Cambridge Isotope Laboratories, Inc.
isotope.com

RESEARCH PRODUCTS

Stable Isotope-Labeled
Mixtures, Sets, and Kits

For Mass Spectrometric Applications

Cambridge Isotope Laboratories, Inc.
North America: 1.800.322.1174 cilsales@isotope.com | International: +1.978.749.8000 intlsales@isotope.com | fax: 1.978.749.2768 | isotope.com

| Stable Isotope-Labeled Mixtures, Sets, and Kits

Cambridge Isotope Laboratories, Inc.

Table of Contents
Mixtures and Sets
Pathway-Specific Mixes

Class-Specific Mixes

3
25
TCA Mixes and Sets  ­Amino Acid Mixes and Sets  ­10
Kits
Bile Acid Mixes  ­Carnitine/Acylcarnitine Mixes and Sets

­11

27
Metabolomic QC Kits  ­Fatty Acid/Lipid Extracts­14

Extract Kits  ­32

Nucleic Acid Sets

­15

PeptiQuant™ Plus Assay Kits  ­39

17
INLIGHT® Glycan Tagging Kit  ­42
Organic Acid Mixes  ­19
Q&As  ­
43
Other Mixes  ­23
Steroid Mixes and Sets  ­Please visit isotope.com for pricing and availability.

For research use only. Not for use in diagnostic procedures.

The global and targeted measurement of biomolecules continues to be two areas of growing focus in analytical chemistry and biomedicine.
Drivers for this research include efforts to better understand the underlying mechanisms of disease pathogenesis and to improve precision
medicine through the qualitative/quantitative screening of candidate biomarkers. To address these objectives, mass spectrometry (MS)-based
approaches are increasingly utilized and have been aided by advancements in experimental methodologies, instrumentation technologies,
and bioinformatic tools.
To help advance research in MS ‘omics and MS/MS screening, Cambridge Isotope Laboratories, Inc. (CIL) is pleased to offer the largest
variety of stable isotope-labeled mixtures, sets, and kits. The mixes are formulated neat and/or as solutions and are readily available for
immediate use. In addition to the packaged mix(es), the kits include a user manual and shipping documents (e.g., certificate of analysis,
safety data sheet). The manual outlines general procedures and processing examples for user reference, as well as troubleshooting notes
and analysis results. Please see isotope.com for product details, pricing, and to inquire about customized mixes.

2

Benefits

Features Overview• Easy to implement• Offers end user flexibility• Reduces development time and cost
• Enhances data quality• Improves reproducibility• Renders confidence in analytical results• Mixes supplied in neat and/or solution form
• Preferentially 13C- and/or 15N-enriched• Site-specific or uniform labeling• Established specification guidelines• Procedural guides in kit manuals• Broad-spanning applications (from QC to quantification)tel: +1.978.749.8000 | 1.800.322.1174 (North America) | fax: +1.978.749.2768

| Stable Isotope-Labeled Mixtures, Sets, and Kits

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Mixtures and Sets
The mixtures available off-the-shelf are class- or pathway-specific for ‘omics and MS/MS screening applications. These mixes are amenable
for use in quality control and qualification/quantification exercises using targeted, semi-targeted, or untargeted MS-based methodologies.
Outlined below is an overview of our current mix offerings, as well as details into their compositions and usage specifications (i.e.,reconstitution guidelines, storage, and stability). For reference purposes, example results and published manuscripts are also provided.

Class-Specific Mixes
Amino Acid Mixes and Sets

RESEARCH PRODUCTS

Cambridge Isotope Laboratories, Inc.
isotope.com

Amino Acid Mixtures

Amino acids (AAs) play critical roles in biological functions as both building blocks of peptides/proteins andintermediates of various metabolic pathways (e.g., citric acid cycle, urea cycle). These compounds are also reported
to influence the pathogenesis and propagation of metabolic disorders/disease. To aid continued development andapplication, CIL has formulated a number of stable isotope-labeled (and unlabeled) AA mixtures. These include mixesof the canonical amino acids (MSK-A2 and MSK-CAA), the rare or unnatural non-canonical amino acids (MSK-NCAA),
and a series of reference standard AA mixes (e.g., NSK-A).

For Identification and Quantification
Amino acids play critical roles in biological functions as bothbuilding blocks of peptides/proteins and intermediates of various
metabolic pathways (e.g., citric acid cycle, urea cycle). These
compounds are also reported to influence the pathogenesis andpropagation of metabolic disorders/disease. Other areas of research
investigate amino acids in biomarker and drug-discovery studies.
Catalog No.

Unlabeled mixes
may be available.

Description

To aid research development and application, Cambridge
Please inquire.
MSK-A2
Metabolomics Amino Acid Mix
Isotope Laboratories, Inc. (CIL) has formulated various
MSK-CAA
Canonical Amino Acid Mix
mixtures of stable isotope-labeled amino acids. These
MSK-NCAA
Non-canonical Amino Acid Mix
include mixes of canonical and branched-chain amino acids
MSK-CNCAA
Canonical / Non-canonical Amino Acid Mix Sets
(MSK-CAA and NSK-BCAA), a mix of rare or unnatural nonNEW! MSK-MSM1
Metabolomics Screening and Monitoring Standards Mix 1
canonical amino acids (MSK-NCAA), and mixes of amino acids/
NSK-AA3 (-10X) 3-Plex Amino Acid Mix (or 10X)
derivatives (e.g., NSK-A, NSK-AA3). All mixes are well characterized
NSK-A (-A1)
Amino Acid Standards Mix Set A (or A1)
for use in quality control and qualification/quantification studies
NSK-BCAA
Branched-chain Amino Acid Standards Mix
using targeted or untargeted LC-MS methodologies.

MSK Mixes
MSK-CAA Stable isotope-labeled canonical amino acidmix composition. Reconstituting with 1 mL solvent results inconcentrations of 2.5 mM (exception L-cystine: 1.25 mM).
CompoundGlycine
L-Alanine
L-Arginine·HCl
L-Asparagine·H2O*
L-Aspartic Acid
L-Cystine
L-Glutamic Acid
L-Glutamine*
L-Histidine·HCl·H2O
L-Isoleucine
L-Leucine
L-Lysine·2HCl
L-Methionine
L-Phenylalanine
L-Proline
L-Serine
L-Threonine
L-Tryptophan*
L-Tyrosine
L-Valine

Abbrev.
Gly
Ala
Arg
Asn
Asp
Cys-Cys
Glu
Gln
His
Iso
Leu
Lys
Met
Phe
Pro
Ser
Thr
Trp
Tyr
Val

Label and Enrichment
C2, 99%; 15N, 99%
13
C3, 99%; 15N, 99%
13
C6, 99%; 15N4, 99%
13
C4, 99%; 15N2, 99%
13
C4, 99%; 15N, 99%
13
C6, 99%; 15N2, 99%
13
C5, 99%; 15N, 99%
13
C5, 99%; 15N2, 99%
13
C6, 97%; 15N3, 97%
13
C6, 99%; 15N, 99%
13
C6, 99%; 15N, 99%
13
C6, 99%; 15N2, 99%
13
C5, 99%; 15N, 99%
13
C9, 99%; 15N, 99%
13
C5, 99%; 15N, 99%
13
C3, 99%; 15N, 99%
13
C4, 97%; 15N, 97%
13
C11, 99%; 15N2, 99%
13
C9, 99%; 15N, 99%
13
C5, 99%; 15N, 99%
13*Compounds absent in MSK-A2-1.2. This amino acid mix comprises 17 compounds
and is supplied as a 1.2 mL solution (in 0.1 M HCl).

MSK-NCAA Stable isotope-labeled non-canonical amino acidmix composition. Reconstituting with 1 mL solvent results inconcentrations of 2.5 mM.
Compoundβ-Alanine
L-Azidohomoalanine·HCl
L-Citrulline
L-Dihydroxyphenylalanine
L-Homoarginine·HCl
L-Ornithine·HCl
Sarcosine·HCl

Abbrev.
β-Ala
hAHA
Cit
DOPA
Harg
Orn
Sar

Label and Enrichment
13
C3, 98%; 15N, 96-99%
1,2,3,4-13C4, 99%; 2,4-15N2, 98%
1,2,3,4,5-13C5, 98%
1-13C, ring-13C6, 99%
13
C7, 98%; 15N4, 98%
13
C5, 98%13
C3, 99%; 15N, 98%

Example References
Weber, P. 2022. Determination of amino acids in food and feed by microwave hydrolysis and UHPLCMS/MS. J Chromatogr B Analyt Technol Biomed Life Sci, 1209, 123429-123437.
Rom, O.; Liu, Y.; Finney, A.C.; et al. 2022. Induction of glutathione biosynthesis by glycine-based
treatment mitigates atherosclerosis. Redox Biol, 52, 102313-102326.
Li, S.; Huang, B.; Liu, M; et al. 2022. The association between leucine and diabetic retinopathy indifferent genders: a cross-sectional study in Chinese patients with type 2 diabetes. Front Endocrinol,
13, 806807-806815.
Lim, E.W.; Handzlik, M.K.; Trefts, E.; et al. 2021. Progressive alterations in amino acid and lipid
metabolism correlate with peripheral neuropathy in PolgD257A mice. Sci Adv, 7(42), eabj4077-eabj4091.
Gauthier-Coles, G.; Vennitti, J.; Zhang, Z.; et al. 2021. Quantitative modelling of amino acid transportand homeostasis in mammalian cells. Nat Commun, 12(1), 5282-5300.
Liu, Y.; Zhao, Y.; Shukha, Y.; et al. 2021. Dysregulated oxalate metabolism is a driver and therapeutictarget in atherosclerosis. Cell Rep, 36(4), 109420-109456.
Gray, N.; Lawler, N.G.; Yang, R.; et al. 2021. A simultaneous exploratory and quantitative amino acid andbiogenic amine metabolic profiling platform for rapid disease phenotyping via UPLC-QToF-MS. Talanta,
223(Pt 2), 121872-121883.
van Gastel, N.; Spinelli, J.B.; Sharda, A., et al. 2020. Induction of a timed metabolic collapse to overcome
cancer chemoresistance. Cell Metab, 32(3), 391-403.

Continued ➤Cambridge Isotope Laboratories, Inc.

Overview
Catalog No.
MSK-A2
MSK-CAA
MSK-NCAA
MSK-CNCAAMSK-MSM1
NSK-AA3
NSK-AA3-10X
NSK-A
NSK-A1
NSK-AB
NSK-BCAA

North America: 1.800.322.1174 cilsales@isotope.com | International: +1.978.749.8000 intlsales@isotope.com | fax: 1.978.749.2768 | isotope.com

Description
Metabolomics Amino Acid Mix
Canonical Amino Acid Mix
Non-canonical Amino Acid Mix
Canonical/Non-canonical Amino Acid Mix Sets
Metabolomics Screening and Monitoring Standards Mix 1
3-Plex Amino Acid Standards Mix
3-Plex Amino Acid Standards Mix (10X)
Amino Acid Standards Mix Set A
Amino Acid Standards Mix Set A1
Standards Mix Sets A and B
Branched-chain Amino Acid Standards Mix

No. of Metabolites
1720
720 (in CAA), 7 (in NCAA)
14
3
3
12
12
12 (in NSK-A), 8 (in NSK-B)
4

Unit Size
1.2 mL
1 vial1 vial2 × 1 vial
1 vial1 vial, 10 vials
1 vial, 10 vials
1 vial, 10 vials
1 vial, 10 vials
2 × 10 vials
1 vialCambridge Isotope Laboratories, Inc.
isotope.com

MS/MS STANDARDS

MS/MS Screening
Mixtures and Standards

Cambridge Isotope Laboratories, Inc.
North America: 1.800.322.1174 cilsales@isotope.com | International: +1.978.749.8000 intlsales@isotope.com | fax: 1.978.749.2768 | isotope.com

Click on the thumbnails or visit
isotope.com for more information.

MSK-A2 and MSK-CAA Mixes
Compositions
The A2 mix (in 0.1 M HCl) consists of 17 amino acids (in solution), and the CAA mix (dried down) comprises 20. Reconstituting the CAA
mix in 1 mL solvent (e.g., water) results in 2.5 mM concentrations (exception: L-cystine at 1.25 mM). Note: This CAA mix should not bereconstituted in 0.1 M HCl as Asn and Gln are unstable in acid.
Compound
L-Alanine
L-Arginine·HCl
L-Asparagine*
L-Aspartic acid
L-Cystine
L-Glutamic acid
L-Glutamine*
Glycine
L-Histidine·HCl
L-Isoleucine
L-Leucine
L-Lysine·2HCl
L-Methionine
L-Phenylalanine
L-Proline
L-Serine
L-Threonine
L-Tryptophan*
L-Tyrosine
L-Valine

Abbrev.
Ala
Arg
Asn
Asp
Cys-Cys
Glu
Gln
Gly
His
Ile
Leu
Lys
Met
Phe
Pro
Ser
Thr
Trp
Tyr
Val

Label and Enrichment
13
C3, 99%; 15N, 99%
13
C6, 99%; 15N4, 99%
13
C4, 99%; 15N2, 99%
13
C4, 99%; 15N, 99%
13
C6, 99%; 15N2, 99%
13
C5, 99%; 15N, 99%
13
C5, 99%; 15N2, 99%
13
C2, 99%; 15N, 99%
13
C6, 97%; 15N3, 97%
13
C6, 99%; 15N, 99%
13
C6, 99%; 15N, 99%
13
C6, 99%; 15N2, 99%
13
C5, 99%; 15N, 99%
13
C9, 99%; 15N, 99%
13
C5, 99%; 15N, 99%
13
C3, 99%; 15N, 99%
13
C4, 97%; 15N, 97%
13
C11, 99%; 15N2, 99%
13
C9, 99%; 15N, 99%
13
C5, 99%; 15N, 99%

Conc. (mM)
2.5
2.5
2.5
2.5
1.25
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5

*Compounds absent in MSK-A2.
Chemical purity (CP) is 98% or greater, unless otherwise indicated.For research use only. Not for use in diagnostic procedures.

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| Stable Isotope-Labeled Mixtures, Sets, and Kits

Example Results

Figure. Application of the MSK-A2 mix to compare the quantitative profiles of cow serum and standard media (see PMID: 28826492 for additional information).
Example References
Morley, S.A.; Ma, F.; Alazem, M.; et al. 2023. Expression of malic enzyme reveals subcellular carbon partitioning for storage reserve production in soybeans. New Phytol,
in press.
McNabney, D.W.G.; Mangal, V.; Kirkwood, A.E.; et al. 2023. Phytoplankton metabolite profiles from two Lake Ontario areas of concern reveal differences associated withtaxonomic community composition. Sci Total Environ, 871, 162042-162053.
Handzlik, M.K.; Gengatharan, J.M.; Frizzi, K.E.; et al. 2023. Insulin-regulated serine and lipid metabolism drive peripheral neuropathy. Nature, 614(7946), 118-124.
Shen, J.L.; Doherty, J.; Allen, E.; et al. 2022. Atg6 promotes organismal health by suppression of cell stress and inflammation. Cell Death Differ, 29(11), 2275-2287.
Fuenzalida, K.; Leal-Witt, M.J.; Guerrero, P.; et al. 2021. NTBC treatment monitoring in Chilean patients with tyrosinemia type 1 and its association with biochemical parameters andliver biomarkers. J Clin Med, 10(24), 5832-5845.
Gao, J.; Chung, T-S. 2021. Membranes made from nonsolvent-thermally induced phase separation (N-TIPS) for decellularization of blood in dry plasma spot (DPS) applications.
Chem Eng Sci, 229. doi.org/10.1016/j.ces.2020.116010.
van Gastel, N.; Spinelli, J.B.; Sharda, A.; et al. 2020. Induction of a timed metabolic collapse to overcome cancer chemoresistance. Cell Metab, 32(3), 391-403.
Røst, L.M.; Thorfinnsdottir, L.B.; Kumar, K.; et al. 2020. Absolute quantification of the central carbon metabolome in eight commonly applied prokaryotic and eukaryotic
model systems. Metabolites, 10(2), 74.
Alcock, R.D.; Shaw, G.C.; Tee, N.; et al. 2019. Plasma amino acid concentrations after the ingestion of dairy and collagen proteins, in healthy active males. Front Nutr, 6, 163.
Sullivan, M.R; Danai, L.V.; Lewis, C.A.; et al. 2019. Quantification of microenvironmental metabolites in murine cancers reveals determinants of tumor nutrient availability.
Elife, 8, e44235.
Chen, W.W.; Freinkman, E.; Sabatini, D.M. 2017. Rapid immunopurification of mitochondria for metabolite profiling and absolute quantification of matrix metabolites.
Nat Protoc, 12(10), 2215-2231.
Muir, A.; Danai, L.V.; Gui, D.Y.; et al. 2017. Environmental cystine drives glutamine anaplerosis and sensitizes cancer cells to glutaminase inhibition. Elife, 6, e27713.

