bioanalytical approaches for early assessment of ...jun 14, 2018 · bioanalytical approaches for...

Bioanalytical Approaches for Early Assessment of Metabolite Exposures in Human and Animals to Address

MIST Fundamentals Strategy and Perspectives

Hongying Gao

Innovo Bioanalysis LLC

June 14 2018

Acknowledgements

R Scott Obach PDM Pfizer Inc

Shibing Deng Biostatistics Pfizer Inc

Safety Testing of Drug Metabolites Guidance

bull First FDA guidance was issued in Feb 2008 followed by ICH M3 (R2) in Jan 2010

bull Recent revision of FDA guidance was issued in Nov 2016

ndash Supersedes previous version and aligns with ICH M3(R2)

ndash 10 of total drug related exposure at steady state

ndash Approximately equal or greater exposure in animals

ndash Acknowledges the difficulties associated with metabolite synthesis

ndash Suggests early assessment of metabolite exposure in human and animals

Bioanalytical Challenges to Address MIST

bull Synthetic standards of metabolites

bull Simultaneous analysis of multiple analytes

bull Radiolabel ADME Studiesmdashonly single dose no steady state data

bull NMR-based standardsmdashbiosynthesis and isolation

Bioanalytical Approaches

Mixed Matrix Peak Area

Approaches (1)

Radiometric Calibrants (2)

NMR-Based Methods (3)

Conventional GLP Bioanalysis

Data Output

Animal vs Human Ratios

Absolute Concentrations



Reagent Needs

None Radio-labeled

Drug None

Metabolite Standards

Special Equipment Needs

None None NMR None

Resource Investment

Low Moderate High Very High

Uses MIST Coverage PK Data

MIST Coverage of total

PK Data MIST Coverage


MIST Tiered Approach

bull Use screening qualified or validated methods as function of stage of development and study type

Timmerman P et al 2010 Bioanalysis 2(7) 1185ndash1194

MIST Coverage is a Comparison Question

Animals have exposures of metabolites ge human

Not the exposure multiples

A simple LCMSMS method can provide the data to ascertain the exposure coverage without developing

and validating standard bioanalytical methods for every metabolite

Simple LCMSMS Peak Area Comparison

bull Principle Hamilton pool the time points Inject animal and human sample side-by-side on HPLC-MS compare peak area ratios of metabolites

bull Whatrsquos known bull HPLC retention time and fragmentation pattern for the

metabolite

bull Mixed matrix approach Routinely used in bioanalysis

Matrix effect is the same in samples provided the matrices are identical

bull Mix animal sample with control human matrix

bull Mix human sample with control animal matrix

bull Mix control animal and control human matrix to ensure no background interference

Time-Averaged Pooling (ldquoHamilton Poolsrdquo)

bull Plasma samples from different time points are mixed to provide a single sample in which the analyte concentration Cavg is AUC over the sampling interval

-0avg

AUCC

Hamilton Garnett Kline Clin Pharmacol Ther 1981

total

1-2-nn130201

V)t-(t)t-(t)t-(t)t-(t)t-(t

)t-(tV

010t

total

1-2-nn130201

1-n1nnt V

)t-(t)t-(t)t-(t)t-(t)t-(t

)t-(tV

total

1-2-nn130201

1-t V


)t-(tV

bull Can be used to reduce the number of samples to be analyzed and still yield exposure comparisons across species

Mixed Matrix Peak Area Comparison

Pooled Human Sample

Pooled Animal Sample

Mix with Opposite

Matrix Control

Control Mix

Add IS and

Solvent

Recover Supernatant Analyze by HPLC-MS

IS IS IS

Met

Met

Human Animal Matrix Control

(Human) AreaPeak IS

(Human) AreaPeak Metabolite

(Animal) AreaPeak IS

(Animal) AreaPeak Metabolite

Ratio Human to Animal

Control Mix

Mixed Human Sample

Mixed Animal Sample

Mixed Human Sample

Mixed Animal Sample

Gao et al DMD 2010 Gao and Obach DMD 2012

Protein Precipitation

bull Selectivity minus High separation resolution chromatography (a long gradient on

a large column) + MRM or high resolution MS (ge30K for parent ion) w product ion scan for major human metabolites (ge10 of total drug related material)

minus Low throughput is offset by the limited of samples

bull Sensitivity Typically not an issue minus Major human metabolites (ge10 of drug related total material)

and the animal is dosed at NOAEL minus MRM or high accurate MS for parent (plusmn 10ppm) is more

sensitive than full scan MSMS for metabolite ID

bull Matrix effect Normalized by mixed matrix approach

bull Linearity Serial dilutions of the samples in blank mixed plasma

How to Establish BA Parameters without Synthetic Standards

bull Stabilize the metabolites if procedures are known

bull Repetitive analysis of samples to cover the storage duration

bull Profile metabolitesstability reference (usually the parent drug) vs the storage time

bull Auto sampler stability re-injecting the supernatant 12-24 hr later

1day

dayX

sample) (Same AreaPeak reference Stable

sample) Same ( AreaPeak Metabolite

sample) (same AreaPeak reference Stable

species)(Any AreaPeak Metabolite

RatioStability

How to Establish Stability without Synthetic Standards

SN

N N

N

O

Cl

NH

Cl

Cl

O

OH

N

OCH3

CH3

NOH

OH

N

NH2

NH N O OH

N3

O

O

ziprasidone

diclofenac tacrine

terfenadine

dextromethorphan

zidovudine

S-Oxidation Reduction+Methylation N-Dealkylation

Hydroxylation Hydroxylation

Oxidation to Carboxylic Acid

Demethylation Glucuronidation

Variety of reaction types alterations in pKa multiple regioisomers

(AZT)

Blind Test 1 Stable Metabolites

M1 236gt134

M2 252gt209

Ziprasidonemetabolites MRM TIC in rat plasma 1

IS

Ziprasidone 413gt194

M9

M5

M10

Dilution Curve M1 236gt134


2 B

80 B

Column Kinetex C18 150 X 46 mm 26 um

Parent Metabolite Potential Instability

Naproxen Naproxen Acyl Glucuronide

Acyl bond migration

Ester hydrolysis

Carbamazepine Carbamazepine-1011-Epoxide

Epoxide hydrolysis

Salicylic Acid 23-Dihydroxybenzoic Acid

Catechol oxidation

Sulfadiazine N-Acetylsulfadiazine

Amide hydrolysis

Methylphenidate Ritalinic Acid

Ester hydrolysis

Nicotine Nicotine-Δ1rsquo5rsquo-Iminium Ion and Cotinine

Imine Oxidation

N

O NH2

MeO

O

O

OH

OH

OH

O OH

N

N+

N

NO

N

N

NH

OMeO

NH

OHO

OH

O

OH

OH

NH

SNH

N

N

O

O

O

NH2

SNH

N

N

O

O

OH

O

OH

N

O NH2

O

MeO

OH

O

Blind Test 2 Unstable Metabolites

Long Term Storage Stability of Naproxen Acyl-Glucuronide Conjugate

0

05

1

15

2

0 50 100 150

(Nap

roxe

n-G

luN

apro

xen

) D

ay X

Day

1

Storage Time at -20ordmC (Day)

Naproxpen-GluNaproxen normalized byDay1 response

Stabilized Naproxen-GluNaproxenNormalized by Day 1

Linear (Naproxpen-GluNaproxen normalizedby Day1 response)

Linear (Stabilized Naproxen-GluNaproxenNormalized by Day 1)

n = 8

Untreated samples

A ratio between 08-12 is deemed stable

Stabilized samples

Gao H and Obach RS (2012) DMD 401290-1296

Correlation of Experimental MS Ratio vs Nominal Ratio

R 0994plusmn0005 Slope 099plusmn0082

MSNM Ratio

Metabolites

MS

NM

Ra

tio

10

15

20

1_O

H_T

acrine

2_O

H_T

acrine

3_M

eth

oxym

orp

hin

4_O

H_D

iclo

fenc

4_O

H_T

acrine

5_O

H_D

iclo

fenc

AZ

T

AZ

T_glu

c

Dextr

om

eth

orp

han

Dextr

orp

han

Dic

lofe

nac

Fexofe

nadin

e

M1

M10

M2

M5

M9

tacrine

Terf

enadin

e

Zip

rasid

one

Geo FC

Geo FC 95

Geo FC 99

Average 125

95 confidence 183

99 confidence 207

Precision Accuracy and Confidence Level bull Showed that a peak area ratio of 2 means that it is at least equal or

greater exposure

bull Increased replicates will increase confidence

95 Confidence Interval (~2SD)

Gao H Deng S and Obach RS (2010) DMD 382147-2156

Confidence Level Increases Dramatically with Measured Ratio

Ratio=4 plt2 ppm

Ratio=2 plt001

Measured animalhuman ratio by LCMSMS


Probability of Nominal AnimalHuman ge1

What Drives High Confidence

bull Fundamental BA parameters are addressed Selectivity sensitivity linearity matrix effect stability precision and accuracy

bull SignalNoise Ratio Measured animalhuman 2

Measurement error (without cancellation) 025

SN (2-1)025=4

Limit of Detection A signal is real if SN=3

bull Minimal 3 independent animalhuman ratio measurements

bull Analysis reproducibility RSD of the ratio measurements (n=3) 33

bull Stability ratio for each metabolite plusmn20 (08-12) over the storage period

How to Evaluate Data Quality



Comparison 1 Giraffe vs Moose

Which one is taller

All that one needs to do is to have them stand next to each other The giraffe is much taller No need to know exact heights

Which one is taller

Too close to call just by looking at them -- The height measurements are needed

Comparison 2 Moose vs Whitetail Deer

If the animalhuman peak area ratio is ge 2 animal has exposure ge human with high confidence (plt001)

How often does it happen Most cases as animal is dosed at much higher

dose when corrected by body weight

A simple peak area ratio comparison is sufficient for most cases

Case Study

bull Drug A in project X at phase 1 14 days 35 mg BID in 6 human volunteers vs 5 days 300 mgkg in Sprague Dawley rats

bull Pooled plasma samples (Hamilton) across time points across subjects

bull 3 human metabolites Parent drug A 396gt203

m-hydroxy 412 ring opened 414 de-alkylated 304

bull HPLC method and major daughter ions were transferred from Biotransformation

bull Compare results with concentration measurements using the synthetic standard

MIST Coverage Evaluation Using RatHuman Ratio of Peak Area Ratios

Definitive Concentration Measurements Using Synthetic Standard

Species Parent De-alkylated Ring-opened m-Hydroxy

Human 0220 00406 000788 00210 Rat 770 0502 0292 000762

RatHuman ratio 1 35 124 371 0363

Human repeat 0184 00290 000506 00164

Rat repeat 734 0472 0266 000618 RatHuman

ratio 2 399 163 526 0378

Species m-Hydroxy (ngmL)

Human 743 plusmn 153

Rat 269 plusmn 258

Cavg RatHuman ratio 0362

Human Metabolite gt10 Drug-Related Material from a Human Radiolabel

ADME Study (or Metabolite Anticipated to Be gt10 from Other Human Studies)

Metabolite With Known Activity of

Interest

Use Standard Bioanalytical

Method

ICH Exempt Metabolite

Peak Area Ratio Method

Method Fails Criteria

Method Succeeds

Animal lt 2X Human

Animal ge 2X Human

No Further Work Needed

Coverage is Assured No Further Work

Needed

Prepare Authentic Standard of Metabolite

MIST Decision Tree

Gao H and Obach RS (2014) Bioanalysis 6(5) 641-650

Data Driven Strategy

bull Rigor of the analysis increases based on data Synthetic standard is needed only when the method fails or animal has less exposure than human

bull Conservative evaluation of MIST coverage

bull Assess MIST coverage at steady state without radio-label or synthetic standards enabling early assessment eg FIH

bull Time- and resource-saving strategy

Summary

bull LCMSMS peak area ratio comparison is scientifically sound and defensible method that can

ndash Ascertain the steady-state exposure of animals compared to humans (ie ldquoMIST coveragerdquo)

ndash Assess the MIST coverage without radio-label or synthetic standard

bull These methods are fit-for-purpose and can offer decision-making data useful for reliable estimation of metabolite exposure

bull Discussions with regulatory experts were positive and helpful

Gao et al (2013) AAPS Journal15(4) 970-973

Prerequisite for Current Methodology

bull Animal and human samples for side-by-side injection

minus In drug development first in human samples are typically ~1 year later than the toxicology study is done

bull Prior knowledge of human metabolites transferred from metabolite identification and scouting

minus Knowledge of major human metabolites may evolve only after in vivo human samples become available

minus Radio-labeled ADME is the most definitive study may take place late in the drug development process

Is it feasible to scan the samplesbank data and compare the data generated at different dates

SN

N N

N

O

Cl

NH

Cl

Cl

O

OH

N

OCH3

CH3

NOH

OH

N

NH2

NH N O OH

N3

O

O

ziprasidone

diclofenac tacrine

terfenadine

dextromethorphan

zidovudine

MeO

O

O

OH

OH

OH

O OHNaproxen Acyl Glucuronide conjugate





(AZT)

