Analytical and Bioanalytical Chemistry

, Volume 377, Issue 1, pp 208–213 | Cite as

Separation and determination of the antitumor drug piritrexim by molecularly imprinted microspheres in high-performance liquid chromatography

  • Jia-Ping Lai
  • Xi-Wen He
  • Feng ChenEmail author
Original Paper


Molecularly imprinted microspheres were synthesised using the antitumor drug piritrexim (PTX) as a template molecule by aqueous microsuspension polymerisation and were used as a high-performance liquid chromatographic stationary phase. The molecularly imprinted column exhibited strong retention behaviour to the template molecule. The influences of pH of the buffer and the ratio of methanol to buffer on the retention behaviour were investigated in detail. Results indicated that the baseline separation of PTX, trimetrexate (TMX), trimethoprim (TMP) and sulfamethazine (SMZ) was achieved when the pH value of the acetate buffer was above pH 3.5 and the ratio of methanol to the buffer was 6:4 (v/v). A gradient elution programme was employed to enhance the separation, which led to an improvement in sensitivity and a reduction in determination errors. The method developed was used to analyse urine samples supplemented with PTX. The recoveries of 5 μg mL−1 PTX in the urine sample were 99.1±3.0% and 93.3±2.8% at the beginning and 24 h later, respectively.


Molecularly imprinted microspheres Aqueous microsuspension polymerisation Chromatographic stationary phase Piritrexim Trimetrexate 



This research was supported by the CRCG (The University of Hong Kong Committee on Research and Conference Grants), RGC (the Hong Kong Research Grants Council) and the National Natural Science Foundation of China (No. 29775014, No. 20175009).


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Copyright information

© Springer-Verlag 2003

Authors and Affiliations

  1. 1.Department of BotanyThe University of Hong KongHong KongChina
  2. 2.Department of ChemistryNankai UniversityTianjinChina

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