Abstract
Gushuling (GSL), a well-known hospital preparation composed of traditional Chinese medicine (TCM), has been widely used in the clinical treatment of osteoporosis (OP) for decades due to its remarkable therapeutic effect. However, the chemical constituents of GSL are still unclear so far, which limits the in-depth study of its pharmacodynamic material basis and further restricts its clinical application. In this study, we developed a strategy for qualitative analysis of the chemical constituents of GSL in vitro and in vivo. Based on the results of ultra-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS) and the UNIFI informatics platform, the chemical constituents of GSL can be determined quickly and effectively. By comparing the retention time, accurate mass, and fragmentation spectrum of the compounds in GSL, a total of 93 compounds were identified or preliminarily identified, including flavonoids, terpenoids, phenylpropanoids, steroids, etc. Among them, nine compounds have been confirmed by standard substances, namely epimedin A, epimedin B, epimedin C, icariin, ecdysterone, calycosin, calycosin-7-glucoside, ononin, and ginsenoside Ro. Fragment patterns and characteristic ions of representative compounds with different chemical structure types were analyzed. At the same time, 20 prototype compounds and 42 metabolites were detected in rat serum. Oxidation, hydration, reduction, dehydration, glutathione S-conjugation, and acetylcysteine conjugation were the main transformation reactions of GSL in rat serum. In this research, the rapid method to characterize the in vitro and in vivo chemical constituents of GSL can not only be used for the standardization and quality control of GSL but also be helpful for further research on its pharmacodynamic material basis.
Highlights
Osteoporosis (OP) is a complex bone disease characterized by low bone density and impaired microstructure of bone tissue, leading to increased bone fragility and being prone to fractures [1]
The importance of traditional Chinese medicine (TCM) and Chinese herbal compound prescription that have a therapeutic effect on OP were gradually been recognized because of their safety and effectiveness [7,8,9]
High-resolution mass spectrometry data of GSL were rapidly acquired by the UPLC-Q-TOF-MS method. e total ion chromatography (TIC) of GSL in positive and negative ion modes is portrayed in Figure 1. e UNIFI screening platform was used to process and analyze its mass spectrometry data, and the data were spontaneously matched with the fragment information
Summary
Osteoporosis (OP) is a complex bone disease characterized by low bone density and impaired microstructure of bone tissue, leading to increased bone fragility and being prone to fractures [1]. The treatment drugs for OP mainly include ingredients that inhibit bone resorption, such as bisphosphonates, calcitonin, and estrogen, and ingredients that promote bone formation, such as parathyroid hormone [2]. Taking bisphosphonates for a long time will increase the incidence of complications such as osteonecrosis of the jaw [3,4]. Long-term use of estrogen will increase the incidence of breast cancer, endometrial cancer, and other diseases [5,6]. The importance of traditional Chinese medicine (TCM) and Chinese herbal compound prescription that have a therapeutic effect on OP were gradually been recognized because of their safety and effectiveness [7,8,9].
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