Negative Control

Blind Test 3 Metabolite Cocktail

Gao et al Anal Chem 2015 87 11771-11776

Scanning and Banking DataSamples

bull Blind Test samples 22 metabolitesdrugs were spiked in human and rat plasmas (n=4 each)

bull No knowledge of the identities of the analytes and their concentrations in plasma were provided before sample analysis

bull Multiple analysis of the same samples (rat or human) using mixed matrix approach at Days 1 30 105 254 and 314

bull Treat the samples using the same IS solution in acetonitrile minus Freshly prepared to maintain the same concentration

bull Acquity UPLC generic gradient on C18 run time 25 min

bull Full scan and AIF scan on QExactive 70K at positive and negative polarity

bull Compare rat vs human samples analyzed at different dates

0 4 8 12 16 20 24 Time (min)

0

100 R

elat

ive

Ab

un

dan

ce

508

478 682

1132 1435 1450 763

773 243 1835 2120 2138 1164 1312 1638 288 534 1473 2338 802 2045 335 1062

42 43 44 45 46 47 48 49 50 51 52 53 54 55 Time (min)

0

100

Rel

ativ

e In

ten

sity

RT 508

RT 478

RT 495

RT 539

4 8 12 16 20 24 Time (min)

0

100 RT 624

RT 1313

RT 1113

RT 1626

RT 275 RT 879

RT 2008 RT 422

548

165 808

1 2

3

3

4

5

5

4

678

9 10 111213

14 16 15

1

2

3

4

56 7

Rel

ativ

e A

bu

nd

ance

Migrated naproxen acyl glucuronide

EIC 15 drugsmetabolites Resolution 70000 Extraction window 5ppm Polarity Positive

EIC 6 drugsmetabolites Resolution 70000 Extraction window 10 ppm Polarity Negative

2-OH Tacrine

4-OH Tacrine

1-OH Tacrine

Naproxen-Glu

Peak Area Ratio Measurement Variation

n=4 per species 5 independent analysis total n=40 per analyte

95 tile 037

Median 021

M1

0

DrugsMetabolites

Bank and Compare Data at Different Days


Separation and Quantitative Analysis of Isomers

2 4 6 8 10 12 14 16 18 20 22 24

Time (min)

0

100

Rela

tive I

nte

nsity

RT 1626

RT 275

RT 422 RT 1188 RT 632

Full Scan 4rsquo (or 5)-OH Diclofenac

XIC 3100032

Would high resolution ion mobility provide the orthogonal separation we need

Ideal Paradigm to Address MIST

Un-biased scan the sample using super LCMS

Process post-acquisition data when the knowledge of analytes is available

Bank Data

Address biological questions using peak

area ratio measurements at different dates

Calibrate the ion response using a synthetic standard

when concentration measurement is needed

Samples may degrade data will not

Necessity is the mother of invention

Laziness is the father of invention

Efficiency

Challenges in real life applications drive innovation and technology advances

Next generation LCMS will change how we do analysis in pharmabiotech industry research labs

Gao et al (2017) JASMS 281248-1249

Looking for innovative BA approaches to address DMPK issues

Contact hongyinggaoinnovobioanalysiscom

Backups

Rat Day1

Human Day 1 Rat same day (n=5)

Human

Rat Day105

Human Day 1

Rat Day 314

Human Day 1

1

2

3

4

5

6

Tre

ate

d n

apro

xe

n

Tre

ate

d n

apro

xe

n a

cyl

glu

cu

ron

ide

Untr

eate

d n

ap

roxen

Untr

eate

d n

ap

roxen

acyl

glu

cu

ron

ide

Tre

ate

d s

alic

ylic

acid

Tre

ate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Un

treate

d s

alic

ylic

acid

Untr

eate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Carb

am

aze

pin

e

Carb

am

aze

pin

e-1

01

1-

epo

xid

e

Sulfa

dia

zin

e

N-A

ce

tyl sulfa

dia

zin

e

Tre

ate

d m

eth

ylp

he

nid

ate

Tre

ate

d r

italin

ic a

cid

Untr

eate

dm

eth

ylp

hen

ida

te

Untr

eate

d r

ita

linic

acid

Nic

otin

e I

min

ium

io

n

Cotinin

e

MS

NM

Ra

tio

Measurement Error for Unstable Metabolites

Acknowledgements

R Scott Obach PDM Pfizer Inc

Shibing Deng Biostatistics Pfizer Inc
















Approaches (1)




Data Output





Reagent Needs

None Radio-labeled

Drug None



None None NMR None

Resource Investment

















metabolite








-0avg

AUCC


total

1-2-nn130201


)t-(tV

010t

total

1-2-nn130201

1-n1nnt V


)t-(tV

total

1-2-nn130201

1-t V


)t-(tV



Pooled Human Sample


Mix with Opposite

Matrix Control

Control Mix

Add IS and

Solvent


IS IS IS

Met

Met


(Human) AreaPeak IS





Control Mix

Mixed Human Sample

Mixed Animal Sample

Mixed Human Sample

Mixed Animal Sample
















1day

dayX





RatioStability


SN

N N

N

O

Cl

NH

Cl

Cl

O

OH

N

OCH3

CH3

NOH

OH

N

NH2

NH N O OH

N3

O

O

ziprasidone

diclofenac tacrine

terfenadine

dextromethorphan

zidovudine






(AZT)


M1 236gt134

M2 252gt209


IS


M9

M5

M10



2 B

80 B




Acyl bond migration

Ester hydrolysis


Epoxide hydrolysis


Catechol oxidation


Amide hydrolysis


Ester hydrolysis


Imine Oxidation

N

O NH2

MeO

O

O

OH

OH

OH

O OH

N

N+

N

NO

N

N

NH

OMeO

NH

OHO

OH

O

OH

OH

NH

SNH

N

N

O

O

O

NH2

SNH

N

N

O

O

OH

O

OH

N

O NH2

O

MeO

OH

O



0

05

1

15

2

0 50 100 150

(Nap

roxe

n-G

luN

apro

xen

) D

ay X

Day

1






n = 8

Untreated samples


Stabilized samples




MSNM Ratio

Metabolites

MS

NM

Ra

tio

10

15

20

1_O

H_T

acrine

2_O

H_T

acrine

3_M

eth

oxym

orp

hin

4_O

H_D

iclo

fenc

4_O

H_T

acrine

5_O

H_D

iclo

fenc

AZ

T

AZ

T_glu

c

Dextr

om

eth

orp

han

Dextr

orp

han

Dic

lofe

nac

Fexofe

nadin

e

M1

M10

M2

M5

M9

tacrine

Terf

enadin

e

Zip

rasid

one

Geo FC

Geo FC 95

Geo FC 99

Average 125

95 confidence 183

99 confidence 207


greater exposure





Ratio=4 plt2 ppm

Ratio=2 plt001








SN (2-1)025=4









Which one is taller


Which one is taller







Case Study










Human 0220 00406 000788 00210 Rat 770 0502 0292 000762


Human repeat 0184 00290 000506 00164


ratio 2 399 163 526 0378



Rat 269 plusmn 258





Interest


Method




Method Succeeds

Animal lt 2X Human

Animal ge 2X Human



Needed


MIST Decision Tree







Summary














SN

N N

N

O

Cl

NH

Cl

Cl

O

OH

N

OCH3

CH3

NOH

OH

N

NH2

NH N O OH

N3

O

O

ziprasidone

diclofenac tacrine

terfenadine

dextromethorphan

zidovudine

MeO

O

O

OH

OH

OH






(AZT)

Negative Control











0 4 8 12 16 20 24 Time (min)

0

100 R

elat

ive

Ab

un

dan

ce

508

478 682

1132 1435 1450 763

773 243 1835 2120 2138 1164 1312 1638 288 534 1473 2338 802 2045 335 1062

42 43 44 45 46 47 48 49 50 51 52 53 54 55 Time (min)

0

100

Rel

ativ

e In

ten

sity

RT 508

RT 478

RT 495

RT 539

4 8 12 16 20 24 Time (min)

0

100 RT 624

RT 1313

RT 1113

RT 1626

RT 275 RT 879

RT 2008 RT 422

548

165 808

1 2

3

3

4

5

5

4

678

9 10 111213

14 16 15

1

2

3

4

56 7

Rel

ativ

e A

bu

nd

ance




2-OH Tacrine

4-OH Tacrine

1-OH Tacrine

Naproxen-Glu



95 tile 037

Median 021

M1

0

DrugsMetabolites




2 4 6 8 10 12 14 16 18 20 22 24

Time (min)

0

100

Rela

tive I

nte

nsity

RT 1626

RT 275

RT 422 RT 1188 RT 632


XIC 3100032





Bank Data








Efficiency



Gao et al (2017) JASMS 281248-1249



Backups

Rat Day1


Human

Rat Day105

Human Day 1

Rat Day 314

Human Day 1

1

2

3

4

5

6

Tre

ate

d n

apro

xe

n

Tre

ate

d n

apro

xe

n a

cyl

glu

cu

ron

ide

Untr

eate

d n

ap

roxen

Untr

eate

d n

ap

roxen

acyl

glu

cu

ron

ide

Tre

ate

d s

alic

ylic

acid

Tre

ate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Un

treate

d s

alic

ylic

acid

Untr

eate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Carb

am

aze

pin

e

Carb

am

aze

pin

e-1

01

1-

epo

xid

e

Sulfa

dia

zin

e

N-A

ce

tyl sulfa

dia

zin

e

Tre

ate

d m

eth

ylp

he

nid

ate

Tre

ate

d r

italin

ic a

cid

Untr

eate

dm

eth

ylp

hen

ida

te

Untr

eate

d r

ita

linic

acid

Nic

otin

e I

min

ium

io

n

Cotinin

e

MS

NM

Ra

tio

















Approaches (1)




Data Output





Reagent Needs

None Radio-labeled

Drug None



None None NMR None

Resource Investment

















metabolite








-0avg

AUCC


total

1-2-nn130201


)t-(tV

010t

total

1-2-nn130201

1-n1nnt V


)t-(tV

total

1-2-nn130201

1-t V


)t-(tV



Pooled Human Sample


Mix with Opposite

Matrix Control

Control Mix

Add IS and

Solvent


IS IS IS

Met

Met


(Human) AreaPeak IS





Control Mix

Mixed Human Sample

Mixed Animal Sample

Mixed Human Sample

Mixed Animal Sample
















1day

dayX





RatioStability


SN

N N

N

O

Cl

NH

Cl

Cl

O

OH

N

OCH3

CH3

NOH

OH

N

NH2

NH N O OH

N3

O

O

ziprasidone

diclofenac tacrine

terfenadine

dextromethorphan

zidovudine






(AZT)


M1 236gt134

M2 252gt209


IS


M9

M5

M10



2 B

80 B




Acyl bond migration

Ester hydrolysis


Epoxide hydrolysis


Catechol oxidation


Amide hydrolysis


Ester hydrolysis


Imine Oxidation

N

O NH2

MeO

O

O

OH

OH

OH

O OH

N

N+

N

NO

N

N

NH

OMeO

NH

OHO

OH

O

OH

OH

NH

SNH

N

N

O

O

O

NH2

SNH

N

N

O

O

OH

O

OH

N

O NH2

O

MeO

OH

O



0

05

1

15

2

0 50 100 150

(Nap

roxe

n-G

luN

apro

xen

) D

ay X

Day

1






n = 8

Untreated samples


Stabilized samples




MSNM Ratio

Metabolites

MS

NM

Ra

tio

10

15

20

1_O

H_T

acrine

2_O

H_T

acrine

3_M

eth

oxym

orp

hin

4_O

H_D

iclo

fenc

4_O

H_T

acrine

5_O

H_D

iclo

fenc

AZ

T

AZ

T_glu

c

Dextr

om

eth

orp

han

Dextr

orp

han

Dic

lofe

nac

Fexofe

nadin

e

M1

M10

M2

M5

M9

tacrine

Terf

enadin

e

Zip

rasid

one

Geo FC

Geo FC 95

Geo FC 99

Average 125

95 confidence 183

99 confidence 207


greater exposure





Ratio=4 plt2 ppm

Ratio=2 plt001








SN (2-1)025=4









Which one is taller


Which one is taller







Case Study










Human 0220 00406 000788 00210 Rat 770 0502 0292 000762


Human repeat 0184 00290 000506 00164


ratio 2 399 163 526 0378



Rat 269 plusmn 258





Interest


Method




Method Succeeds

Animal lt 2X Human

Animal ge 2X Human



Needed


MIST Decision Tree







Summary














SN

N N

N

O

Cl

NH

Cl

Cl

O

OH

N

OCH3

CH3

NOH

OH

N

NH2

NH N O OH

N3

O

O

ziprasidone

diclofenac tacrine

terfenadine

dextromethorphan

zidovudine

MeO

O

O

OH

OH

OH






(AZT)

Negative Control











0 4 8 12 16 20 24 Time (min)

0

100 R

elat

ive

Ab

un

dan

ce

508

478 682

1132 1435 1450 763

773 243 1835 2120 2138 1164 1312 1638 288 534 1473 2338 802 2045 335 1062

42 43 44 45 46 47 48 49 50 51 52 53 54 55 Time (min)

0

100

Rel

ativ

e In

ten

sity

RT 508

RT 478

RT 495

RT 539

4 8 12 16 20 24 Time (min)

