Abstract
Traditional Chinese medicine (TCM) has been practiced for thousands of years, but only within the last few decades has its use become more widespread outside of Asia. Concerns continue to be raised about the efficacy, legality, and safety of many popular complementary alternative medicines, including TCMs. Ingredients of some TCMs are known to include derivatives of endangered, trade-restricted species of plants and animals, and therefore contravene the Convention on International Trade in Endangered Species (CITES) legislation. Chromatographic studies have detected the presence of heavy metals and plant toxins within some TCMs, and there are numerous cases of adverse reactions. It is in the interests of both biodiversity conservation and public safety that techniques are developed to screen medicinals like TCMs. Targeting both the p-loop region of the plastid trnL gene and the mitochondrial 16S ribosomal RNA gene, over 49,000 amplicon sequence reads were generated from 15 TCM samples presented in the form of powders, tablets, capsules, bile flakes, and herbal teas. Here we show that second-generation, high-throughput sequencing (HTS) of DNA represents an effective means to genetically audit organic ingredients within complex TCMs. Comparison of DNA sequence data to reference databases revealed the presence of 68 different plant families and included genera, such as Ephedra and Asarum, that are potentially toxic. Similarly, animal families were identified that include genera that are classified as vulnerable, endangered, or critically endangered, including Asiatic black bear (Ursus thibetanus) and Saiga antelope (Saiga tatarica). Bovidae, Cervidae, and Bufonidae DNA were also detected in many of the TCM samples and were rarely declared on the product packaging. This study demonstrates that deep sequencing via HTS is an efficient and cost-effective way to audit highly processed TCM products and will assist in monitoring their legality and safety especially when plant reference databases become better established.
Highlights
Traditional Chinese medicines (TCMs) have been an integral part of Chinese culture and the primary medicinal treatment for a large portion of the population for more than 3000 years [1,2]
The increasing popularity of TCM products has seen the monetary value of the industry increase to hundreds of millions of dollars per annum [5], its growth paralleled by the global increase in the use of complementary and alternative medicines
Some of the most common plant genera identified in the TCM samples were Glycyrrhiza, found in 62% of samples, Mentha, found in 46% of samples and Asarum found in 31% of samples
Summary
Traditional Chinese medicines (TCMs) have been an integral part of Chinese culture and the primary medicinal treatment for a large portion of the population for more than 3000 years [1,2]. The increasing popularity of TCM products has seen the monetary value of the industry increase to hundreds of millions of dollars per annum [5], its growth paralleled by the global increase in the use of complementary and alternative medicines. Undeclared or misidentified TCM ingredients and adulterants can pose serious health risks to consumers [10,12,13]. These include: allergenic substances [14], plant toxins [7], heavy metals such as mercury, lead, copper and arsenic [15], and pharmaceutically active compounds of undetermined concentration [5]. In the early 1990s the misidentification of the toxic herb Aristolochia fangchi for the anti-inflammatory agent Stephania tetrandra led more than a hundred women to suffer kidney failure, with many later developing cancer of the urinary system [13]
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