In LC operation for LC-MS, the preferred option is a reversed-pha

In LC operation for LC-MS, the preferred option is a reversed-phase system using a gradient or isocratic solvent mixture of water, ACN, or MeOH. Small amounts of acetic acid, formic acid, ammonium hydroxide/ammonia solution, or ammonium acetate can also be used in the mobile phase. In conjunction with these selleck chem inhibitor interfaces, different types of analyzers, e.g., quadrupole, ion trap, or TOF, can be used, and they offer various degrees of mass accuracy and MS-MS possibilities. LC-MS systems do not allow a complete and unambiguous on-line identification of a component, unless it is a well-known natural product, and complementary on-line spectroscopic information is available in databases. One of the main problems associated with LC-MS is that the quality of response strongly depends on various factors, e.

g., nature of the compounds to be analyzed, the solvent and buffer used as the mobile phase, the flow rate and, of course, the type of interface used. For example, a crude natural product extract generally contains a number of various types of compounds that differ considerably in their physicochemical properties, solubilities, molecular size and stability. It is therefore extremely difficult, if not impossible, to optimize the ionization conditions that can be suitable for all those different types of compounds. One way to get around this difficulty is to analyze the extract in different ionization modes.[10] LC-NMR Among the spectroscopic techniques available to date, NMR is probably the least sensitive, and yet it provides the most useful structural 240 Sarker and Nahar information toward the structure elucidation of natural products.

Technological developments have allowed the direct parallel coupling of HPLC systems to NMR, giving rise to the new practical technique HPLC-NMR or LC-NMR, which has been widely known for more than last 15 years. The first on-line HPLC-NMR experiment using superconducting magnets was reported in the early 1980s. However, the use of this hyphenated technique in the analytical laboratories started in the latter part of the 1990s only. LC-NMR promises to be of great value in the analysis of complex mixtures of all types, particularly the analysis of natural products and drug-related metabolites in biofluids. LC-NMR experiments can be performed in both continuous-flow and stop-flow modes.

A wide range of bioanalytical problems can be addressed using 500, 600, and 800 MHz systems with 1H, 13C, 2H, 19F, and 31P probes. The main prerequisites for on-line LC-NMR, in addition to the NMR and HPLC instrumentation, are the continuous-flow Cilengitide probe and a valve installed before the probe for recording either continuous-flow or stopped-flow NMR spectra.[12] A UV�Cvis detector is also used as a primary detector for LC operation. Magnetic field strengths higher than 9.

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