Novel method for screening traditional Chinese medicine anti-osteoporosis active component with zebra fish osteoporosis model
A technology for osteoporosis and application methods, which is applied in the analysis of materials, material analysis by optical means, and material inspection products, etc., can solve the problems of large sample consumption, high labor intensity and high cost, and achieves less compound consumption and labor. Strong and efficient separation effect
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Embodiment 1
[0054] Anti-prednisolone-induced zebrafish osteoporosis activity evaluation of component groups captured by high performance liquid chromatography with different retention time periods within 0 to 80 min.
[0055] 1. Materials and instruments
[0056] Drug under test: Dipsacia
[0057] Experimental animals: adult zebrafish (Institute of Model Animals, Nanjing University).
[0058] Reagents: Prednisolone (Suzhou Yake Chemical Reagent Co., Ltd., batch number: YK2012020101); Alizarin Red S (Zhengzhou Siji Chemical Products Co., Ltd., batch number: Sj20110806); Paraformaldehyde (Chengdu Kelong Chemical Reagent Factory, Lot: 20100504); MS-222 (Acros Organics, Lot: A0288328).
[0059] Instruments: ZEISS fluorescence inverted microscope Axioobserver.A1 (Zeiss Optical Instrument International Trade Co., Ltd.); biochemical incubator SPX-80 (Ningbo Haishu Saifu Experimental Instrument Factory); Agilent1100 series high performance liquid chromatography (Agilent Corporation of the Unite...
Embodiment 2
[0087] Anti-prednisolone-induced zebrafish osteoporosis activity evaluation of different components or component groups captured by HPLC within 27-46 min.
[0088] 1. Materials and instruments
[0089] Tested drug: components within 27-46 minutes of dicontinuation
[0090] Experimental animals and instruments are the same as in Example 1.
[0091] 2. Experimental method
[0092] 2.1 Method
[0093] Dipsacus sample preparation: high performance liquid chromatography condition is the same as embodiment one " 2.1 method ", by 27min-46min (such as Figure 5 Shown) the Dipsacus component is further separated and captured by high performance liquid phase. Inject 50 μL of Dipsacus extract (0.1g crude drug / mL) from Example 1 for high-performance liquid chromatography separation, capture the component peaks of Dipsacus at 27-30min, 30-35min, 35-40min, 40-43min and 43-46min respectively , which are denoted as S 1 , S 2 , S 3 , S 4 and S 5 , where S 5 The reference substance w...
Embodiment 3
[0114] Anti-prednisolone-induced zebrafish osteoporosis activity evaluation of different components or component groups captured by HPLC within 24-60 min.
[0115] 1 Materials and Instruments
[0116] Test drug: components within 24-60 minutes of dicontinuation
[0117] Experimental animals and instruments are the same as in Example 1.
[0118] 2 Experimental methods
[0119] 2.1 Method
[0120] Dipsacus sample preparation: high performance liquid chromatography condition is with embodiment one " 2.1 method ", by 24-60min (such as Figure 12 Shown) the Dipsacus component is further separated and captured by high performance liquid phase. Inject 50 μL of the Dipsacaceae extract (0.1g crude drug / mL) in Example 1 for high-performance liquid chromatography separation, and capture the component peaks of Dipsacaceae at 24-40min and 40-60min respectively. According to our previous research (Wei Yingjie, Jia Xiaobin, Fan Hongwei, Peng Yunru, Zhan Wen. China Journal of Traditional...
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