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Identification of common horsetail (Equisetum arvense L.; Equisetaceae) using Thin Layer Chromatography versus DNA barcoding.

Saslis-Lagoudakis CH, Bruun-Lund S, Iwanycki NE, Seberg O, Petersen G, Jäger AK, Rønsted N - Sci Rep (2015)

Bottom Line: We find that both methods can discriminate between the two species and positively identify E. arvense.The TLC-test is more cost- and time-efficient, but DNA barcoding is more powerful in determining the identity of adulterant species.Our study shows that, although DNA barcoding presents certain advantages, other established laboratory methods can perform as well or even better in confirming species' identity in herbal products.

View Article: PubMed Central - PubMed

Affiliation: Evolutionary Genomics Section, Natural History Museum of Denmark, Sølvgade 83S, Copenhagen, DK-1307, Denmark.

ABSTRACT
The global herbal products market has grown in recent years, making regulation of these products paramount for public healthcare. For instance, the common horsetail (Equisetum arvense L.) is used in numerous herbal products, but it can be adulterated with closely related species, especially E. palustre L. that can produce toxic alkaloids. As morphology-based identification is often difficult or impossible, the identification of processed material can be aided by molecular techniques. In this study, we explore two molecular identification techniques as methods of testing the purity of these products: a Thin Layer Chromatography approach (TLC-test) included in the European Pharmacopoeia and a DNA barcoding approach, used in recent years to identify material in herbal products. We test the potential of these methods for distinguishing and identifying these species using material from herbarium collections and commercial herbal products. We find that both methods can discriminate between the two species and positively identify E. arvense. The TLC-test is more cost- and time-efficient, but DNA barcoding is more powerful in determining the identity of adulterant species. Our study shows that, although DNA barcoding presents certain advantages, other established laboratory methods can perform as well or even better in confirming species' identity in herbal products.

No MeSH data available.


TLC chromatogram of exemplar accessions of all Equisetum species.
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f6: TLC chromatogram of exemplar accessions of all Equisetum species.

Mentions: We analysed one exemplar specimen of all Equisetum species using the TLC-test recommended by the European Pharmacopoeia (Fig. 6). For E. diffusum and E. sylvaticum, which were not monophyletic in the DNA analysis, we could test only one sample, as the other sample did not come from our study. The TLC-test (Identification C) of the European Pharmacopoeia can positively identify E. arvense. Although some of the marker bands outlined in the TLC identification test for E. arvense (Fig. 2) are seen in the chromatograms of other Equisetum species, E. arvense is the only species with the combination of all these markers bands (Fig. 6). As shown in Fig. 6, one or two of the greenish-blue fluorescent zones used in the TLC-test to detect E. palustre were not detected in any other species within subg. Equisetum, but were present in all species in subg. Hippochaete. Therefore, the TLC-test of the European Pharmacopoeia cannot be used to identify E. palustre, because the trait of this species (Fig. 2) is shared with other Equisetum species as well (Fig. 6).


Identification of common horsetail (Equisetum arvense L.; Equisetaceae) using Thin Layer Chromatography versus DNA barcoding.

Saslis-Lagoudakis CH, Bruun-Lund S, Iwanycki NE, Seberg O, Petersen G, Jäger AK, Rønsted N - Sci Rep (2015)

TLC chromatogram of exemplar accessions of all Equisetum species.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4499799&req=5

f6: TLC chromatogram of exemplar accessions of all Equisetum species.
Mentions: We analysed one exemplar specimen of all Equisetum species using the TLC-test recommended by the European Pharmacopoeia (Fig. 6). For E. diffusum and E. sylvaticum, which were not monophyletic in the DNA analysis, we could test only one sample, as the other sample did not come from our study. The TLC-test (Identification C) of the European Pharmacopoeia can positively identify E. arvense. Although some of the marker bands outlined in the TLC identification test for E. arvense (Fig. 2) are seen in the chromatograms of other Equisetum species, E. arvense is the only species with the combination of all these markers bands (Fig. 6). As shown in Fig. 6, one or two of the greenish-blue fluorescent zones used in the TLC-test to detect E. palustre were not detected in any other species within subg. Equisetum, but were present in all species in subg. Hippochaete. Therefore, the TLC-test of the European Pharmacopoeia cannot be used to identify E. palustre, because the trait of this species (Fig. 2) is shared with other Equisetum species as well (Fig. 6).

Bottom Line: We find that both methods can discriminate between the two species and positively identify E. arvense.The TLC-test is more cost- and time-efficient, but DNA barcoding is more powerful in determining the identity of adulterant species.Our study shows that, although DNA barcoding presents certain advantages, other established laboratory methods can perform as well or even better in confirming species' identity in herbal products.

View Article: PubMed Central - PubMed

Affiliation: Evolutionary Genomics Section, Natural History Museum of Denmark, Sølvgade 83S, Copenhagen, DK-1307, Denmark.

ABSTRACT
The global herbal products market has grown in recent years, making regulation of these products paramount for public healthcare. For instance, the common horsetail (Equisetum arvense L.) is used in numerous herbal products, but it can be adulterated with closely related species, especially E. palustre L. that can produce toxic alkaloids. As morphology-based identification is often difficult or impossible, the identification of processed material can be aided by molecular techniques. In this study, we explore two molecular identification techniques as methods of testing the purity of these products: a Thin Layer Chromatography approach (TLC-test) included in the European Pharmacopoeia and a DNA barcoding approach, used in recent years to identify material in herbal products. We test the potential of these methods for distinguishing and identifying these species using material from herbarium collections and commercial herbal products. We find that both methods can discriminate between the two species and positively identify E. arvense. The TLC-test is more cost- and time-efficient, but DNA barcoding is more powerful in determining the identity of adulterant species. Our study shows that, although DNA barcoding presents certain advantages, other established laboratory methods can perform as well or even better in confirming species' identity in herbal products.

No MeSH data available.