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Determination of Phenolic and Flavonoid Contents, Antioxidant 335 AGRIVITA Journal of Agricultural Science. 2017. 39(3): 335-344 Determination of Phenolic and Flavonoid Contents, Antioxidant Activities and GC-MS Analysis of Clinacanthus nutans (Acanthaceae) in Different Locations Noor Zafirah Ismail1), Hasni Arsad1*), Mohammed Razip Samian2) and Mohammad Razak Hamdan3) 1) Advanced Medical and Dental Institute, Universiti Sains Malaysia, Bertam, 13200 Kepala Batas, Penang, Malaysia 2) School of Biological Sciences, Universiti Sains Malaysia, 11800 Gelugor, Penang, Malaysia 3) Central Drug Research Institute, Universiti Sains Malaysia, 11800 Gelugor, Penang, Malaysia *) Corresponding author E-mail: [email protected] Received: September 28, 2016 /Accepted: July 28, 2017 ABSTRACT things same goes Clinacanthusto nutans that have its own role in pharmaceutical remedies in South- Clinacanthus nutans is an essential medicinal plant East Asia such as Malaysia, Thailand, Indonesia, that had been used in various local remedies to Vietnam and China (Ismail, Arsad, Samian, Ab. treat many illnesses. A study had been conducted Majid, & Hamdan, 2016). C. nutans belongs to to determine the phenolic, flavonoid, antioxidant family Acanthaceae that can be described as herbs activities and phytochemical compounds of or perennial shrubs that can grow up to 1 m tall. In C. nutans in different locations. C. nutans were Malaysia, it is well-known as ‘Belalai Gajah’ because harvested from eight locations and the leaves of the curve stems appearances that resemble the were extracted with 80 % methanol by maceration curve of elephant’s trunk that means Belalai Gajah process. Then, the phytochemical screening using in Malay word. Confirming to the people who used Gas Chromatography-Mass Spectrometry (GC-MS), leaves of C. nutans, they used it as raw vegetable 2,2 diphenyl-2-picrylhydrazyl hydrate (DPPH) assay and sometimes mixed with apple juice, sugarcane method, total phenolic content by Folin-Ciocalteu’s juice, orange juice, green tea, and served as a fresh assay method and total flavonoid content by drink or a refreshing beverage. They have been aluminium chloride (AlCl) were carried out. TheC. 3 practiced drinking C. nutans extracts due to their nutans extracts showed higher antioxidant activities history of the disease in cancers, diabetes, fever, than phenolic and flavonoid content. The neutral jaundice, pneumonia, and hepatitis. They also pH sandy clay soil from location KKK (Kuala Ketil, claimed that they have recovered from the illness Kedah, Malaysia) had higher antioxidant activities especially cancer disease (Shim, Aziana, & Khoo, (58.0 %), phenolic (44.1 mg GA.100 g-1) and 2013). The scientific information on benefitsC. of flavonoid content (30.8 mg QE.100 g-1) compared nutans had been proved to treat many diseases such to other locations. The GC-MS analysis showed as varicella-zoster virus and herpes-simplex virus, the presence of phytochemicals constituents anti-snake venom, analgesic and anti-inflammatory of 20 compounds. The results revealed that and anti-proliferative in cancer cells (Ismail, Arsad, environmental factors (light intensity, temperature Samian, Ab. Majid, & Hamdan, 2016). and soil characteristics) of eight locations were Plant of the same species may differ responsible for variations of phenolic, flavonoids, significantly for their secondary metabolite contents antioxidants and GC-MS analysis C.in nutans. The within different environments. This is because findings of this study will provide baseline data for the quality and quantity of secondary metabolites future breeding programs for commercial cultivation. in medicinal plants are largely influenced by the Keywords: antioxidant; C. nutans; environment; environmental conditions. In some cases, secondary flavonoid; phenolic; phytochemistry metabolites accumulation has been affected by geographical altitude and local ecological INTRODUCTION conditions such as rainfall, humidity, temperature, Around 85 % to 90 % of the world’s populationwater variability, exposure to soil microorganisms consumes medicinal plants (Krishnamoorthy &and variations in soil pH and nutrients. Furthermore, Subramaniam, 2014). Plant secondary metabolitesenvironmental factors also interact with the genetic have their own roles in the human and also otherof living the plants resulting in a variable of gene Cite this as: Ismail, N. Z., Arsad, H., Samian, M. R., & Hamdan, M. R. (2017). Determination of phenolic and flavonoid contents, antioxidant activities and GC-MS analysis of Clinacanthus nutans (Acanthaceae) in different locations. AGRIVITA Journal of Agricultural Science, 39(3), 335–344. http://doi.org/10.17503/agrivita.v39i3.1076 Accredited: SK No. 60/E/KPT/2016 336 Noor Zafirah Ismail at al.: Phytochemical of