SUBCRITICAL WATER EXTRACTION OF CAFFEINE FROM

BLACK TEA LEAF OF IRAN

ANVAR SHALMASHI, FERESHTEH GOLMOHAMMAD and

MOHAMMAD H. EIKANI1

Department of Chemical Industries

Iranian Research Organization for Science and Technology
PO Box 15815-3538, Tehran, Iran

Accepted for Publication June 1, 2007

ABSTRACT

This study examines subcritical water extraction (SCWE) of caffeine from

black tea leaf. The effects of various operating conditions such as water
temperature (100, 125, 150 and 175C), water flow rate (1, 2 and 4 g/min),
mean particle size (0.5, 1 and 2 mm) on extraction yield and rate were
determined. SCWE at 175C, water flow rate of 2 g/min and mean particle size
of 0.5 mm were found to be able to recover 3.82% (w/w) of caffeine present in
the black tea leaf within 3 h of extraction. In comparison to the SCWE,
conventional hot water extraction showed 3.30% (w/w) extraction yield. It was
found also that pressure had no effect on extraction yield and rate.

PRACTICAL APPLICATIONS

Recently, subcritical water has become of great interest as an alternative

solvent for extraction of natural active compounds. Subcritical water, as a
green solvent, can be used in many different fields of applications. In recent
years, extraction of flavors, fragrances and antioxidant components from plant
materials, and hydrolysis of carbohydrates, vegetable oils and fatty acids have
been widely investigated by many researchers. Using subcritical water for
analytical purposes, for soil remediation and applying it as a reaction media
are some other interesting fields for practical applications. Subcritical water is
an excellent solvent for caffeine as well as many other organic compounds but
is safer than the organic solvents that are used for caffeine extraction.

1
Corresponding author. TEL/FAX: +9821-88838324; EMAIL: eikani@irost.org

Journal of Food Process Engineering 31 (2008) 330–338. All Rights Reserved.

330 © 2008, The Author(s)
Journal compilation © 2008, Blackwell Publishing
DOI: 10.1111/j.1745-4530.2007.00156.x
 SUBCRITICAL WATER EXTRACTION OF CAFFEINE 331

INTRODUCTION

Caffeine is classified as a natural product belonging to a group of alka-

loids, i.e., nitrogen-containing ring compounds of plant origin that usually
have a bitter taste and some biological activity. As a weak organic base,
caffeine stimulates respiration and the central nervous system. In general, a tea
leaf contains 2–4% caffeine. Isolation of caffeine from tea leaf may use a
series of techniques, including both solid–liquid and liquid–liquid extraction.
It is usually extracted from tea on multistage countercurrent extraction tech-
nique (Treybal 1968). The current commercially available methods for decaf-
feinating black tea are solvent-based extraction using ethyl acetate or
methylene chloride, and extraction using supercritical carbon dioxide under
pressure. Two basic decaffeination processes are used recently, i.e., direct and
indirect solvent extraction methods (Zubrick 2001). In the direct solvent
extraction process, decaffeination is accomplished by direct application of
methylene chloride, ethyl acetate or carbon dioxide to the softened tea leaves.
In the indirect extraction process, the initial polar solvent used in the extraction
of caffeine is hot water often with carbonate salts added to control the pH and
polarity. The extraction of caffeine from aqueous solution was generally done
using chloroform (Helmkamp and Johnson 1968; Birdwhistell and O’Connor
1971; Pavia et al. 1976) or methylene chloride (Mayo et al. 1989; Nimitz
1991; Landgrebe 1993). Recently, supercritical techniques under pressure that
combine solvent extraction and stripping operations in a single process are
gaining considerable attention for separating and recovering caffeine from tea
leaves. Carbon dioxide, which has a low critical temperature (304.3 K) and
moderate critical pressure (73.8 bar), has become a universally attractive alter-
nate solvent in the extraction of natural products (Saldana et al. 2002). There
are several studies on decaffeination of tea leaves by a supercritical carbon
dioxide extraction (Chang et al. 2000).
Generally, the extraction processes depend upon the plants and solvent
selected. A microwave-assisted extraction (MAE) method has been reported
for the extraction of tea polyphenols and tea caffeine from green tea leaf using
a solvent mixture of ethanol and water (Pan et al. 2003). A solid–liquid
extraction of caffeine from tea waste using a battery-type extraction system
using different in nature pure extractant, water and chloroform has been
reported (Senol and Aydin 2006).
Recently, subcritical, superheated or pressurized hot water has become of
great interest as an alternative solvent for extraction of natural active com-
pounds (Herrero et al. 2005). Water is termed subcritical when its temperature
is between its boiling point 100C and its critical temperature 374C and its
pressure high enough to maintain it in the liquid state. In this region, water’s
dielectric constant is greatly reduced from that at ambient conditions, thereby
332 A. SHALMASHI, F. GOLMOHAMMAD and M.H. EIKANI

decreasing its polarity. This therefore results in an increase in its solvent power
for organic compound dissolution. Bioactive compounds have been extracted
by this technique. They are hypercin and pseudohypercin from St. John’s wort
(Mannila et al. 2002), iridoid glycosides from Veronica lonifolia (Soumi et al.
2000).
In this article, the subcritical water extraction (SCWE) of caffeine from
black tea leaf of northern Iran has been investigated. The aim of this study is
to determine the effects of extraction temperature, water flow rate, mean
particle size and extraction pressure on extraction yield and rate of extraction.

