The objectives of this study were to determine the suitability of total serum lipid (TSL) concentrations, which were calculated by three different formulae, for pregnant women and to optimize the improved matrix solid-phase dispersion (MSPD) method. The results showed that the TSL predicted by the three formulae were significantly correlated to our measured values (sums of the total cholesterol, triglycerides and phospholipids contents). In particular, one of formulae was the most statistically suitable for pregnant women. Meanwhile, an improved MSPD method was developed to extract 22 POPs. The developed MSPD method was compared with SPE and EPA (8081 & 8082) to evaluate the performance of each extract method. The method validation showed that the results obtained using the improved MSPD method were closest to the actual concentration (adjusted by lipids), and the dispersion of the data was minimal.