Investigation of Chemiluminescence Behavior of Flavonoids with Cerium (IV)‐Rhodamine B System
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Abstract The chemiluminescence (CL) behavior of five major flavonoid types in cerium (IV)‐rhodamine B system was investigated by flow‐injection. Strong CL was observed when cerium (IV) reacted with rhodamine B in sulfuric acid medium in the presence of flavonoids. This reaction system has been established as a simple, rapid, and highly sensitive flow injection CL analysis for quercetin and kaempferol, and their detection limit (3σ) was 2.7 and 0.22 nmol/L, respectively. The relative standard deviation (n=8) was 1.2% for 1.0 µmol/L quercetin and 1.9% for 0.5 µmol/L kaempferol. This method was successfully applied to the determination of quercetin in the hydrolysate of rutin and compared well with the high performance liquid chromatography (HPLC) method. From a comparison of several related flavonoids, it was concluded that only flavonoids that contain a free 3‐hydroxyl and 2, 3‐double bond in conjugation with 4‐oxo function could produce a relatively strong CL emission.Keywords:
Rhodamine B
Rhodamine
Abstract The aim of the study was to develop a rapid, efficient, and cheap chromatographic method for determining four selected antihypertensive active flavonoid compounds in medicinal plants in Botswana. The determination of rutin, quercetin, and kaempferol in selected medicinal plants was conducted in less than 6 min using the developed reverse phase-high performance liquid chromatography (RP-HPLC) method with a 2.7 µm Ascentis C18 express column (150 × 4.60 mm i.d) at 340, 360, and 368 nm detection wavelengths and mobile phase of methanol and 0.068% of formic acid solution in isocratic elution. Validation results showed good selectivity, linearity ( r 2 > 0.99), high percentage recoveries (90.2–104.7%), and precision (% RSD < 2) for n = 3, confirming suitability of the method for determination of the investigated flavonoids in Zingiber officinale (ginger). Application of the developed RP-HPLC method was performed in selected medicinal plants ( Lippia javanica ) (mosukujane) , Myrothanmus flabellious (galalatshwene) , and Elephantorrhiza elephantina (mositsana) used to manage hypertension by herbalists in Botswana . M. flabellious a very commonly used plant for managing hypertension was found to contain highest amounts of rutin and myricetin, whereas nothing was detected for E. elephantina .
Myricetin
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Rhodamine B
Flow injection analysis
Relative standard deviation
Rhodamine
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Steaming
Roasting
Phytochemical
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A new flow injection chemiluminescence method for the determination of rutin was established.The method was based on the chemiluminescence reaction of rutin with cerium(Ⅳ) sulfate in sulfuric acid condition sensitized by rhodamine 6G.Various factors affecting the chemiluminescence intensity of the system were investigated,and the condition of sulfuric acid,cerium(Ⅳ) sulfate and Rhodamine 6G etc was optimized.We have done research on all kinds of possible disturbers in the sample.The possible mechanism of the proposed method was also studied.The method is simple,convenient and sensitive.The linear relationship between the intensity of chemiluminescence and the concentration of rutin is over the range of 1.0×10-7-1.0×10-4 mol·L-1.The detection limit is 8.1×10-8 mol·L-1,the relative standard deviation is 1.5% for 11 measurements of 1.0×10-5 mol·L-1 standard solution.This method has been successfully applied to the determination of rutin in tablets with recoveries between 99.0% and 104.0%.
