Abstract
As one of the high-incidence diseases in the world, pharyngitis seriously affects the lives of those with the condition. Qingguo Wan is a herbal medicine used for treating pharyngitis, and its quality evaluation is currently only accomplished via traditional identification. However, precise identification becomes challenging with fake products on the market or fungal contamination during the production process. This study used the Illumina NovaSeq platform for targeting the ITS2, psbA-trnH, matK, and rbcL sequences to survey the species composition of lab-made and commercial samples. The results showed that a total of 34.56 Gb of raw data that was obtained represented more than 0.23 billion reads. After assembly, annotation, and operational taxonomic unit clustering, 103, 12, 10, and 12 OTUs were obtained, which belonged to the ITS2, psbA-trnH, matK, and rbcL sequences of the mock lab-made and commercial samples. The analytical results indicated that the sequences of all the prescription ingredients were successfully obtained in the two lab-made samples. The positive control medicinal Panax quinquefolius L. sequence was obtained in HSZY175, while Scutellaria baicalensis Georgi, Lonicera japonica Thunb. Menispermum dauricum DC. and Paeonia lactiflora Pall. were detected in the three commercial samples. The detection results of the other four herbs in different fragments were not all the same. In addition, a total of 28 fungi OTUs, representing 19 families and 20 genera, were obtained from both the commercial and mock lab-made samples. Aspergillus, Cladosporium, and Penicillium dominated among the 20 genera. This study demonstrated that the shotgun metabarcoding method is a powerful tool for the molecular identification of the biological ingredients in Qingguo Wan. It can be used to effectively supplement traditional methods while providing a new technique for the quality evaluation of Qingguo Wan.
Highlights
Pharyngitis is caused by the direct infection of the pharynx and is one of the most common diseases in the world
In 2021, a new approaches combining DNA barcoding and shotgun sequencing was employed for the species identification of Wuhu San and the results showed that this method was potential to effectively complement traditional identification methods (Liu et al, 2021)
The total DNA of lab-made and commercial samples were sequenced according to a shotgun metabarcoding strategy, and a total of 34.56 Gb of raw data, including more than 230 million paired-end reads, were obtained, which was an average of 6.91 Gb of raw data for each sample
Summary
Pharyngitis is caused by the direct infection of the pharynx and is one of the most common diseases in the world. Qinguo Wan is made from eight kinds of medicinal materials, namely Canarii Fructus (Qingguo), Lonicerae Japonicae Flos (Jinyinhua), Scutellariae Radix (Huangqin), Menispermi Rhizoma (Beidougen), Ophiopogonis Radix (Maidong), Scrophulariae Radix (Xuanshen), Paeoniae Radix Alba (Baishao), and Platycodonis Radix (Jiegeng). It eliminates pharyngeal pain, reducing swelling and relieving pain (Commission, 2020). Canarii Fructus represents the primary curative ingredient in relieving throat pain, while displaying various pharmacological activities that include antibacterial and anti-inflammatory properties It has an excellent therapeutic effect on acute pharyngitis (Geng et al, 2011; Jia et al, 2016). The remaining five medicinal herbs can increase the efficacy according to the monarch and the minister of traditional Chinese medicine
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