Determination of quercetin and kaempferal in rabbit plasma by HPLC
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OBJECTIVE A method was developed for the determination of quercetin (Q) and kaempferol ( K) in rabbit plasma.METHODS Plasma samples were treated with acid hydrolyzation and ether extraction, the organic layer was evaporated to dryness, the residues were reconstituted in 100μL mobile phase prior to injection onto the high-performance liquid chr omatography(HPLC). A YWG C 18 column was used with a mobile phase of 0.0 1 mol·L -1 phosphate buffer(pH 2)-tetrahydrofuran-me thanol-isopropynol (60∶15∶10∶ 15). The elution was performed at the flow rate of 0.5 mL· min -1,detected at 380 nm.RESULTS There was a good linear relationship over the concentration range of 16.66 - 156.8 μg·L -1 for quercetin and kaempferol with coefficient of 0.991 2 and 0.994 1 respectively. Limits of detection were 4.2 μg·L -1 for quercetin and kaempferol. The average recoveries of the method were 101.0 % with RSD of 5.05%, and 103.0% with RSD 4.72% for quercetin and kaempferol, respectively.CONCLUSION This method is simple and accurate, it could be applied to the determination of quercetin and kaempferol in rabbit plasma after oral administration of Ginkgo bilaoba extract.Cite
AIM:A rapid,sensitive and highly selective liquid chromatography-tandem mass spectrometry method was developed and validated for simultaneous determination of quercetin,kaempferol and isorhamnetin in dog plasma.METHODS:Quercetin,kaempferol and isorhamnetin conjugates were hydrolysed chemically.The analytes were extracted from plasma samples by liquid-liquid extraction,separated on a Luna ODS-2 column(150 mm×2.1 mm I.D.,5 μm particle size),and detected by tandem mass spectrometry with a Finnigan IonSpray ionization interface.The mobile phase consisted of 0.1% aqueous formic acid(A)and gradient-grade acetonitrile(B).RESULTS:The calibration curves for quercetin,kaempferol and isorhamnetin were linear in concentration ranges of 0.5-100.0 ng/mL in dog plasma.The method has a lower limit of quantification(LLOQ)of 0.5 ng/mL for all the three flavonols.The intra-and inter-day precisions,expressed as the R.S.D.,were less than 7.3%,6.2% and 6.4% for quercetin,kaempferol and isorhamnetin,respectively,and the recovery was more than 70%,66% and 70%,respectively.The application of this assay was examined in a preliminary pharmacokinetic study of quercetin,kaempferol and isorhamnetin in beagle dogs after oral administration of 6 Ginkgo biloba tablets.CONCLUSION:The present method was suitable for determining quetcetin,kaempferol and isorhamnetin in dog plasma.
Isorhamnetin
Flavonols
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Objective: To establish a HPLC method for quantification of quercetin and kaempferol in compound Ganyan granules. Method: Samples were extracted in 80% methanol with ultrasonic process and hydrolyzed for 1 h. The HPLC was performed on an ODS-C 18 column(4. 6 mm × 250 mm,5 μm) with methanol-0. 4% H 3 PO 4(50∶ 50) as mobile phase. The flow rate was 1. 0 mL·min- 1and the column tempreture was kept at 30 ℃. The eluate was detected at 360 nm. Result: The linear ranges of quercetin and kaempferol were 0. 050 1-0. 501 μg(r = 0. 999 9) and 0. 024 2-0. 242 μg(r = 0. 999 5) respectively. The average recoveries of quercetin and kaempferol were 99. 69%(RSD 0. 22%) and 99. 30%(RSD 0. 82%). Conclusion: The method can be used for quality control of Ganyan granules. It is simple,rapid and accurate for simultancous determination of two constituents in compound Ganyan granules.
Content determination
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OBJECTIVE To determine the kaempferol in rat plasma.METHODS A liquid-liquid extraction method with absolute ether was used and baicalein was chosen as internal standard.The analytes were separated on a Shimm-pack VP-ODS column at 40℃ and detected at 370 nm.The mobile phase consisted of acetonitrile-0.5% glacial acetic acid(33.3:66.7) with a flow rate of 1.0 ml·min-1.RESULTS The calibration curve of kaempferol in plasma showed a good linearity over the concentration range of 0.050-5.301 μg·ml-1 and the lower limit of quantitation was 0.020 μg·ml-1.The intra-and inter-assay precision for this analysis were no more than 7.39% and 5.88%,respectively and all the samples inaccuracy(bias) was less than 14.67%.The average extraction recovery of kaempferol from plasma was 96.63%.CONCLUSION The method is simple,sensitive and accurate,and could be applied to the pharmacokinetic study of kaempferol in rats.
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Objective: To develop a HPLC method for the determination of quercetin in Quercetin capsule.Method: the method has been carried out by using the ultrasphere Kromasil column (4.6 mm×250 mm, 5μm) and methanol phosphate buffer solution(1 000 ml H 2O+4 ml H 3PO 4+2 ml TEA)(60∶40) as mobile phase.A flow rate of 1.0 ml/min and the detection wavelength of 373 nm were adopted. Result: A satisfactory separation among Quercetin and related impurities was obtained. The calibration curve was linear in the range of 7～70 μg/ml for Quercetin .The average recovery of Quercetin was 99.47%(RSD=0.97%). Conclusion: This method is simple, rapid,specific,and can be used for the preparation as a Quercetin quality control standard.
