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[Molecular authentication of calcined oyster (Ostrea gigas) and its processed products]
Zhi-Yang Xiao1, Li Hu1, Yun-Jun Bai1
1State Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences Beijing 100700, China.
Abstract:
Calcined oyster is a commonly used shellfish traditional Chinese medicine in clinical practice in China. During the processing of oysters, their microscopic characteristics are destroyed, and open-fire calcination can damage the DNA of oysters, making it difficult to identify the primary source. The establishment of a specific polymerase chain reaction(PCR) method for the identification of calcined oysters can provide a guarantee for the safety and clinical efficacy of the medicine and its processed products. With Ostrea gigas as an example, the DNA extraction method of decoction pieces and formula particles of calcined oysters was improved, and high-quality DNA was obtained. Based on the specific single nucleotide polymorphism(SNP) sites of O. gigas and the other two species, the specific identification primers were designed, and the site-specific identification method of formula granules of calcined oyster(O. gigas) was established. The specificity and applicability of the method were investigated. The results showed that when the annealing temperature was 54 ℃, and the cycle was 44 times, the PCR amplified products of calcined oyster(O. gigas) and its formula granules produced a single bright identification band at 102 bp, while the other two species of oysters, O. talienwhanensis Crosse and O. rivularis Gould, had no band. In this study, DNA extraction and PCR identification of animal medicinal materials by calcination were established for the first time, which provided a tool for solving the difficult identification of calcined decoction pieces and ensuring drug safety.
Insights
A new polymerase chain reaction (PCR) method accurately identifies calcined oysters, ensuring the safety and efficacy of traditional Chinese medicine. This DNA-based technique overcomes challenges posed by traditional processing methods.
Area of Science:
- Pharmacology
- Molecular Biology
- Traditional Chinese Medicine
Background:
- Calcined oyster is a traditional Chinese medicine derived from shellfish.
- Traditional processing destroys microscopic oyster characteristics, hindering source identification.
- Calcination can damage oyster DNA, complicating authentication and potentially affecting medicinal efficacy.
Purpose of the Study:
- To develop a specific polymerase chain reaction (PCR) method for identifying calcined oysters.
- To ensure the safety and clinical efficacy of calcined oyster-based traditional Chinese medicines.
- To establish a reliable method for identifying the primary source of calcined oysters and their processed products.
Main Methods:
- Improved DNA extraction from decoction pieces and formula particles of calcined oysters.
- Designed species-specific primers based on single nucleotide polymorphism (SNP) sites.
- Established a site-specific identification method using PCR for calcined oyster formula granules (Ostrea gigas).
Main Results:
- High-quality DNA was successfully extracted from calcined oyster samples.
- The developed PCR method produced a distinct 102 bp band for Ostrea gigas at 54°C annealing temperature and 44 cycles.
- The method showed high specificity, with no amplification observed for O. talienwhanensis and O. rivularis.
Conclusions:
- A novel DNA extraction and PCR identification method for calcined animal medicinal materials was established.
- This technique provides a reliable tool for identifying calcined decoction pieces and formula granules.
- The study ensures drug safety and authenticity in traditional Chinese medicine applications.
