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Mal J Nutr 5:47-54, 1999

Effect of different drying methods on concentrations of several


phytochemicals in herbal preparation of 8 medicinal plants leaves
Mahanom, H. , Azizah, AH and Dzulkifly, M.H.

Department of Food Science, Faculty of Food Science and Biotechnology, Universiti


Putra Malaysia, 43400 Serdang, Selangor, Malaysia

ABSTRACT

The effect of oven drying at 50°C ± 1°C for 9 hour, 70°C ± 1°C for 5 hour
and freeze drying on retention of chlorophyll, riboflavin, niacin, ascorbic
acid and carotenoids in herbal preparation consisting of 8 medicinal plants
was evaluated. The medicinal plants selected were leaves of Apium
graveolens (saderi), Averrhoa bilimbi (belimbing buluh), Centella asiatica
(pegaga), Mentha arvensis (pudina), Psidium guajava (jambu batu ),
Sauropus androgynous (cekor manis), Solanum nigrum (terung meranti) and
Polygonum minus ( kesum ). Results revealed that both type and conditions
of the drying treatments affected retention of all phytochemicals analysed.
Herbal preparation developed using oven drying was found to have inferior
phytochemicals content compared to that obtained by freeze dryer.
Nevertheless, the herbal preparation developed using all treatments still
retain appreciable amount of phytochemicals studied, especially
carotenoids, ascorbic acid, niacin and riboflavin and thus have potential for
commercial purposes.

INTRODUCTION

Malaysia is rich in natural phytochemicals is often used to indicate


resources and a large segment of the the disease preventing compounds
local community use these natural available from them (Mohamed, 1997).
products for medicinal purposes Over Examples of phytochemicals that have
the past decade, medical and scientific been reported to provide medical or
knowledge on the role of various health benefits and therefore why they
nutrients in specific disease processes were studied, are the use of beta carotene
has advanced at an accelerating pace and to prevent lung and skin cancer, niacin to
create an exciting and explosive new prevent recurrent heart attacks, ascorbic
area of research, resulting in increasing acid in improving immune system,
numbers of potential nutritional products riboflavin for lesion treatment, lycopene
with medical and health benefits. The in the prevention of cancer and
majority of these health promoting food chlorophyll to study the symptoms of
are from plants, hence the term senescene in
48 Mahonom H, Azizah AH & Dzulkifly MH

attempts to maintain the green colour for plants were obtained from Sri Serdang
producing higher quality medicinal wet market and “herbal unit” of
plants products. The current concept Universiti Putra Malaysia.
and trend of today is towards total health
management, the emphasis being on Drying Treatments
prevention rather than cure and since the
connection between diet and some Three drying methods used are
chronic diseases like cancer and oven drying at 50°C ± 1°C for 9 hour,
cardiovascular diseases are inrefutable, 70°C ± 1°C for 5 hour and freeze drying.
there is a real need for development of Portions of the leaves were dried to a
disease-fighting foods (Mohamed, 1997; final moisture content of below 10% and
Shukla, 1993). In view of these, aim of then ground in an electric grinder.
the study is to evaluate different drying Different types of the dried medicinal
methods in retaining several leaves were then mixed manually
phytochemicals in herbal preparation according to a specified ratio and stored
consisting of 8 medicinal plants. in an airtight amber bottle at
temperature of 4° ± 1 °C, until analysis
MATERIALS AND METHODS were performed.

Eight types of medicinal plants Chemical Analysis


leaves were used in this study, namely
Apium graveolens (saderi), Averrhoa Chlorophyll
bilimbi (belimbing buluh), Hydroctyle
asiatica (pegaga), Mentha arvensis Total chlorophyll was determined
(pudina), Psidium guajava (jambu batu), by spectrophotometric method described
Sauropus androgynous ( cekor manis), by Amar Singh (1977). A small amount
Solanum nigrum ( terung meranti ) and of CaCO3 was added to 1g sample and
Polygonum minus ( kesum ). The plants then blended with 80% acetone for 3
were selected based on their long term minutes. The mixture was then filtered
usage in Malaysian traditional medicine. and chlorophyll extracted with 80%
. For example, Apium graveolens has acetone Absorbance reading was done
been used for asthma, bronchitis and at 645nm and 663nm using
chest pain, Averrhoa bilimbi used for its spectrophotometer.
antibacterial property, hydrocotyle
asiatica for wound healing and memory, Riboflavin
Mentha arvensis used in cough mixtures,
Psidium guajava for curing stomach Riboflavin content of the sample
ache and gastroentitis, Sauropus was determined using AOAC method
androgynous for treating nose ulceration (1984). About 5g fresh or 2g dry sample
and hypertension, Solanum nigrum as was extracted with 65ml 0.1N sulphuric
laxative and diuretic and Polygonum acid in boiling water bath for 30
minus used for indigestion and after minutes. Protein and other interfering
childbirth (Burkill, 1966; Goh et al., substances from the cool extract are then
1995; Zakaria and Mohd., 1994). The precipitated
Effect of different drying methods on medicinal plants 49

at pH 6.0. and 4.5. Potassium anhydrous sodium sulphate to dry.


