Objective To establish the method for simultaneous determination of epimedium glycoside and other 4 kinds of active ingredients in Xianling Gubao Capsules by three wavelength switching method. Methods An Waters Atlantis T3 C18 column (4.6 mm × 250 mm, 5 μm) was used with the mixture of acetonitrile-0.05％ formic acid solution as the mobile phase in gradient elution (0–5 min, 12％–20％ A; 5–15 min, 20％–55％ A; 15–35 min, 55％ A;35–55 min, 55％–76％ A; 55.1 min, 12％ A; 55.1–60 min, 12％ A). Detection wavelength was as follow: 0–30 min, 212 nm; 30–42 min, 246 nm; 42–60 min, 270 nm. The flow rate was 1.0 mL/min. The column temperature was 30 ℃. Results The calibration curves of asperosaponin Ⅵ, psoralen, angelicin, epimedin C, and icariin were in good linearity among the ranges of 101.6–3048 ng, 8.7–261 ng, 7.9–237 ng, 117.2–3516 ng, 78–2340 ng, respectively. In the instrument precision test, stability test, repeatability test, the RSD was less than 3％. The average recoveries were 99.83％, 100.35％, 100.59％, 100.60％, 99.72％, respectively, and all the RSD were less than 3％. Conclusion The method is sensitive, accurate, and separation effect is good, which can provide a basis for quality evaluation standard of Xianling Gubao Capsules.

Objective To establish a multi-component quantitative control method for Shenhuang Yangyin Capsules;To explore the application of chemical pattern recognition and grey relational analysis(GRA)model in the quality evaluation of Shenhuang Yangyin Capsules.Methods The contents of ginsenoside Rg1,ginsenoside Rb1,ginsenoside Rb3,ginsenoside Rd,calycosin 7-O-β-D-glucopyranoside,ononin,calycosin,medicarpin,sodium danshensu,salvianolic acid B,tanshinone ⅡA,methylophiopogonanone A and methylophiopogonanone B in Shenhuang Yangyin Capsules were determined by HPLC.Agilent Eclipse Plus C18 column was adopted with acetonitrile as mobile phase A and 0.5％phosphate acid solution as mobile phase B,gradient elution.The detection wavelengths were 203,280 and 296 nm.Through chemical pattern recognition analysis and GRA,the results of multi-component content were analyzed,and the comprehensive quality evaluation system of Shenhuang Yangyin Capsules was constructed.Results The results of methodological verification of external standard method met the requirements.Chemometric analysis showed that salvianolic acid B,ononin,tanshinone ⅡA,calycosin,ginsenoside Rb3 and ginsenoside Rb1 were the main factors affecting the quality of Shenhuang Yangyin Capsules.The relative correlation degree of GRA were 0.319 8-0.642 1,and it was helpful for enterprises to analyze the differences between products.Conclusion Multi-component quantitative combined chemical mode recognition and GRA can be used for quality evaluation of Shenhuang Yangyin Capsules.

OBJECTIVE: To assess the value of Ploton silver staining and phalloidin-iFlour 488 staining in observation of the morphology of osteocyte dendrites of mice at different developmental stages. METHODS: The humerus and femurs were harvested from mice at 0 (P0), 5 (P5), 15 (P15), 21 (P21), 28 (P28), and 35 days (P35) after birth to prepare cryo-sections and paraffin sections. HE staining of P35 mouse femur sections served as a reference for observing osteocytes in the trabecular bone and cortical bone. The humeral sections at different developmental stages were stained with Ploton silver staining to observe the morphology of osteocytes and canaliculi, and the canalicular lengths in the cortical and trabecular bones of the humerus of the mice in each developmental stage were recorded. The cryo-sections of the humerus from P10 and P15 mice were stained with phalloidin iFlour-488 to observe the morphology of osteocytes and measurement of the length of osteocyte dendrites in the cortical bone. RESULTS: In the trabecular bone of the humerus of P0-P15 mice, Ploton silver staining only visualized the outline of the osteocytes, and the morphology of the canaliculi was poorly defined. In P21 or older mice, Ploton silver staining revealed the morphology of the trabecular bone osteocytes and the canaliculi, which were neatly arranged and whose lengths increased significantly with age (P21 CONCLUSIONS Mouse osteocyte dendrites elongate progressively and their arrangement gradually becomes regular with age. Ploton silver staining can clearly visualize the morphology of the osteocytes and the canaliculi in adult mice but not in mice in early stages of development. Phalloidin iFlour-488 staining for labeling the cytoskeleton can be applied for mouse osteocytes at all developmental stages and allows morphological observation of mouse osteocytes in early developmental stages.
Animals ; Bone and Bones ; Dendrites ; Mice ; Osteocytes ; Phalloidine ; Silver Staining