Companion unlabeled standard mixes and kits may be available; please inquire.
Chemical purity (CP) is 98% or greater, unless otherwise indicated.

4

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For research use only. Not for use in diagnostic procedures.

| Stable Isotope-Labeled Mixtures, Sets, and Kits

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MSK-NCAA Mix
Composition
Reconstituting the dried-down NCAA mix with 1 mL solvent (e.g., 1:1 v/ v methanol:water) results in 2.5 mM concentrations for eachcompound.
Compoundβ-Alanine
L-Azidohomoalanine·HCl
L-Citrulline
L-Dihydroxyphenylalanine
L-Homoarginine·HCl
L-Ornithine·HCl
Sarcosine·HCl

Abbrev.
β-Ala
hAHA
Cit
DOPA
Harg
Orn
Sar

Label and Enrichment
13
C3, 98%; 15N, 96%
1,2,3,4-13C4, 99%; 2,4-15N2, 98%
1,2,3,4,5-13C5, 98%
1-13C, ring-13C6, 99%
13
C7, 98%; 15N4, 98%
13
C5, 98%13
C3, 99%; 15N, 98%

Usage Specifications
Mix Type
Form
Before reconstitution:
Storage
Recommended retest
Upon reconstitution:
Storage
Recommended retest

MSK-A2
1.2 mL solution

MSK-CAA
MSK-NCAA
dried down

use several of CIL’s metabolomics mixes in my LC-MS“ Ianalysis.
They make quantitative metabolomic work
convenient with a single internal standard mix spike andprovide the corresponding unlabeled mix for absolute
quantitation. The ready mixes save time from the tedioustask of making up individual solutions of each analyte forquantitation. I find the different mixes applicable to avariety of analyses in complicated matrices that provide
metabolic insight to my studies.
– Andrew Downey
Senior Scientist, Axcella Health Inc. (USA)”-5 to 5°C; protect from light
2 years from date of manufacture
N/A
N/A-5 to 5°C; protect from light
4 weeks

MSK-MSM1 Mix
Composition
Reconstituting a labeled MSM1 mix in 10 mL of solvent (e.g., water) yields an equimolar concentration of 500 µM. Note: Corresponding
unlabeled mix has identical compound composition but yields a 100 mM equimolar concentration upon a 1 mL solvent addition.
CompoundL-allo-Isoleucine
DL-2-Aminoadipic acidDL-2-Aminobutyric acid
4-Aminobutyric acid
L-Argininosuccinic acid
DL-Cystathionine
L-Cysteine, N-acetyl
Ethanolamine
π-Methyl-L-histidine
τ-Methyl-L-histidine
DL-Homocystineo-Phospho-L-serineo-Phosphorylethanolamine
TaurineAbbrev.
allo-Ile
Aad
AABA
GABA
ASA
Cyst
NAC
MEA
1-MH
3-MH
Hcys
SEP
PE
T

Label and Enrichment
13
C6, 97%
2,5,5-D3, 98%
D6, 98%13
C4, 97%
arginine-13C6, 99%; 15N4, 99%
3,3,4,4-D4, 98%
cysteine-13C3, 97%; 15N, 97%
1,1,2,2-D4, 98%
methyl-D3, 98%
methyl-D3, 98%
3,3,3',3',4,4,4',4'-D8, 98%
13
C3, 99%; 15N, 98%
1,1,2,2-D4, 98%
13
C2, 99%; 15N, 98%

Usage Specifications
Criteria
Recommendation
Before reconstitution:
Storage-5 to 5°C; protect from light
Recommended retest
2 years from date of manufacture
Upon reconstitution:
Storage-80°C; protect from light
Recommended retest
10 months**Metabolite stability demonstrated through 10 months with MSM1 mix solubilized in water stored at -80°C.

Companion unlabeled standard mixes and kits may be available; please inquire.
Chemical purity (CP) is 98% or greater, unless otherwise indicated.For research use only. Not for use in diagnostic procedures.

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NSK-AA3 Mix
Composition
Reconstituting the AA3 mix in 1 mL of solvent (e.g., 50% acetonitrile) will produce the tabulated concentrations below. Note: NSK-AA310X has 10× the specified concentrations.
Standard (Abbreviation)
Creatine (Cre)Label and Enrichment
N-Methyl-D3; glycine-2,2-D2, 99%

MW (Da)154.18Conc. (µM)
500

Structure
CD3 O

H 2N

N

NH D D

Guanidinoacetic acid (GAA) 1,2-13C2, 97%; 3-15N, 97% (CP 97%)120.09

NH

50
H 2N

L-Proline (Pro)D7, 97%122.17

OH

500

D
D

D

N
H
D

D

N
H

OH
O
D
D

OH

O

Usage Specifications
Criteria
Recommendation
Before reconstitution:
Storage-5 to 5ºC; protect from light
Recommended retest
3 years from date of manufacture
After reconstitution:*
Storage-20ºC
Recommended retest
3 months*Represents minimum stability period when AA3 mix is reconstituted with 1:1 purified water:acetonitrile.

Companion unlabeled standard mixes and kits may be available; please inquire.
Chemical purity (CP) is 98% or greater, unless otherwise indicated.

6

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For research use only. Not for use in diagnostic procedures.

| Stable Isotope-Labeled Mixtures, Sets, and Kits

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NSK-A Mix
Composition
Reconstituting a given NSK-A vial’s contents in 1 mL of high-purity solvent (1:1 water:methanol recommended) will produce the tabulated
concentrations below. To facilitate complete dissolution, it is recommended to vortex manually for 1 minute then auto-vortex for a minimumof 30 seconds. Note: A combined set of NSK-A and NSK-B – the carnitine/acylcarnitine reference standard mix – is also available (NSK-AB).
Compound
L-Alanine
L-Arginine·HCl
L-Aspartatic acid
L-Citrulline
DL-Glutamic acid
Glycine
L-Leucine
L-Methionine
L-Ornithine·HCl*
L-Phenylalanine
L-Tyrosine
L-Valine

Abbrev.
Ala
Arg
Asp
Cit
Glu
Gly
Leu
Met
Orn
Phe
Tyr
Val

Label and Enrichment
2,3,3,3-D4, 98%
5-13C, 99%; 4,4,5,5-D4, 95%
2,3,3-D3, 98%
5,5-D2, 98%
2,4,4-D3, 98%
2-13C, 99%; 15N, 98%
5,5,5-D3, 99%
methyl-D3, 98%
5,5-D2, 98%
ring-13C6, 99%
ring-13C6, 99%
D8, 98%Conc. (µM)
500
500
500
500
500
2500
500
500
500
500
500
500

*NSK-A1 contains Orn 3,3,4,4,5,5,-D6, 98% instead of 5,5-D2, 98%. The remaining components and concentrations areequivalent.

Usage Specifications
Criteria
Recommendation
No. of uses
960 samples/vial
Before reconstitution:
Storage≤25ºC; protect from light
Recommended retest
4 years from date of manufacture
Upon reconstitution:
Storage5±3ºC in a tightly sealed vial
Note: Storing the sealed vial in a second sealed container helps maintain the
integrity of the solution.
Recommended retest
4 weeks

Companion unlabeled standard mixes and kits may be available; please inquire.
Chemical purity (CP) is 98% or greater, unless otherwise indicated.For research use only. Not for use in diagnostic procedures.

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Example Results

A

StandardAla (D4)
Arg (13C/D4)
Asp (D3)
Cit (D2)
Glu (D3)
Gly (13C/15N)
Leu (D3)
Met (D3)
Orn (D2)
Phe (13C6)
Tyr (13C6)
Val (D8)SUAC (13C5)
C0 (D9)C2 (D3)C3 (D3)C4 (D3)C5 (D9)C5-DC (D3)C5-OH (D3)C8 (D3)C12 (D9)
C14 (D9)
C16 (D3)
C18 (D3)

B

Precursor m/z
Derivatized
Nonderivatized
150.2094.07
236.22180.22249.23137.20234.22
178.14263.23151.15134.0978.07
191.22135.19209.23153.12
191.22135.19228.27172.20244.22
188.16182.17126.17216.20160.18227.31
171.24263.23207.21
277.28221.18291.28235.21
311.23255.27391.36279.14321.27265.24347.33291.29409.47353.36437.46381.42459.47403.41487.46431.42

Figure. Full scan MS1 spectra of the DBS internal standards, with the derivatized standards shown in A and the nonderivatized in B. The standard mixture consists of stable
isotope-labeled amino acids (NSK-A and -T) and carnitine/acylcarnitines (NSK-B and NSK-B-G). Please refer to Thermo Scientific technical note #73398 for method andapplication details.
Example References
Li, S.; Huang, B.; Liu, M; et al. 2022. The association between leucine and diabetic retinopathy in different genders: a cross-sectional study in Chinese patients with type 2 diabetes.
Front Endocrinol, 13, 806807-806815.
Habib, A.; Azize, N.A.A.; Rahman, S.A.; et al. 2021. Novel mutations associated with carnitine-acylcarnitine translocase and carnitine palmitoyl transferase 2 deficiencies in Malaysia.
Clin Biochem, 98, 48-53.
Zhang, S.; Li, X.; Luo, H.; et al. 2020. Role of aromatic amino acids in pathogeneses of diabetic nephropathy in Chinese patients with type 2 diabetes. J Diabetes Complications,34(10), 107667.
Wang, L.; Liu, D.; Shen, H.; et al. 2020. Analysis of amino acid patterns with nutrition regimens in preterm infants with extrauterine growth retardation. Front Pediatr, 8, 184.
Brennenstuhl, H.; Kohlmüller, D.; Gramer, G.; et al. 2020. High throughput newborn screening for aromatic L-amino-acid decarboxylase deficiency by analysis of concentrationsof 3-O-methyldopa from dried blood spots. J Inherit Metab Dis, 43(3), 602-610.
Cao, Y-F.; Li , J.; Zhang, Z; et al. 2019. Plasma levels of amino acids related to urea cycle and risk of type 2 diabetes mellitus in Chinese adults. Front Endocrinol, 10, 50.
Jing, F.; Hu, X.; Cao, Y.; et al. 2018. Discriminating gastric cancer and gastric ulcer using human plasma amino acid metabolic profile. UBMB Life, 70(6), 553-562.
Technical Note
Xie, X.; Kozak, M. 2020. Simultaneous analysis of amino acids, acylcarnitines, and succinylacetone in dried blood spots for research using nonderivatized and derivatized methods.
(Thermo Scientific technical note #73398)

been using CIL’s NSK-A and NSK-B reference standards in our LC-MS/MS method for newbom screening tests since 2001. From that“ Wetimehavemore than 4,000,000 newborns have been screened. The products maintain good stability after dissolution and the high analysis
repeatability enabled us to stabilize their use as internal standards. We appreciate the high-quality products, timely deliveries, and excellent
customer relations.
– Mariusz Oltarzewski
Head of Department of Screening Tests and Metabolic Diagnostics, Institute of Mother and Child (Poland)”Companion unlabeled standard mixes and kits may be available; please inquire.
Chemical purity (CP) is 98% or greater, unless otherwise indicated.

8

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For research use only. Not for use in diagnostic procedures.

| Stable Isotope-Labeled Mixtures, Sets, and Kits

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NSK-BCAA Mix
Composition
Reconstituting the BCAA mix in 1 mL of solvent (e.g., 0.1 M HCl) will produce the tabulated concentrations below.
Standard (Abbreviation)
L-Allo-isoleucine (Alle)
L-Isoleucine (Ile)
L-Leucine (Leu)
L-Valine (Val)Label and Enrichment
13
C6, 97%; 15N, 97%
D10, 98%
5,5,5-D3, 99%
13
C5, 99%; 15N, 99%

Conc. (µM)
400
400
400
400

Q1 m/z
139.2
142.2
135.2
124.1

Q3 m/z
92.2
96.2
89.2
77.1

Usage Specifications
Criteria
Recommendation
Before reconstitution:
Storage≤25ºC; protect from light and moisture
Recommended retest
5 years from date of manufacture
After reconstitution:*
Storage
4°C
Recommended retest
5 weeks*Represents minimum stability period when the BCAA mix is reconstituted with 100% water.

Intensity (cps)

2.4e7
2.2e7
2.0e7

13

C5/15N Val
RT 10.80

13

1.8e7
1.6e7

C6/15N Alle
RT 10.21

D3 LeuRT 11.11

D10 IleRT 11.90

1.4e7
1.2e7
1.0e7
8.0e6
6.0e6
4.0e6
2.0e6
6.0

6.5

7.0

7.5

8.0

8.5

9.0

9.5

10.0 10.5 11.0 11.5 12.0 12.5 13.0 13.5

Time (min)Figure. Representative XICs of the BCAA mix metabolites measured by RPLC-MRM/MS (ESI+, 5500+ QqQ). Separationof the isobaric metabolites is shown in the main plot, with Val in the inset. Displayed are the [M+H]+ ions (see Table forprecursor and product ion values).

Companion unlabeled standard mixes and kits may be available; please inquire.
Chemical purity (CP) is 98% or greater, unless otherwise indicated.For research use only. Not for use in diagnostic procedures.

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| Stable Isotope-Labeled Mixtures, Sets, and Kits

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Bile Acid Mixes

RESEARCH PRODUCTS

Cambridge Isotope Laboratories, Inc.
isotope.com

Bile acids (BAs) are steroid-like compounds that act as a detergent in the breakdown of fats. This family ofcompounds comprises primary BAs (synthesized in the liver) and secondary BAs (produced in the gut by bacteria).
These are essential regulatory compounds that are involved in various metabolic processes (e.g., cholesterol and lipid
metabolism) and signaling interactions (e.g., in glucose and energy homeostasis). Investigations into their synthesis/
metabolism, disease linkage, and biomarker potential are examples of the type of research studies being undertaken.
To aid further research and development efforts in this space, CIL has formulated stable isotope-labeled (andunlabeled) BA mixes. These dried-down mixes are constructed with the unconjugated BAs in one vial and the
conjugated BAs in a second.

Metabolomics
Bile Acid Mixtures
For Qualification and Quantification
Bile acids (BAs) are steroid-like compounds that act as a detergent in the breakdown of fats. This family of compounds comprises primary
BAs (synthesized in the liver) and secondary BAs (produced in the gut via primary BA modification). These are essential regulatory compounds
that are involved in various metabolic processes (e.g., cholesterol and lipid metabolism) and signaling interactions (e.g., in glucose and energy
homeostasis). Investigations into their synthesis /metabolism, disease linkage, and biomarker potential are examples of the studied explorations.
To aid further research and development efforts in this space, Cambridge Isotope Laboratories, Inc. (CIL) is pleased to offer stable
isotope-labeled and unlabeled BA mixes. These formulations are dried down, with the unconjugated BAs in one vial and the conjugatedBAs in a second (see table for mix compositions). The mixes can be used in untargeted and targeted LC-MS methods for a variety of purposes(from quality control to quantification). Please inquire for pricing.
Catalog No.
MSK-BA1
MSK-BA1-25X

Description
Bile Acid Standards Mix 1 – UnconjugatedBile Acid Standards Mix 1 (25X) Unconjugated

Catalog No.
MSK-BA2MSK-BA2-20X

Description
Bile Acid Standards Mix 2 – ConjugatedBile Acid Standards Mix 2 (20X) – Conjugated

Table. Composition of stable isotope-labeled BA mixes. Reconstituting each standard vial in 1 mL of solvent (e.g., 50% methanol) willproduce an equimolar concentration of 100 μM (0.04 mg/mL BA1 and 0.05 mg/mL BA2). The concentrated BA mixes are formulated
to equal mass of 1 mg/mL, which is rendered upon a 1 mL solvent addition.
Compound (label, enrichment)
Chenodeoxycholic acid (2,2,4,4-D4, 98%)
Cholic acid (2,2,4,4-D4, 98%)
Deoxycholic acid (2,2,4,4-D4, 98%)
Lithocholic acid (2,2,4,4-D4, 98%)
β-Muricholic acid (2,2,3,4,4-D5, 99%)
Ursodeoxycholic acid (2,2,4,4-D4, 98%) CP 95%
Glycochenodeoxycholic acid (2,2,4,4-D4, 98%) CP 97%
Glycocholic acid (2,2,4,4-D4, 98%) CP 96%
Glycodeoxycholic acid (2,2,4,4-D4, 98%)
Glycolithocholic acid (2,2,4,4-D4, 98%)
Glycoursodeoxycholic acid (2,2,4,4-D4, 98%) CP 97%
Taurochenodeoxycholic acid, sodium salt (2,2,4,4-D4, 98%) CP 97%
Taurocholic acid, sodium salt (2,2,4,4-D4, 98%)
Taurodeoxycholic acid, sodium salt (2,2,4,4-D4, 98%)
Taurolithocholic acid, sodium salt (2,2,4,4-D4, 98%)
Tauroursodeoxycholic acid, sodium salt (2,2,4,4-D4, 98%)

Abbrev.