MATERIALS AND METHODS

Sample Preparation
Black tea leaf was obtained from Gilan province of northern Iran from a
local tea manufacturer. Five grams of samples was used for SCWEs and hot
water extractions. The samples were ground immediately prior to extraction.
Two replications of the extraction were performed for each of the runs. The
percentage relative standard deviation (% RSD) values for the extraction
efficiencies were calculated on the basis of the obtained purified caffeine
weights. The % RSD values ranged from 5 to 10%.
Chemicals
Chloroform (Merck, Darmstadt, Germany) was used as solvent in the
liquid–liquid extraction step; ethyl acetate (Merck) was used in the crystalli-
zation of caffeine. Doubly distilled, degassed water purified through a Milli-Q
deionizing unit (Millipore, Bedford, MA) was used as extractant.
Conventional Hot Water Extraction Method
Five grams of ground black tea leaf was extracted with 240 mL of water
for 3 h in a round-bottom flask equipped with a condenser. Temperature of
extraction was kept at boiling point temperature. The extraction mixture was
constantly stirred with a magnetic stirrer. The extraction mixture was cooled
and vacuum filtered. The solvent was removed, and the crude caffeine obtained
was dissolved in hot water and decolorized with activated charcoal and fil-
tered. The caffeine solution was then concentrated and allowed to crystallize.
Finally, the caffeine was purified by crystallization with ethyl acetate.

SCWES

SCWEs were carried out in a laboratory-built apparatus shown in Fig. 1.

Detailed description of the apparatus has been presented elsewhere (Eikani
 SUBCRITICAL WATER EXTRACTION OF CAFFEINE 333

FIG. 1. SCHEMATIC DIAGRAM OF SUBCRITICAL WATER EXTRACTION SYSTEM

B, burette; BV, ball valve; CV, check valve, DN, drain; EC, extraction cell; F, fan; HX, heat
exchanger; MF, micro filter; NV, needle valve; OV, oven; P, pump; PI, pressure indicator; PR,
pressure regulator; RV, relief valve; TI, temperature indicator; TIC, temperature indicator controller;
WI, water inlet; WO, water outlet.

et al. 2007). In brief, it mainly consists of a deionized water feed tank, a

high-pressure metering pump (H9 series, Dosapro Milton Roy, Pont-Saint-
Pierre, France), a fan-equipped temperature-controlled oven (Teb Azma Co.,
Tehran, Iran), a preheating coil (3-m length), a cylindrical extraction chamber
(103-mm ¥ 16-mm i.d.) and a double pipe heat exchanger (cooling surface area:
240 cm2). The flow of extractor direction was top to bottom.After preheating the
coil, a three-way line was made by using three high-pressure heat-resistant
needle valves. Needle valves 4 and 6 were inserted on the inlet and outlet line of
the extractor, respectively. Needle valve 5 was used as the bypassing line. All
parts in contact with the extractant water were made from stainless steel 316L.
The extractant water was first purged for 2 h with N2 to remove dissolved
O2. For all SCWEs, the extractor was filled with 5.0 g ground tea leaf. To
prevent moving of particles from the fixed bed, fiber glass wool sandwiched in
between two stainless steel filters was inserted in both sides of the fixed bed.
The extractor was assembled in an oven and pressurized by closing valves 4
and 6, closing the end line regulator and opening valve 5. Then, valve 4 was
opened and pumping of water continued to pressurize the system again to the
desired pressure. After that, the pump was turned off, valve 5 was closed,
valves 4 and 6 were fully opened and the oven was heated to the required
temperature. At that time, the pressure regulator was opened and the pump was
334 A. SHALMASHI, F. GOLMOHAMMAD and M.H. EIKANI

turned on, and the flow rate was adjusted to the desired rate. Regarding the
selected flow rate and a system void volume between extractor and collection
vessel (~60 mL), in all runs, around 30 mL of water coming out of the system
was discarded. By analyzing this amount of extract, it was observed that it
contains no caffeine. At this time, the collection of the extract was started. The
extraction process was supposed to be started at that time (extraction time = 0).
After collecting the required volume of extract, a liquid–liquid extraction step
using chloroform was carried out. The volumetric ratio of chloroform to
extract was 1:4 in all experiments, and the extraction was completed by two
equal volumes of solvent in two steps. The organic phases were concentrated
under a N2 stream to dryness. The crude caffeine was first dissolved in hot
water to separate it from wax, and then decolorized with activated charcoal and
filtered. The caffeine solution was then concentrated and allowed to crystal-
lize. Finally, the caffeine was purified by crystallization with ethyl acetate.

