Sample preparation: Metabolite extraction (tutorial 3/5)

The metabolome of a biological system can change within a few seconds or minutes. Sample preparation is one of the most critical parts of a successful metabolomics experiment.
Sample collection, processing and storage can affect concentrations of metabolites changes, resulting in an inaccurate data.
Here we demonstrate a step-by-step procedure for aqueous methanol extraction of polar metabolites from HeLa cell culture. This general protocol can be adapted for other samples considering: 1) type of biological matrix 2) sample collection 3) quenching of cellular metabolism 4) extraction and recovery of metabolites 5) drying 6) storage and shipment.
Read more about protocols via www.embl.de/mc....

[@lizhang2849]

Today I have 90 samples prepared for targeted metabolomic assay. thank you.

[@faithogbonna5484]

Please what did you use to aspirate? Or can you please explain the aspiration part?

[@Ouekkaphot]

I put my cell lysate in -80. Can I use them for metabolomic? Too late to add methanol to quench metabolism?

[@Etatdespritdecroissance]

GREAT video. Thanks.

[@shakirshamas6286]

superb, really awsome and informative lecture

[@emmanuelrios6832]

super. Thank you

[@violettracey]

Thanks!

[@leewilliam3417]

Thankyou 😊

[@faithogbonna5484]

Please can you explain the quenching procedure? Aspiration was done . What was used for aspiration? Am currently working on extraction of metabolites from Enterococcus species and I need more guides

[@shakirshamas6286]

same question and same procedure and guidance I needed. i am working on primary metabolites of chickpeas

[@azizabed7692]

Hi, Thanks for this video. Please I have 2 questions: 1- I why you stored the extraction solution for 1 hour before use? 2- What is the red solution? Is it bacteria cells with its extracts? Could you please please tell me more about the bacteria cells sources , preparation , how you stimulate them to excrete secondary metabolite? Thanks

[@EMBLorg]

Information about the protocols can be found here: www.embl.de/mcf/metabolomics-core-facility/protocols/index.html including contact details for the EMBL Metabolomics Core Facility, if you have further questions.

[@PrasadPhapale]

Hi Aziz, 1) the storage is required to have cold methanol for extraction. The cold methanol is necessary for quenching the metabolism before extraction 2) This protocol is for adherent mammalian cells; the red color is from DMEM medium in which cell are grown.

[@jarrysc]

Here I can't find the subtitles too, can you help us please.

[@EMBLorg]

There are no subtitles, because there is no speaking in the video, it is just background music.

[@swathireddy8807]

Sir I want extract metabolite from actinobacteria and Its not yielding good amount of extractant

[@swathireddy8807]

And I'm following protocol from one of the article and they will say to repeat the extraction 3 times should I do this

[@abhijitmandal3245]

if you read my reply to your mail kindly response . i am working on antibiotic producing bacillus from . can you help me on this? ... from India

[@Risperi1]

0:21

[@purplerain365]

hi, can I get a softcopy of this protocol of metabolite extraction for my final year project? Thank you.

[@EMBLorg]

Please contact the Metabolomics Core Facility team, clearly stating the tutorial you are referring to and what you would like them to send you. You can contact them through the EMBL website: www.embl.de/mcf/metabolomics-core-facility/contact/index.html

[@PrasadPhapale]

Hi Jennifer, soft copy is available on our webpage under protocols. Thanks!

[@jarrysc]

Hi, Jenn, Did you find the protocols?, can we speak in a private conversation by mail... [e-mail], will write whoever worked with extraction of metabolites... I'm really trying to get the way to extract from Trichoderma sp. and I have actually lot of questions about.