0

100 RT 624

RT 1313

RT 1113

RT 1626

RT 275 RT 879

RT 2008 RT 422

548

165 808

1 2

3

3

4

5

5

4

678

9 10 111213

14 16 15

1

2

3

4

56 7

Rel

ativ

e A

bu

nd

ance




2-OH Tacrine

4-OH Tacrine

1-OH Tacrine

Naproxen-Glu



95 tile 037

Median 021

M1

0

DrugsMetabolites




2 4 6 8 10 12 14 16 18 20 22 24

Time (min)

0

100

Rela

tive I

nte

nsity

RT 1626

RT 275

RT 422 RT 1188 RT 632


XIC 3100032





Bank Data








Efficiency



Gao et al (2017) JASMS 281248-1249



Backups

Rat Day1


Human

Rat Day105

Human Day 1

Rat Day 314

Human Day 1

1

2

3

4

5

6

Tre

ate

d n

apro

xe

n

Tre

ate

d n

apro

xe

n a

cyl

glu

cu

ron

ide

Untr

eate

d n

ap

roxen

Untr

eate

d n

ap

roxen

acyl

glu

cu

ron

ide

Tre

ate

d s

alic

ylic

acid

Tre

ate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Un

treate

d s

alic

ylic

acid

Untr

eate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Carb

am

aze

pin

e

Carb

am

aze

pin

e-1

01

1-

epo

xid

e

Sulfa

dia

zin

e

N-A

ce

tyl sulfa

dia

zin

e

Tre

ate

d m

eth

ylp

he

nid

ate

Tre

ate

d r

italin

ic a

cid

Untr

eate

dm

eth

ylp

hen

ida

te

Untr

eate

d r

ita

linic

acid

Nic

otin

e I

min

ium

io

n

Cotinin

e

MS

NM

Ra

tio













metabolite








-0avg

AUCC


total

1-2-nn130201


)t-(tV

010t

total

1-2-nn130201

1-n1nnt V


)t-(tV

total

1-2-nn130201

1-t V


)t-(tV



Pooled Human Sample


Mix with Opposite

Matrix Control

Control Mix

Add IS and

Solvent


IS IS IS

Met

Met


(Human) AreaPeak IS





Control Mix

Mixed Human Sample

Mixed Animal Sample

Mixed Human Sample

Mixed Animal Sample
















1day

dayX





RatioStability


SN

N N

N

O

Cl

NH

Cl

Cl

O

OH

N

OCH3

CH3

NOH

OH

N

NH2

NH N O OH

N3

O

O

ziprasidone

diclofenac tacrine

terfenadine

dextromethorphan

zidovudine






(AZT)


M1 236gt134

M2 252gt209


IS


M9

M5

M10



2 B

80 B




Acyl bond migration

Ester hydrolysis


Epoxide hydrolysis


Catechol oxidation


Amide hydrolysis


Ester hydrolysis


Imine Oxidation

N

O NH2

MeO

O

O

OH

OH

OH

O OH

N

N+

N

NO

N

N

NH

OMeO

NH

OHO

OH

O

OH

OH

NH

SNH

N

N

O

O

O

NH2

SNH

N

N

O

O

OH

O

OH

N

O NH2

O

MeO

OH

O



0

05

1

15

2

0 50 100 150

(Nap

roxe

n-G

luN

apro

xen

) D

ay X

Day

1






n = 8

Untreated samples


Stabilized samples




MSNM Ratio

Metabolites

MS

NM

Ra

tio

10

15

20

1_O

H_T

acrine

2_O

H_T

acrine

3_M

eth

oxym

orp

hin

4_O

H_D

iclo

fenc

4_O

H_T

acrine

5_O

H_D

iclo

fenc

AZ

T

AZ

T_glu

c

Dextr

om

eth

orp

han

Dextr

orp

han

Dic

lofe

nac

Fexofe

nadin

e

M1

M10

M2

M5

M9

tacrine

Terf

enadin

e

Zip

rasid

one

Geo FC

Geo FC 95

Geo FC 99

Average 125

95 confidence 183

99 confidence 207


greater exposure





Ratio=4 plt2 ppm

Ratio=2 plt001








SN (2-1)025=4









Which one is taller


Which one is taller







Case Study










Human 0220 00406 000788 00210 Rat 770 0502 0292 000762


Human repeat 0184 00290 000506 00164


ratio 2 399 163 526 0378



Rat 269 plusmn 258





Interest


Method




Method Succeeds

Animal lt 2X Human

Animal ge 2X Human



Needed


MIST Decision Tree







Summary














SN

N N

N

O

Cl

NH

Cl

Cl

O

OH

N

OCH3

CH3

NOH

OH

N

NH2

NH N O OH

N3

O

O

ziprasidone

diclofenac tacrine

terfenadine

dextromethorphan

zidovudine

MeO

O

O

OH

OH

OH






(AZT)

Negative Control











0 4 8 12 16 20 24 Time (min)

0

100 R

elat

ive

Ab

un

dan

ce

508

478 682

1132 1435 1450 763

773 243 1835 2120 2138 1164 1312 1638 288 534 1473 2338 802 2045 335 1062

42 43 44 45 46 47 48 49 50 51 52 53 54 55 Time (min)

0

100

Rel

ativ

e In

ten

sity

RT 508

RT 478

RT 495

RT 539

4 8 12 16 20 24 Time (min)

0

100 RT 624

RT 1313

RT 1113

RT 1626

RT 275 RT 879

RT 2008 RT 422

548

165 808

1 2

3

3

4

5

5

4

678

9 10 111213

14 16 15

1

2

3

4

56 7

Rel

ativ

e A

bu

nd

ance




2-OH Tacrine

4-OH Tacrine

1-OH Tacrine

Naproxen-Glu



95 tile 037

Median 021

M1

0

DrugsMetabolites




2 4 6 8 10 12 14 16 18 20 22 24

Time (min)

0

100

Rela

tive I

nte

nsity

RT 1626

RT 275

RT 422 RT 1188 RT 632


XIC 3100032





Bank Data








Efficiency



Gao et al (2017) JASMS 281248-1249



Backups

Rat Day1


Human

Rat Day105

Human Day 1

Rat Day 314

Human Day 1

1

2

3

4

5

6

Tre

ate

d n

apro

xe

n

Tre

ate

d n

apro

xe

n a

cyl

glu

cu

ron

ide

Untr

eate

d n

ap

roxen

Untr

eate

d n

ap

roxen

acyl

glu

cu

ron

ide

Tre

ate

d s

alic

ylic

acid

Tre

ate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Un

treate

d s

alic

ylic

acid

Untr

eate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Carb

am

aze

pin

e

Carb

am

aze

pin

e-1

01

1-

epo

xid

e

Sulfa

dia

zin

e

N-A

ce

tyl sulfa

dia

zin

e

Tre

ate

d m

eth

ylp

he

nid

ate

Tre

ate

d r

italin

ic a

cid

Untr

eate

dm

eth

ylp

hen

ida

te

Untr

eate

d r

ita

linic

acid

Nic

otin

e I

min

ium

io

n

Cotinin

e

MS

NM

Ra

tio




-0avg

AUCC


total

1-2-nn130201


)t-(tV

010t

total

1-2-nn130201

1-n1nnt V


)t-(tV

total

1-2-nn130201

1-t V


)t-(tV



Pooled Human Sample


Mix with Opposite

Matrix Control

Control Mix

Add IS and

Solvent


IS IS IS

Met

Met


(Human) AreaPeak IS





Control Mix

Mixed Human Sample

Mixed Animal Sample

Mixed Human Sample

Mixed Animal Sample
















1day

dayX





RatioStability


SN

N N

N

O

Cl

NH

Cl

Cl

O

OH

N

OCH3

CH3

NOH

OH

N

NH2

NH N O OH

N3

O

O

ziprasidone

diclofenac tacrine

terfenadine

dextromethorphan

zidovudine






(AZT)


M1 236gt134

M2 252gt209


IS


M9

M5

M10



2 B

80 B




Acyl bond migration

Ester hydrolysis


Epoxide hydrolysis


Catechol oxidation


Amide hydrolysis


Ester hydrolysis


Imine Oxidation

N

O NH2

MeO

O

O

OH

OH

OH

O OH

N

N+

N

NO

N

N

NH

OMeO

NH

OHO

OH

O

OH

OH

NH

SNH

N

N

O

O

O

NH2

SNH

N

N

O

O

OH

O

OH

N

O NH2

O

MeO

OH

O



0

05

1

15

2

0 50 100 150

(Nap

roxe

n-G

luN

apro

xen

) D

ay X

Day

1






n = 8

Untreated samples


Stabilized samples




MSNM Ratio

Metabolites

MS

NM

Ra

tio

10

15

20

1_O

H_T

acrine

2_O

H_T

acrine

3_M

eth

oxym

orp

hin

4_O

H_D

iclo

fenc

4_O

H_T

acrine

5_O

H_D

iclo

fenc

AZ

T

AZ

T_glu

c

Dextr

om

eth

orp

han

Dextr

orp

han

Dic

lofe

nac

Fexofe

nadin

e

M1

M10

M2

M5

M9

tacrine

Terf

enadin

e

Zip

rasid

one

Geo FC

Geo FC 95

Geo FC 99

Average 125

95 confidence 183

99 confidence 207


greater exposure





Ratio=4 plt2 ppm

Ratio=2 plt001








SN (2-1)025=4









Which one is taller


Which one is taller







Case Study










Human 0220 00406 000788 00210 Rat 770 0502 0292 000762


Human repeat 0184 00290 000506 00164


ratio 2 399 163 526 0378



Rat 269 plusmn 258





Interest


Method




Method Succeeds

Animal lt 2X Human

Animal ge 2X Human



Needed


MIST Decision Tree







Summary














SN

N N

N

O

Cl

NH

Cl

Cl

O

OH

N

OCH3

CH3

NOH

OH

N

NH2

NH N O OH

N3

O

O

ziprasidone

diclofenac tacrine

terfenadine

dextromethorphan

zidovudine

MeO

O

O

OH

OH

OH






(AZT)

Negative Control











0 4 8 12 16 20 24 Time (min)

0

100 R

elat

ive

Ab

un

dan

ce

508

478 682

1132 1435 1450 763

773 243 1835 2120 2138 1164 1312 1638 288 534 1473 2338 802 2045 335 1062

42 43 44 45 46 47 48 49 50 51 52 53 54 55 Time (min)

0

100

Rel

ativ

e In

ten

sity

RT 508

RT 478

RT 495

RT 539

4 8 12 16 20 24 Time (min)

0

100 RT 624

RT 1313

RT 1113

RT 1626

RT 275 RT 879

RT 2008 RT 422

548

165 808

1 2

3

3

4

5

5

4

678

9 10 111213

14 16 15

1

2

3

4

56 7

Rel

ativ

e A

bu

nd

ance




2-OH Tacrine

4-OH Tacrine

1-OH Tacrine

Naproxen-Glu



95 tile 037

Median 021

M1

0

DrugsMetabolites




2 4 6 8 10 12 14 16 18 20 22 24

Time (min)

0

100

Rela

tive I

nte

nsity

RT 1626

RT 275

RT 422 RT 1188 RT 632


XIC 3100032





Bank Data








Efficiency



Gao et al (2017) JASMS 281248-1249



Backups

Rat Day1


Human

Rat Day105

Human Day 1

Rat Day 314

Human Day 1

1

2

3

4

5

6

Tre

ate

d n

apro

xe

n

Tre

ate

d n

apro

xe

n a

cyl

glu

cu

ron

ide

Untr

eate

d n

ap

roxen

Untr

eate

d n

ap

roxen

acyl

glu

cu

ron

ide

Tre

ate

d s

alic

ylic

acid

Tre

ate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Un

treate

d s

alic

ylic

acid

Untr

eate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Carb

am

aze

pin

e

Carb

am

aze

pin

e-1

01

1-

epo

xid

e

Sulfa

dia

zin

e

N-A

ce

tyl sulfa

dia

zin

e

Tre

ate

d m

eth

ylp

he

nid

ate

Tre

ate

d r

italin

ic a

cid

Untr

eate

dm

eth

ylp

hen

ida

te

Untr

eate

d r

ita

linic

acid

Nic

otin

e I

min

ium

io

n

Cotinin

e

MS

NM

Ra

tio



Pooled Human Sample


Mix with Opposite

Matrix Control

Control Mix

Add IS and

Solvent


IS IS IS

Met

Met


(Human) AreaPeak IS





Control Mix

Mixed Human Sample

Mixed Animal Sample

Mixed Human Sample

Mixed Animal Sample
















1day

dayX





RatioStability


SN

N N

N

O

Cl

NH

Cl

Cl

O

OH

N

OCH3

CH3

NOH

OH

N

NH2

NH N O OH

N3

O

O

ziprasidone

diclofenac tacrine

terfenadine

dextromethorphan

zidovudine






(AZT)


