High pressure liquid chormatography determination of the concentration and integrity of L-thyroxine in free T4 stock solution

Characterization studies were designed to evaluate the concentration and integrity of the L-thyroxine (T4) molecule (3,5,3',5'-tetraiodothyronine) in the free T4 stock solution (FT4SS) (code 99544). The determination of the concentration of T4 in FT4SS is critical to ensure that the free T4 calibrators and controls are manufactured with the least number of adjustments possible. The most significant conclusions drawn from these characterization studies are the following: (1) An accurate and sensitive HPLC method has been developed to measure the T4 concentration in FT4SS. The root cause of the failure of FT4SS to pass retest/ review is the presence of an unknown T4 degradation product with significantly higher molar extinction coefficient at 230 nm than T4 itself. The L-thyroxine concentration reference comparison spectrophotometric test with the current 43 to 58 ug/ml specification range (as per scp.99544, ed. 13A) is adequate to monitor the generation of the unknown T4 degradation product. The characterized T4 degradation product is not 3,5,3'-triiodo-thyronine (T3) and it is suspected that the identity of the degradation product is reverse T3 (3,3',5'-triiodothyronine). The use of sodium l-thyroxine pentahydrate (Na- T4-5H2O) as the equivalent of T4 (free base) is adequate provided that an excess of 15over the desired amount of T4 is weighed.

Effect of three extraction techniques on submitochondrial particle and Microtox bioassays for airborne particulate matter

The effect that three extraction techniques (e.g., Soxhlet, ultrasound and microwave-assisted extraction) have on the toxicity, as measured by submitochondrial particle (SMP) and Microtox assays, of organic extracts was compared from three sources of airborne particulate matter (APM). The extraction technique influenced the toxicity response of APM extracts and it was dependent on the bioassay method, and APM sample source. APM extracts from microwave-assisted extraction (MAE) were similar or more toxic than the conventional extraction techniques of Soxhlet and ultrasound, thus, providing an alternate extraction method. The microwave extraction technique has the advantage of using less solvent volume, less extraction time, and the capacity to simultaneously extract twelve samples. The ordering of APM toxicity was generally urban dust > diesel dust > PM10 (particles with diameter < 10 microm), thus, reflecting different chemical composition of the samples. This study is the first to report the suitability of two standard in-vitro bioassays for the future toxicological characterization of APM collected from Puerto Rico, with the SMP generally showing better sensitivity to the well-known Microtox bioassay.