Supplementary Material

Clinical Relevance of Liquid Chromatography Tandem Mass Spectrometry as an Analytical Method in Microdose Clinical Studies

Naoe Yamane • Zenzaburo Tozuka • Makiko Kusama • Kazuya Maeda • Toshihiko Ikeda • Yuichi Sugiyama

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Fig. S1a Chemical structures separated by sample treatment procedure. The positions for fragmentation from precursor ion to product ion are shown by dotted line. Used SPE cartridge: InertSep MC-1.

Fig. S1b Chemical structures separated by sample treatment procedure. The positions for fragmentation from precursor ion to product ion are shown by dotted line. Sample treatment procedure: SPE, used SPE cartridge; InertSep Pharma.

Fig. S1c Chemical structures separated by sample treatment procedure. The positions for fragmentation from precursor ion to product ion are shown by dotted line. Sample treatment procedure: SPE, used SPE cartridge; Oasis HLB.

Fig. S1d Chemical structures separated by sample treatment procedure. The positions for fragmentation from precursor ion to product ion are shown by dotted line. Sample treatment procedure: LLE under alkali.

Fig. S1e Chemical structures separated by sample treatment procedure. The positions for fragmentation from precursor ion to product ion are shown by dotted line. Sample treatment procedure: LLE under acid.

* It is considered that the elimination of SO2 and rearrangement of NH are simultaneously occurred while the yield from precursor ion to product ion.

Table S1 Mass spectrometric conditions for 31 compounds

Analyte / Polarity / Precursor ion / Product ion / Collision energy / Nebulizer gas / Turbo gas / Curtain gas / Collision gas / Ion spray voltage / Turbo gas
Temperature
(m/z) / (m/z) / (V) / (psi) / (psi) / (psi) / (no unit) / (V) / (°C)
Amlodipine / Positive / 409.1 / 238.0 / 15 / 50 / 30 / 20 / 5 / 5000 / 600
Atorvastatin / Positive / 559.3 / 440.1 / 31 / 40 / 70 / 10 / 7 / 5000 / 650
Bupropion / Positive / 240.1 / 184.0 / 17 / 70 / 70 / 20 / 5 / 5000 / 650
Candesartan / Positive / 441.2 / 263.1 / 19 / 30 / 70 / 30 / 5 / 5000 / 550
Celecoxib / Negative / 380.1 / 316.0 / –30 / 70 / 70 / 10 / 7 / –3500 / 600
Cetirizine / Positive / 389.2 / 201.1 / 25 / 30 / 70 / 10 / 7 / 5000 / 600
(S)-Citalopram / Positive / 325.1 / 109.0 / 35 / 70 / 70 / 10 / 7 / 5000 / 600
Clopidogrel / Positive / 322.1 / 211.9 / 23 / 70 / 70 / 20 / 6 / 4000 / 600
Docetaxel / Positive / 808.4 / 527.3 / 11 / 30 / 30 / 20 / 8 / 5500 / 500
Donepezil / Positive / 380.3 / 91.2 / 53 / 30 / 70 / 10 / 10 / 5000 / 600
Esomeprazole / Positive / 346.2 / 198.0 / 19 / 70 / 30 / 40 / 5 / 5000 / 600
Imatinib / Positive / 494.2 / 394.1 / 37 / 70 / 70 / 10 / 7 / 4500 / 600
Lansoprazole / Positive / 370.1 / 252.0 / 17 / 40 / 70 / 20 / 4 / 5000 / 600
Levofloxacin / Positive / 362.2 / 318.3 / 29 / 70 / 50 / 10 / 6 / 5500 / 600
Losartan / Positive / 423.2 / 207.0 / 33 / 30 / 70 / 10 / 5 / 5000 / 600
Montelukast / Positive / 586.1 / 422.1 / 35 / 70 / 70 / 10 / 4 / 4500 / 500
Oseltamivir / Positive / 313.2 / 166.1 / 27 / 50 / 70 / 10 / 4 / 4500 / 600
Olanzapine / Positive / 313.2 / 256.0 / 33 / 70 / 30 / 10 / 5 / 4500 / 600
Pantoprazole / Positive / 384.1 / 200.0 / 19 / 40 / 70 / 20 / 4 / 5000 / 600
Pioglitazone / Positive / 357.1 / 134.2 / 41 / 70 / 70 / 20 / 6 / 4000 / 600
Quetiapine / Positive / 384.1 / 253.1 / 31 / 40 / 70 / 10 / 6 / 4500 / 600
Risperidone / Positive / 411.2 / 191.1 / 39 / 70 / 70 / 20 / 4 / 4500 / 600
Rosiglitazone / Positive / 358.2 / 135.2 / 39 / 60 / 70 / 10 / 9 / 5000 / 550
Rosuvastatin / Positive / 482.1 / 258.2 / 49 / 70 / 60 / 10 / 7 / 5500 / 600
Sertraline / Positive / 306.2 / 158.9 / 41 / 40 / 70 / 20 / 7 / 4500 / 600
Simvastatin / Positive / 436.4 / 199.2 / 21 / 70 / 50 / 10 / 4 / 5500 / 550
Tamusulosin / Positive / 409.1 / 228.1 / 35 / 60 / 70 / 20 / 7 / 4000 / 600
Topiramate / Negative / 337.9 / 78.0 / –40 / 70 / 70 / 20 / 7 / –4000 / 600
Valsartan / Positive / 436.2 / 291.2 / 25 / 30 / 70 / 10 / 5 / 4500 / 500
Venlafaxine / Positive / 278.2 / 58.1 / 35 / 50 / 70 / 20 / 6 / 5000 / 600
Zolpidem / Positive / 308.0 / 235.0 / 47 / 50 / 70 / 10 / 7 / 4000 / 600

