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Combination of DNA isolation and RP-HPLC analysis method for bark samples of Saraca asoca and its adulterant

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      Abstract

      Abstract

      DNA fingerprinting singly or in combination with phytochemical analysis is ideal for quality control of crude plant-based drugs. However, when the source material is tannin rich stem bark, extraction of DNA by conventional methods becomes challenging. In such cases, phytochemical profiling serves as very useful tool for its identification. The work herein described a method for simultaneous DNA isolation and phytochemical extraction for downstream analysis and applications from dried bark powder of Saraca asoca and commercial samples of this crude drug as well as from those of Polyalthia longifolia, its most common adulterant. It is a modified CTAB-based method which involves a pre-extraction step by soaking samples overnight in de-ionized water followed by filtration. The residues in the filter paper were used for DNA isolation and dried filtrate was used for Reverse Phase-High-Performance Liquid Chromatography analysis. Results revealed that genomic DNA isolated was PCR amplifiable with Inter Simple Sequence Repeat and Start Codon Targeted markers. Phenolic compounds of catechin, epicatechin, and gallic acid were detected from the above dried filtrate. The method is simple, reliable and it requires small amount of sample with an option of integrating both phytochemical and DNA-based profiling, from the same starting material. Therefore, the present method could be useful for further potential applications such as quality control assessment of S. asoca products.

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      Protein measurement with the Folin phenol reagent.

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        Survey of commercial Rhodiola products revealed species diversity and potential safety issues

        The adulteration of herbal products is a threat to consumer safety. Here we surveyed the species composition of commercial Rhodiola products using DNA barcoding as a supervisory method. A Rhodiola dietary supplement DNA barcode database was successfully constructed using 82 voucher samples from 10 Rhodiola species. Based on the DNA barcoding standard operating procedure (SOP), we used this database to identify 100 Rhodiolae Crenulatae Radix et Rhizoma decoction piece samples that were purchased from drug stores and hospitals. The results showed that only 36 decoction piece sequences (40%) were authentic R. crenulata, which is recorded in Chinese Pharmacopeia, whereas the other samples were all adulterants and may indicate a potential safety issue. Among the adulterants, 35 sequences (38.9%) were authenticated as R. serrata, nine sequences (10%) were authenticated as R. rosea, which is documented in the United States Pharmacopeia, and the remaining samples were authenticated as other three Rhodiola species. This result indicates decoction pieces that are available in the market have complex origins and DNA barcoding is a convenient tool for market supervision.
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          Start Codon Targeted (SCoT) Polymorphism: A Simple, Novel DNA Marker Technique for Generating Gene-Targeted Markers in Plants

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            Author and article information

            Affiliations
            [1 ]ISNI 0000 0004 1767 225X, GRID grid.19096.37, ICMR-National Institute of Traditional Medicine (Formerly Regional Medical Research Centre), , Indian Council of Medical Research, Department of Health Research, Govt. of India, ; Belagavi, Karnataka 590 010 India
            [2 ]ISNI 0000 0001 1889 7360, GRID grid.411053.2, , KLE Academy of Higher Education and Research (KLE University), ; Belagavi, Karnataka 590 010 India
            [3 ]ISNI 0000 0001 1889 7360, GRID grid.411053.2, Dr. Prabhakar Kore Basic Science Research Centre, , KLE Academy of Higher Education and Research (KLE University), ; Belagavi, Karnataka 590010 India
            [4 ]ISNI 0000 0004 1805 0217, GRID grid.444644.2, Amity Institute of Biotechnology (AIB), , Amity University, ; Mumbai, Maharashtra 410206 India
            Contributors
            +91-831-2475477/78 , drsubarnaroy@gmail.com , roys@icmr.gov.in
            Journal
            3 Biotech
            3 Biotech
            3 Biotech
            Springer Berlin Heidelberg (Berlin/Heidelberg )
            2190-572X
            2190-5738
            30 June 2017
            July 2017
            : 7
            : 3
            28667648
            5493565
            791
            10.1007/s13205-017-0791-9
            © Springer-Verlag GmbH Germany 2017
            Funding
            Funded by: FundRef http://dx.doi.org/10.13039/501100001411, Indian Council of Medical Research;
            Award ID: 45/53/2013/BMS/TRM
            Award Recipient :
            Funded by: KLE University
            Award ID: KLEU/Accs/12-13/D-1552
            Award Recipient :
            Categories
            Original Article
            Custom metadata
            © Springer-Verlag GmbH Germany 2017

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