In the last two decades, there has been a tremendous increase in the global use of herbal medicinal products (HMPs) due to their claimed health benefits. This has led to increase in their demand and consequently, also, resulted in massive adulteration. This is due to the fact that most of the traditional methods cannot identify closely related species in a process product form. Therefore the urgent need for simple and rapid identification methods resulted in the discovery of a novel technique. DNA barcoding is a process that uses short DNA sequence from the standard genome for species identification. This technique is reliable and is not affected by external factors such as climates, age, or plant part. The difficulties in isolation of DNA of high quality in addition to other factors are among the challenges encountered using the DNA barcoding in the authentication of HMP. These limitations indicated that using DNA barcoding alone may ineffectively authenticate the HMP. Therefore, the combination of DNA barcoding with chromatographic fingerprint, a popular and generally accepted technique for the assessment and quality control of HMP, will offer an efficient solution to effectively evaluate the authenticity and quality consistency of HMP. Detailed and quality information about the main composition of the HMPs will help to ascertain their efficacy and safety as these are very important for quality control.
Context:
Eurycoma longifolia Jack (Simaroubaceae) commonly known as Tongkat Ali is one of the most important plants in Malaysia. The plant extracts (particularly roots) are widely used for the treatment of cough and fever besides having antimalarial, antidiabetic, anticancer and aphrodisiac activities.
Objectives: This study assesses the extent of adulteration of E. longifolia herbal medicinal products (HMPs) using DNA barcoding validated by HPLC analysis.
Materials and methods: Chloroplastic rbcL and nuclear ITS2 barcode regions were used in the present study. The sequences generated from E. longifolia HMPs were compared to sequences in the GenBank using MEGABLAST to verify their taxonomic identity. These results were verified by neighbor-joining tree analysis in which branches of unknown specimen are compared to the reference sequences established from this study and other retrieved from the GenBank. The HMPs were also analysed using HPLC analysis for the presence of eurycomanone bioactive marker.
Results: Identification using DNA barcoding revealed that 37% of the tested HMPs were authentic while 27% were adulterated with the ITS2 barcode region proven to be the ideal marker. The validation of the authenticity using HPLC analysis showed a situation in which a species which was identified as authentic was found not to contain the expected chemical compound.
Discussion and conclusions: DNA barcoding should be used as the first screening step for testing of HMPs raw materials. However, integration of DNA barcoding with HPLC analysis will help to provide detailed knowledge about the safety and efficacy of the HMPs.
Antioxidative compounds play a vital role to inhibit the oxidation of oxidizable products and influences the body defense system against Reactive Oxygen Species (ROS), which are harmful byproducts engendered during normal aerobic cellular respiration. ROS are specialized class of highly reactive molecules that originates during oxygen metabolism, whereas extensive evidences indicate the involvement of ROS in the development of degenerative diseases in humans. Despite the fact that synthetic antioxidants such as butylated hydroxyanisole (BHA) and butylated hydroxytoluene (BHT) are threat to human's health; they are widely employed as chemical preservatives to avert potential deterioration of food products. Several toxicological findings reveal that natural antioxidants are capable of providing protection against free radical-related diseases, cancer and other cardiovascular infections. In cognizance with these facts, natural antioxidant attracts a great deal of research interest; therefore, this review provides a comprehensive and an up-to-date overview of the three most essential natural plant antioxidants; their antioxidative properties, methods of analysis and evaluation. These Polyphenols comprise: Phenolic acid, an essential polyphenol that are broadly dispersed in plants to exert positive influence on oxidative stress; Flavonoid, an ubiquitous antioxidant that functions in free radical scavenging, metal ion chelation and enzymes inhibition to suppress the synthesis of free radicals; and Tannic acid, a water soluble anti-nutrient that acts to precipitate alkaloids & proteins, reduce mineral & vitamin utilization and restrict the activities of digestive enzymes. In conclusion, a variety of plant species were characterized to indicate in vitro antioxidative properties.
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