permanganate is then added to the Hexane was then evaporated and
extract to remove interfering fluorescent transferred to a 10ml volumetric and
substances, followed by addition of make up to volume. Extracted samples
hydrogen peroxide to remove excess were then filtered through 0.45µm nylon
permanganate. The fluorescence was membrane filter and analyzed by a non
measured with excitation wavelength of aqueous reverse-phase HPLC using a
440nm and emission wavelength of µBondapak C18 ( 3.9 x 300mm ) column
565nm. and a ternary mixture of acetonitrile,
methanol and ethyl acetate ( 88 : 10 : 2,
Niacin v/v ) as the mobile phase. The standards
used were α-carotene, β-carotene and
Niacin was determined using lycopene. The carotenoids eluted
cyanogen bromide colorimeter method detected and quantitated using a UV-
(AOAC, 1984). Fresh sample (5g) or dry Visible detector set at 450nm .
form (2g) was homogenized in sodium
hydroxide and distilled water. The Statistical Analysis
mixture was heated for 1 hour over a
boiling water-bath, cooled and pH Experimental data were analyzed
adjusted to 4.5. 17g of ammonium using analysis of variance (ANOVA)
sulphate were added. Color and significant differences among means
development is achieved by reacting the at p<0.05 were determined by Duncan’s
extract with cyanogen bromide and then multiple range test (DMRT) using the
measured at 450nm. Statistical Analysis System (SAS, 1987)
computing programme.
Ascorbic acid
Ascorbic acid was determined RESULTS AND DISCUSSIONS
using phenolindolphenol dye method Overall Content of Phytochemicals
(AOAC, 1984). 10g of sample were
blended with metaphosphoric acid-acetic Table 1 showed the levels of
acid extracting solution to a homogenous phytochemicals analysed in fresh and
slurry. 5ml of the filtrate extract were dried herbal preparation. The drying
then titrated with the standard treatments caused significant (p< 0.05)
indophenol to a pink end point. reduction in the levels of all
phytochemicals analysed. Oven drying
Carotenoids caused greater reduction in the levels of
Total carotenoids were determined phytochemicals compared to freeze
according to Tee, E.S. and Lim, C.L. drying. This results confirmed previous
(1992). 5g of sample was hydrolysed reports in the literature and supports the
with 95% alcohol and 5ml of 100% advantages of freeze drying over oven
KOH, by water-cooled refluxing drying (Salunkhe, 1974). Freeze drying
apparatus for 30 minutes. The resulted in the lowest % losses of all the
hydrolysate was extracted with 50ml phytochemicals analysed, followed by
hexane for 3 times and passed through oven drying at 70°C ± 1°C
50 Mohanom H, Azizah AH & Dzulkifly MH

Table 1. Phytochemicals composition of fresh and dried herbal tea 1,2

Drying Treatment
Phytochemicals Fresh (control) Oven drying Oven drying Freeze drying
50 ºC ± 1 ºC, 70 ºC ± 1 ºC,
9 hour 5 hour
a
Chlorophyll 1.230 ± 0.008 0.435 ± 0.013c 0.963 ± 0.036b 0.991 ± 0.010b
Ascorbic Acid 58.200±1.760a 14.200± 1.429d 38.300± 0.942c 45.900± 0.963b
Niacin 0.359 ± 0.008a 0.153 ± 0.003d 0.198 ± 0.003c 0.304 ± 0.003b
Riboflavin 0.400 ± 0.000a 0.191 ± 0.047d 0.196 ± 0.002c 0.200 ± 0.000b
3829.37 2795.44 3063.50 3277.90
Carotenoids
± 58.10a ± 22.5d ± 22.1c ± 22.40b

1
Means ± standard deviation based on three observations, data reported on mg/100g
samples except for carotenoids, data reported on µg/100g samples.
2
Means within a row with different superscripts are significantly different using Duncan’s
Multiple Range Test at P<0.05

for 5 hour and oven drying at 50°C ± attributes of foods is color, especially
1°C for 9 hour. appearance of green in dried herbs
leaves. Therefore, adverse changes in
Effect of different drying treatments colour may alienate potential customer
on phytochemicals content due to suggestion of possibility of poorly
controlled processing and this must be
Chlorophyll prevented or minimized (Ramada et al.,
1988).
In fresh herbal preparation, the
total chlorophyll content was Ascorbic acid
1.230mg/100g and the value ranged
from 0.435 - 0.991mg/100g after The different treatments resulted in
different drying treatments was applied. a significant (p< 0.05) loss in ascorbic
Drying treatment affect the chlorophyll acid contents of the samples (Table 1).
content significantly (p< 0.05) in all As expected, the loss of ascorbic acid
treatments (Table 1). The loss of total was found to be highest in samples dried
chlorophyll was found to be greatest in at 50°C ± 1°C for 9 hour (75.60%),
oven drying at 50°C ± 1°C for 9 hour followed by that of 70°C ± 1°C for 5
(64.63%), followed by oven drying at hour ( 34.19%) and minimum loss in
70°C ± 1°C for 5 hour (21.70%) and freeze drying ( 21.13% ) (Table 2). The
least reduction in freeze drying samples losses of ascorbic acid maybe attributed
(19.43%) (Table 2). This is reflected in to the thermal treatment applied (Yang
the visual observations of the various and Atallah, 1985). In addition,
samples. One of the most pleasing oxidation to diketogulonic acid when
Effect of difference drying methods on medicinal plants 51

