Comparative Evaluation of Different DNA Extraction Methods from E. Longifolia Herbal Medicinal Product

Abstract

The aphrodisiac property of Eurycoma longifolia has led to an increase in the demand for its Herbal Medicinal Products (HMPs). However, the efficiency of such HMPs depends on the usage of their genuine raw materials. The conventional methods cannot identify species in processed form. The authentication of HMPs can be achieved effectively using DNA barcoding as the method species-specific. However, the use of this method solely relied on the extraction of high-quality DNA from the HMPs. Therefore, it is necessary to establish a satisfactory method for extracting high-quality DNA from the HMPs. Here, four DNA extraction methods were compared to evaluate the best protocol in yield, purity, polymerase chain reaction (PCR) amplification, sequencing, and species identification. The spectrophotometer analysis showed that the Nucleospin Plant II extraction kit has the best purity as this can be severely affected by the presence of various contaminants in the HMPs. Our findings reveal that DNA purity was more important as a predictor for PCR amplification than yield. Therefore, the present study results demonstrate that the Nucleospin Plant II extraction kit is the best because it produces the purest, amplifiable, and sequenceable DNA for identification and authentication of E. Longifolia HMPs.

Rapid identification of Amanita citrinoannulata poisoning using colorimetric and real-time fluorescence and loop-mediated isothermal amplification (LAMP) based on the nuclear ITS region

“…This is consistent with the research findings of Njiru et al ( Njiru et al, 2008 ), and may be due to a fact that mushroom fruiting bodies contain more polysaccharides and phenolic compounds. These secondary metabolites easily affect the DNA amplification results ( Abubakar et al, 2021 ). The amplification efficiency could be improved by constantly purifying the DNA to be used in the experiment.…”

Floristic composition and structure of Eurycoma longifolia habitat in Muka Kuning Nature Tourism Park, Riau Islands, Indonesia

“…Because the aphrodisiac properties and high portion of urycomanine are contained in the root part of E. longifolia (Jusoh et al 2015;Abubakar et al 2018), the harvesting of E. longifolia is carried out destructively by uprooting the plant, leading to a significant population decline in the wild (Susilowati et al 2021). Further, the sustainability of E. longifolia is increasingly pressured with the emergence of various products that use E. longifolia as the main ingredient such as capsules, powders, and herbal teas (Abubakar et al 2021).…”

EVALUATION OF DNA QUALITY FROM MODIFIED DNA EXTRACTION AND AMPLIFICATION OF ITS2 FROM Eurycoma longifolia CAPSULE HERBAL PRODUCTS

“…Production of high-quality DNA is characterized predominantly by high molecular weight fragments with an A260/280 ratio between 1.8 to 2.0 and lacking contaminating substances such as polysaccharides and other secondary metabolites [24]. In a previous study, the commercial kit, NucleoSpin Plant II kit, was the best method for DNA extraction from Eurycoma longifolia products [1]. An economical DNA extraction method for dried herbal powder has been developed to retrieve DNA with successful PCR amplification [3].…”