title: Development and validation of a sensitive quantification method for hematoporphyrin monomethyl ether in plasma using high-performance liquid chromatography with fluorescence detection.
abstract: A rapid, sensitive, precise and specific method for determination of hematoporphyrin monomethyl ether (HMME), a novel photodynamic therapy (PDT) drug, was developed and validated using high-performance liquid chromatography (HPLC) with fluorescence detection. HMME was isolated from the plasma by a single-step liquid-liquid extraction with ethyl acetate. The analyte and internal standard fluorescein were baseline separated on a Diamonsil C(18) analytical column (4.6 x 150 mm, 5 microm) and analyzed using a fluorescence detector with the excitation and emission wavelengths set at 395 and 613 nm, respectively. The method was linear in the concentration range 0.025-5 microg/mL with a lower limit of quantitation (LLOQ) of 10 ng/mL. The inter- and intra-day accuracies and precisions were all within 10% and the mean recoveries of HMME and fluorescein were 95 +/- 3.7 and 90 +/- 2.3%, respectively. The analyte was stable during all sample storage, preparation and analysis periods. This method was successfully applied to a pharmacokinetic study after a single-dose intravenous administration of HMME (5 mg/kg) to beagle dogs. This method was reproducible and sensitive enough for the pharmacokinetic study of HMME. Based on the results of the pharmacokinetic study, we suggest that a rather long light-avoiding time is essential for patients under HMME therapy.
authors.affiliation: Key Laboratory of Drug Metabolism and Pharmacokinetics, China Pharmaceutical University, Nanjing 210009, People's Republic of China.
authors.forename: Peng
authors.initials: P
authors.lastname: Li
authors.forename: Jian-guo
authors.initials: JG
authors.lastname: Sun
authors.forename: Chen-rong
authors.initials: CR
authors.lastname: Huang
authors.forename: Guo-Yu
authors.initials: GY
authors.lastname: Pan
authors.forename: Mei-juan
authors.initials: MJ
authors.lastname: Xu
authors.forename: Jing
authors.initials: J
authors.lastname: Li
authors.forename: Guang-ji
authors.initials: GJ
authors.lastname: Wang
authors.forename: Ji-ning
authors.initials: JN
authors.lastname: Tao
mesh_terms: D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004285:Dogs; D004355:Drug Stability; D005453:Fluorescence; D006415:Hematoporphyrins / Q000097:blood* / Q000493:pharmacokinetics; D008297:Male; D010778:Photochemotherapy; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D006415:Hematoporphyrins; C494379:hematoporphyrin monomethyl ether