Table 2. Stability of Phytochemicals in relation to the drying treatment applied

Losses during processing ( drying ), ( % )


Phytochemicals Oven drying Oven drying Freeze drying
50 ºC ± 1 ºC, 70 ºC ± 1 ºC,
9 hour 5 hour
Chlorophyll 64.63 21.70 19.43
Ascorbic Acid 75.60 34.19 21.13
Niacin 57.38 44.85 15.32
Riboflavin 52.25 51.00 50.00
Carotenoids 27.00 20.00 14.40

samples were exposed to air, light, and freeze drying showed losses of
alkaline conditions and metal ions may riboflavin of 52.25%, 51.00% and
also contribute to the losses encountered 50.00% respectively (Table 2).
(Addo, 1981; Harris, 1975; IFT, 1986) Riboflavin in fresh herbal preparation
were 0.4mg/100g and reduced to 0.20-
Niacin 0.19mg/100g after various drying
treatments. Thermal processing by oven
Niacin content in fresh samples drying and low temperature processing
were 0.369mg/100g and ranged from by freeze drying resulted in significant
0.153 - 0.34mg/100g after different (p<0.05) decrease in riboflavin content
drying treatments were employed. There (Table 1). Among the various drying
were significant (p < 0.05) differences treatments applied, the retention levels
amongst niacin contents of the various was found to be highest in freeze drying
treatments (Table 1). Freeze dried (50.00%) followed by oven drying at
herbal preparation retained the highest 70°C ± 1°C for 5 hour (43.50 - 44.50%)
niacin (83.84-85.52%) followed by oven and lowest in oven drying at 50°C ± 1°C
drying at 70°C ± 1°C for 5 hour (54.32- for 9 hour (36.00-59.50%).
55.99%) and the lowest occurred in oven
drying at 50°C ± 1°C for 9 hour (41.78- Carotenoids
43.45%). Result revealed that herbal
preparation obtained by freeze drying During the three methods of
and oven drying at 70°C for 5 hours still drying, a significant (p<0.05) decrease in
retain appreciable amount of niacin. carotenoid content were observed (Table
1). Carotenoid content of fresh herbal
Riboflavin preparation was 3829.37µg/100g and
decrease due to oven drying at 50°C ±
Analysis of fresh samples and 1°C for 9 hour, 70°C ± 1°C for 5 hour
samples after different drying at 50°C ± and
1°C for 9 hour, 70°C ± 1°C for 5 hour
52 Mahonom H, Azizah AH & Dzulkifly MH

freeze drying were 2795.44µg/100g, plants because it is important to retain as


3063.50µg/100g and 3277.90µg/100g much as possible the active ingredients
respectively. It was seen that in the dried herbal preparation. This is
dehydration at higher temperatures of crucial in order to preserve the biological
50°C-70°C led to greater destruction of benefits that the herbs can do.
carotenoid than losses encountered by Nevertheless, the study revealed that
dehydration by freeze drying (Table 2). dried medicinal plants leaves still retain
Retention levels in carotenoid content appreciable amount of chlorophyll,
for the various drying treatments were ascorbic acid, niacin, riboflavin and
72.41-3.59% (oven drying at 50°C ± 1°C carotenoids in all the treatments except
for 9 hour ), 79.42 - 80.58% (oven for the sample undergone oven drying at
drying at 70°C ± 1°C for 5 hour ) and 50°C for 9 hours. Drying medicinal
85.01-86.18% for freeze drying. The plants by oven at 70°C for 5 hours
principle cause of deterioration of the warrant further research based on level
carotenoid is oxidation, this being more of phytochemicals remain in the treated
severe once cellular integrity has been samples and relatively low cost
lost due to the high degree of involved. Further study is required to
unsaturation of the carotenoid (De la carry out effectiveness of the herbal
Mar and Francis, 1969; Kanner and preparation in reducing risks for chronic
Mandel, 1976; Carnevale et al., 1980; diseases especially coronary heart
Malchev et al., 1982). Nevertheless, disease, cancer and diabetes using rabbit
appreciable amount of carotenoid still or mouse before the product can be
remain in all the samples. Carotenoids, recommended to human.
especially β-carotene has been found to
be a potent antioxidant that can aid ACKNOWLEDGEMENTS
endogenous tocoferol in trapping free
radicals produced during normal The authors would like to thank
biological metabolism. Free radicals The Ministry of Science, Technology
have been shown to be involved in and Environment of Malaysia for
etiology of various chronic diseases like financing the project and the laboratory
coronary heart diseases, cancer, arthritis facilities of University Putra Malaysia.
and premature aging.

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