Clinacanthus nutans extract......................................................................... expression that ultimately leads to the alteration quantity of phenolic, flavonoid, antioxidant activities of secondary metabolites concentration and thus and secondary metabolites from GC-MS analysis affects the quality of medicinal plants (Sampaio, were varied in different locations. Therefore, it is Edrada-ebel, & Batista Da Costa, 2016). Plant important to understand that the chemical properties secondary metabolites have been divided into of the plant in different locations as they might many classes, one of the general classes are contain powerful compounds in certain locations that phenolic that can be divided into two which are can benefit to human health and also other living flavonoids and non-flavonoids. Non-flavonoids things. Moreover, the selected C. nutans locations consist of secoiridoids, stilbenes, simple phenols, can be used as breeding programs and commercial benzoic acids, lignans, hydrolysable tannins, cultivation for medicinal purposes. acetophenones, cinnamic acids, phenylacetic MATERIALS AND METHODS acids, benzophenones, coumarins, and xanthones. While flavonoid compounds assemble from a large In this study, eight locations had been selected group of polyphenolic compounds which consist of in the northern region of Peninsular Malaysia. benzo-γ-pyrone structure that present all over the The code names for locations of C. nutans were secondary metabolite of the plant. The structure shown in Table 1 and the map was shown in Fig. of flavonoids is categorised by a 15-carbon (C6- 1. At Penang, four locations ofC. nutans cultivation C3-C6) backbone comprising of two aromatic and had been identified which were Pongsu Seribu one oxygenated ring. Both of them are grouped (PBF) and Tasek Gelugor (PTG) (situated at north together into subclasses based on their basic Seberang Perai), Batu Maung (PBM) (situated at chemical structures which are flavanones, flavones, the south-eastern part of Penang Island) and Batu chalcones, flavanols, flavonols, isoflavones and Ferringhi (PBF) (situated at the hilly northern side anthocyanins (Kumar & Pandey, 2013). of Penang Island). In Kedah, three locations of C. The research was conducted to study the nutans cultivation had been identified which were amount of phenolic, flavonoid, antioxidant activities Jeniang (KJN) (a small town that situated between as well as GC-MS analysis of C. nutans in different Gurun and Sik), Sungai Petani (KSP) and Kuala location sites. Most of the researchers had been Ketil (KKK). At Perlis, it had been identified one using colorimetric methods to obtain the total location site which was at Taman Herba Perlis at contents of phenolic, flavonoid and antioxidant Sungai Batu Pahat (SBP). It lies at the northern assay from Folin-Ciocalteu assay, AlCl3 assay, part of the west coast of Peninsular Malaysia. C. and DPPH assay respectively. To investigate the nutans and are distributed at the elevation between phytochemical compounds of C. nutans in different 8 m to 44 m above sea level. Temperature among locations, GC–MS had been successfully applied. eight locations showed averaged of 32 oC between The GC-MS analysis had been used for plants 30 oC to 34 oC. The soil characteristics and light such as Ziziphus spina-christi (Moustafa, Hesham, intensity were determined using soil and light tester Quraishi, & Alrumman, 2016), Cannabis sativa meter (soil moisture, soil pH and light) while the (Tayyab & Shahwar, 2015), and Justicia adhatoda amount of nitrogen, potassium and phosphorus (Gilani, Fujii, Kikuchi, Shinwari, & Watanabe, 2011) level in the soil were determined using advanced to determined variations of chemical compounds soil test kit. The KSP and KKK showed the highest found in different locations. In recent years, the value of pH of 7.0 (neutrality) while the lowest screening of phytochemical compounds of C. nutans value came from location KJN with a pH value of has already been studied such as stigmasterol, 5.0 (acidity). The mean rainfall in different location belutin, C-glycosyl flavones, vitexin, isovitexin, sites was collected from the nearest Malaysian orientin, isoorientin, sulfur-containing glycosides, Meteorological Department. This research was monoacylmonogalatosylglycerol, lupeol, β-sitosterol, conducted for three months (February 2015 to April shaftoside, isomollupentin-7-O-β-glucopyranoside, 2015) in Integrative Medicine Cluster Laboratory, glycoglycerolipids and cerebrosides (Sakdarat, Advanced Medical, and Dental Institute, Universiti Shuyprom, Pientong, Ekalaksananan, & Thongchai, Sains Malaysia. The identification of plant was 2009). Until the scientific research have informed, confirmed by the herbarium of School of Biological the diversity of phenolic,
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