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Integrating macro-microscopy, DNA barcoding and HPTLC for quality assessment of berberine containing botanicals
Sunil Kumar Koppala Narayana1, Pushkar Kaira1, Mahima Karthikeyan1
1Department of Pharmacognosy, Siddha Central Research Institute (Central Council for Research in Siddha, Ministry of AYUSH, Govt. of India), Arumbakkam, Chennai, Tamil Nadu, 600106, India.
Background:
Daruharidra/Maramanjal is one of the most popular shrub used in Ayurveda, Siddha and other Indian medicinal systems. More than one botanical source is traded under this name, predominantly Berberis aristata and Coscinium fenestratum with an annual trade of 1000-2000 metric tonnes. The herbal drug trade is often reported with misidentification, adulteration and/or substitution issues due to morphological resemblance and confusion in vernacular names. This work aimed to integrate macro-microscopic, DNA marker strategies and phytochemical assay to differentiate Berberis aristata from its traded sources.
Material And Methods:
Thirteen marketed samples and one authentic field sample from natural habitat were collected from various regions of the Indian market under the trade name Maramanjal/Daruharidra. The traditional identification methods included macro-microscopic and phytochemical screening by High-Performance Thin Layer Chromatography (HPTLC). Additionally, DNA barcode-based molecular identification and phylogenetic analysis were done using the ITS2 (Internal Transcribed Spacer 2) marker.
Results:
The macroscopic observations revealed 80 % ad-mixing of various allied botanicals in addition to accepted north Indian and south Indian sources such as B. aristata and C. fenestratum respectively. DNA barcoding enabled the identification of genuine and adulterated raw drugs from the collected samples. The HPTLC quantification revealed the presence of berberine in all 14 samples varying from 1.12 % to 26.33 %.
Conclusions:
The macro-micro, HPTLC, and DNA barcoding helped in the identification of adulteration and substitution practices in this highly traded botanical drug. DNA barcoding can prove an effective tool for discovering the adulteration and substitution of Maramanjal/Daruharidra and this is its first report on the application of morphology, microscopy, phytochemical analysis, and DNA markers in differentiating these traded species.
