Solid phase extraction (SPE) experiments were performed for the digested samples from the Ambi/urea extracts in order to purify and enrich analytical samples (peptides) prior the MS/MS analysis. Briefly, columns containing 300 mg of C18 material (Chromabond® sorbent C18 ec, Macherey-Nagel GmbH &CO. KG, Düren, Germany) were activated with 6 mL of buffer A (50% ACN/50% distilled water containing 0.1% formic acid). A total of 6 mL of distilled water was then run for conditioning the solid phase. Samples were applied on the columns and thereafter, 6 mL of distilled water was run as the washing step. Peptides were finally eluted using 1 mL of buffer B (100% acetonitrile containing 0.1% formic acid) and the eluates were filled up to 5 mL with distilled water by giving the water through the columns. After that step, 100 µL of each sample was mixed with 10 µL of the internal standard and transferred to the vials for the analysis.
Chromabond sorbent c18 ec
Manufactured by Macherey-Nagel
Sourced in Germany
Chromabond® sorbent C18 ec is a solid-phase extraction (SPE) material composed of octadecylsilyl (C18) modified silica. It is designed for the extraction and purification of a wide range of organic compounds from various sample matrices.
Automatically generated - may contain errors
3 protocols using chromabond sorbent c18 ec
1
Solid-Phase Extraction for Peptide Purification
2
Optimized Powder Extraction and Purification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Powder samples were extracted by maceration at optimized conditions descripted in our research paper (Albuquerque et al., 2017) . The extracts were filtrated through a Whatman paper filter n°4, and after that were evaporated at 40 °C using a rotary evaporator (Büchi R-210, Flawil, Switzerland) to remove the ethanol (Fisher Scientific, Lisbon, Portugal) . The purification to clear away sugars and more polar substance of extracts was performed using a C-18 solid phase column (Chromabond sorbent C18 ec, Macherey-Nagel, Duren, Germany), as described by Albuquerque, Prieto, Barros and Ferreira (2017) .
3
Purification of Bioactive Compounds
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Each sample (1 g) was extracted by maceration with 30 mL of ethanol:water 20:80 (v/v) at 80 • C at 150 rpm during 90 min (Albuquerque et al., n.d.) . The extract was filtered through Whatman no. 4 paper. The obtained extracts were evaporated at 40 • C (rotary evaporator Büchi R-210) to remove ethanol. For purification, the aqueous phase was deposited onto a C-18 solid phase column (Chromabond sorbent C 18 ec, Macherey-Nagel, Duren, Germany), previously activated with ethanol followed by water; sugars and more polar substances were removed by passing through 50-80 mL of water and the purified extract was further eluted with 20-40 mL of ethanol. The purified extract was evaporated at 40 • C to remove ethanol.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!