M1 236gt134

M2 252gt209


IS


M9

M5

M10



2 B

80 B




Acyl bond migration

Ester hydrolysis


Epoxide hydrolysis


Catechol oxidation


Amide hydrolysis


Ester hydrolysis


Imine Oxidation

N

O NH2

MeO

O

O

OH

OH

OH

O OH

N

N+

N

NO

N

N

NH

OMeO

NH

OHO

OH

O

OH

OH

NH

SNH

N

N

O

O

O

NH2

SNH

N

N

O

O

OH

O

OH

N

O NH2

O

MeO

OH

O



0

05

1

15

2

0 50 100 150

(Nap

roxe

n-G

luN

apro

xen

) D

ay X

Day

1






n = 8

Untreated samples


Stabilized samples




MSNM Ratio

Metabolites

MS

NM

Ra

tio

10

15

20

1_O

H_T

acrine

2_O

H_T

acrine

3_M

eth

oxym

orp

hin

4_O

H_D

iclo

fenc

4_O

H_T

acrine

5_O

H_D

iclo

fenc

AZ

T

AZ

T_glu

c

Dextr

om

eth

orp

han

Dextr

orp

han

Dic

lofe

nac

Fexofe

nadin

e

M1

M10

M2

M5

M9

tacrine

Terf

enadin

e

Zip

rasid

one

Geo FC

Geo FC 95

Geo FC 99

Average 125

95 confidence 183

99 confidence 207


greater exposure





Ratio=4 plt2 ppm

Ratio=2 plt001








SN (2-1)025=4









Which one is taller


Which one is taller







Case Study










Human 0220 00406 000788 00210 Rat 770 0502 0292 000762


Human repeat 0184 00290 000506 00164


ratio 2 399 163 526 0378



Rat 269 plusmn 258





Interest


Method




Method Succeeds

Animal lt 2X Human

Animal ge 2X Human



Needed


MIST Decision Tree







Summary














SN

N N

N

O

Cl

NH

Cl

Cl

O

OH

N

OCH3

CH3

NOH

OH

N

NH2

NH N O OH

N3

O

O

ziprasidone

diclofenac tacrine

terfenadine

dextromethorphan

zidovudine

MeO

O

O

OH

OH

OH






(AZT)

Negative Control











0 4 8 12 16 20 24 Time (min)

0

100 R

elat

ive

Ab

un

dan

ce

508

478 682

1132 1435 1450 763

773 243 1835 2120 2138 1164 1312 1638 288 534 1473 2338 802 2045 335 1062

42 43 44 45 46 47 48 49 50 51 52 53 54 55 Time (min)

0

100

Rel

ativ

e In

ten

sity

RT 508

RT 478

RT 495

RT 539

4 8 12 16 20 24 Time (min)

0

100 RT 624

RT 1313

RT 1113

RT 1626

RT 275 RT 879

RT 2008 RT 422

548

165 808

1 2

3

3

4

5

5

4

678

9 10 111213

14 16 15

1

2

3

4

56 7

Rel

ativ

e A

bu

nd

ance




2-OH Tacrine

4-OH Tacrine

1-OH Tacrine

Naproxen-Glu



95 tile 037

Median 021

M1

0

DrugsMetabolites




2 4 6 8 10 12 14 16 18 20 22 24

Time (min)

0

100

Rela

tive I

nte

nsity

RT 1626

RT 275

RT 422 RT 1188 RT 632


XIC 3100032





Bank Data








Efficiency



Gao et al (2017) JASMS 281248-1249



Backups

Rat Day1


Human

Rat Day105

Human Day 1

Rat Day 314

Human Day 1

1

2

3

4

5

6

Tre

ate

d n

apro

xe

n

Tre

ate

d n

apro

xe

n a

cyl

glu

cu

ron

ide

Untr

eate

d n

ap

roxen

Untr

eate

d n

ap

roxen

acyl

glu

cu

ron

ide

Tre

ate

d s

alic

ylic

acid

Tre

ate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Un

treate

d s

alic

ylic

acid

Untr

eate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Carb

am

aze

pin

e

Carb

am

aze

pin

e-1

01

1-

epo

xid

e

Sulfa

dia

zin

e

N-A

ce

tyl sulfa

dia

zin

e

Tre

ate

d m

eth

ylp

he

nid

ate

Tre

ate

d r

italin

ic a

cid

Untr

eate

dm

eth

ylp

hen

ida

te

Untr

eate

d r

ita

linic

acid

Nic

otin

e I

min

ium

io

n

Cotinin

e

MS

NM

Ra

tio















1day

dayX





RatioStability


SN

N N

N

O

Cl

NH

Cl

Cl

O

OH

N

OCH3

CH3

NOH

OH

N

NH2

NH N O OH

N3

O

O

ziprasidone

diclofenac tacrine

terfenadine

dextromethorphan

zidovudine






(AZT)


M1 236gt134

M2 252gt209


IS


M9

M5

M10



2 B

80 B




Acyl bond migration

Ester hydrolysis


Epoxide hydrolysis


Catechol oxidation


Amide hydrolysis


Ester hydrolysis


Imine Oxidation

N

O NH2

MeO

O

O

OH

OH

OH

O OH

N

N+

N

NO

N

N

NH

OMeO

NH

OHO

OH

O

OH

OH

NH

SNH

N

N

O

O

O

NH2

SNH

N

N

O

O

OH

O

OH

N

O NH2

O

MeO

OH

O



0

05

1

15

2

0 50 100 150

(Nap

roxe

n-G

luN

apro

xen

) D

ay X

Day

1






n = 8

Untreated samples


Stabilized samples




MSNM Ratio

Metabolites

MS

NM

Ra

tio

10

15

20

1_O

H_T

acrine

2_O

H_T

acrine

3_M

eth

oxym

orp

hin

4_O

H_D

iclo

fenc

4_O

H_T

acrine

5_O

H_D

iclo

fenc

AZ

T

AZ

T_glu

c

Dextr

om

eth

orp

han

Dextr

orp

han

Dic

lofe

nac

Fexofe

nadin

e

M1

M10

M2

M5

M9

tacrine

Terf

enadin

e

Zip

rasid

one

Geo FC

Geo FC 95

Geo FC 99

Average 125

95 confidence 183

99 confidence 207


greater exposure





Ratio=4 plt2 ppm

Ratio=2 plt001








SN (2-1)025=4









Which one is taller


Which one is taller







Case Study










Human 0220 00406 000788 00210 Rat 770 0502 0292 000762


Human repeat 0184 00290 000506 00164


ratio 2 399 163 526 0378



Rat 269 plusmn 258





Interest


Method




Method Succeeds

Animal lt 2X Human

Animal ge 2X Human



Needed


MIST Decision Tree







Summary














SN

N N

N

O

Cl

NH

Cl

Cl

O

OH

N

OCH3

CH3

NOH

OH

N

NH2

NH N O OH

N3

O

O

ziprasidone

diclofenac tacrine

terfenadine

dextromethorphan

zidovudine

MeO

O

O

OH

OH

OH






(AZT)

Negative Control











0 4 8 12 16 20 24 Time (min)

0

100 R

elat

ive

Ab

un

dan

ce

508

478 682

1132 1435 1450 763

773 243 1835 2120 2138 1164 1312 1638 288 534 1473 2338 802 2045 335 1062

42 43 44 45 46 47 48 49 50 51 52 53 54 55 Time (min)

0

100

Rel

ativ

e In

ten

sity

RT 508

RT 478

RT 495

RT 539

4 8 12 16 20 24 Time (min)

0

100 RT 624

RT 1313

RT 1113

RT 1626

RT 275 RT 879

RT 2008 RT 422

548

165 808

1 2

3

3

4

5

5

4

678

9 10 111213

14 16 15

1

2

3

4

56 7

Rel

ativ

e A

bu

nd

ance




2-OH Tacrine

4-OH Tacrine

1-OH Tacrine

Naproxen-Glu



95 tile 037

Median 021

M1

0

DrugsMetabolites




2 4 6 8 10 12 14 16 18 20 22 24

Time (min)

0

100

Rela

tive I

nte

nsity

RT 1626

RT 275

RT 422 RT 1188 RT 632


XIC 3100032





Bank Data








Efficiency



Gao et al (2017) JASMS 281248-1249



Backups

Rat Day1


Human

Rat Day105

Human Day 1

Rat Day 314

Human Day 1

1

2

3

4

5

6

Tre

ate

d n

apro

xe

n

Tre

ate

d n

apro

xe

n a

cyl

glu

cu

ron

ide

Untr

eate

d n

ap

roxen

Untr

eate

d n

ap

roxen

acyl

glu

cu

ron

ide

Tre

ate

d s

alic

ylic

acid

Tre

ate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Un

treate

d s

alic

ylic

acid

Untr

eate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Carb

am

aze

pin

e

Carb

am

aze

pin

e-1

01

1-

epo

xid

e

Sulfa

dia

zin

e

N-A

ce

tyl sulfa

dia

zin

e

Tre

ate

d m

eth

ylp

he

nid

ate

Tre

ate

d r

italin

ic a

cid

Untr

eate

dm

eth

ylp

hen

ida

te

Untr

eate

d r

ita

linic

acid

Nic

otin

e I

min

ium

io

n

Cotinin

e

MS

NM

Ra

tio


SN

N N

N

O

Cl

NH

Cl

Cl

O

OH

N

OCH3

CH3

NOH

OH

N

NH2

NH N O OH

N3

O

O

ziprasidone

diclofenac tacrine

terfenadine

dextromethorphan

zidovudine






(AZT)


M1 236gt134

M2 252gt209


IS


M9

M5

M10



2 B

80 B




Acyl bond migration

Ester hydrolysis


Epoxide hydrolysis


Catechol oxidation


Amide hydrolysis


Ester hydrolysis


Imine Oxidation

N

O NH2

MeO

O

O

OH

OH

OH

O OH

N

N+

N

NO

N

N

NH

OMeO

NH

OHO

OH

O

OH

OH

NH

SNH

N

N

O

O

O

NH2

SNH

N

N

O

O

OH

O

OH

N

O NH2

O

MeO

OH

O



0

05

1

15

2

0 50 100 150

(Nap

roxe

n-G

luN

apro

xen

) D

ay X

Day

1






n = 8

Untreated samples


Stabilized samples




MSNM Ratio

Metabolites

MS

NM

Ra

tio

10

15

20

1_O

H_T

acrine

2_O

H_T

acrine

3_M

eth

oxym

orp

hin

4_O

H_D

iclo

fenc

4_O

H_T

acrine

5_O

H_D

iclo

fenc

AZ

T

AZ

T_glu

c

Dextr

om

eth

orp

han

Dextr

orp

han

Dic

lofe

nac

Fexofe

nadin

e

M1

M10

M2

M5

M9

tacrine

Terf

enadin

e

Zip

rasid

one

Geo FC

Geo FC 95

Geo FC 99

Average 125

95 confidence 183

99 confidence 207


greater exposure





Ratio=4 plt2 ppm

Ratio=2 plt001








SN (2-1)025=4









Which one is taller


Which one is taller







Case Study










Human 0220 00406 000788 00210 Rat 770 0502 0292 000762


Human repeat 0184 00290 000506 00164


ratio 2 399 163 526 0378



Rat 269 plusmn 258





Interest


Method




Method Succeeds

Animal lt 2X Human

Animal ge 2X Human



Needed


MIST Decision Tree







Summary














SN

N N

N

O

Cl

NH

Cl

Cl

O

OH

N

OCH3

CH3

NOH

OH

N

NH2

NH N O OH

N3

O

O

ziprasidone

diclofenac tacrine

terfenadine

dextromethorphan

zidovudine

MeO

O

O

OH

OH

OH






(AZT)

Negative Control











0 4 8 12 16 20 24 Time (min)

0

100 R

elat

ive

Ab

un

dan

ce

508

478 682

1132 1435 1450 763

773 243 1835 2120 2138 1164 1312 1638 288 534 1473 2338 802 2045 335 1062

42 43 44 45 46 47 48 49 50 51 52 53 54 55 Time (min)

0

100

Rel

ativ

e In

ten

sity

RT 508

RT 478

RT 495

RT 539

4 8 12 16 20 24 Time (min)

0

100 RT 624

RT 1313

RT 1113

RT 1626

RT 275 RT 879

RT 2008 RT 422

548

165 808

1 2

3

3

4

5

5

4

678

9 10 111213

14 16 15

1

2

3

4

56 7

Rel

ativ

e A

bu

nd

ance




2-OH Tacrine

4-OH Tacrine

1-OH Tacrine

Naproxen-Glu



95 tile 037

Median 021

M1

0

DrugsMetabolites




2 4 6 8 10 12 14 16 18 20 22 24

Time (min)

