Sains Malaysiana 43(2)(2014): 185–194
In
vitro Evaluation of Biological Activities of Citrus Leaf Extracts
(Penilaian in vitro Aktiviti Biologi Ekstrak Daun Limau)
MUHAMMAD ADNAN1, ATIKA
UMER2, IMTIAZ
AHMAD1, KHIZAR
HAYAT3
&
SAMINA
N.
SHAKEEL*1
1Department
of Biochemistry, Quaid-i-Azam University, Islamabad, Pakistan
2National
Institute of Health, Islamabad, Pakistan
3Department
of Chemistry, COMSATS Institute of Information Technology, Abbotabad,
22060,
Pakistan
Diserahkan:
6 Januari 2013/Diterima: 18 Mei 2013
ABSTRACT
Leaves extracts of three different citrus species namely Citrus sinensis (Malta), Citrus paradisi (Grape
fruit) and Citrus jambhiri (Khatti) were evaluated for their
antioxidant, cytotoxic, antitumor, antimicrobial activities and finally the
mineral contents were determined. Four types of extraction solvents (100, 80
and 50% methanol and distilled water heated at 50oC)
were used for extraction of antioxidant compounds. Extracts yield increased
with the elevated levels of aqueous component of organic solvent and our data
showed the highest yield in aqueous extracts. All extraction regimes showed in
vitro antioxidant activity. Citrus paradisi showed the highest total
flavonoid content (TFC), total phenolic content (TPC),
reducing power and 1,1-Diphenyl-2- picrylhydrazil free radical scavenging
activity in all combinations of solvents followed by Citrus sinensis and Citrus jambhiri. Antioxidant activity was also increased with higher
aqueous component of organic solvents in each case. While Citrus sinensis (in
100% methanolic solvent) and Citrus paradisi (in 50% methanolic extract)
showed significant cytotoxicity (LD50 values
<1000). Antitumor activity was observed in all extracts, however Citrus
sinensis and Citrus paradisi (in 50% aqueous methanolic) extracts had
the highest antitumor activity from the selected citrus species whereas no
antimicrobial activity was observed at higher concentrations (4000 μgmL-1) against specified strains. We found high
calcium contents in all three citrus species tested by atomic absorption
method. The results showed that the leaves of selected Citrus species possess
significant antioxidant, antitumor and cytotoxic activities. Citrus leaves
extracts can be potentially helpful in antioxidant protection in food as well
as in human body against lipid peroxidation and free radicals damage. It can
further be evaluated after in vivo studies using animal models or
identifications of high throughput methods for enhanced biological activities.
Keywords: Biological activities; citrus; in vitro evaluation; mineral micronutrients
ABSTRAK
Ekstrak daun tiga spesies limau berbeza iaitu Citrus sinensis (Malta), Citrus paradisi (Grape
fruit) dan Citrus jambhiri (Khatti) telah dinilai untuk aktiviti
antioksidan, sitotoksik, antitumor dan agen antimikrob serta menentukan
kandungan mineral mereka. Empat jenis pelarut pengekstrakan (100, 80 dan 50%
metanol dan air suling yang dipanaskan pada 50oC)
telah digunakan untuk pengekstrakan sebatian antioksidan. Hasil ekstrak
meningkat dengan peningkatan tahap komponen akueus pelarut organik dan data
kami menunjukkan hasil tertinggi dalam ekstrak akueus. Semua rejim
pengekstrakan menunjukkan aktiviti antioksidan in vitro. Citrus
paradisi menunjukkan kandungan flavonoid keseluruhan (TFC),
kandungan fenolik keseluruhan (TPC), kuasa penurunan dan
aktiviti skaveng radikal bebas 1,1-difenil-2-pikrilhidrazil tertinggi dalam
semua kombinasi pelarut diikuti oleh Citrus sinensis dan Citrus
jambhiri. Aktiviti antioksidan juga ditingkatkan dengan peningkatan komponen
akueus pelarut organik untuk setiap kes. Manakala Citrus sinensis (dalam
pelarut metanol 100%) dan Citrus paradisi (dalam ekstrak metanol 50%)
menunjukkan kesitotoksikan yang signifikan (nilai LD50 <
1000). Aktiviti antitumor telah diperhatikan dalam semua ekstrak. Walau
bagaimanapun, ekstrak Citrus sinensis dan Citrus paradisi (dalam
akueus metanol 50%) didapati mempunyai aktiviti antitumor tertinggi daripada
spesies limau terpilih manakala tiada aktiviti antimikrob telah diperhatikan
pada kepekatan yang lebih tinggi (4000 μgmL-1)
terhadap strain khusus. Kami mendapati kandungan kalsium adalah tinggi dalam
kesemua spesies limau yang diuji oleh kaedah penyerapan atom. Hasil menunjukkan
bahawa daun spesies limau terpilih memiliki aktiviti antioksidan, antitumor dan
sitotoksik yang signifikan. Ekstrak daun limau berpotensi membantu dalam
perlindungan antioksidan dalam makanan serta dalam badan manusia terhadap peroksidasi
lipid dan kerosakan oleh radikal bebas. Ia boleh dinilai selanjutnya berikutan
kajian in vivo menggunakan model haiwan atau pengecaman kaedah berdaya
pemprosesan tinggi untuk aktiviti biologi tertingkat.
Kata kunci: Aktiviti biologi; limau; mikronutrien
mineral; penilaian in vitro
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*Pengarang
untuk surat-menyurat; email: snq28@yahoo.com
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