Hi everyone,
I'm looking to find a way to determine the concentration of the ketone body Beta-hydroxybutyrate (BHB) within cell culture media. Preferably, a method that would employ the use of analytical techniques like HPLC or LCMS. I am not sure about what solvents to use for LCMS. According to literature, it says:
water+0.1% acetic acid (eluent A) and acetonitrile/isopropanol (1:1)+0.1% acetic acid (eluent B)
Hi Iqra Pervaiz ,
for determination of BHB in biological metarial (e.g. blood) as a mobile phase I'm using a mixture of 10 mM ammonium formate and 0.1% formic acid in water (A) and 0.1% formic acid in methanol (B) (the method is UHPLC-QqQ-MS/MS ). For determination of BHB I'm also using GS-MS (after derivatisation of sample with N-tert_Buthyldimethylsilyl-N-methyltrifluoroacetamide with 1 % tert-Butyldimethylchlorosilane).
Best regards,
Karolina Nowak
Dear Iqra;
I agree with Karolina Nowak this is true. I usually using GS-MS (after derivatization of the sample with N-tert_Buthyldimethylsilyl-N-methyltrifluoroacetamide with 1 % tert-Butyldimethylchlorosilane).
Karolina Nowak and Ladan Ajdanian thankyou for your response. So I assume, I could use same for my experiment where I need to detect ketone bodies in cell culture media. I am using stem cells for this.
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