0

100

Rela

tive I

nte

nsity

RT 1626

RT 275

RT 422 RT 1188 RT 632


XIC 3100032





Bank Data








Efficiency



Gao et al (2017) JASMS 281248-1249



Backups

Rat Day1


Human

Rat Day105

Human Day 1

Rat Day 314

Human Day 1

1

2

3

4

5

6

Tre

ate

d n

apro

xe

n

Tre

ate

d n

apro

xe

n a

cyl

glu

cu

ron

ide

Untr

eate

d n

ap

roxen

Untr

eate

d n

ap

roxen

acyl

glu

cu

ron

ide

Tre

ate

d s

alic

ylic

acid

Tre

ate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Un

treate

d s

alic

ylic

acid

Untr

eate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Carb

am

aze

pin

e

Carb

am

aze

pin

e-1

01

1-

epo

xid

e

Sulfa

dia

zin

e

N-A

ce

tyl sulfa

dia

zin

e

Tre

ate

d m

eth

ylp

he

nid

ate

Tre

ate

d r

italin

ic a

cid

Untr

eate

dm

eth

ylp

hen

ida

te

Untr

eate

d r

ita

linic

acid

Nic

otin

e I

min

ium

io

n

Cotinin

e

MS

NM

Ra

tio


M1 236gt134

M2 252gt209


IS


M9

M5

M10



2 B

80 B




Acyl bond migration

Ester hydrolysis


Epoxide hydrolysis


Catechol oxidation


Amide hydrolysis


Ester hydrolysis


Imine Oxidation

N

O NH2

MeO

O

O

OH

OH

OH

O OH

N

N+

N

NO

N

N

NH

OMeO

NH

OHO

OH

O

OH

OH

NH

SNH

N

N

O

O

O

NH2

SNH

N

N

O

O

OH

O

OH

N

O NH2

O

MeO

OH

O



0

05

1

15

2

0 50 100 150

(Nap

roxe

n-G

luN

apro

xen

) D

ay X

Day

1






n = 8

Untreated samples


Stabilized samples




MSNM Ratio

Metabolites

MS

NM

Ra

tio

10

15

20

1_O

H_T

acrine

2_O

H_T

acrine

3_M

eth

oxym

orp

hin

4_O

H_D

iclo

fenc

4_O

H_T

acrine

5_O

H_D

iclo

fenc

AZ

T

AZ

T_glu

c

Dextr

om

eth

orp

han

Dextr

orp

han

Dic

lofe

nac

Fexofe

nadin

e

M1

M10

M2

M5

M9

tacrine

Terf

enadin

e

Zip

rasid

one

Geo FC

Geo FC 95

Geo FC 99

Average 125

95 confidence 183

99 confidence 207


greater exposure





Ratio=4 plt2 ppm

Ratio=2 plt001








SN (2-1)025=4









Which one is taller


Which one is taller







Case Study










Human 0220 00406 000788 00210 Rat 770 0502 0292 000762


Human repeat 0184 00290 000506 00164


ratio 2 399 163 526 0378



Rat 269 plusmn 258





Interest


Method




Method Succeeds

Animal lt 2X Human

Animal ge 2X Human



Needed


MIST Decision Tree







Summary














SN

N N

N

O

Cl

NH

Cl

Cl

O

OH

N

OCH3

CH3

NOH

OH

N

NH2

NH N O OH

N3

O

O

ziprasidone

diclofenac tacrine

terfenadine

dextromethorphan

zidovudine

MeO

O

O

OH

OH

OH






(AZT)

Negative Control











0 4 8 12 16 20 24 Time (min)

0

100 R

elat

ive

Ab

un

dan

ce

508

478 682

1132 1435 1450 763

773 243 1835 2120 2138 1164 1312 1638 288 534 1473 2338 802 2045 335 1062

42 43 44 45 46 47 48 49 50 51 52 53 54 55 Time (min)

0

100

Rel

ativ

e In

ten

sity

RT 508

RT 478

RT 495

RT 539

4 8 12 16 20 24 Time (min)

0

100 RT 624

RT 1313

RT 1113

RT 1626

RT 275 RT 879

RT 2008 RT 422

548

165 808

1 2

3

3

4

5

5

4

678

9 10 111213

14 16 15

1

2

3

4

56 7

Rel

ativ

e A

bu

nd

ance




2-OH Tacrine

4-OH Tacrine

1-OH Tacrine

Naproxen-Glu



95 tile 037

Median 021

M1

0

DrugsMetabolites




2 4 6 8 10 12 14 16 18 20 22 24

Time (min)

0

100

Rela

tive I

nte

nsity

RT 1626

RT 275

RT 422 RT 1188 RT 632


XIC 3100032





Bank Data








Efficiency



Gao et al (2017) JASMS 281248-1249



Backups

Rat Day1


Human

Rat Day105

Human Day 1

Rat Day 314

Human Day 1

1

2

3

4

5

6

Tre

ate

d n

apro

xe

n

Tre

ate

d n

apro

xe

n a

cyl

glu

cu

ron

ide

Untr

eate

d n

ap

roxen

Untr

eate

d n

ap

roxen

acyl

glu

cu

ron

ide

Tre

ate

d s

alic

ylic

acid

Tre

ate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Un

treate

d s

alic

ylic

acid

Untr

eate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Carb

am

aze

pin

e

Carb

am

aze

pin

e-1

01

1-

epo

xid

e

Sulfa

dia

zin

e

N-A

ce

tyl sulfa

dia

zin

e

Tre

ate

d m

eth

ylp

he

nid

ate

Tre

ate

d r

italin

ic a

cid

Untr

eate

dm

eth

ylp

hen

ida

te

Untr

eate

d r

ita

linic

acid

Nic

otin

e I

min

ium

io

n

Cotinin

e

MS

NM

Ra

tio




Acyl bond migration

Ester hydrolysis


Epoxide hydrolysis


Catechol oxidation


Amide hydrolysis


Ester hydrolysis


Imine Oxidation

N

O NH2

MeO

O

O

OH

OH

OH

O OH

N

N+

N

NO

N

N

NH

OMeO

NH

OHO

OH

O

OH

OH

NH

SNH

N

N

O

O

O

NH2

SNH

N

N

O

O

OH

O

OH

N

O NH2

O

MeO

OH

O



0

05

1

15

2

0 50 100 150

(Nap

roxe

n-G

luN

apro

xen

) D

ay X

Day

1






n = 8

Untreated samples


Stabilized samples




MSNM Ratio

Metabolites

MS

NM

Ra

tio

10

15

20

1_O

H_T

acrine

2_O

H_T

acrine

3_M

eth

oxym

orp

hin

4_O

H_D

iclo

fenc

4_O

H_T

acrine

5_O

H_D

iclo

fenc

AZ

T

AZ

T_glu

c

Dextr

om

eth

orp

han

Dextr

orp

han

Dic

lofe

nac

Fexofe

nadin

e

M1

M10

M2

M5

M9

tacrine

Terf

enadin

e

Zip

rasid

one

Geo FC

Geo FC 95

Geo FC 99

Average 125

95 confidence 183

99 confidence 207


greater exposure





Ratio=4 plt2 ppm

Ratio=2 plt001








SN (2-1)025=4









Which one is taller


Which one is taller







Case Study










Human 0220 00406 000788 00210 Rat 770 0502 0292 000762


Human repeat 0184 00290 000506 00164


ratio 2 399 163 526 0378



Rat 269 plusmn 258





Interest


Method




Method Succeeds

Animal lt 2X Human

Animal ge 2X Human



Needed


MIST Decision Tree







Summary














SN

N N

N

O

Cl

NH

Cl

Cl

O

OH

N

OCH3

CH3

NOH

OH

N

NH2

NH N O OH

N3

O

O

ziprasidone

diclofenac tacrine

terfenadine

dextromethorphan

zidovudine

MeO

O

O

OH

OH

OH






(AZT)

Negative Control











0 4 8 12 16 20 24 Time (min)

0

100 R

elat

ive

Ab

un

dan

ce

508

478 682

1132 1435 1450 763

773 243 1835 2120 2138 1164 1312 1638 288 534 1473 2338 802 2045 335 1062

42 43 44 45 46 47 48 49 50 51 52 53 54 55 Time (min)

0

100

Rel

ativ

e In

ten

sity

RT 508

RT 478

RT 495

RT 539

4 8 12 16 20 24 Time (min)

0

100 RT 624

RT 1313

RT 1113

RT 1626

RT 275 RT 879

RT 2008 RT 422

548

165 808

1 2

3

3

4

5

5

4

678

9 10 111213

14 16 15

1

2

3

4

56 7

Rel

ativ

e A

bu

nd

ance




2-OH Tacrine

4-OH Tacrine

1-OH Tacrine

Naproxen-Glu



95 tile 037

Median 021

M1

0

DrugsMetabolites




2 4 6 8 10 12 14 16 18 20 22 24

Time (min)

0

100

Rela

tive I

nte

nsity

RT 1626

RT 275

RT 422 RT 1188 RT 632


XIC 3100032





Bank Data








Efficiency



Gao et al (2017) JASMS 281248-1249



Backups

Rat Day1


Human

Rat Day105

Human Day 1

Rat Day 314

Human Day 1

1

2

3

4

5

6

Tre

ate

d n

apro

xe

n

Tre

ate

d n

apro

xe

n a

cyl

glu

cu

ron

ide

Untr

eate

d n

ap

roxen

Untr

eate

d n

ap

roxen

acyl

glu

cu

ron

ide

Tre

ate

d s

alic

ylic

acid

Tre

ate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Un

treate

d s

alic

ylic

acid

Untr

eate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Carb

am

aze

pin

e

Carb

am

aze

pin

e-1

01

1-

epo

xid

e

Sulfa

dia

zin

e

N-A

ce

tyl sulfa

dia

zin

e

Tre

ate

d m

eth

ylp

he

nid

ate

Tre

ate

d r

italin

ic a

cid

Untr

eate

dm

eth

ylp

hen

ida

te

Untr

eate

d r

ita

linic

acid

Nic

otin

e I

min

ium

io

n

Cotinin

e

MS

NM

Ra

tio



0

05

1

15

2

0 50 100 150

(Nap

roxe

n-G

luN

apro

xen

) D

ay X

Day

1






n = 8

Untreated samples


Stabilized samples




MSNM Ratio

Metabolites

MS

NM

Ra

tio

10

15

20

1_O

H_T

acrine

2_O

H_T

acrine

3_M

eth

oxym

orp

hin

4_O

H_D

iclo

fenc

4_O

H_T

acrine

5_O

H_D

iclo

fenc

AZ

T

AZ

T_glu

c

Dextr

om

eth

orp

han

Dextr

orp

han

Dic

lofe

nac

Fexofe

nadin

e

M1

M10

M2

M5

M9

tacrine

Terf

enadin

e

Zip

rasid

one

Geo FC

Geo FC 95

Geo FC 99

Average 125

95 confidence 183

99 confidence 207


greater exposure





Ratio=4 plt2 ppm

Ratio=2 plt001








SN (2-1)025=4









Which one is taller


Which one is taller







Case Study










Human 0220 00406 000788 00210 Rat 770 0502 0292 000762


Human repeat 0184 00290 000506 00164


ratio 2 399 163 526 0378



Rat 269 plusmn 258





Interest


Method




Method Succeeds

Animal lt 2X Human

Animal ge 2X Human



Needed


MIST Decision Tree







Summary














SN

N N

N

O

Cl

NH

Cl

Cl

O

OH

N

OCH3

CH3

NOH

OH

N

NH2

NH N O OH

N3

O

O

ziprasidone

diclofenac tacrine

terfenadine

dextromethorphan

zidovudine

MeO

O

O

OH

OH

OH






(AZT)

Negative Control











0 4 8 12 16 20 24 Time (min)

0

100 R

elat

ive

Ab

un

dan

ce

508

478 682

1132 1435 1450 763

773 243 1835 2120 2138 1164 1312 1638 288 534 1473 2338 802 2045 335 1062

42 43 44 45 46 47 48 49 50 51 52 53 54 55 Time (min)

0

100

Rel

ativ

e In

ten

sity

RT 508

RT 478

RT 495

RT 539

4 8 12 16 20 24 Time (min)

0

100 RT 624

RT 1313

RT 1113

RT 1626

RT 275 RT 879

RT 2008 RT 422

548

165 808

1 2

3

3

4

5

5

4

678

9 10 111213

14 16 15

1

2

3

4

56 7

Rel

ativ

e A

bu

nd

ance




2-OH Tacrine

4-OH Tacrine

1-OH Tacrine

Naproxen-Glu



95 tile 037

Median 021

M1

0

DrugsMetabolites




2 4 6 8 10 12 14 16 18 20 22 24

Time (min)

