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I've seen something similar after extraction with dichloromethane for urine cortisol. We did look at the protein conc but for the life of me I can't remember the outcome! Gareth Gareth Llewelyn ap Huw Jones Principal Clinical Scientist Clinical Biochemistry UCLH 3rd Floor, 60 Whitfield Street London W1T 4EU (e) [log in to unmask] (t) 0845 155 5000 x2972 (f) 0203 447 9584 www.uclh.nhs.uk/biochemistry ________________________________ From: Clinical biochemistry discussion list [mailto:[log in to unmask]] On Behalf Of Salter, Simon Sent: 20 May 2011 11:32 To: [log in to unmask] Subject: A Friday problem Dear brain trust, Yesterday I was doing an extraction in blood using 90:10 Hexane:ethyl acetate (4ml). After mixing (pretty vigorously for 10 minutes) and spinning I had a lovely layer of cells at the bottom of my tube and a large layer of jelly above. The hexane/ethyl acetate mixture had formed a gel with something in the blood. My question is this: what is it in the blood that causes the gel formation and why does it only happen in some samples (it happened in 4 of 40 samples yesterday but normally nothing happens at all, I just end up with a nice layer of hexane/ethyl acetate and a layer of blood at the bottom). All samples are treated the same and mixed at the same time on a vibramixer. I have noticed something similar when extracting with diethyl ether too. My only thoughts is that the hydrophobic solvent crashes the proteins out of solution which then all bind to each other in a random disorganised manner thus forming a gel. The samples are post mortem and in fluoride oxalate tubes. Anyway, I thought someone out there might have a proper answer. Simon Disclaimer This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. Any views or opinions presented are solely those of the author and do not necessarily represent those of the Trust unless explicitly stated otherwise. If you have received this e-mail in error please delete it and contact the Southampton University Hospitals NHS Trust Helpdesk on:- 023 80796000 The information contained in this e-mail may be subject to public disclosure under the Freedom of Information Act 2000. Unless the Information is legally exempt from disclosure, the confidentiality of this e-mail and your reply cannot be guaranteed. This footnote also confirms that this email message has been checked for computer viruses. Please visit our website at http://www.suht.nhs.uk Think of the environment. Please avoid printing this e-mail unnecessarily ------ACB discussion List Information-------- This is an open discussion list for the academic and clinical community working in clinical biochemistry. Please note, archived messages are public and can be viewed via the internet. Views expressed are those of the individual and they are responsible for all message content. ACB Web Site http://www.acb.org.uk Green Laboratories Work http://www.laboratorymedicine.nhs.uk List Archives http://www.jiscmail.ac.uk/lists/ACB-CLIN-CHEM-GEN.html List Instructions (How to leave etc.) http://www.jiscmail.ac.uk/ ------------------------------------------------------------------------ This email is confidential and is intended solely for the person or Entity to whom it is addressed. If this is not you, please forward the Message to [log in to unmask] We have scanned this email before sending it, but cannot guarantee that malicious software is absent and we shall carry no liability in this regard. We advise that information intended to be kept confidential should not Be sent by email. We also advise that health concerns should be Discussed with a medical professional in person or by telephone. NHS Direct can also provide advice. We shall not be liable for any failure to follow this advice. University College London Hospitals NHS Foundation Trust (UCLH). ------ACB discussion List Information-------- This is an open discussion list for the academic and clinical community working in clinical biochemistry. Please note, archived messages are public and can be viewed via the internet. Views expressed are those of the individual and they are responsible for all message content. ACB Web Site http://www.acb.org.uk Green Laboratories Work http://www.laboratorymedicine.nhs.uk List Archives http://www.jiscmail.ac.uk/lists/ACB-CLIN-CHEM-GEN.html List Instructions (How to leave etc.) http://www.jiscmail.ac.uk/