Rhodamine 6G
Linear range
Relative standard deviation
Flow injection analysis
Rhodamine B
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Abstract Flavonoids present in the leaves of lotus ( Nelumbo nucifera ) grown in different regions of South Korea (Yeongcheon, Haenam, and Seocheon) and at different harvest times (July to September) were determined. Flavonoid contents in lotus extracts were identified and analyzed using high-performance liquid chromatography (HPLC). The HPLC results revealed that the flavonoid contents of the lotus extracts varied at different harvesting times, with the highest content in July. Analysis of the flavonoid content in the leaves from the different regions showed the highest contents of isorhamnetin-3- O -glucoside, quercetin 3- O -glucuronide, and quercetin 3- O -glucoside in Yeongcheon, Korea, and highest contnts of rutin, myricetin, kaempferol 3- O -glucoside, and quercetin in Haenam, Korea. The HPLC method was validated and optimized to quantify quercetin 3- O -glucuronide; it showed good linearity (1000–62.5 µg/mL, r 2 = 0.9999), accuracy (106%–108%), and precision (RSD ≤ 1.70%). Determination of flavonoid content in lotus is valuable for producing medicinal crops and identifying the optimal sources to increase the quantity of clinically available medicines.
Isorhamnetin
Myricetin
Lotus effect
Aglycone
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To establish a sensitive and specific high performance liquid chromatography( HPLC) method for simultaneous determination of flavone glycosides( Rutin,Quercetin,Kaempferol) in Chestnut involucre. The HPLC separation was performed on an Agilent Extend-C18column( 4. 6 mm × 250 mm,5 μm) and detected with an UV detector at the wavelength of 360 nm. Gradient elution was carried out with acetonitrile-phosphoric buffer( pH = 3) at the flow rate of 1. 0mL / min. The column temperature was set at 25 ℃. The calibration curves were linear in the range of 0. 067 ~ 0. 67μg /mL for rutin,0. 046 ~ 0. 46 μg / mL for quercetin and 0. 044 ~ 0. 44 μg / mL for kaempferol( r ≥ 0. 9990),respectively. The average recoveries of rutin,kaempferol and quercetin were 99. 60%( RSD = 3. 20%),99. 20%( RSD =3. 30%) and 97. 90%( RSD = 4. 00%),respectively. The content of rutin,quercetin,kaempferol were 5. 68,0. 62,1. 94 mg / g respectively. This method is simple,accurate,reliable and hence can be used as a method for the determination of flavone in Chestnut involucre.
Phosphoric acid
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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.
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Summary A simple, rapid, and specific reversed-phase HPLC method with DAD detection has been used for analysis of two flavonoids, quercetin and kaempferol, in Cissus quadrangularis Linn. The flavonoids were well resolved within 10 min, and quantification was achieved, on an endcapped C18 column at 370 nm with acetonitrile-phosphate buffer (pH 3.4, adjusted with glacial acetic acid) 60:40 (v/v) as isocratic mobile phase at a flow rate of 1.0 mL min−1. The method was validated for limit of detection, limit of quantification, linearity, precision, accuracy, and recovery. Linearity was demonstrated over the range 0.75 to 10 μg mL−1 for quercetin and 1.0 to 10 μg mL−1 for kaempferol with good correlation coefficients (r2 > 0.998). Detection limits were 0.075 and 0.10 μg mL−1 for quercetin and kaempferol respectively. Recovery was 98.0–105.3% for quercetin and 95.0–101.1% for kaempferol. The method was successfully applied to analysis of two flavonoids in four samples of Cissus quadrangularis Linn.
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Objective: The present study was carried out to estimate the phenolics compound, Rutin, Quercetin and Kaempferol in Brassica oleracea L.var capitata. by high-performance liquid chromatography (HPLC). Methods: For the estimation of Rutin, Quercetin and Kaempferol by HPLC method, the mobile phase used are, Acetonitrile and Phosphate buffer (pH=5.8) in ratio of 55: 45.Quantification of Rutin, Quercetin and Kaempferol was carried by Athena C18 column and absorbance was measured at 254 nm with flow rate of 1 ml/min. Results: In HPLC analysis the retention time(Rt) of standards, Rutin, Quercetin and Kaempferol were found to be 2.357, 6.093 and 9.373 respectively, while the Retention times of Rutin and Kaempferol in Brassica oleracea L.var capitata are 2.387, 6.060 and 9.113 which are found to be matching with standards retention time values respectively. Conclusion: Thus this HPLC method was found to be simple and rapid for quantitative determination of Rutin, Quercetin and Kaempferol in Brassica oleracea L.var capitata.
Capitata
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