J Sri Food Agric 1992.

58, 49-53

Steam Distillation of Tea Tree (Melaleuca

alternifolia) Oil
Michael R Johns,* Julia E Johns and Victor Rudolph
D e p a r t m e n t of C h e m i c a l Engineering, The University of Queensland, Queensland 4072, Australia
(Received 31 July 1991 ; accepted 21 August 1991)

Abstract: The steam distillation of tea tree (Melaleuca alternifolia Cheel) oil was
investigated in a small batch still to ascertain the effect of operating variables on
the yield, composition and rate of recovery of oil. Provided suitable distillation
times (2-3 h) were used, the oil yield and composition did not vary with steam rate
or pretreatment of the leaves by maceration. However, the rates of recovery of
individual oil components were markedly affected.
The results demonstrate that the oil components can be divided into two main
groups according to their distillation behaviour. Oxygenated components,
particularly terpinen-4-01 and 1,8-cineole, are recovered the most rapidly, despite
their high boiling points relative to other components. Their rates of recovery are
increased with increasing steam rate, but are insensitive to leaf maceration. A
second group of components, characterised by either a hydrophobic structure (the
monoterpenes) or a larger molecular size (sesquiterpenes), exhibited the opposite
behaviour. The rates of recovery of these components were unaffected by steam
rate and improved by leaf maceration. It is hypothesised that the rate-limiting
processes for oxygenated and hydrophobic components are different. For the
former, recovery is mass-transfer film controlled, whereas the latter are diffusion
controlled. This understanding provides the basis for manipulating oil quality by
process conditions.
Key words : Tea tree, essential oil, steam distillation, terpinen-4-01,

INTRODUCTION properties (Penfold a n d G r a n t 1925; Lassak and

Tea tree oil is present subcutaneously in the leaves of
Melaleucu species, particularly M alternifolia Cheel, a n d
may be recovered by steam distillation. Tea tree oil has
powerful antiseptic properties, penetrates infected tissue
and is non-irritant and non-corrosive (Laakso 1965). It is
used in perfumes, in the control of wood-rot fungi, as
an antiseptic in water-gel fire blankets, in combating
Legionella in air-conditioning systems and as a nutmeg
substitute (Southwell 1988). The oil has bactericidal
properties against a wide range of gram-negative and
gram-positive bacteria, yeast and fungi (Altman 1988).
The composition of tea tree oil has been reported to
be a mixture of terpenes, terpene alcohols and ses-
quiterpenes (Laakso 1965; Guenther 1968; Swords and
Hunter 1978). The major component, terpinen-4-01, is
considered to be responsible for its antimicrobial
* To whom correspondence should be addressed.
49
J Sci Food Agric 0022-5142/92/$05.00 0 1992 SCI. Printed in Great Britain
McCarthy 1983). A recent study concluded that it is
derived from thujane precursors either as a n artefact of
steam distillation o r biogenetically (Southwell and Stiff
1989). The Australian Standard for oil of Meluleuca
specifies that the oil must contain a minimum of 30% v
of terpinen-4-01 a n d not more than 15 YOv of cineole.
However, the concentrations required for optimal
efficacy are not known.
M alternifolia leaves are harvested either manually
from natural forest stands o r by mechanised harvesters
working plantations of cultivars selected for their high
oil content. In the latter case, steam distillation of the
biomass, which requires as long as 3 h per batch, can
become the rate-limiting step in the processing of the oil.
There are few reports concerning the steam distillation
of tea tree oil in the literature. Current practice entails
packing freshly harvested twigs containing the leaves
into a batch still and steaming until oil recovery
diminishes. The oil-bearing steam is condensed and the

