Evaluation of Medicinal Categorization of Atractylodes japonica Koidz. by Using Internal Transcribed Spacer Sequencing Analysis and HPLC Fingerprinting Combined with Statistical Tools

Evidence-Based Complementary and Alternative Medicine, Apr 2016

Atractylodes rhizomes have been used as the herbal medicine “Changchul” or “Baekchul,” according to their clinical purpose, in Korea, China, and Japan. Among the Atractylodes species, the medicinal use of Atractylodes japonica has been controversial, as it is categorized as both Changchul and Baekchul in those countries, and, moreover, parts of the rhizome have been differently used, depending on age of the plant, in Korea. Chromatographic fingerprinting by using HPLC combined with chemometric analyses and internal transcribed spacer (ITS) sequencing analysis were conducted to classify and identify 34 crude drugs derived from Atractylodes rhizomes. The identification of the samples, authenticated by their morphological features as A. japonica Koidz. (Changchul and Baekchul), A. chinensis Koidz., and A. macrocephala Koidz., was confirmed as A. japonica, A. chinensis, and A. macrocephala by ITS sequencing. The results from chemometric analyses showed that the chemical components of the crude drugs from A. japonica were significantly different from those from A. macrocephala but were similar to those from A. chinensis. The analyses also suggested that the categorization by age of A. japonica as Changchul or Baekchul is not recommended. The results indicate that A. japonica should be categorized as “Changchul” and should not be further categorized by age.

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Evaluation of Medicinal Categorization of Atractylodes japonica Koidz. by Using Internal Transcribed Spacer Sequencing Analysis and HPLC Fingerprinting Combined with Statistical Tools

Evaluation of Medicinal Categorization of Atractylodes japonica Koidz. by Using Internal Transcribed Spacer Sequencing Analysis and HPLC Fingerprinting Combined with Statistical Tools Jung-Hoon Kim,1 Eui-Jeong Doh,2,3 and Guemsan Lee2 1Division of Pharmacology, School of Korean Medicine, Pusan National University, Yangsan 626-870, Republic of Korea 2Department of Herbology, College of Korean Medicine, Wonkwang University, Iksan 570-749, Republic of Korea 3Center for Metabolic Function Regulation, Wonkwang University, Iksan 570-749, Republic of Korea Received 9 December 2015; Accepted 14 March 2016 Academic Editor: Salih Mollahaliloglu Copyright © 2016 Jung-Hoon Kim et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Abstract Atractylodes rhizomes have been used as the herbal medicine “Changchul” or “Baekchul,” according to their clinical purpose, in Korea, China, and Japan. Among the Atractylodes species, the medicinal use of Atractylodes japonica has been controversial, as it is categorized as both Changchul and Baekchul in those countries, and, moreover, parts of the rhizome have been differently used, depending on age of the plant, in Korea. Chromatographic fingerprinting by using HPLC combined with chemometric analyses and internal transcribed spacer (ITS) sequencing analysis were conducted to classify and identify 34 crude drugs derived from Atractylodes rhizomes. The identification of the samples, authenticated by their morphological features as A. japonica Koidz. (Changchul and Baekchul), A. chinensis Koidz., and A. macrocephala Koidz., was confirmed as A. japonica, A. chinensis, and A. macrocephala by ITS sequencing. The results from chemometric analyses showed that the chemical components of the crude drugs from A. japonica were significantly different from those from A. macrocephala but were similar to those from A. chinensis. The analyses also suggested that the categorization by age of A. japonica as Changchul or Baekchul is not recommended. The results indicate that A. japonica should be categorized as “Changchul” and should not be further categorized by age. 1. Introduction The genus Atractylodes (Asteraceae) are perennial herbs distributed in Korea, China, and Japan. Their dried rhizomes have been classified into two kinds of herbal medicines according to their clinical purpose, “Baekchul” (Baizhu in Chinese, Byakujutsu in Japanese) and “Changchul” (Cangzhu in Chinese, Soujutsu in Japanese) [1]. In medicinal applications in Korea, Japan, and China, the rhizomes of A. lancea DC. and A. chinensis Koidz. have been classified as Changchul, while that of A. macrocephala Koidz. has been classified as Baekchul in the pharmacopeias of Korea, China, and Japan [2–4]. However, there has been disagreement between countries in classifying the rhizome of A. japonica Koidz.: Korean and Japanese pharmacopeias, as well as some studies from Korea and Japan, have classified the rhizome of A. japonica as Baekchul, whereas Chinese studies have classified it as “Gwan-Changchul” (Guan-Cangzhu in Chinese), a type of Changchul, which is not even listed in the Chinese pharmacopeia [5, 6]. Moreover, A. japonica is recorded as a synonym of A. lancea in the Flora of China [7]. Confusion also occurs in local herbal markets in Korea, where the rhizomes of A. japonica have been used as Changchul when the fibrous substance has formed after it has grown over two years [8]. The dried rhizomes of Atractylodes species can be identified by their morphological features; however, it is difficult to discriminate them by macroscopic observation, a subjective method, due to their morphological similarity. Consequently, the misuse of Atractylodes rhizomes may occur when identification is based only on their morphological features. Therefore, strict classification is required for the exact use of Atractylodes rhizomes for medicinal purposes. Genetic identification achieved by hybridization, polymerase chain reaction (PCR), and sequencing techniques is considered more objective, precise, and reliable method for identifying and authenticating herbal species [9, 10]. Internal transcribed spacer (ITS) regions, which are rapidly evolving regions of nuclear ribosomal DNA, have been widely used for the identification of plant materials, and phylogenetic analysis using ITS regions has been conducted to investigate the genetic variability of complicated herbal species [11–13]. It has been reported that the four species of Atractylodes (A. japonica, A. macrocephala, A. lancea, and A. chinensis) can be distinguished by their different genotypes on ITS sequences [14]. In contrast, one study conducted by PCR-restriction fragment length polymorphism (PCR-RFLP) analysis of the ITS region on nucleotide ribosome DNA (nrDNA) reported that A. japonica is not different from A. mac (...truncated)


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Jung-Hoon Kim, Eui-Jeong Doh, Guemsan Lee. Evaluation of Medicinal Categorization of Atractylodes japonica Koidz. by Using Internal Transcribed Spacer Sequencing Analysis and HPLC Fingerprinting Combined with Statistical Tools, Evidence-Based Complementary and Alternative Medicine, 2016, 2016, DOI: 10.1155/2016/2926819