RESUMO
Objective::Because traditional methods are difficult to identify the fermentation mycelium, DNA barcoding technology was used to quickly identify the raw material strain Paecilomyces hepiali of Jinshuibao capsules and related products. Method::A total of 168 samples of 8 species of P. hepiali and its confusable species were identified by internal transcribed spacer (ITS) sequences, and based on the ITS sequences, P. hepiali specific primers were designed to quickly identify the related products. Result::The length of ITS sequences in 44 P. hepiali samples from different sources was 499 bp and there was no mutation site. It was shown that P. hepiali could be distinguished from 7 confusable species based on ITS sequences. The specific primer (ITS-BF/ITS-BR) of P. hepiali designed by ITS sequences could be amplified to obtain a short fragment of 102 bp in length, which could be used to rapidly identify P. hepiali from other confusable species, and to distinguish relevant products in the market. Conclusion::The rapid identification of P. hepiali and its related products can be achieved through the ITS sequences and specific primers, which provides a reference for the production and quality control of Jinshuibao capsules.
RESUMO
Objective::To improve the quality control system in the production of Jinshuibao capsules, and to provide experimental basis for the follow-up research and application of this preparation. Method::High performance liquid chromatography (HPLC) was employed, the analysis was performed on a Ultimate AQ-C18 column (4.6 mm×150 mm, 5 μm). The chromatographic conditions for the determination of adenosine, guanosine and uridine were as following: mobile phase of methanol-0.1%formic acid aqueous solution for gradient elution, the flow rate of 0.4 mL·min-1, column temperature at 30 ℃, sample quantity of 10 μL, detection wavelength at 260 nm. The chromatographic conditions for the determination of ergosterol were as follows: mobile phase of methanol-water (98∶2), the flow rate of 1 mL·min-1, column temperature at 25 ℃, sample quantity of 10 μL, detection wavelength at 283 nm. Result::The main chromatographic peaks of fermented Cordyceps powder samples in different production stages showed little difference. The linear relationships of adenosine, guanosine and uridine were good (R2 >0.999), their recoveries were 106.06%, 101.25%and 105.88%, respectively. The contents of adenosine and ergosterol in 20 batches of samples extracted from 2016 to 2018 were in line with the requirements in the 2015 edition of Chinese Pharmacopoeia, the contents of guanosine and uridine were 0.97-1.36, 0.67-1.38 mg/capsule, respectively. Conclusion::The quality of Jinshuibao capsules in the market is stable. This method can be used to detect the quality of Jinshuibao capsules, and it is simple, stable and reliable.