Type

Vial No.

CDCA
CA
DCA
LCA
β-MCA
UDCA
GCDCA
GCA
GDCA
GLCA
GUDCA
TCDCA
TCA
TDCA
TLCA
TUDCA

Unconjugated (primary)Unconjugated (primary)Unconjugated (secondary)
Unconjugated (secondary)
Unconjugated (primary)Unconjugated (secondary)
Conjugated (primary)
Conjugated (primary)
Conjugated (secondary)
Conjugated (secondary)
Conjugated (secondary)
Conjugated (primary)
Conjugated (primary)
Conjugated (secondary)
Conjugated (secondary)
Conjugated (secondary)

1
1
1
1
1
1
2
2
2
2
2
2
2
2
2
2

Chemical purity (CP) is 98% or greater, unless otherwise indicated. For research use only. Not for use in diagnostic procedures.
Example References
Engevik, M.E.; Thapa, S.; Lillie, I.; et al. 2023. Repurposing dried blood spot (DBS) device technology
to examine bile acid profiles in human dried fecal spot (DFS) samples. Am J Physiol Gastrointest Liver
Physiol, 326(2), G95-G106.
Foley, M.H.; Walker, M.E.; Stewart, A.K.; et al. 2023. Bile salt hydrolases shape the bile acid landscapeand restrict Clostridioides difficile growth in the murine gut. Nat Microbiol, 8(4), 611-628.

Application Notes
Baker, P.R.S.; Proos, R.; Seferovic, M.D.; Horvath, T.D. 2023. Quantitative analysis and structural characterization of bile acids using the ZenoTOF 7600 System. (SCIEX application note MKT-30747-A)
Horvath, T.D.; Engevik, A.C.; Percy, A.J. 2022. Bile acid analysis in mouse samples to study livercholestasis etiology. (CIL application note #52)

Cambridge Isotope Laboratories, Inc.
North America: 1.800.322.1174 cilsales@isotope.com | International: +1.978.749.8000 intlsales@isotope.com | fax: 1.978.749.2768 | isotope.com
MET_RSCH_BAMIX (2/15/24) Supersedes all previously published literature

Click on the thumbnail or visit
isotope.com for more information.

Overview
Catalog No.
MSK-BA1
MSK-BA1-25X
MSK-BA2MSK-BA2-20X

Description
Bile Acid Standards Mix 1 – UnconjugatedBile Acid Standards Mix 1 (25X) – UnconjugatedBile Acid Standards Mix 2 – ConjugatedBile Acid Standards Mix 2 (20X) – Conjugated

No. of Metabolites
6
6
10
10

Unit Size
1 vial1 vial1 vial1 vialNote: The concentrated bile acid mixes are formulated to equal mass (instead of equimolar) concentrations at 1 mg/mL, which is rendered
upon a 1 mL solvent addition.
Compositions
Reconstituting a given BA mix in 1 mL of solvent (e.g., 1:1 v/ v methanol:water) yields an equimolar concentration of 100 µM.
Unconjugated BA Mix (MSK-BA1)
Compound
Chenodeoxycholic acid
Cholic acid
Deoxycholic acid
Lithocholic acid
β-Muricholic acid
Ursodeoxycholic acid

Abbrev.
CDCA
CA
DCA
LCA
β-MCA
UDCA

Type of BA
Primary
Primary
Secondary
SecondaryPrimary
SecondaryLabel and Enrichment
2,2,4,4-D4, 98%
2,2,4,4-D4, 98%
2,2,4,4-D4, 98%
2,2,4,4-D4, 98%
2,2,3,4,4-D5, 99%
2,2,4,4-D4, 98% (CP 95%)

Conjugated BA Mix (MSK-BA2)
CompoundGlycochenodeoxycholic acid
Glycocholic acid
Glycodeoxycholic acid
Glycolithocholic acid
Glycoursodeoxycholic acid
Taurochenodeoxycholic acid, sodium salt
Taurocholic acid, sodium salt
Taurodeoxycholic acid, sodium salt
Taurolithocholic acid, sodium salt
Tauroursodeoxycholic acid, sodium salt

Abbrev.
GCDCA
GCA
GDCA
GLCA
GUDCA
TCDCA
TCA
TDCA
TLCA
TUDCA

Type of BA
Primary
Primary
Secondary
Secondary
SecondaryPrimary
Primary
Secondary
Secondary
SecondaryLabel and Enrichment
2,2,4,4-D4, 98% (CP 97%)
2,2,4,4-D4, 98% (CP 96%)
2,2,4,4-D4, 98%
2,2,4,4-D4, 98%
2,2,4,4-D4, 98% (CP 97%)
2,2,4,4-D4, 98% (CP 97%)
2,2,4,4-D4, 98%
2,2,4,4-D4, 98%
2,2,4,4-D4, 98%
2,2,4,4-D4, 98%

Usage Specifications

Example References

Criteria
Recommendation
Before reconstitution:
Storage-20ºC; protect from light
Recommended retest
2 years from date of manufacture
After reconstitution:
Storage
4ºC
Recommended retest
4 weeks

Engevik, M.E.; Thapa, S.; Lillie, I.; et al. 2024. Repurposing dried blood spot (DBS)
device technology to examine bile acid profiles in human dried fecal spot (DFS)
samples. Am J Physiol Gastrointest Liver Physiol, 326(2), G95-G106.
Foley, M.H.; Walker, M.E.; Stewart, A.K.; et al. 2023. Bile salt hydrolases shape the bile
acid landscape and restrict Clostridioides difficile growth in the murine gut. Nat
Microbiol, 8(4), 611-628.
Application Notes
Baker, P.R.S.; Proos, R.; Seferovic, M.D.; Horvath, T.D. 2023. Quantitative analysis andstructural characterization of bile acids using the ZenoTOF 7600 System. (SCIEX
application note MKT-30747-A)
Horvath, T.D.; Engevik, A.C.; Percy, A.J. 2022. Bile acid analysis in mouse samples tostudy liver cholestasis etiology. (CIL application note #52)

Companion unlabeled standard mixes and kits may be available; please inquire.
Chemical purity (CP) is 98% or greater, unless otherwise indicated.

10

tel: +1.978.749.8000 | 1.800.322.1174 (North America) | fax: +1.978.749.2768

For research use only. Not for use in diagnostic procedures.

| Stable Isotope-Labeled Mixtures, Sets, and Kits

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he materials that Cambridge Isotope“ TLaboratories(CIL) supply are always of the

Example Results
140

LCA

highest quality in terms of chemical and isotopic
purity. Recently, we have embarked on two new
major research areas involving the analyses ofbile acids (BAs) and per- and polyfluorinatedalkylsubstances (PFAS). The standards/mixes from
CIL that enable these, and other, projects are
precisely what we have come to expect –perfection!
– David C. Muddiman, PhD
Jacob and Betty Belin Distinguished Professor,
Department of Chemistry, North Carolina State University
Director of Molecular Education, Technology, andResearch Innovation Center (METRIC)

0

1

DCA

CDCA

2

3

”

GLCA
TLCA

GDCA

TDCA

20
0

GCDCA

GCA
TCA

40

TCDCA

60

CA

80

GUDCA

MCA(b)

100

GUDCA
TUDCA

Intensity (106, cps)

120

4

5

6

Retention Time (min)Figure. Chromatographic overview of a combined BA mix (6 unconjugates and 10 conjugates) analysis
measured by RPLC-MS (Orbitrap ID-X, negative ESI). The m/z displayed are the [M-H]- ions. Procedurally,
the labeled /unlabeled BA mixes were reconstituted in 50% methanol before aliquot mixing and MS1
measurement. Note: The labeled and unlabeled BAs coeluted.

Carnitine/Acylcarnitine Mixes and Sets

Cambridge Isotope Laboratories, Inc.
isotope.com

Carnitine and acylcarnitines play an essential role in fatty acid metabolism. Metabolism disorders of fatty acidoxidation and several organic acidurias impose major clinical manifestations (e.g., hypoketotic hypoglycemia, skeletal
myopathy, liver disease, and/or failure). These are largely attributed to enzymatic deficiencies and can be monitored
through carnitine/acylcarnitine measurement. To aid MS/MS screening studies, CIL has formulated stable isotopelabeled (and unlabeled) standard carnitine/acylcarnitine mixes.

MS/MS STANDARDS

MS/MS Screening
Mixtures and Standards

Overview
Cambridge Isotope Laboratories, Inc.
North America: 1.800.322.1174 cilsales@isotope.com | International: +1.978.749.8000 intlsales@isotope.com | fax: 1.978.749.2768 | isotope.com

Catalog No.
NSK-B
NSK-B-G1
NSK-AB

Description
Carnitine/Acylcarnitine Standards Mix Set B
Carnitine/Acylcarnitine Standards Mix
Supplement to NSK-B
Standards Mix Sets A and B

No. of Metabolites
85

Unit Size
1 vial, 10 vials
1 vial, 10 vials

12 (in NSK-A), 8 (in NSK-B)

2 × 10 vials

Click on the thumbnail or visit
isotope.com for more information.

clinical chemistry-based applications of mass spectrometry, the first lesson the laboratory learns is the requisite nature of stable isotope“ Inenriched
standards for quantification of metabolites in biological fluids. In newborn screening of amino acids and acylcarnitines, Cambridge
Isotope Laboratories, Inc., set the standard for quantification of these metabolites in dried blood spots. As research and development of thenewborn screening analysis by mass spectrometry progressed, it was clear that a half dozen isotope-labeled internal standards would not beadequate for the analysis of an amino acid and acylcarnitine profile, together comprising a range of 500 separate mass units and more than
30 important metabolites, most of which require accurate quantification. When screening began to expand beyond research, it was clear thatweighing out small quantities of individual standards would reduce accuracy and introduce unnecessary error. Therefore, together, we set out
to develop sets of standards for amino acids and acylcarnitine analysis that would enable quantification. We started this development morethan 20 years ago adding, changing and improving these standards. CIL, together with the early developers of tandem mass spectrometrybased newborn screening, set the standard by which all other laboratories follow. CIL’s commitment to supporting the metabolic and newborn
screening community is exceptional. It is our good fortune in the clinical chemistry and mass spectrometry community to have CIL as part ofour laboratory solutions.
– Donald H. Chace, PhD MSFS FACB
Medolac Laboratories (USA)”Companion unlabeled standard mixes and kits may be available; please inquire.
Chemical purity (CP) is 98% or greater, unless otherwise indicated.For research use only. Not for use in diagnostic procedures.

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| Stable Isotope-Labeled Mixtures, Sets, and Kits

Cambridge Isotope Laboratories, Inc.

Compositions
Reconstituting a given NSK-B or NSK-B-G1 vial’s contents in 1 mL of highly pure methanol will produce the tabulated concentrations
below. To facilitate complete dissolution, it is recommended to vortex manually for 1 minute then auto-vortex for a minimum of
30 seconds. Note: A combined set of NSK-B and NSK-A – the amino acid reference standard mix – is also available (NSK-AB).
NSK-B
Compound
L-Carnitine
O-Acetyl-L-carnitine·HCl
O-Propionyl-L-carnitine·HCl
O-Butyryl-L-carnitine·HCl
O-Isovaleryl-L-carnitine·HCl
O-Octanoyl-L-carnitine·HCl
O-Myristoyl-L-carnitine·HCl
O-Palmitoyl-L-carnitine·HCl

Abbrev.
C0
C2
C3
C4
C5
C8
C14
C16

Label and Enrichment
trimethyl-D9, 98%
N-methyl-D3, 98%
N-methyl-D3, 98%
N-methyl-D3, 98%
N,N,N-trimethyl-D9, 98%
N-methyl-D3, 98%
N,N,N-trimethyl-D9, 98%
N-methyl-D3, 98%

Conc. (µM)
152
38
7.6
7.6
7.6
7.6
7.6
15.2

NSK-B-G1
Compound
O-Glutaryl-L-carnitine·ClO4
3-Hydroxyisovaleryl-L-carnitine·ClO4
O-Dodecanoyl-L-carnitine·HCl
O-3-DL-Hydroxypalmitoyl-L-carnitine·ClO4
O-Octadecanoyl-L-carnitine·HCl

Abbrev.
C5-DC
C5-OH
C12
C16-OH
C18

Label and Enrichment
N-methyl-D3, 98% (CP 97%)
N-methyl-D3, 98%
N,N,N-trimethyl-D9, 98%
N-methyl-D3, 98%
N-methyl-D3, 98%

Conc. (µM)
15.2
7.6
7.6
15.2
15.2

Usage Specifications
Mix Type
No. of uses
Before reconstitution:
Storage
Recommended retest
After reconstitution:
Storage

Recommended retest

NSK-B

NSK-B-G1
960 samples/vial≤8°C; protect from light
1 year from date of manufacture
2 years from date of manufacture

5±3ºC in a tightly sealed vial
5±3ºC in a tightly sealed vial
Note: Storing the sealed vial in a second sealed container helps maintain the
integrity of the solution
4 weeks

Companion unlabeled standard mixes and kits may be available; please inquire.
Chemical purity (CP) is 98% or greater, unless otherwise indicated.

12

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Example Results

A

B

StandardAla (D4)
Arg (13C/D4)
Asp (D3)
Cit (D2)
Glu (D3)
Gly (13C/15N)
Leu (D3)
Met (D3)
Orn (D2)
Phe (13C6)
Tyr (13C6)
Val (D8)SUAC (13C5)
C0 (D9)C2 (D3)C3 (D3)C4 (D3)C5 (D9)C5-DC (D3)C5-OH (D3)C8 (D3)C12 (D9)
C14 (D9)
C16 (D3)
C18 (D3)

Precursor m/z
Derivatized
Nonderivatized
150.2094.07
236.22180.22249.23137.20234.22
178.14263.23151.15134.0978.07
191.22135.19209.23153.12
191.22135.19228.27172.20244.22
188.16182.17126.17216.20160.18227.31
171.24263.23207.21
277.28221.18291.28235.21
311.23255.27391.36279.14321.27265.24347.33291.29409.47353.36437.46381.42459.47403.41487.46431.42

Figure. Full scan MS1 spectra of the DBS internal standards, with the derivatized standards shown in A and the nonderivatized in B. The standard mixture consists of stable
isotope-labeled amino acids (NSK-A and -T) and carnitine/acylcarnitines (NSK-B and NSK-B-G). Please refer to Thermo Scientific technical note #73398 for method andapplication details.
Example References
Habib, A.; Nazri, M.I.B.A.; Rahman, S.A.; et al. 2023. Dataset from dried blood spot acylcarnitine for detection of Carnitine-Acylcarnitine Translocase (CACT) deficiency and Carnitine
Palmitoyl Transferase 2 (CPT2) deficiency. Data Brief, 48, 109090-109096.
Mak, J.; Peng, G.; Le, A.; et al. 2023. Validation of a targeted metabolomics panel for improved second-tier newborn screening. J Inherit Metab Dis, 46(2), 194-205.
Daas, S.; Salah, N.A.; Anikster, Y.; et al. 2023. Addition of galactose-1-phosphate measurement enhances newborn screening for classical galactosemia. J Inherit Metab Dis,46(2), 232-242.
Habib, A.; Azize, N.A.A.; Rahman, S.A.; et al. 2021. Novel mutations associated with carnitine-acylcarnitine translocase and carnitine palmitoyl transferase 2 deficiencies in Malaysia.
Clin Biochem, 98, 48-53.
Brailova, M.; Clerfond, G.; Trésorier, R.; et al. 2020. Inherited metabolic diseases and cardiac pathology in adults: diagnosis and prevalence in a cardiometabo study.
J Clin Med, 9(3), 694-707.
Varkey, A.; Devi, S.; Mukhopadhyay, A.; et al. 2020. Metabolome and microbiome alterations related to short-term feeding of a micronutrient-fortified, high-quality legume
protein-based food product to stunted school age children: a randomized controlled pilot trial. Clin Nutr, 39(11), 3251-3261.
Brennenstuhl, H.; Kohlmüller, D.; Gramer, G.; et al. 2020. High throughput newborn screening for aromatic ι-amino-acid decarboxylase deficiency by analysis of concentrationsof 3-O-methyldopa from dried blood spots. J Inherit Metab Dis, 43(3), 602-610.
Cao, B.; Wang, D.; Pan, Z.; et al. 2019. Characterizing acylcarnitine biosignatures for schizophrenia: a longitudinal pre- and post-treatment study. Transl Psychiatry, 9(1), 19.
Puskarich, M.A.; Evans, C.R.; Karnovsky, A.; et al. 2018. Septic shock nonsurvivors have persistently elevated acylcarnitines following carnitine supplementation. Shock, 49(4),412-419.
Technical Note
Xie, X.; Kozak, M. 2020. Simultaneous analysis of amino acids, acylcarnitines, and succinylacetone in dried blood spots for research using nonderivatized and derivatized methods.
(Thermo Scientific technical note #73398)

Chemical purity (CP) is 98% or greater, unless otherwise indicated.For research use only. Not for use in diagnostic procedures.

