Optima lc ms grade water

Hydroxychloroquine sulfate (HCQ) was purchased from TCI Chemicals (Portland, OR, USA) (catalog H1306). Deuterated hydroxychloroquine-d4 (HCQ-d4) were purchased from Toronto Research Chemicals (Toronto, ON, Canada) as the analytical internal standard (IS). 2-Deoxy-D-glucose (2-DG) was purchased from Sigma-Aldrich (St. Louis, MO, USA) (catalog number D6134). ¹³C₆-2-deoxy-D-glucose (¹³C₆-2-DG) and ¹³C₁-2-DG (in initial method testing) were purchased from Toronto Research Chemicals (Toronto, ON, Canada) as IS for 2-DG. Formic acid was purchased from J. T. Baker (Phillipsburg, New Jersey, USA). Optima LC-MS grade water, acetonitrile and methanol and other chemicals were purchased from Fisher Scientific (Fair Lawn, New Jersey, USA).

The separation was achieved using a
Dionex UltiMate 3000 Nano LC system (NCS-3200RS, Thermo Scientific,
Germering, Germany) fitted with a micro-LC flow selector to deliver
a mobile phase. Channel A contained 0.1% trifluoroacetic acid (TFA,
Sigma Aldrich) in Optima LC-MS grade water (Fisher Chemical), and
channel B contained 0.1% TFA in LC-MS grade acetonitrile (Fisher Chemical).
Isocratic elution at 20% channel B was used at a constant flow rate
of 2 μL/min for the entirety of the experiment. The pump was
connected to an externally mounted 6-port two-position Cheminert injection
valve (C72x-669D, VICI Valco, Houston, TX) using a 750 mm × 0.100
mm nanoViper capillary. A 140 mm × 0.100 mm nanoViper capillary
was used as a 1.1 μL sample loop. A 350 mm × 0.025 mm i.d.
× 0.360 mm o.d. fused silica capillary was used to connect the
outlet of the column to a Waters Acquity TUV detector fitted with
a 10 nL nano flow cell (Waters Corporation, Milford, MA) set to 214
nm. An Atlas analog-to-digital converter and Chromeleon version 6.8
software (Thermo) were used to acquire data at 100 Hz.
The yaGfl
standards were injected in triplicate (21 data). The regression results
from data (peak area vs injected concentration) indicated an intercept
indistinguishable from 0 (95% CI of −0.0026 to +0.0049) and
a slope of 0.00575 (95% CI of 0.00566 to 0.00583)

The GluCer isoform mixture from the Gaucher spleen (˃98%), galactosylceramide (GalCer) isoform mixture (˃98%), and GluCer(C16:0)D₃ were purchased from Matreya (Pleasant Gap, PA, USA). Optima LC/MS grade water and A.C.S. reagent grade ammonium formate (amm. form.) were from Fisher Scientific (Fair Lawn, NJ, USA). HPLC grade methanol (MeOH) and acetonitrile (ACN), and A.C.S. grade acetone were supplied by EMD Chemicals Inc. (Darmstadt, Germany). ReagentPlus grade dimethyl sulfoxide (DMSO) (≥99.5%) and terfenadine were from Sigma-Aldrich (St. Louis, MO, USA) and formic acid (FA) (˃99%) from Acros Organics (Morris Plain, NJ, USA).