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Table S2 Liquid chromatographic conditions for 31 compounds

Analyte / Analytical / Mobile / Flow rate / Retention time
column / phase / (mL/min) / (min)
Amlodipine / C / IV / 0.2 / 1.7
Atorvastatin / C / IV / 0.3 / 1.3
Bupropion / F / I / 0.2 / 3.5
Candesartan / F / V / 0.2 / 2.9
Celecoxib / C / I / 0.3 / 1.6
Cetirizine / D / VII / 0.3 / 1.7
(S)-Citalopram / E / II / 0.15 / 3.2
Clopidogrel / A / I / 0.5 / 0.9
Docetaxel / G / VI / 0.3 / 1.2
Donepezil / B / VIII / 0.3 / 1.0
Esomeprazole / F / I / 0.2 / 2.4
Imatinib / F / I / 0.2 / 3.0
Lansoprazole / F / I / 0.2 / 2.5
Levofloxacin / H / V / 0.2 / 1.4
Losartan / F / IV / 0.2 / 2.6
Montelukast / A / IV / 0.6 / 1.0
Oseltamivir / C / IV / 0.2 / 0.95
Olanzapine / G / I / 0.2 / 1.8
Pantoprazole / F / I / 0.2 / 2.3
Pioglitazone / F / I / 0.2 / 3.4
Quetiapine / F / I / 0.2 / 3.3
Risperidone / F / I / 0.2 / 2.8
Rosiglitazone / F / II / 0.2 / 4.0
Rosuvastatin / G / I / 0.2 / 1.8
Sertraline / G / I / 0.2 / 2.5
Simvastatin / G / III / 0.2 / 3.6
Tamusulosin / F / I / 0.2 / 2.6
Topiramate / C / I / 0.3 / 1.0
Valsartan / F / IV / 0.2 / 3.2
Venlafaxine / F / I / 0.2 / 2.8
Zolpidem / F / I / 0.2 / 2.7

A: ACQUITY UPLC BEH C18 2.1×50 mm, 1.7 μm (Waters)

B: ACQUITY UPLC BEH C18 2.1×100 mm, 1.7 μm (Waters)

C: ACQUITY UPLC BEH Shield RP18 2.1×100 mm, 1.7 μm (Waters)

D: Atlantis HILIC Silica 2×150 mm, 3 μm (Waters)

E: Cadenza CD-C18 2×150 mm, 3 μm (Imtakt)

F: Phenomenex Aqua C18 2×150 mm, 5 μm (Phenomenex)

G: Phenomenex Luna C18(2) 2×100 mm, 2.5 μm (Phenomenex)

H: TSK-gel ODS-100V 2×150 mm, 3 μm (TOSOH)

I: Acetonitrile/10 mM ammonium formate (7:3, v/v)

II: Acetonitrile/10 mM ammonium formate (65:35, v/v)

III: Acetonitrile/10 mM ammonium acetate-acetic acid buffer (pH4.5) (85:15, v/v)

IV: Acetonitrile/0.1% formic acid (7:3, v/v)

V: Acetonitrile/0.1% formic acid (65:35, v/v)

VI: Methanol containing 0.1% formic acid/0.1% formic acid (9:1, v/v)

VII: Methanol/0.1% formic acid (98:2, v/v)

VIII: Methanol/10 mM ammonium acetate (87:13, v/v)

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