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| Stable Isotope-Labeled Mixtures, Sets, and Kits

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Fatty Acid/Lipid Extracts

RESEARCH PRODUCTS

Cambridge Isotope Laboratories, Inc.
isotope.com

Fatty acids and lipids are important biological compounds that are essential to the regulation and control of cellularfunctions and metabolic pathways. These biomolecules are also tied to the energetic balance of an organism. Theirqualitative/quantitative analysis has emerged to better understand the underlying pathophysiology, as well as to
identify new biomarkers or diagnose existing ones. To aid such initiatives, CIL offers an array of mixed fatty acids andtriglycerides. These dried-down extracts are uniformly labeled and available in different forms (i.e., free acid, methyl
ester) as research-grade material.

Stable Isotopes
for Lipidomics
For Monitoring Lipid Synthesis
and TurnoverFatty Acids (FAs)
Catalog No.

Description

4:0 Butyric Acid or Butanoic Acid
CLM-9666
Butyric acid 1-13C, 99%)
CLM-9215
Butyric acid (13C4, 99%)
DLM-1110
Butyric acid (3,3,4,4,4-D5, 97%)
DLM-1508
Butyric acid (D7, 98%)
5:0 Pentanoic Acid or Valeric Acid
CLM-10349
Pentanoic acid (3,5-13C2, 99%)
DLM-572
Pentanoic acid (D9, 98%)
6:0 Hexanoic Acid or Caproic Acid
CLM-3519
Hexanoic acid (1-13C, 99%)
DLM-3030
Hexanoic acid (2,2-D2, 98%)
DLM-612Hexanoic acid (methyl-D3, 98%)
DLM-11023
Hexanoic acid (4,4,5,5,6,6,6-D7, 98%)
DLM-277
Hexanoic acid (D11, 98%)
7:0 Heptanoic Acid or Enanthic Acid
DLM-11088
Heptanoic acid (7,7,7-D3, 99%)
DLM-1820
Heptanoic acid (2,2,3,3-D4, 98%)
DLM-2731
Heptanoic acid (D13, 98%)
8:0 Octanoic Acid or Caprylic Acid
CLM-293
Octanoic acid (1-13C, 99%)
CLM-293-MPT
Octanoic acid (1-13C, 99%)
CLM-3827
Octanoic acid (1,2-13C2, 99%)
CLM-2721
Octanoic acid (1,2,3,4-13C4, 99%)
CLM-3981
Octanoic acid (13C8, 99%)
DLM-619
Octanoic acid (D15, 98%)
9:0 Nonanoic Acid or Pelargonic Acid
CLM-8724
Nonanoic acid (U-13C9, 98%)
DLM-7490
Nonanoic acid (9,9,9-D3, 98%)
DLM-9501
Nonanoic acid (D17, 98%)
10:0 Decanoic Acid or Capric Acid
DLM-2006
Decanoic acid (methyl-D3, 98%)
DLM-270
Decanoic acid (D19, 98%)
12:0 Dodecanoic Acid or Lauric Acid
CLM-1586
Lauric acid (1-13C, 99%)
DLM-3062
Lauric acid (methyl-D3, 99%)
DLM-563
Lauric acid (D23, 98%)

Catalog No.

Description

13:0 Tridecanoic Acid or Tridecylic Acid
DLM-1392
Tridecanoic acid (D25, 98%)
14:0 Myristic Acid or Tetradecanoic Acid
DLM-2922
DL-3-Hydroxymyristic acid (2,2,3,4,4-D5, 96%)
CLM-1844
Myristic acid (1-13C, 99%)
CLM-3665
Myristic acid (1,2,3-13C3, 99%)
DLM-1039
Myristic acid (methyl-D3, 98%)
DLM-7487
Myristic acid (13,13,14,14,14-D5, 98%)
DLM-11024
Myristic acid (12,12,13,13,14,14,14-D7, 98%)
DLM-208
Myristic acid (D27, 98%)
15:0 Pentadecanoic Acid or Pentadecylic Acid
DLM-1307
Pentadecanoic acid (methyl-D3, 98%)
16:0 Palmitic Acid or Hexadecanoic Acid
CLM-150
Palmitic acid (1-13C, 99%)
CLM-150-MPT
Palmitic acid (1-13C, 99%)
CLM-2120
Palmitic acid (2-13C, 99%)
CLM-214
Palmitic acid (1,2-13C2, 99%)
CLM-7896
Palmitic acid (1,2,3,4-13C4, 99%)
CLM-7896-MPT
Palmitic acid (1,2,3,4-13C4, 99%)
CLM-10962
Palmitic acid (1,2,3,4,5,6-13C6, 99%)
CLM-409
Palmitic acid (U-13C16, 98%)
CLM-409-MPT
Palmitic acid (U-13C16, 98%)
DLM-8673
Palmitic acid (12-D, 98%)
DLM-1153
Palmitic acid (2,2-D2, 98%)
DLM-2890
Palmitic acid (9,9-D2, 98%)
DLM-2891
Palmitic acid (13,13-D2, 98%)
DLM-611
Palmitic acid (methyl-D3, 98%)
DLM-2893
Palmitic acid (7,7,8,8-D4, 98%)
DLM-2893-MPT
Palmitic acid (7,7,8,8-D4, 98%)
DLM-2894
Palmitic acid (11,11,12,12-D4, 98%)
DLM-9424
Palmitic acid (13,13,14,14,15,15,16,16,16-D9, 98%)
DLM-2895
Palmitic acid (9,9,...16,16,16-D17, 98%) CP 97%
DLM-215
Palmitic acid (D31, 98%)
DLM-215-MPT
Palmitic acid (D31, 98%)
ULM-10680
Palmitic acid (unlabeled)

MPT: microbiological /pyrogen tested

Chemical purity (CP) is 98% or greater, unless otherwise indicated. For research use only. Not for diagnostic procedures.

Continued ➤Cambridge Isotope Laboratories, Inc.
North America: 1.800.322.1174 cilsales@isotope.com | International: +1.978.749.8000 intlsales@isotope.com | fax: 1.978.749.2768 | isotope.com

Overview
Catalog No.
CLM-8455
DLM-8572
CDLM-8376
CLM-8381
DLM-2497
DLM-8375

Description
Fatty Acid Algal Extract (U-13C, 98%)
Fatty Acid Algal Extract (U-D, 96%)
Fatty Acid Algal Extract (U-13C, 98%; U-D, 97%)
Fatty Acid, Methyl Ester Algal Extract (U-13C, 98%)
(unlabeled terminal ester) CP 95%
Fatty Acid, Methyl Ester Algal Extract (U-D, 96%)
Triglyceride Algal Extract (U-D, 97%)

No. of Metabolites
5-10
5-10
5-10
5-10

Unit Size
0.25 g, 1 g
Please inquire
0.25 g, 1 g
0.25 g, 1 g

5-10
5-10

Please inquire
0.25 g, 1 g

Click on the thumbnail or visit
isotope.com for more information.

Compositions
These fatty acid and triglyceride products are derived from the Agmenelum quadriplicatum algal source. Please inquire for details as thecompositions can vary by lot.
Usage Specifications
Mix Type

Catalog No.

Fatty acids

CLM-8455
DLM-8572
CDLM-8376
CLM-8381
DLM-2497
DLM-8375

Fatty acid, methyl esters
Triglycerides

Before Reconstitution
Storage
Recommended Retest-5 to 5ºC; protect from light
2 years-5 to 5ºC; protect from light

5 years-5 to 5ºC; protect from light

5 years

Example References
Xie, J.; Bruggeman, A.; De Nolf, C.; et al. 2023. Gut microbiota regulates blood-cerebrospinal fluid barrier function and Aβ pathology. EMBO J, 42(17), e111515-3111533.
Xia, F.; Guo, L.; Cui, P.; et al. 2023. A sensitive and accurate GC-MS method for analyzing microbial metabolites short chain fatty acids and their hydroxylated derivatives in newborn
fecal samples. J Pharm Biomed Anal, 223, 115148-115152.
Zeng, X.; Xing, X.; Gupta, M.; et al. 2022. Gut bacterial nutrient preferences quantified in vivo. Cell, 185(18), 3441-3456.
Bihan, D.G.; Rydzak, T.; Wyss, M.; et al. 2022. Method for absolute quantification of short chain fatty acids via reverse phase chromatography mass spectrometry. PLoS One, 17(4),e0267093-0267106.
Chen, J.; Zou, L.; Lu, G.; et al. 2022. PFKP alleviates glucose starvation-induced metabolic stress in lung cancer cells via AMPK-ACC2 dependent fatty acid oxidation.
Cell Discov, 8(1), 52-67.
Hernandez-Saavedra, D.; Sanders, L.; Freeman, S.; et al. 2020. Stable isotope metabolomics of pulmonary artery smooth muscle and endothelial cells in pulmonary hypertension
and with TGF-beta treatment. Sci Rep, 10(1), 413-426.
He, C.; Weston, T.A.; Jung, R.S.; et al. 2018. NanoSIMS analysis of intravascular lipolysis and lipid movement across capillaries and into cardiomyocytes. Cell Metab, 27(5),1055-1066.

Chemical purity (CP) is 98% or greater, unless otherwise indicated.

14

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| Stable Isotope-Labeled Mixtures, Sets, and Kits

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Nucleic Acid Sets
Nucleic acids are necessary biomolecules of living systems, being fundamentally important to a multitude of cellular processes. Its basic
building blocks are nucleobases (e.g., adenine, cytosine), nucleosides (e.g., adenosine, guanosine), and nucleotides (e.g., ATP, CDP). The
qualification/quantification of these compounds is conducted for a number of purposes. This includes the screening of metabolic errors andevaluating the efficacy of drug treatments (be it anticancer, antiviral, or immunosuppressive), among other target areas. CIL offers a variety
of stable isotope-labeled nucleic acid sets to help advance MS- and/or NMR-based research. The sets consist of deoxyribonucleosidemonophosphates (dNMPs), deoxyribonucleoside triphosphates (dNTPs), and ribonucleoside triphosphate (rNTPs) in different unit
configurations.
Overview
Catalog No.
CNLM-7871-SL
NLM-7512-SL
DLM-7511-SL
CNLM-7513-SL
NLM-7519-SL
DLM-7518-SL
CNLM-7503-SL
NLM-7519-CA
DLM-7518-CA

Description
Set of 4 2’-deoxyribonucleoside 5’-monophosphates, lithium
salt (13C, 98%; 15N, 98%) (in solution) CP 95%
Set of 4 2’-deoxyribonucleoside 5’-triphosphates, lithium salt
(15N, 98%) (in solution) CP 95%
Set of 4 2’-deoxyribonucleoside 5’-triphosphates, lithium salt
(D, 98%) (in solution) CP 95%
Set of 4 2’-deoxyribonucleoside 5’-triphosphates, lithium salt
(13C, 98%; 15N, 98%) (in solution) CP 95%
Set of 4 ribonucleoside 5’-triphosphates, lithium salt
(15N, 98%) (in solution) CP 95%
Set of 4 ribonucleoside 5’-triphosphates, lithium salt
(D, 98%) (in solution) CP 95%
Set of 4 ribonucleoside 5’-triphosphates, lithium salt
(13C, 98%; 15N, 98%) (in solution) CP 95%
Set of 4 ribonucleoside 5’-triphosphates, ammonium salt
(15N; 98%) (in solution) CP 90%
Set of 4 ribonucleoside 5’-triphosphates, ammonium salt
(D, 98%) (in solution) CP 95%

CNLM-7503-CA Set of 4 ribonucleoside 5’-triphosphates, ammonium salt
(13C, 98%; 15N, 98%) (in solution) CP 95%

Chemical purity (CP) is 98% or greater, unless otherwise indicated.

No. of Metabolites
4

Unit Size10 mg

410 mg, 50 mg

4

10 mg, 50 mg

4

10 mg, 50 mg

4

10 mg, 50 mg

4

10 mg, 50 mg

4

10 mg, 50 mg

4

4 × 20 μmol
4 × 100 μmol
4 × 20 μmol
4 × 50 μmol
4 × 100 μmol
4 × 20 μmol
4 × 50 μmol
4 × 100 μmol

44

For research use only. Not for use in diagnostic procedures.

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| Stable Isotope-Labeled Mixtures, Sets, and Kits

Cambridge Isotope Laboratories, Inc.

dNMP
Composition and Specification
Catalog No.
CNLM-7871-SL

dNMP
AMP, CMP, GMP, TMP

Conc. (mM)
80

• Supplied on dry ice as a solution of 5 mM Tris-HCl (pH 7.5) in water.
• Store below -20ºC; protect from light.

dNTPs
Compositions and Specifications
Catalog No.
NLM-7512-SL
DLM-7511-SL
CNLM-7513-SL

dNTP

Conc. (mM)

dATP, dCTP, dGTP, dTTP

50-100• Supplied on dry ice as a solution of 5 mM Tris-HCl (pH 7.5) in water (for NLM-7512-SL and CNLM-7513-SL) or D2O (for DLM-7511-SL)• Store below -20ºC; protect from light.
Example References
Liu, B.; Winkler, F.; Herde, M.; et al. 2019. A link between deoxyribonucleotide metabolites and embryonic cell-cycle control. Curr Biol, 29(7), 1187-1192.
Song, Y.; Marmion, R.A.; Park, J.O.; et al. 2017. Dynamic control of dNTP synthesis in early embryos. Dev Cell, 42(3), 301-308.

rNTPs
Compositions and Specifications
Catalog No.
NLM-7519-SL
DLM-7518-SL
CNLM-7503-SL

rNTP

Conc. (mM)rATP, rCTP, rGTP, rTTP

50-100• Supplied on dry ice as a solution of 5 mM Tris-HCl (pH 7.5) in water (for NLM-7519-SL and CNLM-7503-SL) or D2O (for DLM-7518-SL)• Store below -20ºC; protect from light.
Catalog No.
NLM-7519-CA
DLM-7518-CA
CNLM-7503-CA

rNTP

Conc. (mM)rATP, rCTP, rGTP, rUTP

20, 50, and/or 100• Supplied on dry ice as a solution (e.g., 100 mM – contains 100 µmol of each rNTP in 1 mL water, pH ~7.5).
• Store at -20°C; protect from light. Recommended retest is 5 years from date of manufacture.

Companion unlabeled standard mixes and kits may be available; please inquire.
Chemical purity (CP) is 98% or greater, unless otherwise indicated.

16

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| Stable Isotope-Labeled Mixtures, Sets, and Kits

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Organic Acid Mixes

Organic Acid Mixtures

Organic acids are byproducts of amino acids and are intermediates in various biochemical pathways. To aid MS-based
metabolomic research endeavors, CIL has formulated an isotope-enriched organic acid mix. This single-vial
mixture comprises 33 organic acids (MSK-OA) and is also available as an unlabeled mix (MSK-OA-US).

For Qualification, Quantification,and Systems Biology

Organic acids are byproducts of amino acids and are intermediates in various biochemical pathways. To aid MS-based metabolomic
research endeavors, Cambridge Isotope Laboratories, Inc. (CIL) has formulated an isotope-enriched organic acid mix.
This single-vial mixture comprises 33 organic acids (MSK-OA) and is also available as an unlabeled mix (MSK-OA-US).
Table. Composition of the stable isotope-labeled organic acid mixture. Reconstituting this dried-down mix in 1 mL of solvent (e.g., water,
50% methanol) yields an equimolar concentration of 250 µM.

