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Effect of heat treatment on curcuminoid, colour value and total polyphenols

of fresh turmeric rhizome

Article  in  International Journal of Food Science & Technology · July 2009

DOI: 10.1111/j.1365-2621.2009.01976.x

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1438 International Journal of Food Science and Technology 2009, 44, 1438–1444

Original article
Effect of heat treatment on curcuminoid, colour value and total
polyphenols of fresh turmeric rhizome

A. Prathapan, M. Lukhman, C. Arumughan, A. Sundaresan & K. G. Raghu*

Agroprocessing and Natural Products Division, National Institute for Interdisciplinary Science and Technology (NIIST), Pappanamcodu,
Thiruvananthapuram 695 019, Kerala, India
(Received 9 February 2009; Accepted in revised form 23 March 2009)

Summary Studies were undertaken to examine the effect of heat treatment on total phenolic content (TPC), colour
value (yellowishness and brightness), polyphenol oxidase (PPO) activity and curcuminoid of fresh turmeric
rhizome. Fresh turmeric rhizomes were subjected to heat treatment at different temperatures (60–100 C) for
different durations (10–60 min), causing a reduction in browning which was evident from the improved
yellowishness and brightness. Activity of PPO was also decreased during heat treatment and PPO was almost
inactivated when heated at 80 C for 30 min. TPC of heat-treated turmeric after drying (powder) is
significantly higher than that after the fresh process. TPC values increased gradually when samples were
heated from 60 to 80 C. At 90 and 100 C, TPC values were almost identical. Maximum brightness and
yellowishness were obtained when the turmeric was heated above 80 C. Quantitation of curcuminoids in the
turmeric sample was made with high performance thin layer chromatography (HPTLC). There was no
significant change in the concentration of curcuminoids among the heat-treated samples. But in the sun-dried
samples, a significant reduction in curcuminoid concentration was observed.
Keywords Browning, curcuminoid, heat treatment, HPTLC, polyphenol oxidase, total polyphenols.

anti-HIV, anti-tumour, anti-carcinogenic and anti-

Introduction
arthritic activities (Chattopadhyay et al., 2004).
Spices are a group of esoteric food adjuncts, which have Cooking or roasting alters the nature of many food
been in use for thousands of years. Some of the spices are constituents such as starches and proteins by changing
also known to possess several medicinal properties their physical, chemical and nutritional characteristics
(Nadkarni & Nadkarni, 1976) and are effectively used (Belitz & Grosch, 1987). Heat processing also changes
in the indigenous systems of medicine. The physiological the bioavailability of proteins, carbohydrates, lipids and
effects of spices in most instances have been attributed to vitamins. It is also evidenced that heat processing of
the main spice active principles present in them. Among turmeric results in a significant loss of their active
the physiological influences the spices are documented to principles (Srinivasan et al., 1992; Suresh et al., 2007).
exhibit, their hypolipidemic and antioxidant properties Little information is available on the extent of the
have far-reaching health implications. destruction of the bioactive principles of spices during
Turmeric (Curcuma longa L.) belongs to the Zingiber- food processing. In addition to that, no systematic study
acea family reported to possess numerous medicinal has been conducted to reveal the effect of heat process-
properties including antioxidant activity (Pulla Reddy & ing on turmeric in relation to their total phenolic
Lokesh, 1992), anti-protozoal activity (Araujo et al., content (TPC), colour value and the concentration of
1999), anti-tumour activity (Huang et al., 1988), curcuminoids.
anti-inflammatory activity (Arora et al., 1971) and anti- In the present study, fresh turmeric rhizomes were
venom activity (Ferreira et al., 1992). The major biolog- evaluated for their TPC, colour value and curcuminoid
ically active component of turmeric is curuminoid which level after heat treatment and conventional sun-drying.
comprises curcumin (C1), demethoxy curcumin (C2) and
bis-demethoxy curcumin (C3). These are reported to
Materials and methods
possess cardioprotective, hypolipidemic, antibacterial,
Plant material
*Correspondent: Fax: +91-(0)471-2491712 ⁄ 2490186; Fresh turmeric (Curcuma longa L.) rhizome (Alleppy
e-mail: raghukgopal@rediffmail.com variety from Kerala, India) was harvested (250 kg)

doi:10.1111/j.1365-2621.2009.01976.x
 2009 The Authors. Journal compilation  2009 Institute of Food Science and Technology
 Effect of heat treatment on fresh turmeric A. Prathapan et al. 1439

