應用巢式PCR-DNA定序方法與巢式PCR-限制片段多型性鑑別中藥製劑中的人參藥材   全文下載

Identification of Ginseng Radix in Chinese Medicine Preparations by Nested PCR-DNA Sequencing Method and Nested PCR-Restriction Fragment Length Polymorphism

人參藥材為植物Panax ginseng的乾燥根，因為外觀相似，容易與Panax屬的植物藥材發生混淆，特別在中藥製劑中，不易鑑別其中人參藥材是否誤用。在本研究中，應用巢式PCR-DNA定序方法鑑別製劑的人參成分， 58件市售製劑檢體中，48件含有P. ginseng，8件含有P. quinguefolius ， 2件含有P. notoginseng。再以限制?BstYI與DdeI處理檢體的巢式PCR產物，做限制片段多型性分析，結果顯示這58件檢體中有23件含有混合的P. ginseng與P. quinguefolius，25件含有單純的P. ginseng，而有10件是誤用P. quinguefolius或P. notoginseng取代P. ginseng入藥。巢式PCR-限制片段多型性的P. ginseng定性結果與巢式PCR-DNA定序結果一致。本研究所建立之巢式PCR-DNA定序方法可鑑別中藥製劑中的人參藥材，同時運用巢式PCR與限制片段多型性分析方法結合，可自Panax屬的植物藥材中鑑別出人參藥材，並且可檢測製劑是否使用不純的人參藥材。

Ginseng Radix, the dried root of Panax ginseng, is easily confused with other herbs of the Panax species by appearance. In Chinese medicine preparations, it is difficult to identify misuse of Ginseng Radix. In this study, the nested PCR and DNA sequencing methods were established for the identification of Ginseng Radix, whereas the combination of nested PCR and restric-tion fragment length polymorphism (RFLP) could differentiate Ginseng Radix from other Panax species and thus certify the use of impure raw material of Ginseng Radix in Chinese medicine preparations. Nested PCR and DNA sequencing methods were applied to verify ginseng ingredients in the preparations. Of the 58 samples analyzed, 48 samples contained P. ginseng, 8 P. quinguefolius, and 2 P. notoginseng. Utilizing restriction enzyme BstYI and DdeI, nested PCR products of the samples were digested for restric-tion fragment lengths of polymorphism (RFLP). It was proven that 23 of the 58 samples contained a mixture of Ginseng Radix and Panacis quinquefolii Radix. Twenty-five samples contained pure raw material of Ginseng Radix. Ginseng components of 10 samples were confirmed using a substitute of P. quinguefolius or P. notoginseng. The RFLP results were consistent with those of the sequencing method results.