Quantitative high-resolution melting analysis for detecting adulterations.
Admixtures of different plant species are a common problem in raw materials for medicinal use. Two exemplary assays were developed to admixtures in Helleborus niger with high-resolution melting analysis. HRM proved to be a very sensitive tool in detecting admixtures, able to detect a ratio of 1:1000 with unknown species, and of 1:200,000 with Veratrum nigrum. The example proves the ability of HRM for quantification in multiplex PCR. The method is not limited to detecting adulterations. It can also be used to quantify a specific target by integrating a second amplicon in the assay as internal standard.
Mader
Eduard
E
Institute for Applied Botany, University of Veterinary Medicine, Veterinärplatz 1, A-1210 Wien, Austria. eduard.mader@vetmeduni.ac.at
Ruzicka
Joana
J
Schmiderer
Corinna
C
Novak
Johannes
J
D017931:DNA Primers / Q000737:chemistry; D018744:DNA, Plant / Q000737:chemistry*; D005838:Genotype; D039322:Helleborus / Q000235:genetics; D009691:Nucleic Acid Denaturation; D016133:Polymerase Chain Reaction / Q000379:methods*; D013050:Spectrometry, Fluorescence; D044366:Transition Temperature; D014703:Veratrum / Q000235:genetics
D017931:DNA Primers; D018744:DNA, Plant