Method development and laboratory intercomparison of an RP-HPLC-UV method for energetic chemicals in marine tissues

Experiments were conducted to develop a method for the determination of a set of 17 military-relevant energetic compounds (including nitroaromatics, nitramines, and nitrate esters) in 5 types of marine tissues (Dungeness crab, Manila clam, starry flounder, sea cucumber, and geoduck) using reversed-phase high performance liquid chromatography with a UV detector (RP-HPLC-UV). Dry-ice grinding was evaluated and found to be an excellent method of sample homogenization prior to sample extraction and determination. An extract cleanup procedure based on solid-phase extraction was assessed. A cleanup procedure using solid phase extraction was adequate for the removal of interferences prior to HPLC analysis for the five marine tissue matrices tested. Mean method detection limits (MDLs) were estimated using two columns at two wavelengths (254 and 210 nm) and ranged from 17 to 293 µg/kg for the five tissue matrices tested. A six-laboratory intercomparison test was conducted to evaluate the performance of the method, each analyzing five marine tissue matrices fortified at three levels. The same marine tissues were used in the laboratory intercomparison study except Pacific halibut was substituted for starry flounder. Overall, USEPA Method 8330B modified for tissue analysis showed suitable detection capability, analytical accuracy, precision, sensitivity, linear range, and robustness for sixteen (16) of the seventeen (17) analytes, for all five (5) of the marine tissue matrices studied. The exception was tetryl that proved to be unstable for all matrices as has been found for soils and sediments.