RESULTS AND DISCUSSION

Effect of Temperature
One of the most important parameters in the SCWE process is the tem-
perature of extraction. SCWE must be carried out at the highest permitted
temperature. The extraction temperature for tea leaf was optimized in order to
maximize caffeine extraction yield. Its influence was studied between 100 and
175C and mean particle size, flow rate, extraction time and pressure were
selected to be 0.5 mm, 2 g/min, 120 min and 20-bar pressure, respectively. As
can be seen in Fig. 2, the amount of caffeine extracted increased with an
increase in extraction temperature. The yields of caffeine extracted with
SCWE from black tea leaf at 100, 125, 150 and 175C were 1.4, 1.64, 1.88 and
3.70% (w/w, dry basis), respectively. This temperature dependence of the yield
is because of the increased solubility of caffeine in subcritical water as the
water temperature increases. This result obviously suggests that SCWE is
greatly affected by extraction temperature. On the other hand, it should be
noted that pressure does not affect the performance of SCWE as long as it is
high enough for the water to maintain the liquid state.

Effect of Particle Size

The black tea leaves were ground and screened by standard sieves. The
mean ground seed particles were 0.5, 1 and 2 mm. The effect of mean particle
size on the caffeine extraction yield at 175C temperature, 2-g/min flow, 20-bar
pressure and 180-min extraction time has been shown in Fig. 3. As seen, the
extraction efficiencies of caffeine for 0.5- and 1.0-mm size particles were
 SUBCRITICAL WATER EXTRACTION OF CAFFEINE 335

3
Extraction yield (%)

0
100 125 150 175

Temperature ( o C)

FIG. 2. EFFECT OF SUBCRITICAL WATER EXTRACTION TEMPERATURE ON CAFFEINE

EXTRACTION YIELD (%, W/W) FROM BLACK TEA LEAF
Operating conditions: sample weight = 5.0 g; flow rate = 4 g/min; particle size = 0.50 mm;
pressure = 20 bar; and extraction time = 120 min.

3
Extraction yield (%)

0.5 mm 1.0 mm 2.0 mm

0
0 30 60 90 120 150 180
time (min)

FIG. 3. EFFECT OF PARTICLE SIZE ON SUBCRITICAL WATER EXTRACTION OF

CAFFEINE FROM BLACK TEA LEAF AS EXTRACTION YIELD (%, W/W)
Operating conditions: sample weight = 5.0 g; flow rate = 2 g/min; temperature = 175C;
and pressure = 20 bar.
336 A. SHALMASHI, F. GOLMOHAMMAD and M.H. EIKANI

almost the same. Regarding the larger 2.0-mm size particles, the yield is
substantially lower. It suggests that the process may be controlled by mass
transfer of caffeine for larger particle sizes. For further experiments, the
optimum value for the mean particle size was 0.5 mm.

Effect of Water Flow Rate

The effect of water flow rate on the caffeine extraction yield at 175C
temperature, 0.5-mm particle size, 20-bar pressure and 180-min extraction
time has been shown in Fig. 4. The water flow rate has been studied in the
range of 1, 2 and 4 g/min. As can be seen, the rate of caffeine extraction was
faster at the higher flow rates. The rate is slower at 2 g/min and even slower at
1 g/min. It means that the mass transfer of caffeine from the surface of the
solid phase into the water phase regulated most of the extraction process.
Increase of flow rate resulted in the increase of superficial velocity and thus
quicker mass transfer. The main disadvantage of applying higher flow rates is
increasing the extract volume and, consequently, lowers the concentration of
the final extract. In practice, the best flow rate must be selected considering
two important factors, including extraction time and extract concentration. It is
clear that shorter extraction time and more concentrated extracts are desirable.