Overview
Catalog No.
MSK-OA

Description
Organic Acid Mix

No. of Metabolites
33

Keto Acids

Hydroxy Acids

α-Ketobutyric acid, sodium salt (13C4, 98%)
α-Ketoglutaric acid, disodium salt (1,2,3,4-13C4, 99%)
α-Ketoisocaproic acid, sodium salt (13C6, 99%)
α-Ketoisovaleric acid, sodium salt (13C5, 98%)
Sodium pyruvate (13C3, 99%)
Diacids
Adipic acid, disodium salt (13C6, 99%)
Fumaric acid, disodium salt (13C4, 99%)
Maleic acid, disodium salt·H2O (13C4, 99%)
Malonic acid, disodium salt (13C3, 99%)
Methylmalonic acid, disodium salt (13C4, 99%)
Oxalic acid, disodium salt (1,2-13C2, 99%)
Sodium isobutyrate (13C4, 99%)
Succinic acid, disodium salt (13C4, 99%)
Monoacids
Sodium acetate (1,2-13C2, 99%)
Sodium butyrate (13C4, 99%)
Sodium propionate (13C3, 99%)

Glycolic acid, sodium salt (1,2-13C2, 99%)
Malic acid, disodium salt·H2O (13C4, 99%)
Sodium D-3-hydroxybutyrate (13C4, 99%) CP 97%
Sodium L-lactate (13C3, 98%)
DL 2-Hydroxyglutarate, disodium salt (13C5, 99%)
Aromatic Acids
Hippuric acid (ring-13C6, 99%)
Homovanillic acid, sodium salt (1,2-13C2, 98%)
Phthalic acid, disodium salt (13C4, 99%)
Sodium benzoate (ring-13C6, 99%)
DL-Vanilmandelic acid (ring-13C6, 99%)
Other Acids
trans-Aconitic acid (triscarboxyl-13C3) CP 95%
L-Ascorbic acid (13C6, 98%)
Creatine (15N3, 98%)
Orotic acid, sodium salt (15N2, 98%)
Sodium D-gluconate (13C6, 99%)
Trisodium citrate, hemihydrate (1,5,6-carboxy-13C3, 99%)
Uric acid, sodium salt (15N2, 98%) CP 95%

Example Reference

Unit Size
1 vialDeArmond, P.D.; Bunch, D.R. 2022. Quantitation of non-derivatized free amino acids for detecting inborn errors of metabolism by incorporating mixed-mode chromatography withtandem mass spectrometry. J Mass Spectrom Adv Clin Lab, 25, 1-11.
Application Note
Petucci, C.*; Percy, A.J.*; Zelenin, A.; Gardell, S.J.; Backiel, K. 2017. Organic acid quantitation in mouse muscle by ion chromatography-mass spectrometry with isotopically labeledstandards. (CIL application note #47)
*These authors contributed equally.
Chemical purity (CP) is 98% or greater, unless otherwise indicated. For research use only. Not for use in diagnostic procedures.

Cambridge Isotope Laboratories, Inc.
North America: 1.800.322.1174 cilsales@isotope.com | International: +1.978.749.8000 intlsales@isotope.com | fax: 1.978.749.2768 | isotope.com
MET_RSCH_OAMIX (8/27/24)
Supersedes all previously published literature

Compositions
Composition of the stable isotope-labeled organic acid mixture. Reconstituting this dried-down mix in 1 mL of solvent(e.g., water, 1:1 v/ v methanol:water) yields an equimolar concentration of 250 µM.
Keto Acids
α-Ketobutyric acid, sodium salt (13C4, 98%)
α-Ketoglutaric acid, disodium salt (1,2,3,4-13C4, 99%)
α-Ketoisocaproic acid, sodium salt (13C6, 99%)
α-Ketoisovaleric acid, sodium salt (13C5, 98%)
Sodium pyruvate (13C3, 99%)
Diacids
Adipic acid, disodium salt (13C6, 99%)
Fumaric acid, disodium salt (13C4, 99%)
Maleic acid, disodium salt·H2O (13C4, 99%)
Malonic acid, disodium salt (13C3, 99%)
Methylmalonic acid, disodium salt (13C4, 99%)
Oxalic acid, disodium salt (1,2-13C2, 99%)
Sodium isobutyrate (13C4, 99%)
Succinic acid, disodium salt (13C4, 99%)
Monoacids
Sodium acetate (1,2-13C2, 99%)
Sodium butyrate (13C4, 99%)
Sodium propionate (13C3, 99%)

RESEARCH PRODUCTS

Cambridge Isotope Laboratories, Inc.
isotope.com

Click on the thumbnail or visit
isotope.com for more information.

Hydroxy Acids
Glycolic acid, sodium salt (1,2-13C2, 99%)
Malic acid, disodium salt·H2O (13C4, 99%)
Sodium D-3-hydroxybutyrate (13C4, 99%) CP 97%
Sodium L-lactate (13C3, 98%)
DL 2-Hydroxyglutarate, disodium salt (13C5, 99%)
Aromatic Acids
Hippuric acid (ring-13C6, 99%)
Homovanillic acid, sodium salt (1,2-13C2, 98%)
Phthalic acid, disodium salt (13C4, 99%)
Sodium benzoate (ring-13C6, 99%)
DL-Vanilmandelic acid (ring-13C6, 99%)
Other Acids
trans-Aconitic acid (triscarboxyl-13C3) CP 95%
L-Ascorbic acid (13C6, 98%)
Creatine (15N3, 98%)
Orotic acid, sodium salt (15N2, 98%)
Sodium D-gluconate (13C6, 99%)
Trisodium citrate, hemihydrate (1,5,6-carboxy-13C3, 99%)
Uric acid, sodium salt (15N2, 98%) CP 95%

Usage Specifications
Criteria
Recommendation
Before reconstitution:
Storage-5 to 5ºC; protect from light
Recommended retest
2 years from date of manufacture
After reconstitution:
Storage
4ºC
Recommended retest
4 weeks

Note: Extended mix stabilities have been demonstrated when solubilized in 50% methanol and stored as specified above.

Companion unlabeled standard mixes and kits may be available; please inquire.
Chemical purity (CP) is 98% or greater, unless otherwise indicated.For research use only. Not for use in diagnostic procedures.

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| Stable Isotope-Labeled Mixtures, Sets, and Kits

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Example Results
Butyrate(13C4, m/z 91.0586)

α-Ketoisovalerate(13C5, m/z 120.0568)

Hippuric acid
(13C6, m/z 184.0709)

Orotate(15N2, m/z 157.0039)

Lactate(13C3, m/z 92.0345)

RT: 1.55

RT: 1.55

RT: 2.20

RT: 2.39

RT: 3.06

Time (min)Time (min)Time (min)Time (min)Time (min)Creatine(15N3, m/z 133.0534)

Malonate(13C2, m/z 106.0138)

2-Hydroxyglutarate(13C5, m/z 152.0466)

Oxalate(13C2, m/z 90.9948)

trans-Aconitate(13C3, m/z 176.0193)

RT: 4.19

RT: 4.86

RT: 4.98

RT: 5.16

RT: 5.60

Time (min)Time (min)Time (min)Time (min)Time (min)Figure. XICs of example metabolites measured in the labeled OA master mix by HILIC-MS (Orbitrap ID-X, negative ESI). Shown are a collection of OAs from a variety of subset
classes. The m/z displayed are the [M-H]- ions. Procedurally, the dried-down master mix was reconstituted in 1 mL of 50% methanol then an aliquot 10-fold diluted before
MS1 analysis.
Example Reference
DeArmond, P.D.; Bunch, D.R. 2022. Quantitation of non-derivatized free amino acids for detecting inborn errors of metabolism by incorporating mixed-mode chromatography withtandem mass spectrometry. J Mass Spectrom Adv Clin Lab, 25, 1-11.
Application Note
Petucci, C.*; Percy, A.J.*; Zelenin, A.; Gardell, S.J.; Backiel, K. 2017. Organic acid quantitation in mouse muscle by ion chromatography-mass spectrometry with isotopically labeledstandards. (CIL application note #47)
*These authors contributed equally.

Companion unlabeled standard mixes and kits may be available; please inquire.
Chemical purity (CP) is 98% or greater, unless otherwise indicated.

18

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For research use only. Not for use in diagnostic procedures.

| Stable Isotope-Labeled Mixtures, Sets, and Kits

isotope.com

Other Mixes

Cambridge Isotope Laboratories, Inc.
isotope.com

A number of additional stable isotope-labeled mixes are also available at CIL (see overview). Included in thiscollection is a lysophosphatidylcholine (LPC or lysoPC) mix (NSK-LPC), a metabolomics standard mix (MSK-MET1), andMS/MS screening mixes for basic and translational MS research. The details of these mix offerings are outlined below.

MS/MS STANDARDS

MS/MS Screening
Mixtures and Standards

Overview
Catalog No.
NSK-LPC
NSK-NI
NSK-KR
NSK-FA
NSK-GA
NSK-MP
NSK-POMSK-MET1

Description
Lysophosphatidylcholine Mix
Acid Sphingomyelinase Substrate and Internal Standard Mix
Galactocerebrosidase Substrate and Internal Standard Mix
α-Galactosidase Substrate and Internal Standard Mix
Glucocerebrosidase Substrate and Internal Standard Mix
α-L-Iduronidase Substrate and Internal Standard Mix
Acid α-Glucosidase Substrate and Internal Standard Mix
Metabolomics Standard Mix 1

No. of Metabolites
4
1 (S + IS)
1 (S + IS)
1 (S + IS)
1 (S + IS)
1 (S + IS)
1 (S + IS)
11

Unit Size
1 vial1 vial1 vial1 vial1 vial1 vial1 vial1 vial, 5 vials, 10 vials

Cambridge Isotope Laboratories, Inc.
North America: 1.800.322.1174 cilsales@isotope.com | International: +1.978.749.8000 intlsales@isotope.com | fax: 1.978.749.2768 | isotope.com

Cambridge Isotope Laboratories, Inc.
isotope.com

Stable Isotope-Labeled Products
For Metabolic Research

NSK-LPC Mix

Cambridge Isotope Laboratories, Inc.
North America: 1.800.322.1174 cilsales@isotope.com | International: +1.978.749.8000 intlsales@isotope.com | fax: 1.978.749.2768 | isotope.com

Composition
Reconstituting a given LysoPC mix (labeled, NSK-LPC; unlabeled, NSK-LPC-US) in 1 mL of high-purity solvent(e.g., 95:5 v/v methanol:water) will produce the concentrations specified below. To facilitate complete dissolution,
it is recommended to sonicate the vial for 3 minutes then auto-vortex for a minimum of 10 seconds.
Standard (Abbreviation)
Lysophosphatidylcholine 20:0 (LysoPC 20:0)
Lysophosphatidylcholine 22:0 (LysoPC 22:0)
Lysophosphatidylcholine 24:0 (LysoPC 24:0)
Lysophosphatidylcholine 26:0 (LysoPC 26:0)

RESEARCH PRODUCTS

Label and Enrichment
eicosanoyl-12,12,13,13-D4, 98%

Conc. (µM)
5.5docosanoyl-1,2,3,4,5,6-13C6, 99%

5.5

tetracosanoyl-1,2,3,4,5,6-13C6, 99%

5.5

hexacosanoyl-1,2,3,4,5,6-13C6, 99%

5.5

Click on the thumbnails or visit
isotope.com for more information.

StructureP

CH3(CH2)6(CD2)2(CH2)10

CH3(CH2)15(CH2)5

CH3(CH2)17(CH2)5

CH3(CH2)19(CH2)5

P

P

P

Usage Specifications
Criteria
Recommendation
No. of uses
~765 samples/vial
Before reconstitution:
Storage-20ºC; protect from light
Recommended retest
3 years from date of manufacture

After reconstitution:
Storage
Recommended retest

5±3ºC or -20±5ºC
6 weeks

to 2,000 newborn specimens every day is a challenging job. In order to ensure high data quality and proper efficacy of an“ TestingMS-based1,200
screening assay (1st and 2nd tier), we utilize CIL’s lyophilized LysoPC mix. This 4-plex cocktail helps reduce significantly ourlaboratory day-to-day variation and preparation time. We also use the LysoPC mixes (both unlabeled and isotope-labeled) in the tuningof our mass spectrometers and in determining response factors. This ensures that the concentrations rendered for each LysoPC is accurately
obtained and can be cross-checked from platform-to-platform.
– Kuldeep Dhillon, Research Scientist Supervisor I
California Department of Public Health (USA)”Companion unlabeled standard mixes and kits may be available; please inquire.
Chemical purity (CP) is 98% or greater, unless otherwise indicated.For research use only. Not for use in diagnostic procedures.

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| Stable Isotope-Labeled Mixtures, Sets, and Kits

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Example Results

Time (min)Time (min)Lysophosphatidylcholine 26:0
LysoPC 26:0; (13C6, m/z 641.85)
RT: 1.44% Intensity

Lysophosphatidylcholine 24:0
LysoPC 24:0; (13C6, m/z 613.81)
RT: 1.27% Intensity

Lysophosphatidylcholine 22:0
LysoPC 22:0; (13C6, m/z 585.75)
RT: 1.09% Intensity% Intensity

Lysophosphatidylcholine 20:0
LysoPC 20:0; (D4, m/z 555.77)
RT: 0.91

Time (min)Time (min)Figure. XICs of four labeled LysoPCs from the NSK-LPC mix measured by LC-MS/MS (positive ESI, Xevo-TQS). These serve as individual calibrators for their unlabeled counterparts insample screening experiments. Data courtesy of CA Department of Public Health.

MS/MS Screening Mixes
α-Galactosidase Substrate and Internal Standard (NSK-FA-1)
Each vial contains the following compounds at a molar ratio of 500:1.
Substrate(6-Benzoylamino-hexyl)-{2-[4-(3,4,5-trihydroxy-6-hydroxymethyl-tetrahydropyran-2-yloxy)-phenylcarbamoyl]-ethyl}-carbamic acid t-butyl ester
C33H47N3O10
MW: 645.7 DaInternal Standard(6-D5-Benzoylamino-hexyl)-[2-(4-hydroxy-phenyl-carbamoyl)-ethyl]-carbamic
acid t-butyl ester
C27H32N3O5D5
MW: 488.5 Da

Glucocerebrosidase Substrate and Internal Standard (NSK-GA-1)
Each vial contains the following compounds at a molar ratio of 50:1.
Substrate
D-Glucosyl-β1-1‘-N-dodecanoyl-D-erythro-sphingosine[C12-glucocerebroside]
C36H69NO8
MW: 643.9 DaInternal Standard
N-Myristoyl-D-erythro-sphingosine [C14-ceramide]
C32H63NO3

MW: 509.8 Da

Galactocerebrosidase Substrate and Internal Standard (NSK-KR-1)
Each vial contains the following compounds at a molar ratio of 150:1.
Substrate
D-Galactosyl-β1-1’-octanoyl-D-erythro-sphingosine [C8-galactosylceramide]
C32H61NO8
MW: 587.8 DaInternal Standard
N-Decanoyl-D-erythro-sphingosine [C10-ceramide]
C28H55NO3
MW: 453.7 Da

α-L-Iduronidase Substrate and Internal Standard (NSK-MP-1)
Each vial contains the following compounds at a molar ratio of 150:1.
Substrate(7-(1-Iduronic acid)-oxycoumarin-4-methylamine-(5’-N-boc-aminopentanoyl)amide)
C26H34N2O12
MW: 566.6 DaInternal Standard(7-Hydroxycoumarin-4-methylamine-(4’-N-boc-aminobutanoyl)-amide)
C19H24N2O6

Chemical purity (CP) is 98% or greater, unless otherwise indicated.

20

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MW: 376.4 Da

For research use only. Not for use in diagnostic procedures.

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Acid Sphingomyelinase Substrate and Internal Standard (NSK-NI-1)
Each vial contains the following compounds at a molar ratio of 50:1.
Substrate
N-Hexanoyl-D-erythro-sphingosylphosphorylcholine [C6-sphingomyelin]
C29H59N2O6P
MW: 562.8 DaInternal Standard
N-Butyroyl-D-erythro-sphingosine [C4-ceramide]
C22H43NO3
MW: 369.6 Da

Acid α-Glucosidase Substrate and Internal Standard (NSK-PO-1)
Each vial contains the following compounds at a molar ratio of 100:1.
Substrate(7-Benzoylamino-heptyl)-{2-[4-(3,4,5-trihydroxy-6-hydroxymethyl-tetrahydropyran-2-yloxy)-phenylcarbamoyl]-ethyl}-carbamic acid t-butyl ester
C34H49N3O10
MW: 659.8 DaInternal Standard(7-D5-Benzoylamino-heptyl)-[2-(4-hydroxy-phenyl-carbamoyl)-ethyl]-carbamic
acid t-butyl ester
C28H34N3O5D5
MW: 502.7 Da

Usage Specifications
Criteria
Recommendation
Use~600 samples/vial
Before reconstitution:
Storage-20°; protect from light
Recommended retest
2 years from date of manufacture
After reconstitution:
Storage5±3ºC or -20±5ºC
Recommended retest
4 weeks
Example References
Ribas, G.; De Mari, J.F.; Civallero, G.; et al. 2017. Validation of a multiplex tandem mass spectrometry method for the detection of selected lysosomal storage diseases in dried blood
spots. JIMES, 5, 1-7.
Tortorelli, S.; Turgeon, C.T.; Gavrilov, D.K.; et al. 2016. Simultaneous testing for 6 lysosomal storage disorders and x-adrenoleukodystrophy in dried blood spots by tandem mass
spectrometry. Clin Chem, 62(9), 1248-1254.
Cho,S.E.; Kwak, J.R.; Lee, H.; et al. 2016. Triplex tandem mass spectrometry assays for the screening of 3 lysosomal storage disorders in a Korean population. Clin Chim Acta, 454,
20-27.

