Identification of Saposhinkoviae Radix in Concentrated Chinese Medicine Preparations by Nested PCR and DNA Sequencing Methods

KANG-TSU LU*, CHI-FANG LO, HSIEN-CHANG CHANG AND JER-HUEI LIN

Division of Pharmacognosy, Bureau of Food and Drug Analysis, Department of Health, Taiwan, R.O.C.

(Received: February 14, 2005; Accepted: April 14, 2005)

ABSTRACT

   Saposhinkoviae Radix, the dried root of Saposhinkovia divaricata (Turcz.) Schischk., is widely used in Chinese medicine preparations.  This paper reports on the development of a nested polymerase chain reaction (nested PCR) and DNA sequencing method able to detect the presence of authentic Saposhikoviae Radix in Chinese medicine preparations. Forty concentrated preparation samples in 17 formulas containing a total of 54 kinds of raw materials were purchased. DNA samples were completely digested with a lysis buffer containing proteinase and sarcosyl, extracted with organic solutions and CTAB, and then purified using a commercial kit (QIAGEN GmbH, Germany) that contained silica membrane to absorb the DNA. The internal transcribed spacer (ITS) regions of 40 preparations were amplified by nested PCR. All PCR products were analyzed by auto-sequencer.  We compared resultant sequences with the ITS sequences obtained from the standard and Genbank databases.  Saposhinkoviae Radix in all 40 preparations tested were identified as Saposhinkovia divaricata. These results indicate the above is a valid method by which to identify the presence of authentic Saposhinkoviae Radix in Chinese medicine preparations.

Key words: Saposhinkovia divaricata, nested PCR, internal transcribed spacer (ITS)