4.2
50
oil is separated from the water and collected. Typical
volumetric yields of oil are 1-3% of the wet weight of
biomass (Penfold and Morrison 1950).
Previous reports have shown that the components in
the tea tree oil distil off in an unexpected sequence not
related to their boiling points (Southwell 1988). No
detailed explanation has been given to date for this
phenomenon. Furthermore, no studies have been pub-
lished demonstrating the effects of leaf age, steam rate or
leaf treatments on oil yield and composition.
This paper describes research performed to examine
the effects of these variables on the distillation of tea
tree oil using steam so as to gain an insight into the
mechanisms controlling the rate of recovery of the oil
and its components.
EXPERIMENTAL
Tea tree supply and charge preparation
Tea tree (Meluleuca ulternifolia) was supplied from a
commercial plantation (Australia Tea Tree Estates,
Mullumbimby, Australia) within 24 h of harvest and was
stored at 4°C in a double thickness of air-tight plastic
bags. An initial supply was used for the first four runs
at high steam flow rate, and a second for subsequent
experiments. The foliage consisted of short twigs (up to
10 mm dia) with leaves attached, and a large proportion
of unattached leaves. These were separated by sieving
(20 mm mesh) and kept separately. Large pieces of twig
and bark were removed from the sample.
All distillation experiments used 2 kg of tea tree
biomass in the mass ratio 1.7 :0.3 leaf: twig unless stated
otherwise. In each instance the twig portion of the
sample was laid over the mesh (20 mm opening) at the
base of the still to prevent the loss of the finer leaf
fraction, which was placed on top, giving a final charge
height of approximately 100 mm.
Grinding of the leaf fraction was accomplished, where
required, by macerating 100-g samples in a Waring
blender at high speed for 30 s. Oil and water released by
grinding prevented size measurement, although from
visual observation leaf size was in the order of 2 mm.
Distillation apparatus
The stainless steel still (Fig 1) was lagged to prevent
steam condensation. Steam flow rate was controlled by
passing steam regulated to 20 kPa g through a needle
valve, which was calibrated against condensate flow. For
lower steam flow rates (less than 8 kg h-'), the still was
modified to reduce the headspace and increase the
number of sparge holes in the steam distributor. This
minimised channelling and the condensation of steam in
the still. The pressure in the still was equivalent to
atmospheric pressure for all runs.
A4 R Johns, J E Johns, V Rudolph
\
@
Steam
Inlet x
still
water
Condenser1
Drain I
Fig 1. Schematic diagram of experimental apparatus,
The condenser consisted of 3 m x 10 mm OD stainless
steel tubing wound in a spiral and immersed in water.
The condensate temperature was maintained at 30°C by
controlling cooling water flow.
The separator used to disengage the condensed oil
from water depended on the steam flow rate. For steam
rates greater than 2 kg h-' it consisted of a glass aspirator
bottle with a graduated burette attached to the top into
which the condensed mixture flowed. A stopcock was
fitted to remove excess condensed water. For lower
steam flow rates, graduated 100-ml burettes were used to
collect and measure condensate and oil volume.
Analysis of oil composition
Oil composition was determined using the method
specified in the Australian Standard AS 2782-1985. A
GC-MS (Hewlett-Packard 5955) equipped with a flame
ionisation detector and a J&W DB5 capillary column
(30 m x 0.25 mm ID) was used with helium carrier gas
and temperature programming of 1 min at 90°C, in-
creasing at 5°C min-' to 220°C. An injection volume of
0.5 PI was employed. Components were identified by
comparison of their retention times against authentic
standards (Aldrich Chemical Co, WI, USA) or, in the
case of minor components, were deduced from their
mass spectral and chromatographic characteristics.
Component concentrations are expressed as % v of oil.
Terpinen-4-01 dissolved in the separator water was
isolated by freeze extraction and the concentration was
obtained by gas chromatography.
RESULTS
Distillation sequence of oil components
Tea tree was steamed at a rate of I kg steam h-' and
the composition of the oil recovered during the dis-
tillation was determined. A total of 27.6ml of oil was
obtained, a yield of 1.4% v/w. The final composition of
the oil (Table 1) was similar to previously reported
Steam di.ytillation of tea tree oil 51

