Abstract
Traditional Chinese Medicine (TCM) preparations have been used in China for thousands of years. Quality evaluation for TCM preparations could be conducted based on chemical ingredients or biological ingredients. To date, the overwhelming majority of researches have focused on chemical ingredients while few studies were reported for biological ingredients. It is only recently that the assessments based on biological ingredients have drawn broader attentions. In this work, we have established a method for quality evaluation of TCM preparations by combination of chemical ingredients determined by HPLC fingerprint and biological ingredients obtained by high-throughput sequencing. This proof-of-concept method has been evaluated and compared with existing methods on Liuwei Dihuang Wan, a classical TCM preparation in China. By comparison of this method with those only based on chemical or biological ingredients, it is suggested that (1) Biological ingredient could complement chemical ingredient in separating TCM preparation from different manufacturers and batches with high accuracy; (2) classification of samples based on selected features would always out-perform those based on all features (either chemical or biological or both). By rationally selecting representative biological and chemical features, we have proven that these two types of features could complement each other for the assessment of ingredient consistencies and differences among various TCM samples, which is helpful to ensure the effectiveness, safety and legality of TCM preparations.
Highlights
Traditional Chinese medicine (TCM) has been used in clinical applications for thousands of years in China, and it has drawn increasing attentions around the world recently due to its proven therapeutic effects with little side effects
Www.nature.com/scientificreports compounds of interests, yet could not assess biological ingredients including prescribed species and unprescribed species that were present in Traditional Chinese Medicine (TCM) preparations
We have established a novel method (M-TCM) for biological ingredient analysis of a classical TCM preparation, Liuwei Dihuang Wan (LDW), based on metagenomic approaches via high-throughput sequencing (HTS), in which the ribosomal internal transcribed spacer 2 (ITS2) and the chloroplast genome trnL (UAA) intron were chosen as biomarkers[16,17]
Summary
Traditional Chinese medicine (TCM) has been used in clinical applications for thousands of years in China, and it has drawn increasing attentions around the world recently due to its proven therapeutic effects with little side effects. Www.nature.com/scientificreports compounds of interests, yet could not assess biological ingredients including prescribed species (species listed on the package) and unprescribed species (species not listed on the package) that were present in TCM preparations. The biological ingredient analysis methods for TCM preparations based on molecular diagnostic assay are widely used currently[10,11,12], which have broken though the limitation of difficulties in discrimination of closely related species with similar morphology or containing the same chemical constituents. The metagenomic approach[13,14,15] has been considered as one of the most effective methods for multiple species analysis from a biological mixture, which would be helpful for the analysis of biological ingredients in TCM preparations. M-TCM could assess the prescribed species and unprescribed species of LDW simultaneously, this method is still limited due to the nature of only using identifiable biological ingredients (that can be sequenced from TCM preparations) as features for differentiating TCM preparations
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