Quality evaluation of Galli Gigerii Endothelium Corneum based on HPLC fingerprints and content determination of nucleosides / 中国中药杂志

Galli Gigerii Endothelium Corneum(GGEC), the dried gizzard membrane of Gallus gallus domesticus is a Chinese medicinal material commonly used for digestion. However, due to the particularity of texture and composition, its active ingre-dients have not been clarified so far, and there is also a lack of quality evaluation indicators. In this study, UPLC-Q-TOF-MS was used to analyze the chemical components from the water extract of GGEC, and ten nucleosides were identified for the first time. HPLC fingerprints of the water extracts of GGEC were established and the content of seven nucleosides was determined. The fingerprint similarities of 40 batches of GGEC samples ranged from 0.765 to 0.959, indicating that there were great differences among the GGEC products processed with different methods. In addition, SPSS 22.0 and SIMCA 14.1 were used for hierarchical cluster analysis(HCA) and principal component analysis(PCA) on the 19 common peaks of the HPLC fingerprints of GGEC, and the 40 batches of samples were divided into three categories: raw GGEC, fried GGEC and vinegar-processed GGEC. Eight differential components in GGEC were marked by orthogonal partial least squares discrimination analysis(OPLS-DA), two of which were adenine and thymine. The results of content determination showed that the total content of the seven nucleosides in raw GGEC, fried GGEC and vinegar-processed GGEC were 182.5-416.8, 205.3-368.7, and 194.2-283.0 μg·g~(-1), respectively. There were significant differences in the content of hypoxanthine, thymine and thymidine among the GGEC products processed with different methods(P<0.05), which were graded in the order of fried GGEC>vinegar-processed GGEC>raw GGEC. This suggested that the content of hypoxanthine, thymine and thymidine tended to increase during the frying process, and the variation range might be related to the degree of heat exposure. The established methods in this study were simple and reproducible, and could be used for qualitative and quantitative analysis of GGEC and its processed pro-ducts. This study also provided reference for the establishment of quality standards of GGEC with chemical components as control index.

Development of a novel and practical bioassay for the quality evaluation and control of Galli Gigerii Endothelium Corneum based on the detection of digestive enzyme activity / 药学学报

Galli Gigerii Endothelium Corneum (GGEC) represents digestion-promoting medicines with measurable effects and extensive clinical application. However, its effective components are not clear. The quality control index in the current edition of Chinese Pharmacopoeia is rather elementary and does not reflect its clinical efficacy. In this study, a bioassay method based on pepsin activity was proposed as a novel quality control method. With pepsin activity as the evaluation index, the extraction of GGEC was optimized and a method for the determination of biological potency was established by using the qualitative reaction parallel line method. The biological potency and consistency of 20 batches of GGEC were investigated. To provide scientific evidence in support of this bioassay method, two validation experiments were designed. One was to study the viscosity-reducing activity of a nutritional semi-solid paste after adding GGEC samples with differing potency. The other was to correlate the gastric residual rate in mice and pepsin activity with the alcohol soluble extract content. The results showed that the optimal preparation method was to dilute crude powder of GGEC with 50 volumes of water and subject to ultrasonic extraction at 300 W and 40 kHz for 0.5 h. The shape of the dose-response curve was similar to that of the positive control drug multienzyme tablets and the precision, intermediate precision and repeatability met the methodology requirements. The results showed that the potency of 20 batches of samples ranged from 13.49 to 34.69 U·mg-1, with an average value of 22.21 U·mg-1. The validation experiment demonstrated that the effect of reducing the viscosity of the nutrient paste became more significant as GGEC sample potency increased. The correlation coefficient R of gastric residual rate with pepsin potency and alcohol soluble extract content was 0.867 and 0.518, respectively, which indicated that the pepsin potency was highly correlated with in vivo activity. This study shows that a bioassay method based on pepsin activity is reliable and reproducible for GGEC and could provide reference method for the quality evaluation of other digestant herbs.

Quality Evaluation of Galli Gigerii Endothelium Corneum by Allele-specific PCR Method / 中国实验方剂学杂志

Objective: To screen the specific reverse primers of Galli Gigerii Endothelium Corneum,duck gizzard membrane and goose gizzard membrane,and establish a specific PCR for molecular identifying Galli Gigerii Endothelium Corneum and its common adulterants. Method: Based on the mutation sites on the 12S rRNA sequence,specific polymerase Chain reaction(PCR) identify primers were designed for chicken,duck and goose gizzard membrane. The specific PCR reaction conditions were optimized,and the PCR identification method was explored and verified in terms of tolerance and feasibility. Thirty batches of Galli Gigerii Endothelium Corneum decoction pieces extracted from the test were identified. Result: Thirty batches of Galli Gigerii Endothelium Corneum decoction pieces were detected using chicken-specific primers, 273 bp of specific bands was amplified and visualized on the agarose electrophoregram. When duck and goose primers were used,no corresponding amplified band was detected. Conclusion: The allele-specific PCR method can be used as a rapid and accurate method to identify Galli Gigerii Endothelium Corneum. It is a promise method for special sampling tasks of Chinese herbal medicine and decoction tablets nationwide.

The Overview of Galli gigerii endothelium Corneum, an Edible Traditional Chinese Medicine / 中国药学杂志

Galli gigerii endothelium corneum is collected from the dried inner wall of the stomach of Gallus gallus domesticus Brisson and it is used for a long history as an edible traditional Chinese medicine. Galli gigerii endothelium corneum contains rich nutrient components such as proteins, polysaccharides, animo acids, and trace elements, and displayed various bioactivities including anti-oxidative effect, improving the level of blood glucose and lipid, as well as hemorheological parameters and gastrointestinal function. Based on the published literatures in the recent years, the advance in the research on the traditional use, chemical constituents, pharmacological activities of Galli gigerii endothelium corneum was summarized, which can provide the scientific basis for the further research and development of Galli gigerii endothelium corneum.