OBJECTIVETo establish a HPLC method for the analysis of Danshensu (DSS), protocatechuic acid (PA) and protocatechuic aldehyde (PAL) of Xiangdan injection in rat's plasma, and to study pharmacokinetic characteristics of Xiangdan injection components in rats with m-hydroxybenzoic acid as internal standard.

METHODprotein was precipitated by 10% trichloroacetic acid and extracted by ethyl acetate. The plasma concentration was detected by HPLC, The pharmacokinetics parameters of DSS, PA and PAL were calculated by DAS2.0 software after iv injection.

RESULTDSS, PA and PAL have a good linear relationship in 0.68-44.0 mg x L(-1), 0.43-14.0 mg x L(-1) and 0.38-12.0 mg x L(-1), respectively. The average recoveries were more than 92% and the RSD of precision and stability of the test were between 0.4%-4.8%. DSS, PA and PAL showed a two-compartment open model, the half-life of absorption (t(1/2alpha)) were (6.26 +/- 4.6), (5.93 +/- 4.9), (18.44 +/- 2.4) min, the half-life of elimination (t(1/2beta)) were (64.11 +/- 8.8), (63.28 +/- 0.13), (69.315 +/- 0) min, the area under curve(AUC(0-infinity)) were (852.98 +/- 175.6), (83.84 +/- 58.8), (147.79 +/- 12.3) mg x min(-1) x L(-1).

CONCLUSIONA method with high recovery rate and good stability was established to determine the blood concentration of DSS, PA, PAL in Xiangdan injection and applied in its pharmacokinetics successfully.

Animals ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Female ; Hydroxybenzoates ; blood ; pharmacokinetics ; Injections ; Limit of Detection ; Linear Models ; Male ; Rats ; Rats, Sprague-Dawley

AIM: To investigate the application of biological amniotic membrane soaked in pirfenidone(PFD)and to evaluate its anti-scarring effect and toxic side effects on glaucoma model of rabbit eyes.METHODS: The right eyes of 72 healthy New Zealand white rabbits were randomly divided into 0.5%PFD+ biological amniotic membrane group, biological amniotic membrane group, mitomycin C(MMC)group and blank control group after the glaucoma model was established by anterior chamber injection of compound carbomer solution, and 18 rabbits in each group underwent trabeculectomy, in which the 0.5% PFD+ biological amniotic membrane group was placed with 0.5% PFD solution-soaked biological amniotic membrane under the scleral flap, and the biological amniotic membrane group was placed with normal saline-soaked rehydrated biological amniotic membrane under the scleral flap. In the MMC group, a cotton pad soaked in MMC was placed under the scleral flap for 3 min and immediately rinsed with normal saline, while the blank control group received no implant after the scleral flap was made. The intraocular pressure(IOP), filtration blebs, toxic side effects and complications were evaluated, and the histopathological changes in the filtration area were observed by hematoxylin-eosin(HE), Masson staining and immunohistochemical staining.RESULTS: The mean IOP at 14, 21 and 28 d after trabeculectomy were 0.5%PFD+ biological amniotic membrane group<MMC group<biological amniotic membrane group<blank control group(all P<0.05). At 28 d after trabeculectomy, 0.5%PFD+ biological amniotic membrane group had the best effect of anti-inflammatory hyperplasia and inhibition of collagen formation, the highest survival rate of filtration blebs, and the inflammatory reaction was mild.CONCLUSION: Biological amniotic membrane soaked in pirfenidone has more obvious anti-scarring effect on glaucoma model, with less toxic side effects and good safety.