ABSTRACT
Objective To establish a method for the simultaneous content determination ofquercetin,kaempferol and pinocembrin in Penthorum Chinense Pursh.Methods The HPLC analysis was carried out on Alltima C18 (250 mm × 4.6 mm,5 μm) with a mixture of methanol-0.2％ phosphate (45:55) as the mobile phase.The determination wavelength was set at 270 nm with the flow rate of 1.0 ml/min.The column temperature was set at 30 ℃.Results The quercetin,kaempferol and pinocembrin showed good linearity in the range of 0.316-10.120 μg (r=0.999 9),0.012-0.397 μg (r=0.999 6) and 0.063-2.016 μg (r=0.999 8) respectively.The average recovery rates of quercetin,kaempferol and pinocembrin were 99.34％ (RSD=1.51％),99.76％ (RSD=l.96％) and 98.34％ (RSD=1.56％) respectively.Conclusions The method is accurate and sensitive,which can be used for the content determination of quercetin,kaempferol and pinocembrin in Penthorum Chinense Pursh.
ABSTRACT
Objective To study the protein binding rates ofgardenioside,paeoniflorin and naringin in Weiyanling granules in bovine serum albumin,rat plasma and human plasma.Methods The equilibrium dialysis method was carried to determine the binding-rates of three components in Weiyanling granules in bovine serum albumin,rat plasma and human plasma.The HPLC method was used to determine the mass concentration of three components in weiyanling granules in inner and outer dialysis and to calculate the plasma protein-binding rates.Results The linear relationship,precision,extraction recovery and stability of gardenoside,paeoniflorin and naringin all conformed to the methodological requirements.Within the low,middle and high concentration (5.084,10.04 and 20.08 g/L) of Weiyanling granules,the protein-binding rates of gardenioside and bovine serum albumin were 23.77％ ± 13.39％,10.75％ ± 1.34％,5.82％ ± 6.53％,respectively;the protein-binding rates of paeoniflorin were 51.81％ ± 6.53％,30.12％ ± 6.61％,21.39％ ± 9.45％,respectively;the protein-binding rates of naringin were 70.21％ ± 3.31％,67.61％ ± 1.31％,56.00％ ± 3.46％,respectively;the protein-binding rates of gardenioside and rat plasma were 71.56％ ± 3.00 ％,43.56％ ± 12.22％,40.63％ ± 4.73％,respectively;the protein-binding rates ofpaeoniflorin and rat plasma were 55.72％ ± 1.60％,47.59％ ± 6.33％,40.07％ ± 0.72％,respectively;the protein-binding rates of naringin and rat plasma were 85.85％ ± 3.53％,85.38％ ± 0.99％,79.99％ ± 2.57％,respectively;the protein-binding rates of gardenioside and human plasma were 13.56％ ± 2.40％,3.02％ ± 2.41％,5.82％ ± 2.08％,respectively;the protein-binding rates of paeoniflorin and human plasma were 9.49％ ± 5.23％,5.01％ ± 1.10％,3.98％ ± 1.54％,respectively;the protein-binding rates of naringin and human plasma were 25.04％ ± 2.61％,26.09％ ± 13.82％,20.20％ ± 2.24％,respectively.Conclusions Within 5.084,10.04 and 20.08 g/L,the protein binding-rate of gardenioside,paeoniflorin and naringin were shown asfollow,rat plasma ＞bovine serum albumin ＞ human plasma.There were significant differences in species.
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OBJECTIVE:To establish a method for simultaneous determination of hyperoside,quercitrin,luteoloside, kaempferol,quercetin,rutin,luteolin and isorhamnetin in total flavanones of Sedum sarmentosum Bunge. METHODS:UPLC-MS/MS method was adopted. The determination was performed on ZOBAX SB C18column with mobile phase consisted of methanol-5 mmol/L ammonium formate aqueous solution(45:55,V/V)at the flow rate of 0.4 mL/min. The column temperature was 30 ℃, and sample size was 2 μL. The electrospray ionization source(ESI)was used;ion source temperature was 400 ℃;desolvation temperature was 300 ℃;desolvation gas flow was 600 L/h;capillary voltage was 3 000 V;nebuliser pressure was 45 psi;the work mode was multiple reaction monitoring mode;detection mode was negative ion mode. The established method was used to determine the contents of 8 components in 3 batches of total flavanones of Sedum sarmentosum Bunge. RESULTS:The linear ranges of hyperoside,quercitrin,luteoloside,kaempferol,quercetin,rutin,luteolin and isorhamnetin were 10.0-640.0,0.5-32.0, 4.5-288.0,8.0-512.0,50.0-3 200.0,2.0-128.0,12.5-800.0 and 25.2-1 612.8 ng/mL(r≥0.991 4),respectively. The limits of detection were 5.0,0.25,2.25,4.0,25.0,1.0,6.25 and 12.6 ng/mL,respectively.The limits of quantitation were 10.0,0.5,4.5, 8.0,50.0,2.0,12.5 and 25.2 ng/mL,separately. RSDs of precision,stability(24 h)and reproducibility tests were no more than 4.3%(n=6). The recoveries were 95.9%-100.6%,and RSDs were 1.5%-3.8%(n=6). The contents of hyperoside,quercitrin, luteoloside,kaempferol,quercetin,rutin,luteolin and isorhamnetin in 3 batches of total flavanones of Sedum sarmentosum Bunge were 507.88-560.37,42.95-50.36,63.52-71.80,1 695.10-1 753.27,10 569.28-10 612.99,25.76-30.13,2 795.22-2 877.43 and 4 869.55-4 971.30 μg/g,respectively. CONCLUSIONS:The established method can be used for simultaneous determination of 8 components in total flavanones of Sedum sarmentosum Bunge.