Looking for protocols to quantify anthocyanins using HPLC ( what standards to use and what column??)
Dear Sir. Concerning your issue about the quantification of anthocyanins from plants using HPLC. Anthocyanins are naturally present in a wide range of plant foods, and are usually associated with the orange, pink, red, violet, and blue colors of fruits, such as berries, plums, cherries, pomegranates, citrus fruits, among others. Juices produced from fruits, especially berries, such as black currants, elderberries, chokeberries, or sour cherries, are a source of vitamins and other compounds, such as anthocyanins. High- or ultra-high-performance liquid chromatography techniques coupled with diode array detection or mass spectrometry detection, are by far the most widely used techniques for the qualitative and quantitative analysis of anthocyanins in fruit juices. Anthocyanin content in fruit juices can be used as quality control parameters to check authenticity. Significant differences can be found for anthocyanin content in the same type of juice, but the profile of anthocyanins should be similar. An HPLC method and an acid hydrolysis HPLC method for the analysis of anthocyanins and anthocyanidins in bilberry extracts have been developed. The HPLC method coupled with a mass detector has identified 11 anthocyanins in bilberry extracts. The method provides anthocyanin profiles that are very useful in verifying the identity of botanical raw materials, monitoring the consistency of the raw material source, and quantitating the total anthocyanins. The acid hydrolysis HPLC method greatly simplifies the anthocyanin profile in bilberry samples and converts anthocyanins to five major anthocyanidin aglycones: delphinidin, cyanidin, petunidin, peonidin, and malvidin. Each of these aglycones can be separated completely and quantitated accurately with external standards. Various extraction and hydrolysis conditions were investigated, and the advantages and disadvantages of the HPLC and acid hydrolysis methods are discussed. I think the following below links and the attached file may help you in your analysis:
https://digitalcommons.library.umaine.edu/cgi/viewcontent.cgi?referer=https://www.google.com.sa/&httpsredir=1&article=1175&context=honors
https://www.sciencedirect.com/science/article/pii/B9780128022306000345
https://pdfs.semanticscholar.org/5eac/5be282ca1a8b65507b1fa59b89f15461573c.pdf
Thanks
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A quick google search is a paper attached. Just a reminder of a new researcher to go to internet and search anything you want to know. It is a rich resouces and is available for you.
Thank you all for the quick response and all the valuable information. I need to find out what HPLC we have and what columns we have first.
You should use reverse HPLC. I recommend you a C18 column using a gradienf water and acetonitrile.
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