Analysis of Nicotine and Related Compounds in Urine Using Raptor Biphenyl

As Applications Chemists in the LC lab, one of the most exciting parts of our jobs is the variety of analyses we are exposed to. One day you are developing a method for potency analysis in cannabis samples, the next you are looking at anti-epileptic drugs in urine.    We’re regularly challenged to think outside the box to provide the best solutions for our customers.

In some recent work, my colleague, Shun-Hsin Liang, was tasked to develop a method for nicotine and related compounds in urine using LC-MS/MS.   The challenge was finding suitable analytical conditions for a rapid, accurate, and reproducible method geared toward high-throughput testing laboratories.    Typically, this analysis uses high-pH chromatography with relatively high concentrations of additives to increase retention, improve peak shape, and reduce peak tailing.   These conditions may lead to some additional non-routine maintenance (like changing pump seals and replacing tubing), and if you’re in a high-throughput laboratory, you know non-routine maintenance events always pop up at the most inopportune time.

Shun-Hsin was able to use the Raptor Biphenyl column and standard, low pH, reversed-phase LC-MS mobile phases that are compatible with a variety of LC-MS instrumentation. The method provided excellent performance for the simultaneous analysis of nicotine, two major metabolites (cotinine and trans-3’-hydroxycotinine), two minor metabolites (nornicotine and norcotinine), and a minor tobacco alkaloid, anabasine, in human urine. Accurate and reproducible analysis was achieved in less than 5 minutes of chromatographic analysis time, making the column and method well suited to low-cost, high-throughput analysis of nicotine-related compounds.