8 months after planting. The rhizomes were selected (250 g · 3) were kept immersed in water in a plastic net
based on their general appearance, size and physical and maintained at 50, 60, 70, 80, 90 and 100 C for
form and organoleptic properties. They were stored at 30 min. After heat treatment, samples were cut into small
4 C until used for the experiments. pieces and ground using a domestic mixer. From this, 5 g
extract was taken in a conical flask and 50 mL of
methanol was added followed by sonication. The remain-
Chemicals
ing sample was kept open for 1 h at ambient temperature
Gallic acid, folin-ciocalteu reagent, curcumin standards (25–27 C). After 1 h, 5 g was again extracted with
were obtained from Sigma (St. Louis, MO, USA). Other methanol as described before. The remaining sample was
chemicals used were all analytical grade obtained from dried in a hot air oven at 50 C for 4–5 h. After drying it
Merck (Darmstadt, Germany). was powdered in a domestic mixer and 5 g was extracted
with methanol as described before.
In another set of experiments the fresh turmeric
Extraction of PPO
rhizome samples (250 g · 3) were subjected to heat
Enzyme extraction was carried out according to the treatment at 60, 70, 80, 90 and 100 C for 10, 20, 30, 40,
method of Yemenicioglu et al. (1997). Fresh turmeric 50 and 60 min at each temperature. After heat treat-
pieces (100 g) were cut into small pieces and homogen- ment, the rhizome samples were ground in a domestic
ised with cold acetone ()20 C). The homogenate was mixer and kept for 1 h at ambient temperature (25–
filtered through a Buchner funnel using Whatman No.1 27 C). After 1 h, 5 g of sample was taken for methanol
filter paper. The residue was washed again with acetone extraction as mentioned before. The remaining sample
and the colourless solid residue was dried overnight at was dried in a hot air oven at 50 C for 4–5 h. The dried
room temperature and then stored at )60 C until use. sample was powdered in a domestic mixer and 5 g was
For polyphenol oxidase (PPO) extraction, 1 g of extracted using methanol as described before. All the
acetone powder was suspended in 0.05 m sodium above experiments were carried out in triplicate.
phosphate buffer (pH 6.8). The suspension was mixed For control, the fresh rhizome samples (250 g · 3)
well for 20 min at 4 C. Then it was centrifuged at were ground, and 5 g sample was taken immediately for
9000 · g for 15 min and the supernatant (crude enzyme extraction. Samples, after exposure for 1 h and after
extract) was separated. All the procedures were carried drying in the oven, were also extracted using methanol.
out in triplicate and at 4 C.
Conventional drying (sun-drying) of fresh turmeric rhizome
Enzyme thermal stability and enzyme assay
Fresh turmeric samples (1000 g · 3) were cleaned and
Samples of 1 mL of total enzymatic extracts were boiled at 100 C for 1 h. The samples were drained off
incubated for 30, 40, 50, 60, 70 and 80 C for 30 min. water and dried in sunlight for 15 days. The conven-
The residual enzymatic activity was then evaluated as tional primary processing used by the farmers in the
described below. region was simulated here. After drying, the samples
The following reaction mixture was employed: 0.02 m were ground in a domestic mixer and 5 g was extracted
sodium phosphate buffer, 20 mm catechol, 400 lL of with methanol as described earlier.
enzyme extract (pH 6.8); total volume 3.0 mL. The
mixture was incubated at 30 C and the enzymatic
Estimation of moisture
activity evaluated by measuring the increase in optical
density (OD) at 410 nm (Serradell et al., 2000). The The moisture content in the ground turmeric sample was
enzymatic activity was calculated from the initial slope determined by co-distilling 10 g of the sample with
of the absorbance vs. time plot activity graph. In all toluene in a Dean stark apparatus (Louli et al., 2004).
experiments, control reactions without enzyme were The sample was transferred quantitatively to the distil-
included and no significant oxidation of catechol was lation flask with toluene. The flask, along with the
observed during the measurement of PPO activity. The contents, was heated under reflux for 2–3 h. At the end
enzymatic activity unit (U) was defined as the amount of of cooling, the distillate was collected in a graduated
enzyme required for an increase of 0.01 OD unit min)1 tube in the Dean stark apparatus.
under test conditions.
Estimation of TPC
Heat treatment of fresh turmeric rhizome
The TPC of the turmeric sample was determined using the
Fresh turmeric rhizomes were selected based on their folin–ciocalteu reagent according to the method of
organoleptic properties and were cleaned before being Singleton & Rossi (1965). Briefly, 60 lL of each methan-
used for the heat treatment. The rhizome samples olic extract of turmeric sample (three replicates) was

