Milli q plus purification equipment

Pooled HLM (M0567) from male donors (20 mg mL⁻¹ in 250 mM sucrose buffer) were procured from Sigma-Aldrich (St. Louis, MO, USA) and stored at −70 °C until use. All solvents used in the study were of HPLC grade. All chemicals and reference powders were of analytical (AR) grade. Pemigatinib (99.88%) and flavopiridol (99.72%) were procured from MedChem Express Company (Princeton, NJ, USA). Acetonitrile and formic acid were purchased from Sigma-Aldrich (St. Louis, MO, USA). Water (HPLC grade) was obtained from an in-house Milli-Q plus purification equipment that was procured from Millipore (Billerica, MA, USA). Waters Acquity UPLC [serial number H10UPH model code UPH] and Acquity TQD MS [serial number QBB1203 and model code TQD] were used for chromatographic separation and mass detection of analyte peaks. The LC-MS/MS system was controlled by MassLynx 4.1 Data acquisition, processing and reporting were automatically performed using ‘QuanLynx’ included in the MassLynx Software package (Version 4.1, SCN 805). Mass tuning was assisted with IntelliStart®. Rotary pump (SV40B; Murrysville, PA, USA) was used for generating vacuum and a nitrogen generator from Peak Scientific (Renfrewshire, Scotland, UK) was used for supplying desolvation gas. Argon gas of 99.999% purity was obtained locally.

The solvents used in the current study were all HPLC grade. The reference powders and other chemicals used were of analytical grade (AR). Hydroxychloroquine sulfate (HCQ) (CAS H916900) and chloroquine diphosphate salt (CLQ) (CAS C379965) were purchased from Toronto Research Chemicals (Toronto, ON, Canada) (Fig. 1). The acetonitrile (ACN) and formic acid (FA) used in the experiment were supplied by Sigma–Aldrich Company (PA, USA). Millipore Milli-Q Plus purification equipment (Millipore, MA, USA) was employed to purify water (HPLC grade). MassLynx 4.1 software was used to control the system. Application manager QuantLynx supplied with MassLynx 4.1 Software (Version 4.1, SCN 805) was utilized to acquire, process, and report data. IntelliStart® assisted in the mass tuning process. Additionally, a rotary pump (Sogevac, SV40B1) was used to assist the vacuum, and a nitrogen generator (Peak Scientific, Scotland) was used to provide the desolvation gas. The purity of argon gas (Ar) was 99.999% and was provided by a local supplier.Fig. 1

Chemical structure for the hydroxychloroquine and chloroquine (IS).