Nil
Nil
79
Nil
Nil
100

IR
Nil
43
38
5
Nil
Nil
31
Nil
Nil
21
7
7
33
21
Nil

NT
Nil
Nil
Nil
Nil
Nil
Nil
Nil
Nil
Nil
Nil
Nil
Nil
Nil
Nil
Nil

*International Contact Dermatitis Research Group scale

? Doubtful reaction
Weak positive reaction
Strong positive reaction
Extreme positive reaction
Negative reaction.
IR Irritation reaction of different types
NT Not tested

Gender based results showed variations as females of 12-16 years age

group showed 25.57%, 20 years age group 31.5% and above 40 years showed
46.6% positive results. Among males 12-16 years age group showed 39.84%, 20

VOL. 58 (1&2)

SKIN IRRITANCY BY ESSENTIAL OILS

165

years age group 33.2% and above 40 years exhibited 37.7% positive results.
These results are in contrast to Rees et al., (1989) and Al-Sheikh & Gad el-Rab,
(1996) which showed that females exhibited a larger response in the
development of contact sensitivity induced by dinitrochlorobenzene (DNCB) at all
challenge doses studied, and the slope of the log-dose response curve was
significantly steeper in females.
Essential oils under study were found composed of variety of chemical
components. Among these, Limonene and its oxidation products are skin and
respiratory irritants (Anonymous, 1999). -cymene is also skin irritant as
determined by Bennett et al., 1982. As per study of Kanikkannan & Singh, 2002
undecanol is irritant to rabbit but not to human. Sax & Lewis (1989) found glycidol
as an irritant of the skin, eyes, mucous membranes, and upper respiratory tract.
Moreover exposure to glycidol may also cause central nervous
system depression, followed by central nervous system stimulation. The possible
mechanism of irritant reaction is due to easy penetration of these substances to
the stratum corneum that covalently bind to keratinocytes in the stratum
spinosum below (Cavani & Luca, 2010; El-Azhary & Yiannias, 2004; Cohen &
Heidary, 2004). Only the superficial regions of the skin are affected in contact
dermatitis (Morris et al., 2002).
Conclusion
Essential oils from some members of family Rutaceae tested in this
study for contact dermatitis can provide information for direct preventive
measures such as exposure reduction to prevent sensitization as well as
elicitation of contact dermatitis as suggested by Nielsen & Menne (1992) and
Nielsen et al., (2002).
REFERENCES
Al-Sheikh, O.A. & Gad el-Rab. M.O., 1996. Allergic contact dermatitis: Clinical
features and profile of sensitizing allergens in Riyadh, Saudi Arabia. Int.
J. Dermatol., 35:493-497.
Anonymous., 1999. WHO, International Agency for Research on Cancer.
pp: 307.
Asilian, A. & Faghihi, G., 2004. Severe irritant contact dermatitis from cypress
spurge. Cont. Dermat., 51: 37-39.
Bennett, M.A., Huang, T.N., Matheson, T.W. & Smith, A. K., 1982. (h 6Hexamethylbenzene)ruthenium complexes. Inorg. Synth., 21: 7478.
Cappiello, P. & Shadow, D. D., 2005. The genus Cornus, Timber Press, Portland,
pp. 78-83.
Cavani, A. & Luca, D., 2010. Anastasia Allergic Contact Dermatitis: Novel
Mechanisms and Therapeutic Perspectives: Current Drug Metabolism,
Bentham Science Publishers, USA, pp. 228-233.
Cohen, D.E. & Heidary, N., 2004. Treatment of irritant and allergic contact
dermatitis. Dermatol. Ther., 17: 334-40.
El-Azhary, R.A. & Yiannias, J.A., 2004. A new patient education approach in
contact allergic dermatitis: The Contact Allergen Replacement database
(CARD). Int. J. Dermatol., 43: 278-80.

