RSC Advances View Article Online PAPER View Journal | View Issue Study on the identification of Pinelliae rhizoma and Pinelliae pedatisectae rhizoma based on the Cite this: RSC Adv.,2019,9, 11774 characteristic component triglochinic acid Yong Jing,a Yueyue Lai,a Hui Chen,a Min Li, *a Juan Zhoub and Zelun Lanc For the first time, a monomeric compound, triglochinic acid, has been isolated from the tubers of Pinellia pedatisecta Schott with its structure analyzed using NMR. Its molecular formula is C7H8O6, with the chemical name (2E)-but-2-ene-1,2,4-tricarboxylic acid. Through HPLC-DAD and HPLC-MS analysis studies, it has concluded that Pinelliae rhizoma does not contain triglochinic acid, while Pinelliae pedatisectae rhizoma Received 4th March 2019 does. This key observation was used as a characteristic component to distinguish these two herbs. We Accepted 1st April 2019 analyzed 39 batches of Pinelliae rhizoma collected in herbal medicine market, among which triglochinic acid DOI: 10.1039/c9ra01626k was detected in 27 batches, resulting in a adulteration ratio of Pinelliae rhizoma reaching 69.2%. Our method rsc.li/rsc-advances demonstrates great potential for authenticating the products, thus ensuring the quality of Pinelliae rhizoma. Creative Commons Attribution 3.0 Unported Licence. 1. Introduction removed during processing, the herbs will be very similar to Pinelliae rhizoma, which makes them difficult to distinguish. Pinelliae rhizoma is the dry tubers of Pinellia ternata (Thunb.) Breit.1 Pinelliae rhizoma enjoys a wide range of clinical applications, being It was documented as Diwen or Shuiyu in Shennong's Materia the raw material in 489 patent traditional Chinese medicine8 and Medica (Shen Nong Ben Cao Jing), and other medical classics in 3029 prescriptions.9 The growing period of Pinellia ternata is 4–5 aerwards. During ancient times, counterfeit or fake products of months with high cost and low yield, and its market price is 85– Pinelliae rhizoma existed, such as Arisaema ringens (Thunb.) Schott 120 yuan per kilogram. On the other hand, the clinical application This article is licensed under a in Tang Ben Cao by Su Gong and Pinellia pedatisecta Schott in Tu of Pinelliae pedatisectae rhizoma is limited, only used in a few JingBenCao.Inmoderntimes,Pinelliae rhizoma in rural areas was prescriptions. Its lifespan is shorter, about 3–4 months with lower replaced by herbs of the same genus like Typhonii agelliformis and cost, higher yield and the price is 40–50 yuan per kilogram. Open Access Article. Published on 16 April 2019. Downloaded 9/29/2021 6:04:14 PM. Arisaema heterophyllum Blume according to Modern Chinese Therefore, the interests drive people to incorporate Pinelliae ped- Materia Medica.2 Recently, Pinelliae rhizoma was discovered to be atisectae rhizoma into Pinelliae rhizoma. mixed with a large amount of Pinelliae pedatisectae rhizoma in the Nowadays, the identications of Pinelliae rhizoma, Typhonii market, which greatly affected the quality of the herbs. Pinelliae agelliformis rhizoma and Arisaematis rhizoma are mostly based pedatisectae rhizoma, also known as the palm leaf Pinellia and on morphology identication, thin layer chromatography – South Star, is the dry tubers of Pinellia pedatisecta Schott, docu- identication, and ngerprint identication.10 12 However, the mented in the Chinese Materia Medica Standards of Shandong, identication studies on Pinelliae rhizoma and Pinelliae ped- Hubei and Jiangsu,3–5 as well as in the Dictionary of Traditional atisectae rhizoma are very limited. In particular, it is hard to Chinese Medicine6 and Chinese Flora.7 identify the incorporation of Pinelliae pedatisectae rhizoma in Pinelliae rhizoma and Pinelliae pedatisectae rhizoma are both of Pinelliae rhizoma. From 2016 to 2017, we collected Pinelliae the Pinellia genus, which can easily be identied by the leaves and rhizoma from several herbal medicine markets: Hehuachi, owers.7 However, their medicinal materials are extremely similar. Anguo, Bozhou, Qingping, Yulin, etc. However, more than 60% Pinelliae rhizoma is spheroidal in shape, while Pinelliae pedatisectae of the samples were counterfeit. This situation urges us to rhizoma is also spheroidal with several small bulbs alongside. If the establish an effective and rapid analytical method that can small bulbs of Pinelliae pedatisectae rhizoma are not yet formed or detect fake herbs and improve the quality control of Pinelliae rhizoma, meanwhile laying the foundation of a standard herbal medicine market. aKey Laboratory of Standardization of Chinese Herbal Medicine, State Key Laboratory Breeding Base of Systematic Research, Development and Utilization of Chinese Medicine Resources, Ministry of Education, College of Pharmacy, Chengdu 2. Material University of