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Objective: To establish a method for rapid screening and identifying antioxidants in natural products by ultra high performance liquid chromatography-(1,1-dipheny l-2-trinitrophenylhydrazine)-photo diode array-electrospray-time-of-flight mass spectrometry (UPLC-DPPH-PDA-ESI-TOF/MS) and apply it to the rapid discovery of antioxidants from Salviamiltiorrhiza extracts. Methods: The Waters ACQUITY UPLC system and a Waters ACQUITY UPLC HSS T3 column (100 mm×3.0mm,1.8 μm; Waters Technologies, USA) were used to elute the sample at a low flow rate of 0.35 mL/min, which could achieve the rapid separation of S. miltiorrhiza extracts. The elution was joined with DPPH solution (0.3 mL/min) and reacted in the PEEK coil, and then flew into the detector. The anti-oxidation components in S. miltiorrhiza extracts were quickly screened by comparing the chromatograms at 517 nm and 254 nm. Results: The UPLC technology was adopted to realize the efficient separation of samples at low volume flow rate, which avoided the baseline noise caused by high volume flow rate, significantly improved the resolution and sensitivity of the online screening method, and reduced the solvent consumption.The results showed that 31 antioxidant active compounds were screened from S. miltiorrhiza extracts, and 11 of them were identified by PAD-ESI-TOF/MS. Activity verification experiments showed that caffeic acid, salvianolic acid B, salvianolic acid A and danshensu had better antioxidant activity. The IC50 value ofcaffeic acid was 29.00 μmol/L, which was significantly better than the positive control (vitamin C). Conclusion: The method is rapid and efficient, which provides methods and technical support for the rapid discovery of antioxidant components from complex natural products.
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[ABSTRACT]AIM:Toobservethelong-termeffectsoftaurine,Danxuetong(Salviamiltiorrhizacombinedwith Xueshuantong ) and breviscapine on contralateral testis with unilateral testicular torsion in prepubertal rats , and to compare their protective effect on testis with ischemia reperfusion injury .METHODS: Four-week-old healthy male SD rats ( n=64) were randomly divided into 8 groups:sham operation group, saline group, single-dose and successive taurine groups, single-dose and successive Danxuetong groups , and single-dose and successive breviscapine groups .The animal model of left testicular torsion reduction (720°for 2 h) was established.After 6 weeks of surgery, the right testes and epididymides from the rats were collected immediately to detect the total antioxidant capacity (T-AOC), the activity of superoxide dis-mutase (SOD), the activity of nitric oxide synthase (NOS) and the content of malondialdehyde (MDA).The sperm de-formity, sperm viability and sperm concentration were measured , and the pathological examination of the testicular tissues was performed.RESULTS:Compared with saline group, the activity of SOD, T-AOC, sperm viability and sperm concen-tration were increased , while MDA content and sperm deformity were decreased in 3 successive groups .SOD activity and sperm viability ( except single-dose breviscapine group ) were increased , but MDA content and sperm deformity were de-creased in 3 single-dose groups .T-AOC activity and sperm concentration were increased in single-dose taurine group .The saline group showed interstitial edema and degeneration of seminiferous tubules , and the rest groups showed a significant improvement in histological morphology .CONCLUSION:The contralateral testis in prepubertal rats was damaged after u-nilateral testicular torsion , and taurine , Danxuetong and breviscapine have long-term protective effect on the tissues under ischemia-reperfusion injury .The protective effect of Danxuetong is better than breviscapine , and taurine is the best of them.The effect of single-dose group is better than the successive application .
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Objiective To evaluate the histocompatibility of gelatin-bletilla/Salvia-miltiorrhiza i material and to discuss the appropriate concentration of salvia-miltiorrhiza in the material. Method Salvia-miltiorrhiza together with gelatin and bletilla was used to make a kind of porous membranes. A total of 175 rats were divided randomly (random number) into 5 groups ( n = 35 in each). The porous membranes containing salvia-miltiorrhiza material in different concentrations of 1, 2, 4, and 5 mL/100 g were implanted subcutaneously into the rats of the four experimental groups, respectively. The gelatin-bletilla material without salvia-miltiorrhiza was implanted into the rats of control group. In 1, 3, 7, 14, 21, 28 and 56 days after implant, the materials were taken out of the bodies of the rats, separately. The material implanted for 1, 3, 7 and 14 days were observed with immunohistoche-mistry. Results In the area of implant, no noticeable inflammation or other was found. The implanted materials were degraded and entirely absorbed in vivo in 8 weeks. The salvia-miltiorrhiza in concentration of 2 mL/100 g in vivo promoted the angiogenesis most significantly. Conclusions The gelatin-bletilla/salvia-miltiorrhiza material has excellent histocompatibility and the appropriate concentration of salvia-miltiorrhiza in material is 2 mL/100 g.