0

100

Rela

tive I

nte

nsity

RT 1626

RT 275

RT 422 RT 1188 RT 632


XIC 3100032





Bank Data








Efficiency



Gao et al (2017) JASMS 281248-1249



Backups

Rat Day1


Human

Rat Day105

Human Day 1

Rat Day 314

Human Day 1

1

2

3

4

5

6

Tre

ate

d n

apro

xe

n

Tre

ate

d n

apro

xe

n a

cyl

glu

cu

ron

ide

Untr

eate

d n

ap

roxen

Untr

eate

d n

ap

roxen

acyl

glu

cu

ron

ide

Tre

ate

d s

alic

ylic

acid

Tre

ate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Un

treate

d s

alic

ylic

acid

Untr

eate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Carb

am

aze

pin

e

Carb

am

aze

pin

e-1

01

1-

epo

xid

e

Sulfa

dia

zin

e

N-A

ce

tyl sulfa

dia

zin

e

Tre

ate

d m

eth

ylp

he

nid

ate

Tre

ate

d r

italin

ic a

cid

Untr

eate

dm

eth

ylp

hen

ida

te

Untr

eate

d r

ita

linic

acid

Nic

otin

e I

min

ium

io

n

Cotinin

e

MS

NM

Ra

tio




MSNM Ratio

Metabolites

MS

NM

Ra

tio

10

15

20

1_O

H_T

acrine

2_O

H_T

acrine

3_M

eth

oxym

orp

hin

4_O

H_D

iclo

fenc

4_O

H_T

acrine

5_O

H_D

iclo

fenc

AZ

T

AZ

T_glu

c

Dextr

om

eth

orp

han

Dextr

orp

han

Dic

lofe

nac

Fexofe

nadin

e

M1

M10

M2

M5

M9

tacrine

Terf

enadin

e

Zip

rasid

one

Geo FC

Geo FC 95

Geo FC 99

Average 125

95 confidence 183

99 confidence 207


greater exposure





Ratio=4 plt2 ppm

Ratio=2 plt001








SN (2-1)025=4









Which one is taller


Which one is taller







Case Study










Human 0220 00406 000788 00210 Rat 770 0502 0292 000762


Human repeat 0184 00290 000506 00164


ratio 2 399 163 526 0378



Rat 269 plusmn 258





Interest


Method




Method Succeeds

Animal lt 2X Human

Animal ge 2X Human



Needed


MIST Decision Tree







Summary














SN

N N

N

O

Cl

NH

Cl

Cl

O

OH

N

OCH3

CH3

NOH

OH

N

NH2

NH N O OH

N3

O

O

ziprasidone

diclofenac tacrine

terfenadine

dextromethorphan

zidovudine

MeO

O

O

OH

OH

OH






(AZT)

Negative Control











0 4 8 12 16 20 24 Time (min)

0

100 R

elat

ive

Ab

un

dan

ce

508

478 682

1132 1435 1450 763

773 243 1835 2120 2138 1164 1312 1638 288 534 1473 2338 802 2045 335 1062

42 43 44 45 46 47 48 49 50 51 52 53 54 55 Time (min)

0

100

Rel

ativ

e In

ten

sity

RT 508

RT 478

RT 495

RT 539

4 8 12 16 20 24 Time (min)

0

100 RT 624

RT 1313

RT 1113

RT 1626

RT 275 RT 879

RT 2008 RT 422

548

165 808

1 2

3

3

4

5

5

4

678

9 10 111213

14 16 15

1

2

3

4

56 7

Rel

ativ

e A

bu

nd

ance




2-OH Tacrine

4-OH Tacrine

1-OH Tacrine

Naproxen-Glu



95 tile 037

Median 021

M1

0

DrugsMetabolites




2 4 6 8 10 12 14 16 18 20 22 24

Time (min)

0

100

Rela

tive I

nte

nsity

RT 1626

RT 275

RT 422 RT 1188 RT 632


XIC 3100032





Bank Data








Efficiency



Gao et al (2017) JASMS 281248-1249



Backups

Rat Day1


Human

Rat Day105

Human Day 1

Rat Day 314

Human Day 1

1

2

3

4

5

6

Tre

ate

d n

apro

xe

n

Tre

ate

d n

apro

xe

n a

cyl

glu

cu

ron

ide

Untr

eate

d n

ap

roxen

Untr

eate

d n

ap

roxen

acyl

glu

cu

ron

ide

Tre

ate

d s

alic

ylic

acid

Tre

ate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Un

treate

d s

alic

ylic

acid

Untr

eate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Carb

am

aze

pin

e

Carb

am

aze

pin

e-1

01

1-

epo

xid

e

Sulfa

dia

zin

e

N-A

ce

tyl sulfa

dia

zin

e

Tre

ate

d m

eth

ylp

he

nid

ate

Tre

ate

d r

italin

ic a

cid

Untr

eate

dm

eth

ylp

hen

ida

te

Untr

eate

d r

ita

linic

acid

Nic

otin

e I

min

ium

io

n

Cotinin

e

MS

NM

Ra

tio



Ratio=4 plt2 ppm

Ratio=2 plt001








SN (2-1)025=4









Which one is taller


Which one is taller







Case Study










Human 0220 00406 000788 00210 Rat 770 0502 0292 000762


Human repeat 0184 00290 000506 00164


ratio 2 399 163 526 0378



Rat 269 plusmn 258





Interest


Method




Method Succeeds

Animal lt 2X Human

Animal ge 2X Human



Needed


MIST Decision Tree







Summary














SN

N N

N

O

Cl

NH

Cl

Cl

O

OH

N

OCH3

CH3

NOH

OH

N

NH2

NH N O OH

N3

O

O

ziprasidone

diclofenac tacrine

terfenadine

dextromethorphan

zidovudine

MeO

O

O

OH

OH

OH






(AZT)

Negative Control











0 4 8 12 16 20 24 Time (min)

0

100 R

elat

ive

Ab

un

dan

ce

508

478 682

1132 1435 1450 763

773 243 1835 2120 2138 1164 1312 1638 288 534 1473 2338 802 2045 335 1062

42 43 44 45 46 47 48 49 50 51 52 53 54 55 Time (min)

0

100

Rel

ativ

e In

ten

sity

RT 508

RT 478

RT 495

RT 539

4 8 12 16 20 24 Time (min)

0

100 RT 624

RT 1313

RT 1113

RT 1626

RT 275 RT 879

RT 2008 RT 422

548

165 808

1 2

3

3

4

5

5

4

678

9 10 111213

14 16 15

1

2

3

4

56 7

Rel

ativ

e A

bu

nd

ance




2-OH Tacrine

4-OH Tacrine

1-OH Tacrine

Naproxen-Glu



95 tile 037

Median 021

M1

0

DrugsMetabolites




2 4 6 8 10 12 14 16 18 20 22 24

Time (min)

0

100

Rela

tive I

nte

nsity

RT 1626

RT 275

RT 422 RT 1188 RT 632


XIC 3100032





Bank Data








Efficiency



Gao et al (2017) JASMS 281248-1249



Backups

Rat Day1


Human

Rat Day105

Human Day 1

Rat Day 314

Human Day 1

1

2

3

4

5

6

Tre

ate

d n

apro

xe

n

Tre

ate

d n

apro

xe

n a

cyl

glu

cu

ron

ide

Untr

eate

d n

ap

roxen

Untr

eate

d n

ap

roxen

acyl

glu

cu

ron

ide

Tre

ate

d s

alic

ylic

acid

Tre

ate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Un

treate

d s

alic

ylic

acid

Untr

eate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Carb

am

aze

pin

e

Carb

am

aze

pin

e-1

01

1-

epo

xid

e

Sulfa

dia

zin

e

N-A

ce

tyl sulfa

dia

zin

e

Tre

ate

d m

eth

ylp

he

nid

ate

Tre

ate

d r

italin

ic a

cid

Untr

eate

dm

eth

ylp

hen

ida

te

Untr

eate

d r

ita

linic

acid

Nic

otin

e I

min

ium

io

n

Cotinin

e

MS

NM

Ra

tio






SN (2-1)025=4









Which one is taller


Which one is taller







Case Study










Human 0220 00406 000788 00210 Rat 770 0502 0292 000762


Human repeat 0184 00290 000506 00164


ratio 2 399 163 526 0378



Rat 269 plusmn 258





Interest


Method




Method Succeeds

Animal lt 2X Human

Animal ge 2X Human



Needed


MIST Decision Tree







Summary














SN

N N

N

O

Cl

NH

Cl

Cl

O

OH

N

OCH3

CH3

NOH

OH

N

NH2

NH N O OH

N3

O

O

ziprasidone

diclofenac tacrine

terfenadine

dextromethorphan

zidovudine

MeO

O

O

OH

OH

OH






(AZT)

Negative Control











0 4 8 12 16 20 24 Time (min)

0

100 R

elat

ive

Ab

un

dan

ce

508

478 682

1132 1435 1450 763

773 243 1835 2120 2138 1164 1312 1638 288 534 1473 2338 802 2045 335 1062

42 43 44 45 46 47 48 49 50 51 52 53 54 55 Time (min)

0

100

Rel

ativ

e In

ten

sity

RT 508

RT 478

RT 495

RT 539

4 8 12 16 20 24 Time (min)

0

100 RT 624

RT 1313

RT 1113

RT 1626

RT 275 RT 879

RT 2008 RT 422

548

165 808

1 2

3

3

4

5

5

4

678

9 10 111213

14 16 15

1

2

3

4

56 7

Rel

ativ

e A

bu

nd

ance




2-OH Tacrine

4-OH Tacrine

1-OH Tacrine

Naproxen-Glu



95 tile 037

Median 021

M1

0

DrugsMetabolites




2 4 6 8 10 12 14 16 18 20 22 24

Time (min)

0

100

Rela

tive I

nte

nsity

RT 1626

RT 275

RT 422 RT 1188 RT 632


XIC 3100032





Bank Data








Efficiency



Gao et al (2017) JASMS 281248-1249



Backups

Rat Day1


Human

Rat Day105

Human Day 1

Rat Day 314

Human Day 1

1

2

3

4

5

6

Tre

ate

d n

apro

xe

n

Tre

ate

d n

apro

xe

n a

cyl

glu

cu

ron

ide

Untr

eate

d n

ap

roxen

Untr

eate

d n

ap

roxen

acyl

glu

cu

ron

ide

Tre

ate

d s

alic

ylic

acid

Tre

ate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Un

treate

d s

alic

ylic

acid

Untr

eate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Carb

am

aze

pin

e

Carb

am

aze

pin

e-1

01

1-

epo

xid

e

Sulfa

dia

zin

e

N-A

ce

tyl sulfa

dia

zin

e

Tre

ate

d m

eth

ylp

he

nid

ate

Tre

ate

d r

italin

ic a

cid

Untr

eate

dm

eth

ylp

hen

ida

te

Untr

eate

d r

ita

linic

acid

Nic

otin

e I

min

ium

io

n

Cotinin

e

MS

NM

Ra

tio









Which one is taller


Which one is taller







Case Study










Human 0220 00406 000788 00210 Rat 770 0502 0292 000762


Human repeat 0184 00290 000506 00164


ratio 2 399 163 526 0378



Rat 269 plusmn 258





Interest


Method




Method Succeeds

Animal lt 2X Human

Animal ge 2X Human



Needed


MIST Decision Tree







Summary














SN

N N

N

O

Cl

NH

Cl

Cl

O

OH

N

OCH3

CH3

NOH

OH

N

NH2

NH N O OH

N3

O

O

ziprasidone

diclofenac tacrine

terfenadine

dextromethorphan

zidovudine

MeO

O

O

OH

OH

OH






(AZT)

Negative Control











0 4 8 12 16 20 24 Time (min)

0

100 R

elat

ive

Ab

un

dan

ce

508

478 682

1132 1435 1450 763

773 243 1835 2120 2138 1164 1312 1638 288 534 1473 2338 802 2045 335 1062

42 43 44 45 46 47 48 49 50 51 52 53 54 55 Time (min)

0

100

Rel

ativ

e In

ten

sity

RT 508

RT 478

RT 495

RT 539

4 8 12 16 20 24 Time (min)

0

100 RT 624

RT 1313

RT 1113

RT 1626

RT 275 RT 879

RT 2008 RT 422

548

165 808

1 2

3

3

4

5

5

4

678

9 10 111213

14 16 15

1

2

3

4

56 7

Rel

ativ

e A

bu

nd

ance




2-OH Tacrine

4-OH Tacrine

1-OH Tacrine

Naproxen-Glu



95 tile 037

Median 021

M1

0

DrugsMetabolites




2 4 6 8 10 12 14 16 18 20 22 24

Time (min)

0

100

Rela

tive I

nte

nsity

RT 1626

RT 275

RT 422 RT 1188 RT 632


XIC 3100032





Bank Data








Efficiency



Gao et al (2017) JASMS 281248-1249



Backups

Rat Day1


Human

Rat Day105

Human Day 1

Rat Day 314

Human Day 1

1

2

3

4

5

6

Tre

ate

d n

apro

xe

n

Tre

ate

d n

apro

xe

n a

cyl

glu

cu

ron

ide

Untr

eate

d n

ap

roxen

Untr

eate

d n

ap

roxen

acyl

glu

cu

ron

ide

Tre

ate

d s

alic

ylic

acid

Tre

ate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Un

treate

d s

alic

ylic

acid

Untr

eate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Carb

am

aze

pin

e

Carb

am

aze

pin

e-1

01

1-

epo

xid

e

Sulfa

dia

zin

e

N-A

ce

tyl sulfa

dia

zin

e

Tre

ate

d m

eth

ylp

he

nid

ate

Tre

ate

d r

italin

ic a

cid

Untr

eate

dm

eth

ylp

hen

ida

te

Untr

eate

d r

ita

linic

acid

Nic

otin

e I

min

ium

io

n

Cotinin

e

MS

NM

Ra

tio


Which one is taller







Case Study










Human 0220 00406 000788 00210 Rat 770 0502 0292 000762


Human repeat 0184 00290 000506 00164


ratio 2 399 163 526 0378



Rat 269 plusmn 258





Interest


Method




Method Succeeds

Animal lt 2X Human

Animal ge 2X Human



Needed


MIST Decision Tree







Summary














SN

N N

N

O

Cl

NH

Cl

Cl

O

OH

N

OCH3

CH3

NOH

OH

N

NH2

NH N O OH

N3

O

O

ziprasidone

diclofenac tacrine

terfenadine

dextromethorphan

zidovudine

MeO

O

O

OH

OH

OH






(AZT)