166

F. MEHMOOD & Z. KHAN

BIOLOGIA (PAKISTAN)

Foussereau, J., Herve-Bazin, B. Meynadier,J., Reuter,G. & Cavelier, C., 1982.

Allergic contact dermatitis to plastic table cloth and phenyl indole. Cont.
Dermat., 8: 73-75.
Fregert, S., 1981. Contact allergy to phenoplastics.Contact Cont. Dermat., 7:
170-73.
Gunjan, M., Modi, C.B., Doherty, R.K. & Orengo, I.F., 2009. Irritant Contact
Dermatitis from Plants. Dermatitis, 20: 63-68.
Heskel, N.S., Amon, R.B., Storrs, F.J. & White, C.R., 1983. Phytophotodermatitis
due to Ruta graveolens. Cont. Dermat., 9: 278-80.
Hughes, T.M., Varma, S. & Stone, N.M., 2002. Occupational contact dermatitis
from a garlic and herb mixture. Cont. Dermat., 47: 48-50.
Kanikkannan, N. & Singh, M., 2002. Skin permeation enhancement effect and
skin irritation of saturated fatty alcohols. Int. J. Pharm., 248: 219-28.
Lovell, C.R., 1993. Plants and the skin. Blackwell Scientific Publications. Oxford
and Boston, pp. 125-26.
Lovell, C.R., 1997. Phytodermatitis. Clin. Dermatol., 15: 607-13.
Morris, J.R., Robertson, S.J. & Ross, J.S., 2002. Dermatitis caused by physical
irritants. Br. J. Dermatol., 147: 270-75.
Nielsen, N.H. & Menne, T., 1992. Allergic contact sensitization in an unselected
Danish population. The Glostrup Allergy Study, Denmark. Acta. Derm.
Venereol., 72: 456-60.
Nielsen, N.H., Linneberg, A., Menne.T, Madsen, F., Frolund, L. & Dirksen, A.,
2002.
Incidence of allergic contact sensitization in Danish adults
between 1990 and 1998: The Copenhagen Allergy Study, Denmark. Br.
J. Dermatol., 147: 487-92.
Opdyke, D.L.J., 1974. Monographs on fragrance raw materials. F. Comset.
Toxicol., 12: 725-66.
Raison, P.N., Roulet, A., Guillot, B. & Guilhou, J. J., 2003. Bromelain: an unusual
cause of allergic contact cheilitis. Cont. Dermat., 49: 218-29.
Rees, J.L., Friedmann, P.S. & Mathews, J.N., 1989. Sex differences in
susceptibility to development of contact hypersensitivity to
dinitrochlorobenzene (DNCB). Br. J. Dermatol., 120: 371-74.
Sams, W.M., 1941 Photodynamic action of lime oil (Citrus aurantifolia). Arch.
derm. Syph., 44: 571-77.
Sawamura, M., 2011. Citrus essential oils: Flavour and Fragrance. John Wiley
and Sons, Inc. Hobken, New Jersey, pp. 180-90.
Sax, N.I. & Lewis, R.J., 1989. Dangerous properties of industrial materials. 7th
ed. Van Nostrand Reinhold Company New York, pp. 240-51.
Seidenari, S., Manzini, B.M., Danese, P. & Motolese, N., 1990. A. Patch and
prick test study of 593 healthy subjects. Cont. Dermat., 23: 162-67.
Suvimol, C & Pranee, A., 2008. Bioactive Compounds and Volatile Compounds
of Thai Bael Fruit (Aegel Marmelos (L.) Correa) as a Valuable Source for
Functional Food Ingredients. Internat. Food Res. J., 15 (3): 287-295.
Yunginger, J.W., 2003. Natural rubber latex allergy: Middleton's allergy principles
and practice. Elsevier. Philadelphia, pp. 110-117.
Yiannias, J.A., 2004. The Principles and Practice of Contact and Occupational
Dermatology in the Asia-Pacific Region. Internat. J. Dermatol., 43: 22931.