100
TABLE 1
Composition of tea tree oil 90

80
Component BP GC Concentration
(“C) RT (% v) 5 70
W
(min) x
3 60
This Ref 1 Ref 2 Ref 3 Ref 4 1
0
>
study W
50
2
4
c 40
a-Pinene 155.0 5.58 2.5 3.6 2.2 0.2 2.8 3
I
P-Pinene 158.3 7.10 1.6 0.9 - 3.8 0.6 3
u
30
P-Myrcene 171.5 6.56 0.3 1.2 - - 0.5 20
a-Terpinene - 7.44 9.4 12.4 7.5 1.9 2.7
p-Cymene 177.2 7.58 2.1 2.6 3.0 8.1 11.4 10

Limonene 175.0 7.71 0.9 0.9 1.0 - 3.1 I I l l , I ,

I$-Cineole 175.0 7.85 2.9 7.2 5.6 2.6 16.5

y-Terpinene - 8.53 20.4 244 17.5 12.0 11.5 TIME (MIN)

Terpinolene 185.0 9.27 3.3 41 3.1 24.5 2.4 Fig 3. Recovery of tea tree oil components from oil added back
Terpinen-4-01 212.0 12.10 38.5 31.2 44.9 32.5 29.4 to previously exhausted leaves. 1,8-Cineole; terpinen-4-
a-Terpineol 217.5 12.26 2.9 2-1 5.2 1.8 3.6 01; 0 p-cymene; V P-pinene; a-pinene; sesquiterpenes.
Sesquiterpene - a 13.7 - 3.3 9.5 -

Components with retention times greater than 12.6 min.

References : 1, Southwell and Stiff (1989) ; 2, Guenther (1968);
3, Laakso (1965); 4, Swords and Hunter (1978).

100

90

80
../
bs
70
W
z
3 60

’
0

W
50
2
+ 40
4 0 20 40 60 80 100 120 140 160
3
30 TIME (MIN)
3
0
20 Fig 4. Comparison of recovery of a-pinene and terpinen-4-01
between normal steam distillation and when oil was added
back. Normal distillation : V a-pinene; terpinen-4-01; oil
0 20 40 60 80 100 120 140 160 added back : a-pinene; V terpinen-4-01,
TIME (MIN)

Fig 2. Recovery of tea tree oil components with time using a

steam rate of 1.5 kg h-l. 0 1,8-Cineole; terpinen-4-01; V p-
cymene; vP-pinene; a-pinene; sesquiterpenes.
analyses of M alternifolia oil with a terpinen-4-01 and
1,8-cineole content of 38.5 YO and 2.9 O h respectively.
There were several unidentified components present in
the oil at concentrations less than 0.5 YO(v).
The rates at which the individual components were
recovered corresponded more to their chemical structure
and size than to their boiling points (Fig 2), as previously
noted by Southwell (1988). The more hydrophilic
compounds (terpinen-4-01, 1,8-cineole and a-terpineol)
were completely recovered within 90 min, whereas 95 YO
recovery of the remaining components required 120 min.
The sesquiterpene fraction exhibited a very slow rate of
recovery.
To examine further the sequence in which components
were recovered from the leaf surface, 30ml of oil
recovered from a previous experiment was sprinkled on
to tea tree material which had been previously steamed
to oil exhaustion. A steam distillation was then per-
formed using operating conditions identical to those of
the original experiment. Less oil (17.3 ml) was recovered
than had been added back, although the composition
was the same. In contrast to the distillation of oil-bearing
leaves, the components were recovered in the sequence
expected from their boiling points and the overall
distillation process was faster (Fig 3).
The distillation behaviour of the components could
again be lumped into two groups. Whereas the rates of
recovery of oxygenated components (terpinen-4-01, I ,8-
cineole and a-terpineol) were unchanged, the rates of
recovery of all the terpene components from the surface
of leaves were greatly increased compared with those in
an oil-in-leaf distillation (Fig 4). The rates of recovery
52 M R Johns, J E Johns, V Rudolph

A
100

90

3 80
v

70
I
3 60
0
' 50

'5
W

40

3 30
x
3
0 20

I I I I I I I

0 20 40 60 80 1 0 0 1 2 0 1 4 0 1 6 0 0 20 40 60 80 100 120 140 160

TIME (MIN) TIME (MIN)

Fig 6. Effect of leaf maceration on oil component recovery.