Chemical purity (CP) is 98% or greater, unless otherwise indicated.For research use only. Not for use in diagnostic procedures.

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| Stable Isotope-Labeled Mixtures, Sets, and Kits

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MSK-MET1 Mix
Composition
Reconstituting a given vial in 1 mL of solvent (e.g., water) will produce the tabulated concentrations below. Note: The unlabeled 2DG was
included to examine its metabolic uptake in tracer studies.
Compound
L-Alanine
L-Carnitine
Creatinine
2-Deoxy-D-glucose
D-Glucose
L-Glutamine
L-Leucine
L-Phenylalanine
Sodium butyrate
Sodium propionate
Urea

Abbrev.
Ala
C0
Crn
2DG
Glc
Gln
Leu
Phe
BTA
PA
UR

Label and Enrichment
2,3,3,3-D4, 98%
trimethyl-D9, 98%
N-methyl-D3, 98%
unlabeled
U-13C6, 99%
2,3,3,4,4-D5, 97%
5,5,5-D3, 99%
ring-D5, 98%
D7, 98%D5, 98%15
N2, 98%Conc. (mM)
20
0.4
0.7
50
400
8
5
0.5
2
2
20

Usage Specifications
Criteria
Recommendation
Before reconstitution:
Storage
4ºC; protect from light
Recommended retest
2 years from date of manufacture

Example Results

A)

B)

L-Alanine(D4, m/z 92.065)

2-Deoxy-D-Glucose(unlabeled, m/z 163.061)

D-Glucose(13C6, m/z 185.076)

RT: 1.32

RT: 2.24

RT: 1.81

Time (min)Time (min)Time (min)L-Carnitine(D9, m/z 171.168)

L-Phenylalanine(D5, m/z 171.117)

Urea(15N2, m/z 63.034)

RT: 0.76

RT: 1.59

RT: 0.76

Time (min)Time (min)Time (min)Figure. Representative XICs of example metabolites in the MET1 mix measured in mouse plasma by LC-MS analysis. A TripleTOF 6600 was fronted by a Hypercarb™ Porous
Graphitic Carbon HPLC column (100 × 4.6 mm, 3 µm) in A) and a Kinetex F5 HPLC column (100 × 2.1 mm, 2.6 µm) in B) operated under negative and positive ESI, respectively.
Data was courtesy of the Kibbey lab at Yale School of Medicine.

Chemical purity (CP) is 98% or greater, unless otherwise indicated.

22

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Steroid Mixes and Sets

Cambridge Isotope Laboratories, Inc.
isotope.com

Steroids play vital roles in the regulation of a diverse array of cellular functions and physiological processes.
These pertain to development, reproduction, homeostasis, and metabolism, among others. Accurate quantificationof this compound class is essential for basic and translation research. To aid MS-based research endeavors in thisspace, CIL is pleased to offer a few different types of stable isotope-labeled steroid mixes.

MS/MS STANDARDS

MS/MS Screening
Mixtures and Standards

Overview
Catalog No.
NSK-S
NSK-S-40X
NSK-S-EXP

Description
Steroid Standards Mix Set S
Steroid Standards Mix Set S (40X)
Expanded Steroid Standards Mix Set S

No. of Metabolites
559

Unit Size
1 vial, 10 vials
1 vial, 10 vials
1 vial, 10 vials

Cambridge Isotope Laboratories, Inc.
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isotope.com for more information.

NSK-S Mix
Composition
Reconstituting a given vial’s contents in 1 mL of highly pure methanol will produce the concentrations listed in the table below. To facilitate
complete dissolution, it is recommended to vortex manually for 1 minute then auto-vortex for a minimum of 30 minutes. Note: NSK-S-40X
has 40× the specified concentrations.
Compound
4-Androstene-3,17-dione
Cortisol
11-Deoxycortisol
21-Deoxycortisol
17α-Hydroxyprogesterone

Abbrev.
A4
F
11-S
21-S
17-OHP

Label and Enrichment
2,2,4,6,6,16,16-D7, 97%
9,11,12,12-D4, 98%
2,2,4,6,6-D5, 98%
2,2,4,6,6,21,21,21-D8, 97%
2,2,4,6,6,21,21,21-D8, 98%

Conc. (nM)
20
100
20
20
20

Usage Specifications
Criteria
Recommendation
No. of uses
48 samples/vial
Before reconstitution:
Storage-5 to 5ºC; protect from light
Recommended retest
5 years from date of manufacture
Upon reconstitution:
Storage
Store in a tightly sealed vial at 5±3ºC.
Note: Storing the sealed vial in a second sealed container helps maintain the
integrity of the solution.
Recommended retest
4 weeks
Example References
Gervasoni, J.; Schiattarella, A.; Primiano, A.; et al. 2016. Simultaneous quantification of 17-hydroxyprogesterone, androstenedione, testosterone and cortisol in human serumby LC-MS/MS using TurboFlow online sample extraction. Clin Biochem, 49(13-14), 998-1003.
Hicks, R.A.; Yee, J.K.; Mao, C.S.; et al. 2014. Precursor-to-product ratios reflect biochemical phenotype in congenital adrenal hyperplasia. Metabolomics, 10(1), 123-131.

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NSK-S-EXP Mix
Composition
Reconstituting a given vial’s contents in 1 mL of highly pure methanol will produce the concentrations listed in the table below.
To facilitate complete dissolution, it is recommended to vortex manually for 1 minute then auto-vortex for a minimum of 30 minutes.
CompoundAldosterone
4-Androstene-3,17-dione
Corticosterone
Cortisol
Dehydroepiandrosterone sulfate·sodium salt·2H2O
11-Deoxycortisol
17-α-Hydroxyprogesterone
Progesterone
Testosterone

Abbrev.
A
A4
B
F
DHEAS
11-S
17-OHPP
T

Label and Enrichment
D7, 98%2,2,4,6,6-D5, 98%
2,2,4,6,6,17α,21,21-D8, 97%
9,12,12-D3, 98%
2,2,3,4,4,6-D6, 95%
2,2,4,6,6-D5, 98% (CP 97%)
2,2,4,6,6,21,21,21-D8, 98%
2,2,4,6,6,17α,21,21,21-D9, 98%
2,2,4,6,6-D5, 98%

Conc. (µM)
0.52
0.12
1.58
2.57
21.69
0.54
0.27
0.14
0.12

Usage Specifications
Criteria
Recommendation
Before reconstitution:
Storage-5 to 5ºC; protect from light
Recommended retest
1 year from date of manufacture

Companion unlabeled standard mixes and kits may be available; please inquire.
Chemical purity (CP) is 98% or greater, unless otherwise indicated.

24

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Pathway-Specific Mixes
TCA Mixes and Sets
The tricarboxylic acid (TCA) cycle plays an essential role in central carbon and energy metabolism. The study of
TCA cycle intermediates (e.g., citrate, α-ketoglutarate, succinate) and its offshoot metabolites (e.g., itaconate and2-hydroxyglutarate), have proven pivotal in not only understanding their impact on metabolism, but also in profilingtheir cellular function/fate in oncogenesis, inflammation, and other pathologies (e.g., necrosis, cirrhosis). To aid theanalysis of TCA cycle-associated compounds in metabolomic studies, we offer stable isotope-labeled and unlabeled
TCA cycle mixes. These mixes are dried down and comprise a collection of TCA cycle and offshoot metabolites.
Overview

RESEARCH PRODUCTS

Cambridge Isotope Laboratories, Inc.
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TCA Cycle
Mixtures
For Qualification, Quantification,and Systems Biology
The tricarboxylic acid (TCA) cycle, also known as the Krebs or citric acid cycle, plays an essential role in central carbon and energy
metabolism. The study of its intermediates, and its ancillary compounds, has proven pivotal in not only understanding their impact
on metabolism but also in profiling their function in oncogenesis, inflammation, and other pathologies.
To aid the analysis of TCA-associated compounds in metabolic studies, Cambridge Isotope Laboratories, Inc. (CIL) is pleased to offer
stable isotope-labeled and unlabeled TCA cycle mixes. These formulations are dried down and comprise a collection of TCA cycle
compounds as well as offshoot metabolites (see composition table below). The mixes can be used in untargeted and targeted LC-MS
methods for applications ranging from quality control to quantification. Please visit isotope.com for pricing and availability.
Catalog No.

Description

MSK-TCA1MSK-TCA2-A
MSK-TCA-A
MSK-TCA1-US
MSK-TCA2-A-US
MSK-TCA-A-USTCA Cycle Standards Mix 1
TCA Cycle Standards Mix 2A
TCA Cycle Standards Mix Sets 1 and 2A
TCA Cycle Standards Mix 1 (unlabeled)
TCA Cycle Standards Mix 2A (unlabeled)
TCA Cycle Standards Mix Sets 1 and 2A (unlabeled)

Unit Size
1 vial1 vial2 × 1 vial
1 vial1 vial2 × 1 vial

Table: Composition of the isotopically labeled TCA mixtures. Reconstituting each vial in 1 mL of solvent (e.g., water) will provide an
equimolar concentration of 100 μM. Note that the unlabeled TCA mix compositions are equivalent.
CompoundFumaric acid, disodium salt
DL-2-Hydroxyglutaric acid, disodium salt
α-Ketoglutaric acid, disodium salt
Malic acid, disodium salt
Sodium L-lactate
Sodium pyruvate
Succinic acid, disodium salt
Trisodium citratecis-Aconitic acid, trisodium salt trihydrate
L-Aspartic acid
L-Glutamic acid
Isocitric acid, trisodium salt
Itaconic acid
Potassium phosphoenol pyruvateLabel and Enrichment
13
C4, 99%13
C5, 99%1,2,3,4-13C4, 99%
13
C4, 99%13
C3, 98%13
C3, 99%13
C4, 99%
1,5,6-carboxy-13C3, 99%
13
C4, 99%13
C4, 99%13
C5, 99%3,4,5,6-13C4, 98% (mixture of diastereomers)
13
C5, 99%2,3-13C2, 99%

Vial No.
1
1
1
1
1
1
1
1
2
2
2
2
2
2

Compounds can also be obtained individually; please inquire.
Chemical purity (CP) is 98% or greater, unless otherwise indicated. For research use only. Not for use in diagnostic procedures.

Cambridge Isotope Laboratories, Inc.
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MET_RSCH_TCAMIX (12/21/23)
Supersedes all previously published literature

Catalog No.
MSK-TCA1MSK-TCA2-A
MSK-TCA-A

Description
TCA Cycle Standards Mix 1
TCA Cycle Standards Mix 2A
TCA Cycle Standards Mix Sets 1 and 2A

No. of Metabolites
86
8 (in mix 1), 6 (in mix 2)

Unit Size
1 vial1 vial2 × 1 vials

Click on the thumbnail or visit
isotope.com for more information.

Compositions
Composition of the stable isotope-labeled TCA mixtures. Reconstituting a given vial in 1 mL of solvent (e.g., water) will produce anequimolar concentration of 100 μM.
CompoundFumaric acid, disodium salt
DL-2-Hydroxyglutaric acid, disodium salt
α-Ketoglutaric acid, disodium salt
Malic acid, disodium salt
Sodium L-lactate
Sodium pyruvate
Succinic acid, disodium salt
Trisodium citratecis-Aconitic acid
L-Aspartic acid
L-Glutamic acid
Isocitric acid, trisodium salt
Itaconic acid
Potassium phosphoenol pyruvate

Abbrev.
Fum
HG
α-KG
Mal
Lac
Pyr
SA
CA
CAA
Asp
Glu
Iso
IA
PEP

Label and Enrichment
13
C4, 99%13
C5, 99%1,2,3,4-13C4, 99%
13
C4, 99%13
C3, 98%13
C3, 99%13
C4, 99%
1,5,6-carboxy-13C3, 99%
13
C4, 99%13
C4, 99%13
C5, 99%3,4,5,6-13C4, 98% (mixture of diastereomers)
13
C5, 99%2,3-13C2, 99%

Vial No.
1
1
1
1
1
1
1
1
2
2
2
2
2
2

Usage Specifications
Criteria
Recommendation
Before reconstitution:
Storageambient temperature; protect from light and moisture
Recommended retest
2 years from date of manufacture
After reconstitution:
Storage
4ºC
Recommended retest
4 weeks

Note: Extended mix stabilities have been demonstrated when solubilized in water and stored as specified above.

Companion unlabeled standard mixes and kits may be available; please inquire.
Chemical purity (CP) is 98% or greater, unless otherwise indicated.For research use only. Not for use in diagnostic procedures.

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Example Results

A

O

B

O

O *

*

*

-

O
-

* O-

*
OH

O

O

OO

HO

O-

O
-

O
*

*

Relative Abundance

O
O- *

* O-

*

Relative Abundance

OH

HO

m/z

O

O

OO
O-

m/z

Figure. Example MS/MS spectra of the TCA mixes. These were obtained from the IC-MS analysis (negative ESI, Orbitrap Fusion Tribrid) of 10 µM working solutions. The compounds
are 13C5 2-hydroxyglutarate (from mix 1) in A) and 13C4 isocitrate (from mix 2) in B). Their complete, unfragmented structures (13C5 hydroxyglutarate, precursor m/z 152.05;
13
C4 isocitrate, precursor m/z 195.03) are illustrated for reference. Data was courtesy of the Metabolomics Facility at MD Anderson Cancer Center.

A

B

O
Si

O *

*

*
O

Si

O

Si

P
Si

O *
O *CH

O

Si

2

Intensity (106, cps)

Intensity (105, cps)

O

* O

O

Si O

m/z

m/z

Figure. Example spectra of the labeled TCA mixes. These were obtained from the analysis of 50 µM working solutions by GC-MS (BSTFA with TMCS, EI ionization, scan mode on
single quadrupole). The compounds are 13C4 malate (from mix 1) in A) and 13C2 phosphenol pyruvate (from mix 2) in B). Their complete, unfragmented structures (13C4 malate,exact mass 354.15; 13C2 phosphenol pyruvate, exact mass 386.11) are illustrated for reference. Data was courtesy of the University of Chicago Metabolomics Facility.

metabolomic compound mixes by Cambridge Isotope Laboratories have expedited several processes in our GC-MS analysis pipeline.
“ TheWe haveused these products, including their novel TCA mixes, to verify metabolite retention times, test derivatization reaction conditions,and to confirm fragmentation patterns detected in biological samples. CIL’s metabolite mixes are ready to go, easy to use, and consistentlyof high quality!
– Ashley M. Sidebottom, PhD
Metabolomics Platform Director, University of Chicago (USA)
Host-Microbe Metabolomics Facility, Duchossois Family Institute”26

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Kits
To help facilitate product application, a number of the CIL mix offerings are accompanied with a user manual. The kit manual provides
a guide to the preparation and processing possibilities for product implementation, as well as supplying further resources for additionaluser reference. The majority of the kits supply the supporting documents through a QR code (via product label on kit materials box); theexception is the PeptiQuant™ Plus kits (documents on USB). The accompanying documents minimally include a user manual, certificate
of analysis (CoA), safety data sheet (SDS), and product flyer. Overall, the mixes are amenable for use in quality control and qualification/
quantification exercises in the MS ‘omics space using targeted, semitargeted, or untargeted LC-MS methodologies. Note that the individual
mixes may be obtained separately without the manual. Please inquire.

Metabolomic QC Kits
To ensure high-quality metabolomics results, the method and instrument platform must be qualified as being fit-forpurpose. This involves testing for losses or errors in the analytical workflow. To aid such performance assessments in
MS metabolomics and potentially other applications (e.g., qualification, quantification), CIL offers three types of QC
kits: MSK-QC-KIT, MSK-QReSS-KIT, and MSK-QReSS-EXP-KIT (see details below). These kits comprise vials of drieddown metabolite mixes (see composition tables below) and have been qualified on both low- and high-resolution
mass spectrometers operated under an untargeted/targeted metabolomics regimen (see example results below).

RESEARCH PRODUCTS

Cambridge Isotope Laboratories, Inc.
isotope.com

Metabolomics
QC Kits
For Untargeted/ Targeted
Mass Spectrometry
Materials for testing assay effectiveness and platform performance are paramountto obtaining quality data. Under routine implementation, such materials can provide
a reliable measure of the efficiency of a specific method, while flagging deficiencies or
errors in an analytical platform (e.g., LC- or GC-MS/MS). To help aid these performanceassessments in MS metabolomics and additionally provide the potential of metabolite
quantification, Cambridge Isotope Laboratories, Inc. (CIL) is pleased to offer threemetabolomics QC kits (MSK-QC-KIT, MSK-QReSS™-KIT, MSK-QReSS-EXP-KIT)
for untargeted/targeted MS-based applications.