3
Extraction yield (%)

4 g/min 2 g/min 1g/min

0
0 30 60 90 120 150 180
time (min)

FIG. 4. EFFECT OF FLOW RATE ON SUBCRITICAL WATER EXTRACTION OF CAFFEINE

FROM BLACK TEA LEAF AS EXTRACTION YIELD (%, W/W)
Operating conditions: sample weight = 5.0 g; particle size = 0.50 mm; temperature = 175C;
and pressure = 20 bar.
 SUBCRITICAL WATER EXTRACTION OF CAFFEINE 337

Comparison with Conventional Method

Comparison of the SCWE at the optimum operating conditions (175C
temperature, 2-g/min water flow rate, 0.5-mm mean particle, 3-h extraction
time) and conventional hot water extraction showed that the extraction yield of
caffeine from black tea leaf by SCWE was higher than the hot water extraction
for 3 h. SCWE was found to be able to recover 3.82% (w/w) of caffeine
present in the black tea leaf and hot water extraction showed 3.30% (w/w)
extraction yield.

CONCLUSIONS

This study investigated the effect of extraction temperature, mean particle

size and water flow rate on extraction yield and rate of extraction. The results
showed that extraction yield increases as the temperature increases as a result
of increased solubility, but on the other hand, the pressure does not affect the
extraction yield. The evaluation of the effect of particle size suggests that the
process may be controlled by mass transfer of caffeine for larger particle sizes.
The extraction rate was faster at higher water flow rates. It means that the mass
transfer of caffeine from the surface of the solid phase into the water phase
regulated most of the extraction process. The results also showed that conven-
tional hot water extraction exhibits lower extraction yields in comparison to
SCWE.

ACKNOWLEDGMENT

The financial support of the Iranian Research Organization for Science

and Technology (IROST) is gratefully acknowledged.

REFERENCES

BIRDWHISTELL, R.K. and O’CONNOR, R.R. 1971. The Freeman Library

of Laboratory Separates in Chemistry, Vol. 2, Freeman, San Francisco,
CA.
CHANG, C.J., CHIU, K.L., CHEN, Y.L. and CHANG, C.Y. 2000. Separation
of cathechins from green tea using carbon dioxide extraction. Food
Chem. 68, 109–113.
EIKANI, M.H., GOLMOHAMMAD, F. and ROWSHANZAMIR, S. 2007.
Subcritical water extraction of essential oils from coriander seeds (Cori-
andrum sativum Mill.). J. Food Eng. 80, 735–740.
338 A. SHALMASHI, F. GOLMOHAMMAD and M.H. EIKANI

HELMKAMP, G.K. and JOHNSON, H.W. 1968. Selected Experiments in

Organic Chemistry, 2nd Ed., pp. 157–158, Freeman, San Francisco, CA.
HERRERO, M., CIFUENTES, A. and IBANEZ, E. 2005. Sub- and supercriti-
cal fluid extraction of functional ingredients from different natural
sources: Plants, food-by-products, algae and microalgae: A review. Food
Chem. 98, 136–148.
LANDGREBE, J.A. 1993. Theory and Practice in Organic Laboratory, 4th
Ed., pp. 381–383, Brooks-Cole, Pacific Grove, CA.
MANNILA, M.H., KIM, H. and WAI, C.M. 2002. Supercritical carbon
dioxide and high-pressure water extraction of bioactive compounds in St.
John’s Wort. Proceedings of Supergreen, November 3–6, 2002, Kyung
Hee University, Suwon, South Korea.
MAYO, D.W., PIKE, R.M. and BUTCHER, S.S. 1989. Microscale Organic
Laboratory, 2nd Ed., pp. 162–164, Wiley, New York, NY.
NIMITZ, J.S. 1991. Experiments in Organic Chemistry, pp. 61–62, Prentice-
Hall, Englewood Cliffs, NJ.
PAN, X., NIU, G. and LIU, H. 2003. Microwave-assisted extraction of tea
polyphenols and tea caffeine from green tea leaves. Chem. Eng. Process.
42, 129–133.
PAVIA, D.L., LAMPMAN, G.M. and KRIZ, G.S. 1976. Introduction to Labo-
ratory Techniques, pp. 58–62, Saunders, Philadelphia, PA.
SALDANA, M.D.A., ZETZL, C., MOHAMED, R.S. and BRUNNER, G.
2002. Decaffeination of guarana seeds in a microextraction column using
water-saturated CO2. J. Supercrit. Fluids 22, 119–127.
SENOL, A. and AYDIN, A. 2006. Solid-liquid extraction of caffeine from tea
waste using battery type extractor. J. Food Eng. 75, 565–573.
SOUMI, J., SIREN, H., HARTONEN, K. and RIEKKOLA, M.L. 2000.
Extraction of iridoid glycosides and their determination by micellar elec-
trokinetic capillary chromatography. J. Chromatogr., A 868, 73–83.
TREYBAL, R.E. 1968. Mass-transfer Operations, 2nd Ed., pp. 406–489,
McGraw-Hill, New York, NY.
ZUBRICK, J.W. 2001. The Organic Chemistry Lab Survival Manual: A Stu-
dent’s Guide to Techniques, 5th Ed., pp. 111–112, Wiley & Sons, New
York, NY.