I am trying to show anthocyanin and its metabolites in porcine plasma using UPLC/PDA detector using aglycone and phenolic acid standards. I want any method for extracting anthocyanins/phenolics from plasma.
I am working on anthocyanins. Use 0.1% CH3COOH in Ethanol for extraction of anthocyanins. Then load on XADs or you can also use HP-20 diaion for preliminary purification. Then on Silica. You can also use prep HPLC or flash chromatography for purification.
For UPLC,
Mobile phase A=ACN in 0.1%CH3CooH
B=0.1%CH3COOHin water
Gud luck
:)
You may need to release anthocyanins from the plasma matrix. How is your analyte transported in the plasma? Surely you need to extract and do SPE before your analysis. What is the detection limit of your method? Have you validated the method?
I tried SPE..........however, when eluting with methanol, I lose out on phenolic acids.......I need both anthocyanins and phenolic acids..........I have tried ethyl acetate to get phenolic acid fraction and methanol for anthocyanin fraction based on an old protocol.........but its a bit time consuming and elevated losses while drying them down, combining and then injecting ...........
I think you should use SPE as said by Sridhar Radhakrishnan because SPE stands for Solid Phase Extraction and SPME for Solid Phase Micro Extraction. There is alot of difference between SPE and SPME.
SPME used for volatiles and SPE used for non-volatiles.
Go with SPE
Hi Guys, Someone of you developed a good tactic to determinate Anthocyanins (Glucoside forms) in human blood plasma?. I have used TC 18 cartrigdes as previous proteins separate SPE process, But when I try to read at UPLC no anthocyanins presence there is. Thank you so much for join your experiences about it.
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