Negative Control











0 4 8 12 16 20 24 Time (min)

0

100 R

elat

ive

Ab

un

dan

ce

508

478 682

1132 1435 1450 763

773 243 1835 2120 2138 1164 1312 1638 288 534 1473 2338 802 2045 335 1062

42 43 44 45 46 47 48 49 50 51 52 53 54 55 Time (min)

0

100

Rel

ativ

e In

ten

sity

RT 508

RT 478

RT 495

RT 539

4 8 12 16 20 24 Time (min)

0

100 RT 624

RT 1313

RT 1113

RT 1626

RT 275 RT 879

RT 2008 RT 422

548

165 808

1 2

3

3

4

5

5

4

678

9 10 111213

14 16 15

1

2

3

4

56 7

Rel

ativ

e A

bu

nd

ance




2-OH Tacrine

4-OH Tacrine

1-OH Tacrine

Naproxen-Glu



95 tile 037

Median 021

M1

0

DrugsMetabolites




2 4 6 8 10 12 14 16 18 20 22 24

Time (min)

0

100

Rela

tive I

nte

nsity

RT 1626

RT 275

RT 422 RT 1188 RT 632


XIC 3100032





Bank Data








Efficiency



Gao et al (2017) JASMS 281248-1249



Backups

Rat Day1


Human

Rat Day105

Human Day 1

Rat Day 314

Human Day 1

1

2

3

4

5

6

Tre

ate

d n

apro

xe

n

Tre

ate

d n

apro

xe

n a

cyl

glu

cu

ron

ide

Untr

eate

d n

ap

roxen

Untr

eate

d n

ap

roxen

acyl

glu

cu

ron

ide

Tre

ate

d s

alic

ylic

acid

Tre

ate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Un

treate

d s

alic

ylic

acid

Untr

eate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Carb

am

aze

pin

e

Carb

am

aze

pin

e-1

01

1-

epo

xid

e

Sulfa

dia

zin

e

N-A

ce

tyl sulfa

dia

zin

e

Tre

ate

d m

eth

ylp

he

nid

ate

Tre

ate

d r

italin

ic a

cid

Untr

eate

dm

eth

ylp

hen

ida

te

Untr

eate

d r

ita

linic

acid

Nic

otin

e I

min

ium

io

n

Cotinin

e

MS

NM

Ra

tio






Case Study










Human 0220 00406 000788 00210 Rat 770 0502 0292 000762


Human repeat 0184 00290 000506 00164


ratio 2 399 163 526 0378



Rat 269 plusmn 258





Interest


Method




Method Succeeds

Animal lt 2X Human

Animal ge 2X Human



Needed


MIST Decision Tree







Summary














SN

N N

N

O

Cl

NH

Cl

Cl

O

OH

N

OCH3

CH3

NOH

OH

N

NH2

NH N O OH

N3

O

O

ziprasidone

diclofenac tacrine

terfenadine

dextromethorphan

zidovudine

MeO

O

O

OH

OH

OH






(AZT)

Negative Control











0 4 8 12 16 20 24 Time (min)

0

100 R

elat

ive

Ab

un

dan

ce

508

478 682

1132 1435 1450 763

773 243 1835 2120 2138 1164 1312 1638 288 534 1473 2338 802 2045 335 1062

42 43 44 45 46 47 48 49 50 51 52 53 54 55 Time (min)

0

100

Rel

ativ

e In

ten

sity

RT 508

RT 478

RT 495

RT 539

4 8 12 16 20 24 Time (min)

0

100 RT 624

RT 1313

RT 1113

RT 1626

RT 275 RT 879

RT 2008 RT 422

548

165 808

1 2

3

3

4

5

5

4

678

9 10 111213

14 16 15

1

2

3

4

56 7

Rel

ativ

e A

bu

nd

ance




2-OH Tacrine

4-OH Tacrine

1-OH Tacrine

Naproxen-Glu



95 tile 037

Median 021

M1

0

DrugsMetabolites




2 4 6 8 10 12 14 16 18 20 22 24

Time (min)

0

100

Rela

tive I

nte

nsity

RT 1626

RT 275

RT 422 RT 1188 RT 632


XIC 3100032





Bank Data








Efficiency



Gao et al (2017) JASMS 281248-1249



Backups

Rat Day1


Human

Rat Day105

Human Day 1

Rat Day 314

Human Day 1

1

2

3

4

5

6

Tre

ate

d n

apro

xe

n

Tre

ate

d n

apro

xe

n a

cyl

glu

cu

ron

ide

Untr

eate

d n

ap

roxen

Untr

eate

d n

ap

roxen

acyl

glu

cu

ron

ide

Tre

ate

d s

alic

ylic

acid

Tre

ate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Un

treate

d s

alic

ylic

acid

Untr

eate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Carb

am

aze

pin

e

Carb

am

aze

pin

e-1

01

1-

epo

xid

e

Sulfa

dia

zin

e

N-A

ce

tyl sulfa

dia

zin

e

Tre

ate

d m

eth

ylp

he

nid

ate

Tre

ate

d r

italin

ic a

cid

Untr

eate

dm

eth

ylp

hen

ida

te

Untr

eate

d r

ita

linic

acid

Nic

otin

e I

min

ium

io

n

Cotinin

e

MS

NM

Ra

tio





Human 0220 00406 000788 00210 Rat 770 0502 0292 000762


Human repeat 0184 00290 000506 00164


ratio 2 399 163 526 0378



Rat 269 plusmn 258





Interest


Method




Method Succeeds

Animal lt 2X Human

Animal ge 2X Human



Needed


MIST Decision Tree







Summary














SN

N N

N

O

Cl

NH

Cl

Cl

O

OH

N

OCH3

CH3

NOH

OH

N

NH2

NH N O OH

N3

O

O

ziprasidone

diclofenac tacrine

terfenadine

dextromethorphan

zidovudine

MeO

O

O

OH

OH

OH






(AZT)

Negative Control











0 4 8 12 16 20 24 Time (min)

0

100 R

elat

ive

Ab

un

dan

ce

508

478 682

1132 1435 1450 763

773 243 1835 2120 2138 1164 1312 1638 288 534 1473 2338 802 2045 335 1062

42 43 44 45 46 47 48 49 50 51 52 53 54 55 Time (min)

0

100

Rel

ativ

e In

ten

sity

RT 508

RT 478

RT 495

RT 539

4 8 12 16 20 24 Time (min)

0

100 RT 624

RT 1313

RT 1113

RT 1626

RT 275 RT 879

RT 2008 RT 422

548

165 808

1 2

3

3

4

5

5

4

678

9 10 111213

14 16 15

1

2

3

4

56 7

Rel

ativ

e A

bu

nd

ance




2-OH Tacrine

4-OH Tacrine

1-OH Tacrine

Naproxen-Glu



95 tile 037

Median 021

M1

0

DrugsMetabolites




2 4 6 8 10 12 14 16 18 20 22 24

Time (min)

0

100

Rela

tive I

nte

nsity

RT 1626

RT 275

RT 422 RT 1188 RT 632


XIC 3100032





Bank Data








Efficiency



Gao et al (2017) JASMS 281248-1249



Backups

Rat Day1


Human

Rat Day105

Human Day 1

Rat Day 314

Human Day 1

1

2

3

4

5

6

Tre

ate

d n

apro

xe

n

Tre

ate

d n

apro

xe

n a

cyl

glu

cu

ron

ide

Untr

eate

d n

ap

roxen

Untr

eate

d n

ap

roxen

acyl

glu

cu

ron

ide

Tre

ate

d s

alic

ylic

acid

Tre

ate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Un

treate

d s

alic

ylic

acid

Untr

eate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Carb

am

aze

pin

e

Carb

am

aze

pin

e-1

01

1-

epo

xid

e

Sulfa

dia

zin

e

N-A

ce

tyl sulfa

dia

zin

e

Tre

ate

d m

eth

ylp

he

nid

ate

Tre

ate

d r

italin

ic a

cid

Untr

eate

dm

eth

ylp

hen

ida

te

Untr

eate

d r

ita

linic

acid

Nic

otin

e I

min

ium

io

n

Cotinin

e

MS

NM

Ra

tio





Interest


Method




Method Succeeds

Animal lt 2X Human

Animal ge 2X Human



Needed


MIST Decision Tree







Summary














SN

N N

N

O

Cl

NH

Cl

Cl

O

OH

N

OCH3

CH3

NOH

OH

N

NH2

NH N O OH

N3

O

O

ziprasidone

diclofenac tacrine

terfenadine

dextromethorphan

zidovudine

MeO

O

O

OH

OH

OH






(AZT)

Negative Control











0 4 8 12 16 20 24 Time (min)

0

100 R

elat

ive

Ab

un

dan

ce

508

478 682

1132 1435 1450 763

773 243 1835 2120 2138 1164 1312 1638 288 534 1473 2338 802 2045 335 1062

42 43 44 45 46 47 48 49 50 51 52 53 54 55 Time (min)

0

100

Rel

ativ

e In

ten

sity

RT 508

RT 478

RT 495

RT 539

4 8 12 16 20 24 Time (min)

0

100 RT 624

RT 1313

RT 1113

RT 1626

RT 275 RT 879

RT 2008 RT 422

548

165 808

1 2

3

3

4

5

5

4

678

9 10 111213

14 16 15

1

2

3

4

56 7

Rel

ativ

e A

bu

nd

ance




2-OH Tacrine

4-OH Tacrine

1-OH Tacrine

Naproxen-Glu



95 tile 037

Median 021

M1

0

DrugsMetabolites




2 4 6 8 10 12 14 16 18 20 22 24

Time (min)

0

100

Rela

tive I

nte

nsity

RT 1626

RT 275

RT 422 RT 1188 RT 632


XIC 3100032





Bank Data








Efficiency



Gao et al (2017) JASMS 281248-1249



Backups

Rat Day1


Human

Rat Day105

Human Day 1

Rat Day 314

Human Day 1

1

2

3

4

5

6

Tre

ate

d n

apro

xe

n

Tre

ate

d n

apro

xe

n a

cyl

glu

cu

ron

ide

Untr

eate

d n

ap

roxen

Untr

eate

d n

ap

roxen

acyl

glu

cu

ron

ide

Tre

ate

d s

alic

ylic

acid

Tre

ate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Un

treate

d s

alic

ylic

acid

Untr

eate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Carb

am

aze

pin

e

Carb

am

aze

pin

e-1

01

1-

epo

xid

e

Sulfa

dia

zin

e

N-A

ce

tyl sulfa

dia

zin

e

Tre

ate

d m

eth

ylp

he

nid

ate

Tre

ate

d r

italin

ic a

cid

Untr

eate

dm

eth

ylp

hen

ida

te

Untr

eate

d r

ita

linic

acid

Nic

otin

e I

min

ium

io

n

Cotinin

e

MS

NM

Ra

tio







Summary














SN

N N

N

O

Cl

NH

Cl

Cl

O

OH

N

OCH3

CH3

NOH

OH

N

NH2

NH N O OH

N3

O

O

ziprasidone

diclofenac tacrine

terfenadine

dextromethorphan

zidovudine

MeO

O

O

OH

OH

OH






(AZT)

Negative Control











0 4 8 12 16 20 24 Time (min)

0

100 R

elat

ive

Ab

un

dan

ce

508

478 682

1132 1435 1450 763

773 243 1835 2120 2138 1164 1312 1638 288 534 1473 2338 802 2045 335 1062

42 43 44 45 46 47 48 49 50 51 52 53 54 55 Time (min)

0

100

Rel

ativ

e In

ten

sity

RT 508

RT 478

RT 495

RT 539

4 8 12 16 20 24 Time (min)

0

100 RT 624

RT 1313

RT 1113

RT 1626

RT 275 RT 879

RT 2008 RT 422

548

165 808

1 2

3

3

4

5

5

4

678

9 10 111213

14 16 15

1

2

3

4

56 7

Rel

ativ

e A

bu

nd

ance




2-OH Tacrine

4-OH Tacrine

1-OH Tacrine

Naproxen-Glu



95 tile 037

Median 021

M1

0

DrugsMetabolites




2 4 6 8 10 12 14 16 18 20 22 24

Time (min)

0

100

Rela

tive I

nte

nsity

RT 1626

RT 275

RT 422 RT 1188 RT 632


XIC 3100032





Bank Data








Efficiency



Gao et al (2017) JASMS 281248-1249



Backups

Rat Day1


Human

Rat Day105

Human Day 1

Rat Day 314

Human Day 1

1

2

3

4

5

6

Tre

ate

d n

apro

xe

n

Tre

ate

d n

apro

xe

n a

cyl

glu

cu

ron

ide

Untr

eate

d n

ap

roxen

Untr

eate

d n

ap

roxen

acyl

glu

cu

ron

ide

Tre

ate

d s

alic

ylic

acid

Tre

ate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Un

treate

d s

alic

ylic

acid

Untr

eate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Carb

am

aze

pin

e

Carb

am

aze

pin

e-1

01

1-

epo

xid

e

Sulfa

dia

zin

e

N-A

ce

tyl sulfa

dia

zin

e

Tre

ate

d m

eth

ylp

he

nid

ate

Tre

ate

d r

italin

ic a

cid

Untr

eate

dm

eth

ylp

hen

ida

te

Untr

eate

d r

ita

linic

acid

Nic

otin

e I

min

ium

io

n

Cotinin

e

MS

NM

Ra

tio









SN

N N

N

O

Cl

NH

Cl

Cl

O

OH

N

OCH3

CH3

NOH

OH

N

NH2

NH N O OH

N3

O

O

ziprasidone

diclofenac tacrine

terfenadine

dextromethorphan

zidovudine

MeO

O

O

OH

OH

OH






(AZT)

