Authentication of Gardeniae Fructus in Chinese Medicine Preparations by Nested PCR and DNA Sequencing Methods

Kang-Tsu Lu^*, Yung-Chuan Hsieh , Hui-Chun Lee, Yi-Chu Liu and Chi-Fang Lo

Food and Drug Administration, Department of Health, Executive Yuan, Taipei, Taiwan, R.O.C.

(Received: December 12, 2011; Accepted: May 30, 2012)

ABSTRACT

Chinese medicinal preparations are usually composed of various raw material components, and the complexity of herbal ingredients therein complicates the identification of constituent species. In this study, nested PCR coupled with DNA sequencing methods were established for Gardeniae Fructus in Chinese medicinal preparation. The botanical origin of Gardeniae Fructus is the dried mature fruits of Gardenia jasminoides Ells. The morphology of the dried mature fruits of G. jasminoides var. grandiflora (Lour.) Nakai is similar to that of G. jasminoides, which results in misuse. In this study, 28 raw materials and 22 preparation samples were collected. The total DNA of all samples were extracted by CTAB method and purified. Authentication was executed based on the sequence of internal transcribed spacer (ITS) which was amplified by nested PCR followed by sequencing. The ITS sequences of all samples were the same, with the exception of position 153. The nucleotide base of 3 raw material samples, identified as G. jasminoides var. grandiflora by morhpology, were base “G”. The raw material samples, identified as G. jasminoides by morphology, had the base “G” or “A” on this position. Gardeniae Fructus could be identified in all preparation samples by nested PCR and DNA sequencing.

Key words: Gardeniae Fructus, Gardenia jasminoides, internal transcribed spacer (ITS), nested PCR, DNA sequencing