Development and Validation of an HPLC Method for Determination of Purity of Sn-ADAM, a Novel Precursor of Serotonin Transporter SPECT Imaging Agent I-123-ADAM

HANG-HSING YANG1, KUNG-TIEN LIU1*, YI-CHIH HSIA1, WEI-HSI CHEN1, CHIA-CHIEH CHEN2, LEE-CHUNG MEN1 AND LIE-HANG SHEN3

1. Chemical Analysis Division, Institute of Nuclear Energy Research,
1000, Wenhua Rd., Jiaan Village, Lungtan, Taoyuan 32546, Taiwan, R.O.C.
2. Isotope Application Division, Institute of Nuclear Energy Research,
1000, Wenhua Rd., Jiaan Village, Lungtan, Taoyuan 32546, Taiwan, R.O.C.
3. Institute of Nuclear Energy Research, 1000, Wenhua Rd., Jiaan Village, Lungtan, Taoyuan 32546, Taiwan, R.O.C.

(Received: February 27, 2010; Accepted: May 21, 2010)

ABSTRACT

A reversed-phase high performance liquid chromatography (RP-HPLC) method for the purity assay of Sn-ADAM (2-((2-((Dimethylamino)methyl)phenyl)thio)-5-(tri-n-butyltin)-phenylamine) was developed and validated. Sn-ADAM is a precursor and free ligand of I-123-ADAM (I-123-2-([2-({dimethylamino}methyl)phenyl]thio)-5-iodophenylamine) that is a SPECT imaging agent for serotonin transporters (SERTs). The chromatographic separation for Sn-ADAM was achieved on a RP-18 column with an eluent consisting of methanol-acetonitrile-ammonium acetate (pH 7.0, 10 mM; 58.8 : 39.2 : 2, v/v/v). The absorbance at 220 nm against the concentration of Sn-ADAM was linear over the range of 9.9-317.8 ppm, with a correlation coefficient (r) of 0.99997. Method validation parameters, including specificity, linearity, precision, accuracy, LOD/LOQ, stability, robustness and suitability were evaluated, indicating the potential of this method in pharmaceutical quality control. Moreover, present results of precursor to product ion transitions of Sn-ADAM and I-127-ADAM by LC/ESI/MS/MS provided evidence for identification of fragmentation ions and proposition of the pathways of Sn-ADAM, I-127-ADAM and I-123-ADAM.

Key words: Sn-ADAM, RP-HPLC, LC/ESI/MS/MS, purity assay, method validation, fragmentation pathway