ABSTRACT
OBJECTIVE: Searching a new molecular method to authenticate Panax ginseng and P. quenquefolium. METHOD: Single primers based on rDNA sequences of Panax species were designed to obtain polymorphic bands of P. ginseng and P. quinquefolius and then sequenced. Four PCR primers (two forword and two reverse primers) specific to P. ginseng and P. quinquefolius were designed. RESULT: Primer Pg-6F, Pg-479R only amplified 474 bp band for P. ginseng and primer Pq-442F, Pq-658R only amplified 217 bp band for P. quinquefolius. It is indicated that the four primers could serve as specific STS primers for Panax species. CONCLUSION: A new way to obtain STS primers of Panax species was established. This method is more quick and efficient than SCAR-PCR method and can serve as a model to obtain molecular markers for other Chinese material medica.
Subject(s)
Panax/genetics , Plants, Medicinal/genetics , Polymorphism, Genetic , Sequence Tagged Sites , Base Sequence , DNA Primers , DNA, Plant/genetics , DNA, Ribosomal/genetics , Genetic Markers/genetics , Molecular Sequence Data , Panax/classification , Plant Roots/genetics , Random Amplified Polymorphic DNA Technique/methods , Sequence Analysis, DNA , Species SpecificityABSTRACT
To build up a stable and easy doing method for molecular identification in traditional Chinese medicine, on basis of RAPD, the new method mainly changed the primer length and PCR annealing temperature. Panax ginseng, Panax quinquefolius and its nine adulterants were used to establish the method and test it using MARMS primers published in 2004. The new method also used to authenticate Chinese Materia Medica of Tian-hua-fen (Radix Trichosanthes) and Bai-zhi (Radix Angelica). Primer Pg-q36F obtained polymorphic bands of P. Ginseng, P. quinquefolius and its adulterants. The identification result is identical to that published before and more stable. Primer TkS1-64F obtained polymorphic bands of Tian-hua-fen and its nine adulterants. Primer AfS1-100F obtained polymorphic bands of Bai-zhi and its three adulterants. The method has good stability and reproducibility and can easily identify authertic medicines from their adulterants. It was a potential molecular method to identify other Chinese Materia Medica. The method was named as anchored primer amplification polymorphism DNA (APAPD).
Subject(s)
Angelica/genetics , Panax/genetics , Plants, Medicinal/genetics , Trichosanthes/genetics , Angelica/classification , DNA Primers , DNA, Plant/analysis , DNA, Plant/genetics , Drug Contamination/prevention & control , Medicine, Chinese Traditional/standards , Panax/classification , Plants, Medicinal/classification , Quality Control , Random Amplified Polymorphic DNA Technique/methods , Reproducibility of Results , Trichosanthes/classificationABSTRACT
OBJECTIVE: To describe the difference between native and nonative herbs by determining contents of seven kinds of flavone for twenty-five samples from seventeen areas. METHODS: HPLC. Fluid phase: MEOH-H2O-CH3COOH(ICE) (41:59:0.2) and (50:50:0.2). Detection wavelength: 275. RESULTS: The contents of baicalin are 6%-9%, wogenin are 2%-8%, baicalein are 0.1%-1.6%, neobaicalein are 0.01%-0.2%, wogonin are 0.01%-0.3%, visidulin and oroxylin are trace amounts or undetected. CONCLUSION: The native and nonative herbs have no distinct differce in absolute component ratio. The ratio of baicalin and wogenin is under three. The ratio of baicalin and baicalein, baicalin and wogonin is between twenty and fifty.
