title: Development and validation of a microfluidic chip-based nano-liquid chromatography-triple quadrupole tandem mass spectrometry method for a sensitive and reliable quantification of 7-ethyl-10-hydroxycamptothecin (SN38) in mouse plasma.
abstract: There is an increasing need for more sensitive analytical methods in pharmacokinetic studies, for example, for phase 0 clinical trials. A novel HPLC Chip-triple quadrupole mass spectrometer method (HPLC Chip-MS/MS method) for the quantification of 7-ethyl-10-hydroxycamptothecin (SN38) was developed, validated, and employed to the pharmacokinetic analysis of SN38 in ICR mice. Protein precipitation with a ratio of plasma/acetonitrile of 1:10 was chosen as the sample processing method. The nano-electrospray inserted in the microfluidic chip operated in positive mode, and selected reaction monitoring was used for quantification. Our bioanalytical method met all essential validation parameters-selectivity, accuracy, precision, dilution integrity, calibration curve, matrix effect, recovery, and different stability tests (benchtop, freeze-thaw, autosampler stability). The calibration curves (weight 1/x (2)) were linear for the range 50-10,000 pg/mL. Clogging was not observed until the end of the lifetime of the microfluidic chip (350-400 injections), and carryover was practically eliminated through the introduction of a step gradient elution program. After intraperitoneal injection of 0.1 mg/kg irinotecan, SN38 concentration could be measured up to 6 h with accuracy and precision. Thus, we developed a new, very sensitive HPLC Chip-MS/MS method for the determination of plasma SN38 that has been validated in compliance with guidelines from different regulation authorities.
authors.affiliation: Biomolecular Function Research Branch, National Cancer Center, 323 Ilsan-ro, Ilsandong-gu, Goyang-si, Gyeonggi-do, 410-769, Republic of Korea.
authors.forename: Gang
authors.initials: G
authors.lastname: Ahn
authors.forename: Dong Min
authors.initials: DM
authors.lastname: Park
authors.forename: Jun Won
authors.initials: JW
authors.lastname: Park
authors.forename: Joo-Youn
authors.initials: JY
authors.lastname: Cho
authors.forename: Su-jin
authors.initials: SJ
authors.lastname: Rhee
authors.forename: Hyo-Young
authors.initials: HY
authors.lastname: Kim
authors.forename: Dae-Seung
authors.initials: DS
authors.lastname: Lee
authors.forename: In-Jin
authors.initials: IJ
authors.lastname: Jang
authors.forename: Hark Kyun
authors.initials: HK
authors.lastname: Kim
mesh_terms: D000818:Animals; D002166:Camptothecin / Q000031:analogs & derivatives* / Q000032:analysis; D002853:Chromatography, Liquid / Q000379:methods*; D000077146:Irinotecan; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D044085:Microfluidics / Q000379:methods*; D036103:Nanotechnology / Q000379:methods*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D000077146:Irinotecan; D002166:Camptothecin