Direct on-line hyphenation of capillary liquid chromatography to nuclear magnetic resonance spectroscopy: practical aspects and application to drug metabolite identification.
In combining the high peak concentrations of capillary liquid chromatography (CapLC) with the high mass sensitivity of micro scale nuclear magnetic resonance (NMR) the hyphenation of CapLC to micro NMR offers a substantial gain in overall sensitivity. This paper deals with our experiences gained using a commercial CapLC-NMR system which has very recently become available. The limits of detection (SNR > 3) for a test compound of a molecular weight of M 318 were found to be approximately 100 ng (0.35 nmol) within an hour acquisition time and approximately 25 ng over night (85 pmol). Practical aspects such as the feasibility of stopped-flow experiments and sample handling issues are discussed in detail and first possible drug metabolite applications to hepatocyte incubations and direct analysis of plasma samples are presented.
DMPK Structural ID Group, GlaxoSmithKline R&D, Park Road, Ware, Herts SG12 0DP, UK. martin.x.sandvoss@gsk.com
Martin
M
Sandvoss
Andy D
AD
Roberts
Ismail M
IM
Ismail
Stephanie E
SE
North
D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000032:analysis / Q000097:blood / Q000378:metabolism; D001711:Biotransformation; D002853:Chromatography, Liquid / Q000379:methods*; D004008:Diclofenac / Q000032:analysis / Q000097:blood / Q000378:metabolism; D020719:Glucuronides / Q000032:analysis / Q000097:blood; D022781:Hepatocytes / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D009862:Online Systems; D004364:Pharmaceutical Preparations / Q000737:chemistry / Q000378:metabolism*; D051381:Rats; D013056:Spectrophotometry, Ultraviolet; D014783:Viscosity
D000894:Anti-Inflammatory Agents, Non-Steroidal; D020719:Glucuronides; D004364:Pharmaceutical Preparations; D004008:Diclofenac