MSK-QC-KIT ➝ Click for additional information
This kit contains the following materials and tools:• 2 vials of stable isotope-labeled metabolites (dried down)• Detailed user manual
Table. Mix composition. Reconstitution in 1 mL of solvent(e.g., 50% methanol) yields the concentrations specified.
Description

Table. Mix composition. Reconstitution in 1 mL of solvent(e.g., 50% methanol) yields the concentrations specified.

Conc. (µg/mL)

Vial

4
4
4
40
4
4
4
4
4
4
4
0.4
4
4

1
1
1
1
1
2
2
2
2
2
2
2
2
2

L-Alanine (13C3, 99%)
L-Leucine (13C6, 99%)
L-Phenylalanine (13C6, 99%)
L-Tryptophan (13C11, 99%)
L-Tyrosine (13C6, 99%)
Caffeine (13C3, 99%)
D-Glucose (13C6, 99%)
Sodium benzoate (13C6, 99%)
Sodium citrate (13C3, 99%)
Sodium octanoate (13C8, 99%)
Sodium propionate (13C3, 99%)
Stearic acid, sodium salt (13C18, 98%)
Succinic acid, disodium salt (13C4, 99%)
D-Sucrose (13C6, 98%)

Individual vials and companion unlabeled mixtures
may also be available. Please inquire.
Example Reference
Barco, S.; Lavarello, C.; Cangelosi, D.; et al. 2022. Untargeted LC-HRMS based-plasma metabolomics
reveals 3-O-methyldopa as a new biomarker of poor prognosis in high-risk neuroblastoma. Front
Oncol, 12, 845936-845946.

Overview

Chemical purity (CP) is 98% or greater, unless otherwise indicated.
For research use only. Not for use in diagnostic procedures.

Example Features and Benefits• Mixes (in neat form) with user manual• Predominantly 13C and /or 15N metabolites• Offers flexibility in use and application• Reduces development time and cost
• Provides enhanced reproducibility

MSK-QReSS-KIT ➝ Click for additional information
This kit contains the following materials and tools:• 2 vials of stable isotope-labeled metabolites (dried down)• Detailed user manual

Description
L-Alanine (13C3, 99%; 15N, 99%)
1,4-Butanediamine·2HCl (13C4, 99%)
Creatinine (N-methyl-D3, 98%)
Ethanolamine·HCl (1,1,2,2-D4, 98%)
Guanosine·2H2O (15N5, 96%)
Hypoxanthine (13C5, 99%)
L-Leucine (13C6, 99%)
L-Phenylalanine (ring-13C6, 99%)
Thymine (1,3-15N2, 98%)
L-Tryptophan (13C11, 99%)
L-Tyrosine (ring-13C6, 99%)
Vitamin B3 (13C6, 99%)
Citric acid (1,5,6-carboxyl-13C3, 99%)
Fumaric acid (13C4, 99%)
Indole-3-acetic acid (phenyl-13C6, 99%)
α-Ketoglutaric acid, disodium salt
(1,2,3,4-13C4, 99%)
Sodium palmitate (U-13C16, 98%)
Sodium pyruvate (13C3, 99%)

Conc. (µg/mL)
100
10
100
10
2
10
5
100
20
100
100
5
10
100
5
100

Vial
1
1
1
1
1
1
1
1
1
1
1
1
2
2
2
2

10
100

2
2

Example Reference: Lippa, K.A.; Aristizabal-Henao, J.J.; Beger, R.D.; et al. 2022. Reference materials
for MS-based untargeted metabolomics and lipidomics: a review by the metabolomics quality
assurance and quality control consortium (mQACC). Metabolomics, 18(4), 24-52.
Example Application Note: Percy, A.J.; Souza, A.; Ntai, I.; et al. 2022. From QC to quantitation:
Utility of QReSS™ metabolites in FBS measurements. (CIL application note #51)

Cambridge Isotope Laboratories, Inc.
North America: 1.800.322.1174 cilsales@isotope.com | International: +1.978.749.8000 intlsales@isotope.com | fax: 1.978.749.2768 | isotope.com

Catalog No.
MSK-QC-KIT

Description
Metabolomics QC Kit

No. of Metabolites
5 (in mix 1)
9 (in mix 2)

MSK-QReSS-KIT*

Metabolomics QReSS Kit

12 (in mix 1)
6 (in mix 2)

MSK-QReSS-EXP-KIT

Expanded Metabolomics QReSS Kit

12 (in mix 1)
6 (in mix 2)
9 (in mix 3)

COMING SOON!

Kit Contents• 2 vials of 13C-labeled metabolites• document package (user manual,CoA, SDS, product flyer)• 2 vials of isotope-labeled metabolites• document package (user manual,CoA, SDS, product flyer)• 3 vials of isotope-labeled metabolites• document package (user manual,CoA, SDS, product flyer)

Click on the thumbnail or visit
isotope.com for more information.

*QReSS™ stands for Quantification, Retention, and System Suitability. This kit was developed and matrix tested in collaboration with SCIEX.

MSK-QC Kit
Mix Compositions
Reconstituting in 1 mL of solvent (e.g., 0.1% FA in 5% ACN) will yield the specified compound concentrations.
Compound
L-Alanine
L-Leucine
L-Phenylalanine
L-Tryptophan
L-Tyrosine
D-Glucose
D-Sucrose
Caffeine
Stearic acid, sodium salt
Sodium octanoate
Sodium propionate
Sodium benzoate
Succinic acid, disodium salt
Trisodium citrate

Abbrev.
Ala
Leu
Phe
Trp
Tyr
Glc
Suc
CAF
18:0
8:0
PA
BZA
SA
CA

Label and Enrichment
13
C3, 99%13
C6, 99%13
C6, 99%13
C11, 99%
13
C6, 99%13
C6, 99%13
C6, 98%13
C3, 99%13
C18, 98%
13
C8, 99%13
C3, 99%13
C6, 99%13
C4, 99%13
C3, 99%Conc. (µg/mL)
4
4
4
40
4
4
4
4
0.4
4
4
4
4
4

Mix No.
1
1
1
1
1
2
2
2
2
2
2
2
2
2

Usage Specifications
Criteria
Recommendation
Before reconstitution:
Storage-20°C (protect from light)
Recommended retest
2 years from date of manufacture

Companion unlabeled standard mixes and kits may be available; please inquire.
Chemical purity (CP) is 98% or greater, unless otherwise indicated.For research use only. Not for use in diagnostic procedures.

cilsales@isotope.com | intlsales@isotope.com | isotope.com 27

| Stable Isotope-Labeled Mixtures, Sets, and Kits

Cambridge Isotope Laboratories, Inc.

Example Results

A

B

Figure. Representative XICs of a subset of kit metabolites – vial 1 in A and vial 2 in B – measured in human plasma by RPLC-MS (negative ESI, Q Exactive). Note that all isotopicallylabeled metabolites in the mixes co-eluted with their endogenous analytes in the plasma analyses and their m/z were predominantly [M-H]-.

have used products from Cambridge Isotope Laboratories (CIL) for well over 15 years because of the high quality provided. I have“ Icollaborated
with CIL to develop a new product that would enable improved quality control in MS metabolomics. This process was excitingand engaging. Through collaboration, I found CIL to be a cutting-edge vendor that seeks opportunities to enable scientific discovery and data
quality. They seek advice from clients to learn how to better serve them. I always look forward to talking with CIL at conferences and often
seek out their booth to visit, not just to talk science, but to also see how their team is doing.
– Timothy J. Garrett, PhD
Associate Professor in Department of Pathology,
Immunology, and Laboratory Medicine, University of Florida (USA)”MSK-QReSS Kit
Mix Compositions
Reconstituting a given QReSS mix in 1 mL of solvent (e.g., 1:1 v/ v methanol:water) will yield the specified
compound concentrations.
Compound
L-Alanine
1,4-Butanediamine·2HCl
Creatinine
Ethanolamine·HCl
Guanosine·2H2O
Hypoxanthine
L-Leucine
L-Phenylalanine
Thymine
L-Tryptophan
L-Tyrosine
Vitamin B3
Citric acid
Fumaric acid
Indole-3-acetic acidα-Ketoglutaric acid, disodium salt
Sodium palmitate
Sodium pyruvate

Abbrev. or Alt. Name
Alaputrescine
Crn
ETA
Guo
HPX
Leu
Phe
T
Trp
Tyr
nicotinamide
CA
FA
IAA
α-KG
16:0
Pyr

Label and Enrichment
13
C3, 99%; 15N, 99%
13
C4, 99%N-methyl-D3, 98%
1,1,2,2-D4, 98%
15
N5, 96%13
C5, 99%13
C6, 99%ring-13C6, 99%
1,3-15N2, 98%
13
C11, 99%
ring-13C6, 99%
13
C6, 99%
1,5,6-carboxyl-13C3, 99%
13
C4, 99%phenyl-13C6, 99%
1,2,3,4-13C4, 99% (CP 97%)
U-13C16, 98%
13
C3, 99%Chemical purity (CP) is 98% or greater, unless otherwise indicated.

28

tel: +1.978.749.8000 | 1.800.322.1174 (North America) | fax: +1.978.749.2768

Conc. (µg/mL)
100
10
100
10
2
10
5
100
20
100
100
5
10
100
5
100
10
100

Mix No.
1
1
1
1
1
1
1
1
1
1
1
1
2
2
2
2
2
2

For research use only. Not for use in diagnostic procedures.

| Stable Isotope-Labeled Mixtures, Sets, and Kits

isotope.com

Usage Specifications
Criteria
Recommendation
Before reconstitution:
Storage-20°C (protect from light)
Recommended retest
2 years from date of manufacture
Upon reconstitution:
Storage
4ºC
Recommended retest
4 weeks

Example ResultsA

B● D4 Ethanolamine● 13C6 Hypoxanthine● 13C6 L-Leucine● 13C6 Nicotinamide● 13C6 L-Phenylalanine● 15N2 Thymine● 13C11 L-Tryptophan● 13C6 L-Tyrosine

Figure.QC results for a subset of stable isotope-labeled QReSS metabolites measured across calibrants, QCs, and FBS samples. Acquisitions were by HILIC-MS (ESI+, Orbitrap
Exploris™ 120). Variability of retention times are shown in A and mass accuracy in B. For additional information on the application, see CIL application note #51.

Metabolite
Hypoxanthine (13C5)
Buffer Day 1

Buffer Day 8

MRM Transition(Precursor → Product Ion m/z)

Metabolite

142 ➝ 124

Phenylalanine (13C6)

Human Plasma

Buffer Day 1

MRM Transition(Precursor → Product Ion m/z)
172 ➝ 126
Buffer Day 8

Human Plasma

Figure. Example XICs for a subset of labeled QReSS metabolites measured in buffer and human plasma by RPLC-MRM/MS (positive ESI, QTRAP 6500+). In the buffer analysis, a vialof a working stock mix was stored in on autosampler rack (maintained at 4°C) and processed eight days apart. Such measurements, when performed and metric tracked routinely,
would constitute a system suitability test. In the human plasma analysis, an aliquot of the working stock mix was matrix spiked and subjected to a metabolomics workflow. The
relative signals to its corresponding endogenous analyte (not shown for simplicity) is within an order of magnitude and can be used for relative quantitative applications, withabsolute quantitation likely involving calibration curves. For additional information and application demonstrations see CIL application note #49.

Chemical purity (CP) is 98% or greater, unless otherwise indicated.For research use only. Not for use in diagnostic procedures.

cilsales@isotope.com | intlsales@isotope.com | isotope.com 29

| Stable Isotope-Labeled Mixtures, Sets, and Kits

Cambridge Isotope Laboratories, Inc.

Fig Part A: MS1 (Putrescine)A

MS1

3.58 Creatinine

Relative Abundance

6.26 L-Tryptophan

2.83 Hypoxanthine
1.94
Indole-3acetic
1.96
acid
Thymine/Nicotinamide
7.41 L-Tyrosine

Relative Abundance

9.08 L-Alanine

14.45 PutrescineTime (min)Fig Part A: MS2 (Putrescine)MS2

5.87 L-Phenylalanine
1.96
Thymine/Nicotinamide
Indole-3-acetic
acid
2.83
Hypoxanthinem/z

6.26 L-Leucine/L-Tryptophan

5.91 L-Guanosine
7.41 L-Tyrosine
m/z

9.08 L-Alanine

13.77 PutrescineTime (min)Fig Part B: MS1 (Indole-3-Acetate)

BMS1

9.14 α-Ketoglutarate

3.15 Indole-3-acetate

Relative Abundance

2.69 Pyruvate

Relative Abundance

1.41 Palmitate

9.96 Fumarate

Time (min)Fig Part B: MS2 (Indole-3-Acetate)

m/z

MS2

1.41 Palmitate
9.10 α-Ketoglutarate
3.15 Indole-3-acetate

9.96 Fumaratem/zTime (min)Figure. XIC overview of stable isotope-labeled QReSS metabolites measured matrix-free by HILIC-MS (Q Exactive), with ESI+ in A and ESI- in B. Chromatographic insets reflectzoomed-in snapshots of lower-abundance metabolites, while the MS1/ MS2 spectra reflect putrescine in A and indole-3-acetate in B. For clarity, the following higher-abundance
metabolites were removed in the insets – creatinine (RT 3.58 min), phenylalanine (RT 5.87 min), and leucine (RT 6.26 min) in A and pyruvate (RT 2.69 min) in B. Data is courtesyof the Jackson Laboratory.

Chemical purity (CP) is 98% or greater, unless otherwise indicated.

30

tel: +1.978.749.8000 | 1.800.322.1174 (North America) | fax: +1.978.749.2768

For research use only. Not for use in diagnostic procedures.

isotope.com

| Stable Isotope-Labeled Mixtures, Sets, and Kits

Example References
MSK-QC-KIT
Kossack, M.E.; Manz, K.E.; Martin, N.R.; et al. 2023. Environmentally relevant uptake, elimination, and metabolic changes following early embryonic exposure to
2,3,7,8-tetrachlorodibenzo-p-dioxin in zebrafish. Chemosphere, 310, 136723-136754.
Barco, S.; Lavarello, C.; Cangelosi, D.; et al. 2022. Untargeted LC-HRMS based-plasma metabolomics reveals 3-O-methyldopa as a new biomarker of poor prognosis in high-risk
neuroblastoma. Front Oncol, 12, 845936-845946.
Strelez, C.; Chilakala, S.; Ghaffarian, K.; et al. 2021. Human colorectal cancer-on-chip model to study the microenvironmental influence on early metastatic spread. iScience, 24(5),102509-102524. 34113836
MSK-QReSS-KIT
Kress, J.; Nandita, E; Jones, E.; et al. 2023. A targeted liquid chromatography mass spectrometry method for routine monitoring of cell culture media components for bioprocessdevelopment. J Chromatogr A, 1706, 464281-464288.
Krusinski, L.; Maciel, I.C.F.; van Vliet, S.; et al. 2023. Measuring the phytochemical richness of meat: effects of grass/grain finishing systems and grapeseed extract supplementationon the fatty acid and phytochemical content of beef. Foods, 12(19), 3547-4566.
Fame, R.M.; Kalugin, P.N.; Petrova, B.; et al. 2023. Defining diurnal fluctuations in mouse choroid plexus and CSF at high molecular, spatial, and temporal resolution. Nat Commun,
14(1), 3720-3739.
Brown, J.I.; Wang, P.; Wong, A.Y.L.; et al. 2023. Cycloguanil and analogues potently target DHFR in cancer cells to elicit anti-cancer activity. Metabolites, 13(2), 151-172.
Lippa, K.A.; Aristizabal-Henao, J.J.; Beger, R.D.; et al. 2022. Reference materials for MS-based untargeted metabolomics and lipidomics: a review by the metabolomics quality
assurance and quality control consortium (mQACC). Metabolomics, 18(4), 24-52.
Heppler, L.N.; Attarha, S.; Persaud, R.; et al. 2022. The antimicrobial drug pyrimethamine inhibits STAT3 transcriptional activity by targeting the enzyme dihydrofolate reductase.
J Biol Chem, 298(2), 101531-10548.
Application Notes
MSK-QReSS Mixes/Kits
Percy, A.J.; Souza, A.; Ntai, I.; et al. 2022. From QC to quantitation: Utility of QReSS™ metabolites in FBS measurements. (CIL application note #51)
Percy, A.J.; Proos, R.; Demianova, Z.; et al. 2021. Standardizing quantitative metabolomics analyses through the QReSS kit. (CIL application note #49)

he unique feature of our media analysis workflow is the use of the QReSS standard mix. We spike this into media samples before extraction,“ Twhichhelps normalize variabilities in metabolite extraction efficiencies and combat matrix effects. The QReSS mix consists of 18 isotopelabeled metabolites, which spans a multitude of metabolic classes and mimics the largely diverse chemical composition of cell culture media.
This serves as an excellent choice for an internal standard mix and helps to improve the accuracy, precision, and robustness of our cell culture
method.
– Hari Kosanam, PhD
Associate Principal Scientist, Vaccine Process Development and Commercialization
Merck & Co. (USA)”

Chemical purity (CP) is 98% or greater, unless otherwise indicated.For research use only. Not for use in diagnostic procedures.

