Use of basic mobile phase to improve chromatography and boost sensitivity for quantifying tetrahydrocurcumin in human plasma by LC-MS/MS.
Tetrahydrocurcumin (THC), a major metabolite of curcumin, is often quantified by LC-MS or LC-MS/MS using acidic mobile phases due to the concern of its instability in a basic medium. However, acidic mobile phases often lead to poor chromatography (e.g. split or double peaks) and reduced detection sensitivity in the commonly used negative ionization mode. To overcome these shortcomings, a basic mobile phase was used for the first time in the LC-MS/MS quantification of THC. In comparison with the acidic mobile phases, a single symmetrical chromatographic peak was obtained and the sensitivity increased by 7-fold or more under the equivalent conditions. The new LC-MS/MS method using the basic mobile phase has been successfully validated for the quantification of THC in human EDTA plasma over the concentration range of 5-2500ng/ml. The within-batch accuracy (% nominal concentration) was between 88.7 and 104.9 and the between-batch accuracy ranged from 96.7 to 108.6. The CVs for within- and between-batch precisions were equal to or less than 5.5% and 9.1%, respectively. No significant matrix interference or matrix effect was observed from normal or lipemic and hemolytic plasma matrices. In addition, the common stabilities with adequate durations were established, including up to 5days of post-preparative stability. Furthermore, when the validated method was applied to a clinical study, the passing rate of ISR samples was 83%, indicating the good reproducibility of the method. The success of the unconventional approach presented in this article demonstrates that a mobile phase could be selected based mainly on its merits to facilitate LC separation and/or MS detection. There is no need for excessive concern about the stability of the compound(s) of interest in the selected mobile phase because the run time of modern LC-MS or LC-MS/MS methods is typically only a few minutes.
Nucro-Technics, Scarborough, ON, Canada. Electronic address: tan@nucro-technics.com.
Aimin
A
Tan
Nucro-Technics, Scarborough, ON, Canada.
Yanxin
Y
Wu
Nucro-Technics, Scarborough, ON, Canada.
Molly
M
Wong
Nucro-Technics, Scarborough, ON, Canada.
Albert
A
Licollari
Nucro-Technics, Scarborough, ON, Canada.
Gordon
G
Bolger
Nucro-Technics, Scarborough, ON, Canada.
John C
JC
Fanaras
Shopp Nonclinical Consulting LLC, Boulder, CO, USA.
George
G
Shopp
SignPath Pharma, Inc., Quakertown, PA, USA.
Lawrence
L
Helson
D000970:Antineoplastic Agents / Q000378:metabolism / Q000493:pharmacokinetics; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003474:Curcumin / Q000031:analogs & derivatives* / Q000378:metabolism / Q000493:pharmacokinetics; D006801:Humans; D057230:Limit of Detection; D010949:Plasma / Q000378:metabolism; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*
D000970:Antineoplastic Agents; C096277:tetrahydrocurcumin; D003474:Curcumin
Basic mobile phase; Bioanalysis; Curcumin; LC–MS; Plasma; Tetrahydrocurcumin