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A Multi-Pronged Technique for Identifying Equisetum palustre and Equisetum arvense-Combining HPTLC, HPLC-ESI-MS/MS and Optimized DNA Barcoding Techniques.

ABSTRACT: The most prominent horsetail species, Equisetum arvense, has an array of different medicinal properties, thus the proper authentication and differentiation of the plant from the more toxic Equisetum palustre is important. This study sought to identify different samples of E. arvense and E. palustre using three analytical methods. The first method involved the use of HPTLC analysis, as proposed by the European Pharmacopoeia. The second, HPLC-ESI-MS/MS, is capable of both identification and quantification and was used to determine the Equisetum alkaloid content in each sample. A third method was DNA barcoding, which identifies the samples based on their genetic make-up. Both HPTLC and HPLC-ESI-MS/MS proved to be suitable methods of identification, with HPLC-ESI-MS/MS proving the more sophisticated method for the quantification of alkaloids in the Equisetum samples and for determining the adulteration of E. arvense. For DNA barcoding, optimal primer pairs were elucidated to allow for the combined use of the rbcL and ITS markers to accurately identify each species. As new DNA marker sequences were added to GenBank, the reference library has been enriched for future work with these horsetail species.

SUBMITTER: Ibi A

PROVIDER: S-EPMC9573248 | biostudies-literature | 2022 Sep

REPOSITORIES: biostudies-literature

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A Multi-Pronged Technique for Identifying Equisetum palustre and Equisetum arvense-Combining HPTLC, HPLC-ESI-MS/MS and Optimized DNA Barcoding Techniques.

Ibi Afoke A Du Min M Beuerle Till T Melchert Dennis D Solnier Julia J Chang Chuck C

Plants (Basel, Switzerland) 20220928 19

The most prominent horsetail species, Equisetum arvense, has an array of different medicinal properties, thus the proper authentication and differentiation of the plant from the more toxic Equisetum palustre is important. This study sought to identify different samples of E. arvense and E. palustre using three analytical methods. The first method involved the use of HPTLC analysis, as proposed by the European Pharmacopoeia. The second, HPLC-ESI-MS/MS, is capable of bo ...[more]

PMID: 36235428

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Project description:The objective of the present work was to develop a simple, specific, and fast high-performance thin-layer chromatographic (HPTLC) method to identify and quantify cordifolioside A, 20-β-hydroxyecdysone and columbin with HPTLC‒electrospray ionization‒tandem mass spectrometry (ESI‒MS/MS) for characterization in Tinospora cordifolia stem extracts. Chromatographic development was performed using a HPTLC aluminum plate, pre-coated with silica gel 60 F254 with hexane‒chloroform‒methanol‒formic acid as the mobile phase. Densitometric quantification for 20-β-hydroxyecdysone and cordifolioside A was performed at 254 nm and for columbin at 600 nm after derivatization with anisaldehyde‒sulfuric acid. The optimized mobile phase resulted in chromatographic separation of peaks for cordifolioside A, 20-β-hydroxyecdysone, and columbin at RF of 0.12, 0.47, and 0.86, respectively. The linear concentration range was found to be 750‒2250 ng/band for 20-β-hydroxyecdysone and cordifolioside A and 675‒1875 ng/band for columbin with (r2 > 0.99). The methodology showed good recoveries as 98.96‒101.43% for cordifolioside A, 98.15‒101.56% for 20-β-hydroxyecdysone, and 98.06‒98.80% for columbin. The limit of detection was found for columbin, 20-β-hydroxyecdysone, and cordifolioside A as 53.86 ng/band, 40.90 ng/band, and 107.05 ng/band, while the limit of quantification was found to be 163.21 ng/band, 123.94 ng/band, and 324.38 ng/band, respectively. The relative standard deviation for precision and robustness study for all the markers was found to be within 2%. Three markers were identified and confirmed in T. cordifolia stem extracts by ESI‒MS/MS. Compounds were assigned as norditerpene furan glycosides, ecdysteroids, and diterpenoid furanolactone: cordifolioside A (m/z = 527 [M + Na]+; UV λmax 221 nm), 20-β-hydroxyecdysone (m/z = 481.30 [M + H]+; UV λmax 247 nm), and columbin (m/z = 359 [M + H]+; UV λmax 210 nm). The optimized method was found accurate, reproducible, robust, and specific and can be applied for the quantification of cordifolioside A, 20-β-hydroxyecdysone, and columbin for quality control of extracts of T. cordifolia. Supplementary Information The online version contains supplementary material available at 10.1007/s00764-021-00115-7.

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Dataset Information

A Multi-Pronged Technique for Identifying Equisetum palustre and Equisetum arvense-Combining HPTLC, HPLC-ESI-MS/MS and Optimized DNA Barcoding Techniques.

Publications

A Multi-Pronged Technique for Identifying <i>Equisetum palustre</i> and <i>Equisetum arvense</i>-Combining HPTLC, HPLC-ESI-MS/MS and Optimized DNA Barcoding Techniques.

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