Simultaneous Determination of Four Effective Components in Rose Oral Liquid by High Performance Liquid Chromatography
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Abstract:
An analytical method for the simultaneous determination of four effective components(Gallic acid,Rutin,Quercetin,Kaempferol)in Rose Oral Liquid was established using high performance liquid chromatography(HPLC).The separation was performed on an Agilent C18 column(250×4.6mm,5μm)at 30℃and 0.5% phosphoric acid-methanol as mobile phases with gradient elution at a flow rate of 0.8mL/min,and the detection wavelength was 260nm.Good linearity was obtained between the peak area and injected amount when the amount was 40.7-244.2μg·mL-1 for Gallic acid,25.0-150.0μg·mL-1for Rutin,13.3-80μg·mL-1 for Quercetin and 2.69-16.2μg·mL-1 for Kaempferol.The correlation coefficient of each component was 0.9997,0.9993,0.9998 and 0.9994,respectively.Average recoveries of the four components were 99.2%,101.2%,98.8%,99.2%.RSD of each group were 0.60%,1.02%,0.62%,0.57%(n=3).Four effective components in Rose Oral Liquid were simultaneously determined,and the method was convenient and accurate.Keywords:
Phosphoric acid
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OBJECTIVE To develop a HPLC method for the determination of total flavonol in Litonglin capsules by HPLC.METHODS The chromatographic analysis was performed on Kromasil C_18(4.6×250mm,5μm)column.The mobile phase consisted of methonal-0.4%phosphoric acid(55∶45).The flow rate was 1.0mL·min-1.The detection wavelength was at 360nm.RESULTS Quercetin:A good linearity was obtained over the range of 4.067μg·mL-1-40.67μg·mL-1(r=0.9998).Kaempferol:A good linearity was obtained over the range of 0.871μg·mL-1-8.71μg·mL-1(r=0.9998).Isorhamnetin:A good linearity was obtained over the range of 0.616μg·mL-1-6.16μg·mL-1(r=0.9996).CONCLUSION The method was simple,accurate,specific and could be used for the quality control of quercetin,kaempferol and isorhamnetin in crude saponins of Litonglin capsules.
Isorhamnetin
Phosphoric acid
Linearity
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OBJECTIVE:To establish RP-HPLC for the determination of the content of Quercetin in the Xiaoyanliniao mixture.METHODS:Samples were Separated on Syltech-500 C18 column(250mm×4.6mm,5μm).The mobile phase consisted of methanol-0.2% phosphoric acid(50∶50)at a flow rate of 1.0mL·min-1.The detection wavelength was 360nm,and the column temperature was 25℃.RESULTS:The linear range of Quercetin was from 19.2 to 96.0μg·mL-1(r=0.999 4).The average recovery was 99.40%(RSD=2.31%,n=6).CONCLUSION:The method was specific,sensitive and reliable in results yet with little interference,and it can be used as quality control for Xiaoyanliniao mixture.
Phosphoric acid
Content determination
Linear range
Content (measure theory)
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Objective: High performance liquid chromatograpghy( HPLC) was used to determine the content of Rutin in Oputia dillenii haw. Method: Chromatograph conditions were as follows: ODS- SP( 250mm × 4. 6mm,5μm),mobile phase was methanol: 0. 4% Phosphate( 40: 60),the flowrate was 1mL / min,column temperature was 25℃,the injection volume was 10μL,λ = 360nm. The detection wavelength was 360 nm. Result: The good liner rang of rutin was within 0. 5 ~ 5μg,r = 0. 999 9; and the recovery rate was 100. 527%( RSD = 1. 970%). Conclusion: The method is simple, sensitive,viable and stable. Moreover,it has a better separative effect.
Recovery rate
Phosphate buffered saline
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Objective To develop a new high performance liquid chromatography(HPLC) method for simultaneous determination of four major components(gallic acid,loganin,paeoniflorin and paeonol) in different dosage forms of Liuweidihuang.Methods The chromatography condition was with Agilent Zorbax SB-C_(18) column(4.6×250 mm,5 μm);mobile phase was A;ACN,B:0.1%formic acid,gradient elution,0~3 min,A:3%~5%;3~18 min,A:5%~22%;18~60 min,A:22%~60%,flow speed was 1.0 ml/min,temperature of column was 25℃,inject volume was 10 μl,detection wavelength was 240 nm.Results The linearity was obtained over 6.296~318.9 μg/ml(r =0.999 8) for gallic acid,1.952~99.04 μg/ml(r =0.999 7) for loganin,6.186~309.3 μg/ml(r =0.999 8)for paeoniflorin and 7.147~214.4 μg/ml(r =0.999 7) for paeonol.The RSDs of precision of the samples were both less than 2%.The average recovery was between 98.2%~102.3%.Conclusion The present method,with satisfactory efficacy,was accurate and simple which could simultaneously determine four components,and could be used for quality control of different dosage forum of Liuweidihuang.
