RESUMO
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.
Assuntos
Panax/genética , Plantas Medicinais/genética , Polimorfismo Genético , Sitios de Sequências Rotuladas , Sequência de Bases , Primers do DNA , DNA de Plantas/genética , DNA Ribossômico/genética , Marcadores Genéticos/genética , Dados de Sequência Molecular , Panax/classificação , Raízes de Plantas/genética , Técnica de Amplificação ao Acaso de DNA Polimórfico/métodos , Análise de Sequência de DNA , Especificidade da EspécieRESUMO
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).
Assuntos
Angelica/genética , Panax/genética , Plantas Medicinais/genética , Trichosanthes/genética , Angelica/classificação , Primers do DNA , DNA de Plantas/análise , DNA de Plantas/genética , Contaminação de Medicamentos/prevenção & controle , Medicina Tradicional Chinesa/normas , Panax/classificação , Plantas Medicinais/classificação , Controle de Qualidade , Técnica de Amplificação ao Acaso de DNA Polimórfico/métodos , Reprodutibilidade dos Testes , Trichosanthes/classificaçãoRESUMO
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.