Traditional Chinese Medicine, Chengdu 611137, China. E-mail: 2.1 Equipment [email protected]; Tel: +86-13980038316 bSichuan Institute for Food and Drug Control, Chengdu 611731, China Agilent 1200 high performance liquid chromatography (with cSichuan Neautus Traditional Chinese Medicine Co., Ltd., Chengdu 611731, China DAD detector), Agilent 1290/6460 LC/MS, Agilent ZORBAX 11774 | RSC Adv.,2019,9, 11774–11780 This journal is © The Royal Society of Chemistry 2019 View Article Online Paper RSC Advances Eclipse Plus C18 column (4.6  250 mm, 5 mm), Agilent ZORBAX Table 1 Market Information of the samples  m Eclipse Plus C18 column (2.1 50 mm, 1.8 m), XB-C18 column ID Sample Origin (80  250 mm, 10 mm), 3–30 K centrifuge (Sigma, Germany), KQ-500DE CNC ultrasonic cleaner (Kunshan Ultrasonic Instru- BX1 Pinelliae rhizoma Muba, Nanchong, Sichuan ment Co., Ltd.); ULUP-IV-10T Youpu series ultrapure water BX2 Pinelliae rhizoma Muba, Nanchong, Sichuan (Chengdu Ultrapure Technology Co., Ltd.), SQP electronic BX3 Pinelliae rhizoma Jinchuan, Aba, Sihuan balance (Sartorious Scientic Instrument Beijing Co., Ltd.). BX4 Pinelliae rhizoma Pengxi, Suining, Sichuan BX5 Pinelliae rhizoma Wenjiang, Chengdu, Sichuan BX6 Pinelliae rhizoma Nanjiang, Bazhong, Sichuan 2.2 Reagents BX7 Pinelliae rhizoma Mianning, Liangshan, Sichuan Triglochinic acid reference substance (96.68% purity, batch BX8 Pinelliae rhizoma Mianning, Liangshan, Sichuan BX9 Pinelliae rhizoma Hongjiang, Suining, Sichuan number 20170201), purchased from Chengdu Push Bio- BX10 Pinelliae rhizoma Changle, Nanchong, Sichuan Technology Co., Ltd. Phosphoric acid (Chengdu Kelon Chem- BX11 Pinelliae rhizoma Shayang, Jinmen, Hubei ical Reagent Factory), formic acid (Tianjin Kemiou Chemical BX12 Pinelliae rhizoma Xihe, Longnan, Gansu Reagent Co., Ltd.) and acetonitrile (Thermo Fisher Scientic BX13 Pinelliae rhizoma Li, Longnan, Gansu (China)) are chromatographically pure, and the water is BX14 Pinelliae rhizoma Qingshui, Tianshui, Gansu BX15 Pinelliae rhizoma Xihe, Longnan, Gansu ultrapure. BX16 Pinelliae rhizoma Qingshui, Tianshui, Gansu BX17 Pinelliae rhizoma Weining, Guizhou 2.3 Medicinal materials BX18 Pinelliae rhizoma Dafang, Guizhou BX19 Pinelliae rhizoma Hechuan, Chongqing From Sichuan, Gansu, Guizhou, Hebei, Shanxi, Chongqing and BX20 Pinelliae rhizoma Shenze, Shijiazhuang, Hebei other provinces and cities, we collected 28 batches of Pinelliae BX21 Pinelliae rhizoma Shenze, Shijiazhuang, Hebei rhizoma. Also from Hebei, Heilongjiang, Gansu, Sichuan and BX22 Pinelliae rhizoma Shenze, Shijiazhuang, Hebei other places, we collected 16 batches of Pinelliae pedatisectae BX23 Pinelliae rhizoma Houma, Shanxi Creative Commons Attribution 3.0 Unported Licence. rhizoma. All the materials were determined by Professor Min Li BX24 Pinelliae rhizoma Houma, Shanxi BX25 Pinelliae rhizoma Houma, Shanxi from College of Pharmacy, Chengdu University of Traditional BX26 Pinelliae rhizoma Weining, Guizhou Chinese Medicine. Information of these samples is shown in BX27 Pinelliae rhizoma Hezhang, Guizhou Table 1. Photos of Pinelliae rhizoma and Pinelliae pedatisectae BX28 Pinelliae rhizoma Hezhang, Guizhou are shown in Fig. 1. HZ1 Pinelliae pedatisectae rhizoma Qizhou, Anguo, Hebei In addition, 39 batches of commercial Pinelliae rhizoma were HZ2 Pinelliae pedatisectae rhizoma Zhengzhang, Anguo, Hebei HZ3 Pinelliae pedatisectae rhizoma Zhengzhang, Anguo, Hebei collected from the herbal medicine markets of Hehuachi HZ4 Pinelliae pedatisectae rhizoma Xifuluo, Anguo, Hebei Chengdu, Anguo Hebei, Yinzhou Anhui, Qingping Guangdong, HZ5 Pinelliae pedatisectae rhizoma Qizhou, Anguo, Hebei This article is licensed under a and Yulin Guangxi. Information of the commercial Pinelliae HZ6 Pinelliae pedatisectae rhizoma Qizhou, Anguo, Hebei rhizoma samples is shown in Table 2. HZ7 Pinelliae pedatisectae rhizoma Tongnan, Nehe, Heilongjia HZ8 Pinelliae pedatisectae rhizoma Anguo, Hebei HZ9 Pinelliae pedatisectae rhizoma Anguo, Hebei Open Access Article. Published on 16 April 2019. Downloaded 9/29/2021 6:04:14 PM. 3. Methods and results HZ10 Pinelliae pedatisectae rhizoma Anguo, Hebei HZ11 Pinelliae pedatisectae rhizoma Tongnan, Nehe, Heilongjia Aer a systematic study on the chemical components of Pine- HZ12 Pinelliae pedatisectae rhizoma Anguo, Hebei lliae rhizoma and Pinelliae pedatisectae rhizoma, we discovered HZ13 Pinelliae pedatisectae rhizoma Muba, Nanchong, Sichuan a characteristic peak of Pinelliae pedatisectae rhizoma in the HZ14 Pinelliae pedatisectae rhizoma Xihe, Longnan, Gansu HZ15 Pinelliae pedatisectae rhizoma Wenjiang, Chengdu, Sichuan HPLC chromatogram of organic acid part, distinguishable from HZ16 Pinelliae pedatisectae rhizoma Li, Longnan, Gansu that of Pinelliae rhizoma. 3.1 Extraction, separation