Macerated leaves : a-pinene; V terpinen-4-01; normal
B distillation : a-pinene; 0 terpinen-4-01; a-pinene added
100 back.
-
u
v
90
80
; 70 Effect of leaf maceration
I3
2 60
0
'
W
50 Pre-treatment of the leaves by maceration was tested to
40 determine whether an improvement in the rate of
2

-
2or
30

10

0
0 20 40 60 80 1 0 0 1 2 0 1 4 0 1 6 0
TIME ( M I N )
Fig 5. Effect of steam rate on recovery of (A), terpinen-4-01,and
(B). a-pinene. Steam rate (kg h-'): H 0.25; 0 1; D 1.5; V 2;
0 8.
decreased slightly with increasing component boiling
point.
Effect of steam flow rate
A series of steam distillations was performed at steam
rates between 0.25 and 8 kg h-l. The lower end of this
range corresponds to specific rates (kg steam kg
charge-' h-l) typically employed by commercial dis-
tillers. The steam rate had little effect on oil composition
or on the total volume of oil recovered, but did affect the
rate of component recovery. Oil recovery varied between
24 and 30 ml as a result of natural differences between
charges.
Tea tree oil can be divided into two groups of
components in terms of their response to steam flow rate.
The members in each group are the same as those
reported above. One group, comprising oxygenated
molecular structures, demonstrated a correlation be-
tween rate of recovery and steam flow rate (Fig 5A). The
second group comprised the terpenes and sesquiterpenes,
whose rate of recovery was independent of steam Bow
rate over the range tested (Fig 5B).
recovery of the pinene fraction might be effected. A
steam flow rate of 1.3 kg h-' was used.
Maceration of the leaves promoted steam channelling
during the distillation, and reduced the overall quantity
of oil recovered (23 ml compared with 30 ml for the
control distillation without maceration), but the oil
composition was similar to that of oil recovered from
standard distillation.
Maceration of the leaves appeared to increase notably
the rate of recovery of the terpene fraction compared
with that in the control distillation (Fig 6), but the
improvement was less than that observed when oil was
added to the leaves. The rate of recovery of oxygenated
components from the macerated leaves was worse than
that from the control. There are two probable explan-
ations for these results. First, the channelling in the
still may account for the reduced rate of oxygenated
component recovery and would suggest that the recovery
of all components was similarly affected. Secondly,
microscopic observation of the macerated leaves revealed
that many oil glands remained intact despite the
treatment.
Effect of storage
Tea tree which had been stored for 35 days at 4°C was
steam distilled at a steam rate of 1.3 kg h-l. Storage was
found to have no effect on the quantity of oil recovered
compared with fresh biomass distilled under identical
process conditions ; however, the rate of recovery of each
of the oil components was slightly reduced for the stored
material.
The composition of the oil recovered from stored
Steam distillutioii of tea tree oil
material was similar to that from fresh leaves, except that
the concentration of p-cymene had increased by 100 YO
during storage. This is consistent with results determined
by Southwell (1988), who stated that p-cymene is
produced from the oxidation of terpinenes during ageing.
DISCUSSION
The steam distillation of tea tree oil is a multi-component
distillation in which the dominant mechanism controlling
the rate of recovery of each component in the oil depends
on the nature of the component. From the results it is
possible to divide the components into two main groups
according to their behaviour during steam distillation.
The first group (terpene group) consists of the terpene
and sesquiterpene components, principally a- and y-
terpinene, a- and 8-pinene, terpinolene and p-cymene.
Whereas these components are the most volatile, they
were recovered more slowly than those with higher
boiling points, except when the oil was added back to the
surface of the charge. Their rate of recovery from tea tree
leaves was unaffected by steam rate and improved by
maceration of the leaves. Since the oil is contained in