Negative Control











0 4 8 12 16 20 24 Time (min)

0

100 R

elat

ive

Ab

un

dan

ce

508

478 682

1132 1435 1450 763

773 243 1835 2120 2138 1164 1312 1638 288 534 1473 2338 802 2045 335 1062

42 43 44 45 46 47 48 49 50 51 52 53 54 55 Time (min)

0

100

Rel

ativ

e In

ten

sity

RT 508

RT 478

RT 495

RT 539

4 8 12 16 20 24 Time (min)

0

100 RT 624

RT 1313

RT 1113

RT 1626

RT 275 RT 879

RT 2008 RT 422

548

165 808

1 2

3

3

4

5

5

4

678

9 10 111213

14 16 15

1

2

3

4

56 7

Rel

ativ

e A

bu

nd

ance




2-OH Tacrine

4-OH Tacrine

1-OH Tacrine

Naproxen-Glu



95 tile 037

Median 021

M1

0

DrugsMetabolites




2 4 6 8 10 12 14 16 18 20 22 24

Time (min)

0

100

Rela

tive I

nte

nsity

RT 1626

RT 275

RT 422 RT 1188 RT 632


XIC 3100032





Bank Data








Efficiency



Gao et al (2017) JASMS 281248-1249



Backups

Rat Day1


Human

Rat Day105

Human Day 1

Rat Day 314

Human Day 1

1

2

3

4

5

6

Tre

ate

d n

apro

xe

n

Tre

ate

d n

apro

xe

n a

cyl

glu

cu

ron

ide

Untr

eate

d n

ap

roxen

Untr

eate

d n

ap

roxen

acyl

glu

cu

ron

ide

Tre

ate

d s

alic

ylic

acid

Tre

ate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Un

treate

d s

alic

ylic

acid

Untr

eate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Carb

am

aze

pin

e

Carb

am

aze

pin

e-1

01

1-

epo

xid

e

Sulfa

dia

zin

e

N-A

ce

tyl sulfa

dia

zin

e

Tre

ate

d m

eth

ylp

he

nid

ate

Tre

ate

d r

italin

ic a

cid

Untr

eate

dm

eth

ylp

hen

ida

te

Untr

eate

d r

ita

linic

acid

Nic

otin

e I

min

ium

io

n

Cotinin

e

MS

NM

Ra

tio










0 4 8 12 16 20 24 Time (min)

0

100 R

elat

ive

Ab

un

dan

ce

508

478 682

1132 1435 1450 763

773 243 1835 2120 2138 1164 1312 1638 288 534 1473 2338 802 2045 335 1062

42 43 44 45 46 47 48 49 50 51 52 53 54 55 Time (min)

0

100

Rel

ativ

e In

ten

sity

RT 508

RT 478

RT 495

RT 539

4 8 12 16 20 24 Time (min)

0

100 RT 624

RT 1313

RT 1113

RT 1626

RT 275 RT 879

RT 2008 RT 422

548

165 808

1 2

3

3

4

5

5

4

678

9 10 111213

14 16 15

1

2

3

4

56 7

Rel

ativ

e A

bu

nd

ance




2-OH Tacrine

4-OH Tacrine

1-OH Tacrine

Naproxen-Glu



95 tile 037

Median 021

M1

0

DrugsMetabolites




2 4 6 8 10 12 14 16 18 20 22 24

Time (min)

0

100

Rela

tive I

nte

nsity

RT 1626

RT 275

RT 422 RT 1188 RT 632


XIC 3100032





Bank Data








Efficiency



Gao et al (2017) JASMS 281248-1249



Backups

Rat Day1


Human

Rat Day105

Human Day 1

Rat Day 314

Human Day 1

1

2

3

4

5

6

Tre

ate

d n

apro

xe

n

Tre

ate

d n

apro

xe

n a

cyl

glu

cu

ron

ide

Untr

eate

d n

ap

roxen

Untr

eate

d n

ap

roxen

acyl

glu

cu

ron

ide

Tre

ate

d s

alic

ylic

acid

Tre

ate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Un

treate

d s

alic

ylic

acid

Untr

eate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Carb

am

aze

pin

e

Carb

am

aze

pin

e-1

01

1-

epo

xid

e

Sulfa

dia

zin

e

N-A

ce

tyl sulfa

dia

zin

e

Tre

ate

d m

eth

ylp

he

nid

ate

Tre

ate

d r

italin

ic a

cid

Untr

eate

dm

eth

ylp

hen

ida

te

Untr

eate

d r

ita

linic

acid

Nic

otin

e I

min

ium

io

n

Cotinin

e

MS

NM

Ra

tio




95 tile 037

Median 021

M1

0

DrugsMetabolites




2 4 6 8 10 12 14 16 18 20 22 24

Time (min)

0

100

Rela

tive I

nte

nsity

RT 1626

RT 275

RT 422 RT 1188 RT 632


XIC 3100032





Bank Data








Efficiency



Gao et al (2017) JASMS 281248-1249



Backups

Rat Day1


Human

Rat Day105

Human Day 1

Rat Day 314

Human Day 1

1

2

3

4

5

6

Tre

ate

d n

apro

xe

n

Tre

ate

d n

apro

xe

n a

cyl

glu

cu

ron

ide

Untr

eate

d n

ap

roxen

Untr

eate

d n

ap

roxen

acyl

glu

cu

ron

ide

Tre

ate

d s

alic

ylic

acid

Tre

ate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Un

treate

d s

alic

ylic

acid

Untr

eate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Carb

am

aze

pin

e

Carb

am

aze

pin

e-1

01

1-

epo

xid

e

Sulfa

dia

zin

e

N-A

ce

tyl sulfa

dia

zin

e

Tre

ate

d m

eth

ylp

he

nid

ate

Tre

ate

d r

italin

ic a

cid

Untr

eate

dm

eth

ylp

hen

ida

te

Untr

eate

d r

ita

linic

acid

Nic

otin

e I

min

ium

io

n

Cotinin

e

MS

NM

Ra

tio





2 4 6 8 10 12 14 16 18 20 22 24

Time (min)

0

100

Rela

tive I

nte

nsity

RT 1626

RT 275

RT 422 RT 1188 RT 632


XIC 3100032





Bank Data








Efficiency



Gao et al (2017) JASMS 281248-1249



Backups

Rat Day1


Human

Rat Day105

Human Day 1

Rat Day 314

Human Day 1

1

2

3

4

5

6

Tre

ate

d n

apro

xe

n

Tre

ate

d n

apro

xe

n a

cyl

glu

cu

ron

ide

Untr

eate

d n

ap

roxen

Untr

eate

d n

ap

roxen

acyl

glu

cu

ron

ide

Tre

ate

d s

alic

ylic

acid

Tre

ate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Un

treate

d s

alic

ylic

acid

Untr

eate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Carb

am

aze

pin

e

Carb

am

aze

pin

e-1

01

1-

epo

xid

e

Sulfa

dia

zin

e

N-A

ce

tyl sulfa

dia

zin

e

Tre

ate

d m

eth

ylp

he

nid

ate

Tre

ate

d r

italin

ic a

cid

Untr

eate

dm

eth

ylp

hen

ida

te

Untr

eate

d r

ita

linic

acid

Nic

otin

e I

min

ium

io

n

Cotinin

e

MS

NM

Ra

tio





Bank Data








Efficiency



Gao et al (2017) JASMS 281248-1249



Backups

Rat Day1


Human

Rat Day105

Human Day 1

Rat Day 314

Human Day 1

1

2

3

4

5

6

Tre

ate

d n

apro

xe

n

Tre

ate

d n

apro

xe

n a

cyl

glu

cu

ron

ide

Untr

eate

d n

ap

roxen

Untr

eate

d n

ap

roxen

acyl

glu

cu

ron

ide

Tre

ate

d s

alic

ylic

acid

Tre

ate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Un

treate

d s

alic

ylic

acid

Untr

eate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Carb

am

aze

pin

e

Carb

am

aze

pin

e-1

01

1-

epo

xid

e

Sulfa

dia

zin

e

N-A

ce

tyl sulfa

dia

zin

e

Tre

ate

d m

eth

ylp

he

nid

ate

Tre

ate

d r

italin

ic a

cid

Untr

eate

dm

eth

ylp

hen

ida

te

Untr

eate

d r

ita

linic

acid

Nic

otin

e I

min

ium

io

n

Cotinin

e

MS

NM

Ra

tio




Efficiency



Gao et al (2017) JASMS 281248-1249



Backups

Rat Day1


Human

Rat Day105

Human Day 1

Rat Day 314

Human Day 1

1

2

3

4

5

6

Tre

ate

d n

apro

xe

n

Tre

ate

d n

apro

xe

n a

cyl

glu

cu

ron

ide

Untr

eate

d n

ap

roxen

Untr

eate

d n

ap

roxen

acyl

glu

cu

ron

ide

Tre

ate

d s

alic

ylic

acid

Tre

ate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Un

treate

d s

alic

ylic

acid

Untr

eate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Carb

am

aze

pin

e

Carb

am

aze

pin

e-1

01

1-

epo

xid

e

Sulfa

dia

zin

e

N-A

ce

tyl sulfa

dia

zin

e

Tre

ate

d m

eth

ylp

he

nid

ate

Tre

ate

d r

italin

ic a

cid

Untr

eate

dm

eth

ylp

hen

ida

te

Untr

eate

d r

ita

linic

acid

Nic

otin

e I

min

ium

io

n

Cotinin

e

MS

NM

Ra

tio




Backups

Rat Day1


Human

Rat Day105

Human Day 1

Rat Day 314

Human Day 1

1

2

3

4

5

6

Tre

ate

d n

apro

xe

n

Tre

ate

d n

apro

xe

n a

cyl

glu

cu

ron

ide

Untr

eate

d n

ap

roxen

Untr

eate

d n

ap

roxen

acyl

glu

cu

ron

ide

Tre

ate

d s

alic

ylic

acid

Tre

ate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Un

treate

d s

alic

ylic

acid

Untr

eate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Carb

am

aze

pin

e

Carb

am

aze

pin

e-1

01

1-

epo

xid

e

Sulfa

dia

zin

e

N-A

ce

tyl sulfa

dia

zin

e

Tre

ate

d m

eth

ylp

he

nid

ate

Tre

ate

d r

italin

ic a

cid

Untr

eate

dm

eth

ylp

hen

ida

te

Untr

eate

d r

ita

linic

acid

Nic

otin

e I

min

ium

io

n

Cotinin

e

MS

NM

Ra

tio


Rat Day1


Human

Rat Day105

Human Day 1

Rat Day 314

Human Day 1

1

2

3

4

5

6

Tre

ate

d n

apro

xe

n

Tre

ate

d n

apro

xe

n a

cyl

glu

cu

ron

ide

Untr

eate

d n

ap

roxen

Untr

eate

d n

ap

roxen

acyl

glu

cu

ron

ide

Tre

ate

d s

alic

ylic

acid

Tre

ate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Un

treate

d s

alic

ylic

acid

Untr

eate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Carb

am

aze

pin

e

Carb

am

aze

pin

e-1

01

1-

epo

xid

e

Sulfa

dia

zin

e

N-A

ce

tyl sulfa

dia

zin

e

Tre

ate

d m

eth

ylp

he

nid

ate

Tre

ate

d r

italin

ic a

cid

Untr

eate

dm

eth

ylp

hen

ida

te

Untr

eate

d r

ita

linic

acid

Nic

otin

e I

min

ium

io

n

Cotinin

e

MS

NM

Ra

tio


1

2

3

4

5

6

Tre

ate

d n

apro

xe

n

Tre

ate

d n

apro

xe

n a

cyl

glu

cu

ron

ide

Untr

eate

d n

ap

roxen

Untr

eate

d n

ap

roxen

acyl

glu

cu

ron

ide

Tre

ate

d s

alic

ylic

acid

Tre

ate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Un

treate

d s

alic

ylic

acid

Untr

eate

d 2

3-

dih

yd

roxyb

en

zo

ic a

cid

Carb

am

aze

pin

e

Carb

am

aze

pin

e-1

01

1-

epo

xid

e

Sulfa

dia

zin

e

N-A

ce

tyl sulfa

dia

zin

e

Tre

ate

d m

eth

ylp

he

nid

ate

Tre

ate

d r

italin

ic a

cid

Untr

eate

dm

eth

ylp

hen

ida

te

Untr

eate

d r

ita

linic

acid

Nic

otin

e I

min

ium

io

n

Cotinin

e

MS

NM

Ra

tio


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