Capsule
Linear range
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Objective To establish an HPLC method for the content determination of quercetin and kaempferol in Wasong Suppository.Methods The HPLC analysis was carried out on C18 analytical column with acetonitrile as mobile phase A and 1.0% HCl solution as mobile phrase B.The detection wavelength was 360 nm.Results The liner ranges of quercetin and kaempferol were 0.006~0.024 mg·mL-1(r=0.990 8) and 0.012~0.047 mg·mL-1(r=0.995 2),respectively.The average recoveries of quercetin and kaempferol were respectively 99.91% and 101.2%.Conclusion The method was simple,accurate and reproducible,and it can be used for the determination of quercetin and kaempferol in Wasong Suppository.
Suppository
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OBJECTIVE:To establish a method for the simultaneous determination of kaempferol and quercetin in Euphorbia sororia.METHODS:HPLC method was adopted.Hypersil ODS-2 C18 column(250 mm×4.6 mm,5 μm)was used with mobile phase consisted of methanol-0.4% phosphonic acid solution(52∶48,V/V)at the flow rate of 1.0 ml/min.The detection wavelength was set at 360 nm,and column temperature was 30 ℃.RESULTS:The linear ranges of kaempferol and quercetin were 5.2-156.0 μg/ml(r=0.999 9)and 3.1-92.1 μg/ml(r=0.999 7),respectively.RSDs of precision,reproducibility and stability were all lower than 3%.The average recoveries were 101.73%(RSD=1.27%,n=6)and 101.39%(RSD=1.59%,n=6),respectively.CONCLUSIONS:The method is quick,simple,rapid and reproducible for the determination of kaempferol and quercetin in E.sororia.
Content determination
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Objective To establish a HPLC method for determination the content of total favonol glycosides in ginkgo tablet.Methods Phenomenex C_18 column was used as chromatographic column,and methanol-0.4% acid water(50:50)as mobile phase.The detection wavelength was set at 360 mm,flow rate at 1.0 ml/min,and column temperature at 25℃.Results There was a good linear relationship when quercetin and kaempferol of total favonol glycosides were in the range of 0.007 5～0.06 mg/ml(r=0.999 9,n=5),and Isorhamnetin in the range of 0.005～0.04 mg/ml(r=0.999 5,n=5).The average recoveries of Quercetin,kaempferol and isorhamnetin were respectively 99.92%(RSD=1.65%,n=6),98.60%(RSD=1.3%,n=6),99.41%(RSD=0.81%,n=6).Conclusion This method is simple in operation,high in sensitivity and good in reproducibility,and can be used for the determination of the preparation.
Isorhamnetin
Content determination
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To better control the quality of U.lobata,an HPLC method was established for simultaneously determining quercetin and kaempferol of U.lobata and the content of quercetin and kaempferol were determined.The column was agilent Eclipse plus C18(100mm×4.6mm,5μm)column.The mobile phase was mixture of methanol and 0.4% phosphoric acid water solution(45∶55).The flow rate was 0.8mL/min,the wavelength was 360 nm.The results showed that the calibration curves of two flavonoids were both linear in the range of 2.500~25.00(g/mL(r = 0.9997 and 0.9999respectively).The precisions test RSD were 0.98% and 0.79% respectively.The stability test RSD were 1.36% and1.13%respectively.The RSD were 2.37% and 2.10%respectively.The method was accurate with high sensitivity and suitable for the determination of quercetin and kaempferol in U.lobata.The highest contents of quercetin and kaempferol in U.lobata were respectively(1.067 0.02)mg/g and(1.232 0.03)mg/g.
Phosphoric acid
Lobata
Content determination
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A high performance liquid chromatography(HPLC) method for simultaneous determination of four effective components(Quercetin;Genistein;Kaempferol;Isorhamnetin) in Sophorajaponica L.was developed.Separating column with Hypersil BDS C18(25 cm×4.6 mm ID,5 μm) was used in the experiment,and CH3OH∶ H2O∶H3PO4(48∶52∶0.3) was used as mobile phase.Detection wavelength was 254 nm and the flow rate was 0.8 mL·min-1.Column temperature was set as 45 ℃.There was good linearity between the peak area and injected amount when the amount is 0.029-0.285 μg for Quercetin,0.030-0.300 μg for Genistein,0.122-1.220 μg for Kaempferol and 0.096-0.960 μg for Isorhamnetin.The correlation coefficient of each component is 0.999 7,0.999 5,0.999 8 and 0.999 8 respectively.Average recoveries of the four components are 98.9%,99.2%,98.3%,98.8%.RSD of each group are 0.62%,0.86%,1.28%,0.82%(n=9).Four effective components in Sophorajaponica L.were simultaneously determinated,and the method is proved to be convenient,accurate,fast,repeatable and of high sensitivity.
Isorhamnetin
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OBJECTIVE To establish a quantitative method for simultaneous determination of rutin,hyperoside,quercetin,kaempferol in Rhododendron athopogoides by RP-HPLC.METHODS The sample was separated on column of Hypersil ODS-2(250 mm×4.6 mm,5 μm)with gradient elution,the mobile phase was comprised of methanol and water(0.04% phosphoric acid).The flow rate was 1mL·min-1.The detection wavelength was at 360 nm and the column temperature was 30 ℃.RESULTS The linear ranges of rutin,hyperoside,quercetin and kaempferol were 0.042-5.95 μg(r=0.9999),0.490-2.450 μg(r=0.9997),0.008-1.600 μg(r=0.9998),0.014-0.142 μg(r=0.9999),respectively.The average recovery were 99.13%(RSD=0.56%),98.95%(RSD=0.29%),100.04%(RSD=1.72%),97.27%(RSD=0.65%),respectively.CONCLUSION The method is rapid and precise.
Hyperoside
Phosphoric acid
Gradient elution
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