 2009 The Authors. Journal compilation  2009 Institute of Food Science and Technology International Journal of Food Science and Technology 2009
1440 Effect of heat treatment on fresh turmeric A. Prathapan et al.

taken and 500 lL of folin–ciocalteu reagent was added. ual curcuminoids in the samples. All the experiments
After 5 min of incubation, 1 mL of sodium carbonate were carried out in triplicate.
(20%) was added and incubated at ambient temperature
(25–27 C) for 90 min. The colour developed was mea-
Statistical analysis
sured at 765 nm against a blank solution (containing
methanol, folin–ciocalteu reagent and sodium carbonate) Statistical analysis was performed using software origin-
using UV-VIS spectrophotometer (UV-2450PC; Shima- 8 (Origin LAB, Origin Lab Corp., MA, USA). Two-
dzu, Kyoto, Japan). Results are expressed as milligram way analysis of variance (anova) was carried out to test
gallic acid equivalents per 100 grams of dry weight of the significant differences and the significance was
turmeric (mg GAE ⁄ 100 g dw). compared by using the Tukey test. Values were deemed
to be significant at P < 0.05.
Determination of colour value
Results and discussion
The colour value (yellowishness, brightness and dark-
ness) of turmeric powder was measured in UV-VIS
Effect of heat treatment to turmeric PPO activity
spectrophotometer (UV-3600 PC, Shimadzu) and results
were expressed in L, a, b scale (CIE, 1986). The Hunter Results obtained after the heat treatment of turmeric
Lab colour coordinate system L*, a* and b* values were PPO activity are shown in Fig. 1. At 30 C, the enzyme
recorded using a CIE L*, a*, b* uniform colour space,
where L* indicates lightness, a* indicates chromaticity PPO activity vs. thermal treatment
120
on a green ()) to red (+) axis and b* indicates
chromaticity on a blue ()) to yellow (+) axis. Exper- PPO activity
100
iments were carried out in triplicate. Relative activity (%)

80
HPTLC determination of curcuminoids
60
Quantification of total and individual curcuminoids in
the methanolic extract of turmeric powder was carried
40
out by using HPTLC (Paramasivam et al., 2008). The
samples were spotted as 6 mm wide bands using
20
Camag 100 ll sample syringe (Hamilton, Bonaduz,
Switzerland) on precoated silica gel aluminium plate
0
60 F-254 (10 cm · 10 cm with 0.2 mm thickness; E. 30 40 50 60 70 80
Merck) using Camag Linomat V auto sampler Treatment temperature (°°C)
(Muttenz, Switzerland). A constant application rate
of 150 nL s)1 was employed and the space between Figure 1 Polyphenol oxidase activity vs. thermal treatment. Values
two bands was kept as 10 mm. The mobile phase plotted on the graph represent relative activity, i.e. the ratio of the
consisted of chloroform:methanol (95:05 v ⁄ v). Linear activity to the maximum activity, expressed as percentage.
ascending development was carried out in a twin
trough glass chamber saturated with the mobile phase. Variation of TPC under different process
The optimised chamber saturation time for mobile 14 conditions
phase was 30 min at ambient temperature (25–27 C). 12
Each plate was developed to a height of 8 cm. The
10
TPC % (DB)

composition of the optimised mobile phase was After grinding

arrived at using different mobile phases with varying 8
After 1 h
solvents. After development, the plates were air-dried 6
and the components quantified by direct densitometric Powder
4
scanning of the developed plate at 425 nm (Camag
2
TLC scanner 3 with software WinCats v. 1.3.0,
Muttenz, Switzerland). Curcumin (C1), demethoxy 0
h
50

