An Optimised Method for Routine Separation and Quantification of Major Alkaloids in Cortex <i>Cinchona</i> by HPLC Coupled with UV and Fluorescence Detection

Holmfred, Else; Cornett, Claus; Maldonado, Carla; Rønsted, Nina; Hansen, Steen Honoré

doi:10.1002/pca.2684

Cited by 10 publications

(10 citation statements)

References 24 publications

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“…They naturally cannot compete with those achievable by fluorescence detection (e.g. LOD for quinine is 2 fmol, [6]); however, they are comparable to conventional HPLC-UV as LOQ values of 5 µg/g are stated in reference [17]. Precision was investigated by repeatedly assaying sample CC-2017-2 under optimized extraction and separation conditions.…”

Section: Resultsmentioning

confidence: 99%

“…The quantitative results were well comparable to published data, which for example report the following values for a drug with Ph.EU. quality: 1.80% quinine, 1.65% cinchonine, and 1.25% cinchonidine [17]. This successful application of SFC, on for the utilized technique “untypical” compounds, should raise further interest to fully explore the potential of this separation technique, which definitely is not limited to the “classics” like carotenoids, fatty acids or terpenes.…”

Section: Resultsmentioning

confidence: 99%

“…For example, Hoffmann et al utilized a chiral strong cation exchange material to excellently resolve eight Cinchona alkaloids in 15 min, yet an application to plant material is missing [16]. The latter was presented in the most recent study, in which Holmfred et al reported on the separation of the four main isomers on 2.6 µm C-18 core shell material (Kinetex XB-C18) in 25 min [17]. Whether Supercritical Fluid Chromatography (SFC) is a possible and equivalent alternative has never been investigated.…”

Section: Introductionmentioning

confidence: 99%

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Quantitative determination of major alkaloids in Cinchona bark by Supercritical Fluid Chromatography

Murauer

Ganzera

2018

Journal of Chromatography A

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Chinoline alkaloids found in Cinchona bark still play an important role in medicine, for example as anti-malarial and antiarrhythmic drugs. For the first time Supercritical Fluid Chromatography has been utilized for their separation. Six respective derivatives (dihydroquinidine, dihydroquinine, quinidine, quinine, cinchonine and cinchonidine) could be resolved in less than 7 min, and three of them quantified in crude plant extracts. The optimum stationary phase showed to be an Acquity UPC2 Torus DEA 1.7 µm column, the mobile phase comprised of CO2, acetonitrile, methanol and diethylamine. Method validation confirmed that the procedure is selective, accurate (recovery rates from 97.2% to 103.7%), precise (intra-day ≤2.2%, inter-day ≤3.0%) and linear (R2 ≥ 0.999); at 275 nm the observed detection limits were always below 2.5 µg/ml. In all of the samples analyzed cinchonine dominated (1.87%–2.30%), followed by quinine and cinchonidine. Their total content ranged from 4.75% to 5.20%. These values are in good agreement with published data, so that due to unmatched speed and environmental friendly character SFC is definitely an excellent alternative for the analysis of these important natural products.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Quantitative determination of major alkaloids in Cinchona bark by Supercritical Fluid Chromatography

Murauer

Ganzera

2018

Journal of Chromatography A

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“…was ACN and ammonium acetate aqueous solution (pH 5.8) adjusted by acetic acid [23]. Major quinoline alkaloids in barks of several Cinchona species were investigated with acidic mobile phase containing ammonium formate [24]. Steroidal alkaloids variation from rhizomes and aerial plants of Veratrum californicum Durand was analyzed with water and ACN, both acidified by FA [25].…”

Section: Chromatographic Separation Methodsmentioning

confidence: 99%

Liquid chromatographic separation of alkaloids in herbal medicines: Current status and perspectives

Zhao

Chen

Yu-qian

et al. 2020

J of Separation Science

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Alkaloids are a widespread group of basic compounds in herbal medicines and have attracted great interest due to various pharmaceutical activities and desirable druggability. Their distinctive structures make chromatographic separation fairly difficult. Peak tailing, poor resolution, and inferior column‐to‐column reproducibility are common obstacles to overcome. In order to provide a valuable reference, the methodologies and/or strategies on liquid chromatographic separation of alkaloids in herbal medicines proposed from 2012 to 2019 are thoroughly summarized.

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“…A few bark samples were covered in lichens, which were brushed off before the bark samples were ground to homogeneity using a custom made grinding device (Hansen et al, 2015 ), and 50.0 mg of the ground bark was dissolved in 1.0 mL DMSO (dimethyl sulfoxide), and ultrasonicated for 10 min. Following Holmfred et al ( in press ), 4.0 mL of 70% methanol containing 0.1% formic acid was added and the mixture was ultrasonicated for 10 min. After centrifugation at 5,000 rpm for 10 min, the clear liquid phase was transferred into a 50.0 mL volumetric flask.…”

Section: Methodsmentioning

confidence: 99%

Phylogeny Predicts the Quantity of Antimalarial Alkaloids within the Iconic Yellow Cinchona Bark (Rubiaceae: Cinchona calisaya)

et al. 2017

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Considerable inter- and intraspecific variation with respect to the quantity and composition of plant natural products exists. The processes that drive this variation remain largely unknown. Understanding which factors determine chemical diversity has the potential to shed light on plant defenses against herbivores and diseases and accelerate drug discovery. For centuries, Cinchona alkaloids were the primary treatment of malaria. Using Cinchona calisaya as a model, we generated genetic profiles of leaf samples from four plastid (trnL-F, matK, rps16, and ndhF) and one nuclear (ITS) DNA regions from twenty-two C. calisaya stands sampled in the Yungas region of Bolivia. Climatic and soil parameters were characterized and bark samples were analyzed for content of the four major alkaloids using HPLC-UV to explore the utility of evolutionary history (phylogeny) in determining variation within species of these compounds under natural conditions. A significant phylogenetic signal was found for the content of two out of four major Cinchona alkaloids (quinine and cinchonidine) and their total content. Climatic parameters, primarily driven by changing altitude, predicted 20.2% of the overall alkaloid variation, and geographical separation accounted for a further 9.7%. A clade of high alkaloid producing trees was identified that spanned a narrow range of altitudes, from 1,100 to 1,350 m. However, climate expressed by altitude was not a significant driver when accounting for phylogeny, suggesting that the chemical diversity is primarily driven by phylogeny. Comparisons of the relative effects of both environmental and genetic variability in determining plant chemical diversity have scarcely been performed at the genotypic level. In this study we demonstrate there is an essential need to do so if the extensive genotypic variation in plant biochemistry is to be fully understood.

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An Optimised Method for Routine Separation and Quantification of Major Alkaloids in Cortex Cinchona by HPLC Coupled with UV and Fluorescence Detection

Abstract: The optimised HPLC method offers a simple and efficient quantification of the four major alkaloids. Copyright © 2017 John Wiley & Sons, Ltd.

Cited by 10 publications

References 24 publications

Quantitative determination of major alkaloids in Cinchona bark by Supercritical Fluid Chromatography

Quantitative determination of major alkaloids in Cinchona bark by Supercritical Fluid Chromatography

Liquid chromatographic separation of alkaloids in herbal medicines: Current status and perspectives

Phylogeny Predicts the Quantity of Antimalarial Alkaloids within the Iconic Yellow Cinchona Bark (Rubiaceae: Cinchona calisaya)

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