subcutaneous glands distributed through the leaf, this
behaviour is consistent with a rate-controlling process
such as a diffusive resistance within the leaf. This has
been classically termed ’ hydrodiffusion ’ in essential oil
literature (Guenther 1948) and has been considered to
predominate during tea tree oil distillation (Southwell
1988).
A quite different situation exists for the second group
of components (oxygenated group) which comprises
terpinen-4-01, cineole and a-terpineol. The results of
experiments performed at different steam rates and with
the oil added back to the leaf surface show that the rate-
controlling process is operating at the leaf surface and
that recovery is mass-transfer film controlled rather than
diffusion controlled. Consequently, increasing steam rate
had a corresponding effect on the rate of recovery of
these components. This group of components experience
a ‘true’ steam distillation and appear to be little affected
by diffusive resistances in the leaf. The component p -
cymene behaved in a manner intermediate between the
two groups.
An examination of the structure of the terpene group
of components shows that they are either relatively
hydrophobic compounds or, in the case of the ses-
quiterpenes, considerably larger than the monoterpene
components. Either of these features might reasonably
be considered to account for a greater resistance to their
diffusion through the aqueous matrix of the leaf
compared with the more oxygenated monoterpenes. The
latter should diffuse more rapidly to the leaf surface and,
despite their higher boiling points, are recovered in the
steam much earlier.
It is clear that by manipulating the operating con-
53
ditions and length of time for the steam distillation
process, oils of varying composition can be produced.
For example, oils containing a high proportion of
terpinen-4-01 can be obtained using a high steam rate and
short times. Furthermore, it would appear that, in order
to increase the rate at which the more hydrophobic
components are recovered, some method of reducing the
diffusive resistance of the leaf to these components is
necessary. Grinding the leaves achieved this to a certain
degree, but also led to the problem of maintaining an
efficient distribution of steam throughout the com-
minuted material. Alternative methods which reduce this
resistance without affecting steam flow through the
material are required.
Finally, the ability to group a multiplicity of tea tree
oil components into two groups according to their
behaviour suggests the possibility that the distillation of
components from any other oils from similar plant
materials may be predicted provided that the oil
composition is known and that the diffusive and mass-
transfer properties of the component groups can be
determined.
ACKNOWLEDGEMENTS
The technical assistance of Mr Kevin Dobson for
GC-MS analyses is gratefully appreciated and our
special thanks go to Australian Tea Tree Estates for the
provision of tea tree foliage. Financial support for this
project from the University of Queensland Foundation
Limited is acknowledged.
REFERENCES
Altman P M 1988 Australian tea tree oil. Aust J Pharm 69
276278.
Guenther E 1948 The Essential Oils, Vol 1. Van Nostrand, New
York, pp 87-185.
Guenther E 1968 Australian tea tree oils. Perjium E.went Oil Rec
59 642-644.
Laakso P V 1965 Fractionation of tea tree oil (Melaleuca
alternifolia). Proc 25th Congr Pharm Sci pp 485492.
Lassak E V. McCarthy T 1983 Australian Medicinal Plants.
Methuen, North Ryde. pp 93-99.
Penfold A R, Grant R 1925 Germicidal values of some
Australian essential oils and their pure constituents. J Proc R
Soc NSW 59 346-350.
Penfold A R, Morrison F R 1950 Tea tree oils. In: The
Essential Oils, Vol 4, ed Guenther E. Van Nostrand. New
York. pp 526548.
Southwell I A 1988 Australian tea tree: oil of Melaleuen,
terpinen-4-01 type. Chem Aust 11 40W02.
Southwell I A , Stiff I A 1989 Ontogenetical changes in
monoterpenoids of Melaleuca alternifolia leaf. Phyto-
chemistry 27 1047-1 05 1.
Swords G. Hunter G L K 1978 Composition of Australian tea
tree oil (Melaleuca alternifolia). J Agric Food Cheni 26
734737.