60

70

80

90

Su 00

curcumin (C2) and bis-demethoxy curcumin (C3) were

es

ie
1

dr
Fr

well separated under the chromatographic conditions

n

optimised. A standard curve was prepared for C1 Temperature (°°C)

(curcumin), C2 (demethoxy curcumin), and C3
(bis-demethoxy curcumin) using reference compounds Figure 2 Variation of total phenolic content (TPC) under different
separately and used for quantification of the individ- process conditions.

International Journal of Food Science and Technology 2009  2009 The Authors. Journal compilation  2009 Institute of Food Science and Technology
 Effect of heat treatment on fresh turmeric A. Prathapan et al. 1441

12
TPC values at 60 °C TPC values at 70 °C
12

After 1 h 10
10 After drying

TPC %
After 1 h
8

TPC %
After drying
8

6 6

4 4
10 20 30 40 50 60 10 20 30 40 50 60
Time (min) Time (min)

TPC values at 80 °C 12 TPC values at 90 °C

12

10 10
After 1 h After 1 h
After drying After drying

TPC %
TPC %

8 8

6 6

4 4
10 20 30 40 50 60 10 20 30 40 50 60
Time (min) Time (min)

12 TPC values at 100 °C

10
After 1 h
TPC %

After drying
8

Figure 3 Variation of total phenolic content 6

(TPC) of turmeric kept open for 1 h and
turmeric powder processed at different
4
temperatures (60–100 C) for different time 10 20 30 40 50 60
periods (10–60 min). Time (min)

remained stable and exhibited complete activity. Treat- After being kept open for 1 h, heat-treated turmeric (at
ment with higher temperature decreased the activity of 70, 80, 90 and 100 C for 30 min) showed higher values
PPO. Thermal treatment at 80 C caused an almost of TPC which is significantly different from those of
complete loss of activity after 30 min. Heat inactivation fresh processed turmeric at 50 and 60 C. In the case of
of PPO intended for avoiding browning reactions is of heat-treated turmeric after drying (turmeric powder), a
fundamental importance for industrial purposes (Serra- significant reduction in TPC values was observed.
dell et al., 2000). From our results it is clear that heat The TPC of heat-treated turmeric rhizome (after
treatment decreased the activity of turmeric PPO, which having been kept open for 1 h) at different temperatures
is thermally unstable. for different time periods are shown in Fig. 3. When
temperature and time of heat treatment increase, TPC
also increased. At 90 and 100 C, TPC values were
Effects of heat treatment on TPC of fresh turmeric rhizome
almost identical. TPC of the turmeric after drying
The TPC of fresh turmeric, heat-treated turmeric and (turmeric powder) decreased when compared with that
sun-dried samples is shown in Fig. 2. The TPC of fresh of samples before drying (Fig. 2). In powder form also,
turmeric without any heat treatment just after grinding TPC increased with increasing temperature and time of
(0 h) is higher than that of heat-treated (for 30 min) and heat treatment.
sun-dried samples. After keeping 1 h, TPC values were After grinding the turmeric and keeping for 1 h,
significantly reduced (P < 0.05) from the 0 h values. browning was observed. The prior art suggests that

 2009 The Authors. Journal compilation  2009 Institute of Food Science and Technology International Journal of Food Science and Technology 2009
1442 Effect of heat treatment on fresh turmeric A. Prathapan et al.