Paeoniflorin
Loganin
Paeonol
Gradient elution
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Objective: To develop the method for quality control of kaempferol in Jianyutangkang tablet.Methods: Diamonsil-C18 column(250 mm × 4.6 mm,5 μm)was used for separation.The mobile phase consisted of acetonitrile-0.1% formic acid(33:67) with the flow rate of 0.8 mL·min-1,and the detection wavelength was set at 258 nm.Results: The kaempferol peak was separated from other peaks without interference by negative sample.The linear range of kaempferol was 2.076-20.76 μg·mL-1(r = 0.999 8).The recovery was 100.39% with RSD of 0.52%.Conclusion:The method is simple,accurate and repeatable,and it is suitable for the content determination of kaempferol in Jianyutangkang tablet.
Content determination
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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.
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Objective To establish the quality control standard for Xuelianhuangtong capsules against high-altitude hypoxia. Methods The Rutin and Chlorogenic Acid were identified by thin layer chromatography(TLC); total flavonoids content were determined by ultraviolet spectrophotometry(UV),and Rutin content was determined by high performance liquid chromatography(HPLC). The Hypersil ODS2 C18(150 mm ×4. 6 mm,5 μm) column was used,and the mobile phase consisted of methanol-water(40∶ 60),and the flow rate was 1 ml /min; the column temperature was room temperature,and the detection wavelength was at 340 nm. Results The spots in the TLC were clear with good disassociation effect; the good linear relationship was achieved within the range of total flavonoids content of 9. 84μg /ml-59.04μg / ml(r = 0. 9997),and its average recovery ratio was 99. 32% with 1. 47% of RSD(n = 6); the good linear relationship was achieved within the range of Rutin content of 4-120 μg /ml(r = 0. 9999),and its average recovery of application of sample was 97. 98% with 1. 35% of RSD(n = 6). Conclusion The method is simple,practical and accurate with good repeatability,so it can be used for the quality control of Xuelianhuangtong capsules.
Repeatability
Chlorogenic Acid
Content determination
Linear relationship
Linear range
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Objective: To establish a method for determination of rutin in shanglucha.Methods: HPLC analysis was carried out using ODS C18 column(4.6 mm×250 mm,5 μm) and acetonitrile-2.5% acetic acid solution(15∶85) as the mobile phase,flow rate 1.0 mL/min,column temperature at 30 ℃.The detection wavelength was 254 nm.Results: Rutin had the good linear relationship with peak area at the range of 0.018 33~0.183 30 g/L(r=0.999 8,n=5).The average recovery was 97.5%(RSD=1.07%,n=6).Conclusion: The method is simple,accurate,reproducible and suitable for determining the rutin in shanglucha.
Linear range
Linear relationship
Recovery rate
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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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Analysis of Quercetin and Kaempferol in an Alcoholic Extract ofConvolvulus pilosellifoliususing HPLC
A simple, rapid, and sensitive high-performance liquid chromatography (HPLC) method was developed and validated for identification and determination of flavonoids in Convolvulus pilosellifolius. The chromatographic separation was achieved in less than 6 min using C18 column (150 × 4.6 mm, 3 μm) with isocratic mixture of methanol and water containing 0.1 percent v/v formic acid in the ration of 80:20 at 258 nm with a flow rate of 0.4 mL/min. The method was validated in the linear calibration curve ranged between 1 and 300 μg/mL with detection limits of 0.39 and 0.26 μg/mL and quantification limits of 1.20 and 0.79 μg/mL for quercetin and kaempferol, respectively. Good repeatability of the method were achieved at percent relative standard deviation (RSD < 2.18 percent) with respect to inter- and intraday repeatability. Recovery values were found to be in the range of 98.2–100.2 percent, indicating high accuracy of the method. The maximum flavonoid contents were 1.07 and 1.54 percent for quercetin and kaempferol, respectively.
Repeatability
Convolvulus
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