browning could be mainly due to the phenolic oxidation Figure 5 shows the colour values of turmeric powder
that contributes significantly to the deterioration of heated at different temperature and time. The brightness
quality (Ding et al., 1998). Lattanzio et al. (1994) (L*) and yellowishness (b*) were increased from 60 to
reported that browning is mainly catalysed by PPO. In 80 C for 30 min. But at 90 and 100 C, L* and b*
the presence of oxygen, PPO catalyses the hydroxylation values were almost identical.
of monophenols to O-diphenols and the oxidation of Browning of raw fruits, vegetables and beverages is
O-diphenols to their corresponding O-quinones. These a major problem in the food industry and is
in turn are polymerised to undesirable brown, red or believed to be one of the main causes of quality
black pigments (Mason, 1955). Peroxidase is another deterioration during post-harvest handling and pro-
oxido-reductase enzyme involved in enzymatic brown- cessing. The mechanism of browning in food is well
ing as diphenol may function as the reducing substrate known and is mainly enzymatic in origin (McEvily
in the reaction (Robinson, 1991). Lee et al. (1990) et al., 1992).
reported that the higher the PPO activity, the higher the In our study, the colour value of heat-treated
rate of browning. turmeric powder improved at higher temperatures
Increase in TPC after heat treatment could be due which is evident from increased brightness. The action
to the inactivation of PPO, as PPO are thermally of PPO promotes the browning reaction, and leads to
unstable and lose activity after 60 C (Chisari et al., the dark brown colour of turmeric. During heating,
2007). In our study also, heat treatment decreased the PPO was inactivated and hence browning decreased.
activity of PPO. At 90 and 100 C, TPC values were Enzymatic discolouration may be correlated with total
almost identical. This could be due to the complete phenolic levels (Mendel, 1997). There were reports
inactivation of PPO after 80 C. Maximum TPC that the higher the degree of browning encountered,
values were observed in turmeric treated at 80 C the lower the L* value of the samples (Rocha &
for 30 min. There was a significant reduction in TPC Morais, 2001). According to Govindarajan (1980),
in turmeric powder obtained after air-drying of heat- heat treatment of the rhizome prior to dehydration
treated turmeric when compared with the samples can inactivate oxidative enzymes, which could cause
before drying. After heat treatment, PPO was inacti- browning of the products. When temperature and time
vated. So the reduction in TPC in powder may be due increase, PPO activity decreases. As a result, browning
to the nonenzymatic oxidation of polyphenols. Kyi decreases and colour value (L* value) increases. Maria
et al. (2005) reported that, during the air drying of Lucia et al. (2002) found that boiling turmeric by
fruits, nonenzymatic oxidation of polyphenols occurs. immersing in water provided high colour value
Air drying also induces oxidative condensation or (brightness and yellow colour) when compared with
decomposition of thermo-labile compounds like cate- fresh turmeric, and that cooking the turmeric prior to
chin. This decomposition may be due to destruction of dehydration is important to obtain a product with
polyphenols or their conversion into non-antioxidant higher intensity of yellow.
forms (Donovan et al., 1998; Ferreira et al., 2000).
Amiot et al. (1993) found that the degree of browning
was closely related to the degradation of polyphenols.
So the overall increase of total phenolics after heat Colour value of turmeric powder
treatment is due to the inactivation of PPO and the 70 L*
reduction during drying may be due to the decompo- a*
60
sition of polyphenols. When temperature and time of b*
heat treatment increase, activity of PPO decreases. 50
L* a* b* scale

Because of the decreased activity of PPO, polymeri-

sation of polyphenols also decreases and thereby TPC 40
increases.
30

Effect of heat treatment on colour values of fresh turmeric 20

rhizome
10
Figure 4 shows the colour values of fresh, heat-treated
(at different temperatures for 30 min) and sun-dried 0
turmeric powder. Heat treatment and sun-drying Fresh 50 60 70 80 90 100 Sun
dried
improved the colour values of turmeric powder than Temperature (°°C)
the fresh sample. Maximum brightness and yellowish-
ness were obtained in the turmeric samples heated at Figure 4 Colour value of turmeric powder under different process
80 C. conditions.

International Journal of Food Science and Technology 2009  2009 The Authors. Journal compilation  2009 Institute of Food Science and Technology
 Effect of heat treatment on fresh turmeric A. Prathapan et al. 1443

Colour value of powder at 60 °C Colour value of powder at 70 °C

70 70

60 60

L*, a*, b*
50 50

L*, a*, b*
40 L*
L* 40
a* a*
30 b* b*
30
20
20
10
10 20 30 40 50 60 10 20 30 40 50 60
Time (min) Time (min)

Colour value of powder at 80 °C

70 Colour value of powder at 90 °C
70
60
60
50
50
L*, a*, b*

L*

L*, a*, b*
40 a*
b* L*
40 a*
b*
30 30

20 20

10 10
10 20 30 40 50 60 10 20 30 40 50 60
Time (min) Time (min)
Colour value of powder at 100 °C
70
L* = brightness
60 a* = + magenta; –ve Green
b* = + yellow; –ve Blue
50
L*, a*, b*

L*
40 a*
b*
30

20
Figure 5 Colour value of turmeric powder
10
process at different temperature (60–100 C) 10 20 30 40 50 60
for different time periods (10–60 min). Time (min)

Effect of heat treatment to fresh turmeric rhizome on

curcuminoid
Variation of total curcuminoids
There were no significant differences in total curcuminoid 7
Total curcuminoids (%)

concentration after drying using different processes Total curcuminoids

6
except in the conventional sun-dried material shown in
Fig. 6. By comparing fresh and heat-treated material, no 5
significant variations in total curcuminoid concentrations 4
were observed. In the conventional sun-dried process the
total curcuminoid concentrations were found to be 3
significantly lower (P < 0.05) than other values. 2
Concentration of total curcuminoids in heat-treated
1
turmeric powder did not vary significantly at any of the
h

50

60

70

80

90

treated temperature and time (data not shown). Con-

es

10

ie
dr
Fr

centration of individual curcuminoids (C1, C2, and C3)

n
Su

was also analysed in all the heat-treated samples and it Temperature (°°C)
was found that, heat treatment does not affect the
concentration of individual curcuminoids also (data not Figure 6 Variation of total curcuminoid concentrations under different
shown). Khurana & Ho (1998) reported that curcumi- process conditions. Significance accepted at P £ 0.05.

 2009 The Authors. Journal compilation  2009 Institute of Food Science and Technology International Journal of Food Science and Technology 2009
1444 Effect of heat treatment on fresh turmeric A. Prathapan et al.

noids were more stable under photooxidation as dry Khurana, A. & Ho, C. (1998). High performance liquid chromato-
powder. Heat treatment of turmeric immersed in water graphic analysis of curcuminoids and their photooxidative decom-
position compounds in Curcuma longa L. Journal of Liquid
produced good-quality turmeric with respect to curc- Chromatography, 11, 2295–2304.
uminoid; and cooked turmeric showed higher unifor- Kyi, T.M., Daud, W.R.W., Mohammad, A.B., Wahid Samsudin, M.,
mity or better pigment distribution (Maria Lucia et al., Kadhum, A.A.H. & Talib, M.Z.M. (2005). The kinetics of
2002). Govindarajan (1980) observed that boiling tur- polyphenol degradation during the drying of malaysian cocoa
beans. International Journal of food Science and Technolology, 40,
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pigments from cells to adjacent tissue, contributing to Lattanzio, V., Cardinali, A. & Palmieri, S. (1994). The role of phenolics
better colour homogenisation. in the post harvest physiology of fruits and vegetables: browning
The present study revealed that the heat treatment of reactions and fungal diseases. Italian Journal of Food Science, 1, 3–22.
fresh turmeric rhizome resulted in good colour value Lee, C.Y., Kagan, V., Jaworski, A.W. & Brown, S.K. (1990).
Enzymatic browning in relation to phenolic compounds and
and high TPC without causing any reduction in curc- polyphenoloxidase activity among various peach cultivars. Journal
uminoid concentrations. Heat treatment at 80 C for of Agricultural and Food Chemistry, 38, 99–101.
30 min showed maximum colour value and TPC. Hence Louli, V., Ragoussis, N. & Magoulas, K. (2004). Recovery of phenolic
it is better to process turmeric rhizome by heat process- antioxidants from wine industry by products. Bio resource Technol-
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International Journal of Food Science and Technology 2009  2009 The Authors. Journal compilation  2009 Institute of Food Science and Technology

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