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| Stable Isotope-Labeled Mixtures, Sets, and Kits

Cambridge Isotope Laboratories, Inc.

Extract Kits
Expanded qualification remains a necessity in MS ‘omics today. Validated identifications are desired that canbe used to springboard in-depth profiling studies and selective quantifications in preclinical screening studies.
To aid method benchmarking (e.g., through a credentialing approach) and improved qualification/quantification
studies, CIL offers two types of cell extracts: yeast (Pichia pastoris, strain CBS 7435) and E. coli (K12 strain MG1655),
as outlined in the overview table below. Upon careful and precise solubilization (see reconstitution guidelines below),
100s of metabolites are potentially observable in their U-13C and unlabeled form. The metabolites span a broad classrange (see example identification tables), having linkage to various biochemical pathways (e.g., citrate and glyoxylate
cycle, nucleotide and lipid metabolism) and cellular/molecular processes (e.g., intracellular signaling, immune system,
blood coagulation, lipolysis). Please refer to the sample results and references below for examples of the productspecific applications.

Description
Crude Lipid Yeast Extract(U-13C, 99%)

No. of Analytes
100s

L-ISO1-UNL

Crude Lipid Yeast Extract(unlabeled)

100s

ISO1

Metabolite Yeast Extract(U-13C, 99%)100s

ISO1-UNL

Metabolite Yeast Extract(unlabeled)

100s

ISO1-KIT

Metabolite Yeast Extract Kit

100s

MSK-CRED-DD-KIT Credentialed E. coli Cell
Extract Kit (dried down)

100s

Credentialed E. coli Cell
Extract Kit (solution)

100s

MSK-CRED-KIT

Metabolite and Lipid
Yeast Extracts
For MS Metabolomics and Lipidomics

Cambridge Isotope Laboratories, Inc. (CIL) is pleased to offer 13C-labeled and unlabeled yeast extracts for use in MS-based
quantitative or profiling studies of various sample types. These extracts have been rigorously characterized by a number ofmethodologies and are amenable to a variety of research uses after simple reconstitution. The components in the extracts span broad
metabolic classes (e.g., amino and organic acids, sugar phosphates, coenzymes, fatty acids and lipids), biochemical pathways (e.g., citrate
and glyoxylate cycle, nucleotide and lipid metabolism), and cellular/molecular processes (e.g., intracellular signaling, immune system, blood
coagulation, lipolysis).

Overview
Catalog No.
ISO1
ISO1-UNL
ISO1-KIT
L-ISO1
L-ISO1-UNL

Uses

Description
Metabolite Yeast Extract (U-13C, 99%)
Metabolite Yeast Extract (unlabeled)
Metabolite Yeast Extract Kit
Crude Lipid Yeast Extract (U-13C, 99%)
Crude Lipid Yeast Extract (unlabeled)• Targeted or untargeted, MS-based analysis• Method and instrument QC• Qualification and /or quantification• Biomarker discovery and verification

Benefits

Dry extract of Pichia pastoris cells (strain CBS 7435).
Produced by ISOtopic Solutions (isotopic-solutions.com).
Please inquire
for pricing.

Kit Contents• 1 vial of U-13C crude lipid yeast extract• document package (user manual, CoA, SDS,
product flyer)• 1 vial of unlabeled crude lipid yeast extract• document package (user manual, CoA, SDS,
product flyer)• 1 vial of U-13C metabolite yeast extract• document package (user manual, CoA, SDS,
product flyer)• 1 vial of unlabeled metabolite yeast extract• document package (user manual, CoA, SDS,
product flyer)• 1 vial of U-13C metabolite yeast extract• 1 vial of unlabeled metabolite yeast extract• document package (user manual, CoA, SDS,
product flyer)• 1 vial (blue cap) of 13C-labeled E. coli cell extract(dried down)• 1 vial (yellow cap) of unlabeled E. coli cell extract(dried down)• document package (user manual, CoA, SDS,
product flyer)• 1 vial (black) of 13C-labeled E. coli cell extract(solution)• 1 vial (red) of unlabeled E. coli cell extract(solution)• document package (user manual, CoA, SDS,
product flyer)• Reduces measurement uncertainty• Improves precision and accuracy• Enhances identification confidence• Decreases development time and cost

Example References
Turtoi, E.; Jeudy, J.; Henry, S.; et al. 2023. Analysis of polar primary metabolites in biological samples
using targeted metabolomics and LC-MS. STAR Protoc, 4(3), 102400-102422.
Serafimov, K.; Lämmerhofer, M. 2022. Metabolic profiling workflow for cell extracts by targetedhydrophilic interaction liquid chromatography-tandem mass spectrometry. J Chromatogr A, 1684,
463556-463555.
Li, P.; Su, M.; Chatterjee, M.; et al. 2022. Targeted analysis of sugar phosphates from glycolysis
pathway by phosphate methylation with liquid chromatography coupled to tandem mass
spectrometry. Anal Chim Acta, 1221, 340099-34109.
Reiter, A.; Asgari, J.; Wiechert, W.; et al. 2022. Metabolic footprinting of microbial systems based oncomprehensive in silico predictions of MS/MS relevant data. Metabolites, 12(3), 257-280.
Zhao, X.; Golic, F.T.; Harrison, B.R,; et al. 2022. The metabolome as a biomarker of aging in
Drosophila melanogaster. Aging Cell, 21(2), e13548.
Rampler, E.; Hermann, G.; Grabmann, G.; et al. 2021. Benchmarking non-targeted metabolomics
using yeast-derived libraries. Metabolites, 11(3), 160-179.
Mairinger, T.; Weiner, T.; Hann, S.; et al. 2020. Selective and accurate quantification of N-acetylglucosamine in biotechnological cell samples via GC-MS/MS and GC-TOFMS. Anal Chem, 92(7),4875-4883.
Galvez, L.; Rusz, M.; Schwaiger-Haber, M.; et al. 2019. Preclinical studies on metal based anticancer
drugs as enabled by integrated metallomics and metabolomics. Metallomics, 11(10), 1716-1728.

Hermann, G.; Schwaiger, M.; Volejnik, P.; et al. 2018. 13C-labelled yeast as internal standard for
LC-MS/MS and LC high resolution MS-based amino acid quantification in human plasma.
J Pharm Biomed Anal, 155, 329-334.
Guijas, C.; Montenegro-Burke, J.R.; Domingo-Almenara, X.; et al. 2018. METLIN: A technology
platform for identifying knowns and unknowns. Anal Chem, 90(5), 3156-3164.
Si-Hung, L.; Causon, T.J.; Hann, S. 2017. Comparison of fully wettable RPLC stationary phases for
LC-MS-based cellular metabolomics. Electrophoresis, 38(18), 2287-2295.
Schwaiger, M.; Rampler, E.; Hermann, G.; et al. 2017. Anion-exchange chromatography coupled tohigh-resolution mass spectrometry: A powerful tool for merging targeted and non-targeted
metabolomics. Anal Chem, 89(14), 7667-7674.
Ortmayr, K.; Hann, S.; Koellensperger, G. 2015. Complementing reversed-phase selectivity with porousgraphitized carbon to increase the metabolome coverage in an on-line two-dimensional LC-MS setup
for metabolomics. Analyst, 140(10), 3465-3473.
Neubauer, S.; Chu, D.B.; Marx, H.; et al. 2015. LC-MS/MS-based analysis of coenzyme A andshort-chain acyl-coenzyme A thioesters. Anal Bioanal Chem, 407(22), 6681-6688.
Application Note
Percy, A.J.; Munjoma, N.; Heywood, D.; et al. 2022. Targeted MRM screening of U-13C lipid yeast
extracts for robust lipidomics applications. (CIL application note #54)

Demarest, T.G.; Truong, G.T.D.; Lovett, J.; et al. 2019. Assessment of NAD+ metabolism in human cell
cultures, erythrocytes, cerebrospinal fluid and primate skeletal muscle. Anal Biochem, 572, 1-8.

Cambridge Isotope Laboratories, Inc.
North America: 1.800.322.1174 cilsales@isotope.com | International: +1.978.749.8000 intlsales@isotope.com | fax: 1.978.749.2768 | isotope.com
MET_RSCH_YEAST (4/23/24)
Supersedes all previously published literature

RESEARCH PRODUCTS

Cambridge Isotope Laboratories, Inc.
isotope.com

Credentialed E. coli
Cell Extract Kits
For Untargeted/ Targeted
Mass Spectrometry
An exceeding challenge in optimizing metabolomic methodologies
toward improved metabolome coverage has been the difficultyin comparing the number of metabolites profiled in each. Thisevaluation is complicated by artifactual signals. Artifacts canarise from sample contamination during metabolite extraction,
background instrument noise, or misannotation of data during
bioinformatic processing. While efforts to minimize artifacts havebeen extended, it is generally not possible to completely remove
these from the features list nor desirable to do so without wellcharacterized extracts and proper software tools designed toeffectively filter biogenics from artifacts.

Overview
Catalog No.
L-ISO1RESEARCH PRODUCTS

Cambridge Isotope Laboratories, Inc.
isotope.com

Cambridge Isotope Laboratories, Inc. (CIL) is pleased tooffer Credentialed E. coli Cell Extract Kits (MSK-CRED-KIT,MSK-CRED-DD-KIT) that can assist the user in developing
and optimizing methodologies for untargeted metabolomic
profiling.
The kit contents are as follows:• 13C-labeled (at 99%) E. coli cell extract (dried down or 100 µL)• Unlabeled E. coli cell extract (dried down or 100 µL)• Detailed user manual (contains instructions on solution/sample
preparation and data analysis with Credentialing software).
The Credentialing tool is also available in Compound Discoverer™
software (version 3.0; Thermo Scientific).
Note: The cells are E. coli K12 strain MG1655; extracts are
generated from 2.5 mg lyophilized cells. Other sizes may beavailable. Please inquire.

Features and Benefits• Credentialing yields 100s of standards• Effective biogenic vs. artifact signal discrimination• Preserves low-intensity signals• Platform-independent metabolite analysis• Enables interlaboratory method comparison• Allows method optimization and benchmarking

The supplied extracts require simple preparation prior to mixing(at defined ratios), LC-MS analysis, and bioinformatic processing.
This latter processing is facilitated by a well-validated, informatic
workflow, which is designed to effectively discriminate biologicalfrom artifactual features on the basis of isotopic spacing andintensity ratios.1-11 Overall, these kits help streamline method
optimization/evaluation in untargeted microbial metabolomics
and provides a robust metric for performance comparisons
between different metabolomic methods and instrument
platforms.
Catalog No.

Description

MSK-CRED-KIT
Credentialed E. coli Cell Extract Kit (solution)
MSK-CRED-DD-KIT Credentialed E. coli Cell Extract Kit (dried down)

Please inquire
for pricing.

References
1. Li, S.; Zheng, S. 2023. Generalized tree structure to annotate untargeted metabolomics and stable isotope
tracing data. Anal Chem, 95(15), 6212-6217.
2. Dodds, J.N.; Wang, L.; Patti, G.J.; et al. 2022. Combining isotopologue workflows and simultaneous
multidimensional separations to detect, identify, and validate metabolites in untargeted analyses.
Anal Chem, 94(5), 2527-2535.
3. Giné, R.; Capellades, J.; Badia, J.M.; et al. 2021. HERMES: a molecular-formula-oriented method to target
the metabolome. Nat Methods, 18(11), 1370-1376.
4. Cho, K.; Schwaiger-Haber, M.; Naser, F.J.; et al. 2021. Targeting unique biological signals on the fly to
improve MS/MS coverage and identification efficiency in metabolomics. Anal Chim Acta, 1149, 338210.
5. Sindelar, M.; Patti, G.J. 2020. Chemical discovery in the era of metabolomics. J Am Chem Soc, 142(20),9097-9105.
6. Wang, L.; Naser, F.J.; Spalding, J.L.; et al. 2019. A protocol to compare methods for untargeted
metabolomics. Methods Mol Biol, 1862, 1-15.

7. Spalding, J.L.; Naser, F.J.; Mahieu, N.G.; et al. 2018. Trace phosphate improves ZIC-pHILIC peak shape,
sensitivity, and coverage for untargeted metabolomics. J Proteome Res, 17(10), 3537-3546.
8. Naser, F.J.; Mahieu, N.G.; Wang, L.; et al. 2018. Two complementary reversed-phase separations forcomprehensive coverage of the semipolar and nonpolar metabolome. Anal Bioanal Chem, 410(4),1287-1297.
9. Mahieu, N.G.; Patti, G.J. 2017. Systems-level annotation of a metabolomics data set reduces 25,000
features to fewer than 1,000 unique metabolites. Anal Chem, 89(19), 10397-10406.
10. Benton, H.P.; Ivanisevic, J.; Mahieu, N.G.; et al. 2015. Autonomous metabolomics for rapid metabolite
identification in global profiling. Anal Chem, 87(2), 884-891.
11. Mahieu, N.G.; Huang, X.; Chen, Y.; et al. 2014. Credentialing features: a platform to benchmark andoptimize untargeted metabolomic methods. Anal Chem, 86(19), 9583-9589.

Cambridge Isotope Laboratories, Inc.
North America: 1.800.322.1174 cilsales@isotope.com | International: +1.978.749.8000 intlsales@isotope.com | fax: 1.978.749.2768 | isotope.com
MET_RSCH_ECOLI (8/19/24)
Supersedes all previously published literature

Click on the thumbnails or visit
isotope.com for more information.

Lipid Yeast Extracts
Compositions
Tabulated is a reproducibly identified panel of fatty acids and lipids measured in the lipid yeast extracts (U-13C, L-ISO1; unlabeled, L-ISO1UNL) using one general RPLC-MS method. Note: Additional analytes have been measured with alternate methods and analysis techniques.
Category
Glycerolipids

Class
Diglyceride (DG)

Triglyceride (TG)

Examples (Fatty Acid Level)
34:1 (16:0/18:1)
34:2 (16:1/18:1)
34:4 (16:2/18:2)
36:1 (18:0/18:1)
36:3 (18:1/18:2)
36:4 (18:1/18:3)
50:1 (16:0/16:0/18:1)
50:2 (16:0/18:2/16:0)
50:4 (16:1/16:1/18:2)
50:5 (16:1/16:1/18:3)
52:2 (16:0/18:1/18:1)
52:3 (16:1/18:1/18:1)
52:5 (16:1/18:2/18:2)
52:6 (16:1/18:2/18:3)
54:1 (18:0/18:1/18:0)
54:2 (18:0/18:1/18:1)
54:4 (18:1/18:1/18:2)
54:5 (18:1/18:2/18:2)
54:7 (18:1/18:3/18:3)
54:8 (18:3/18:2/18:3)

Chemical purity (CP) is 98% or greater, unless otherwise indicated.

32

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34:3 (16:1/18:2)
36:2 (18:0/18:2)
36:5 (18:2/18:3)
50:3 (16:1/16:1/18:1)
52:1 (18:0/16:0/18:1)
52:4 (18:2/18:2/16:0)
52:7 (18:3/18:3/16:1)
54:3 (18:1/18:1/18:1)
54:6 (18:2/18:2/18:2)
54:9 (18:3/18:3/18:3)

For research use only. Not for use in diagnostic procedures.

| Stable Isotope-Labeled Mixtures, Sets, and Kits

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CategoryClass

Examples (Fatty Acid Level)

Glycerophospholipids

Phosphatidic acid (PA)34:1 (16:0/18:1)
36:1 (18:0/18:1)
34:1 (16:0/18:1)
34:4 (16:1/18:3)
36:4 (18:1/18:3)
34:1 (16:0/18:1)
36:1 (18:0/18:1)
36:4 (18:1/18:3)
34:1 (16:0/18:1)
34:1 (16:0/18:1)
36:2 (18:1/18:1)
34:1 (16:0/18:1)
36:1 (18:0/18:1)
16:1
18:1
18:2
18:1
18:2
18:3
d34:1 (d18:1/16:0)
t36:0 (t18:0/18:0)
t36:2 (t18:1/18:1)
16:1
Q8
18:2
18:3

Phosphatidylcholine (PC)

Phosphatidylethanolamine (PE)

Phosphatidylglycerol (PG)
Phosphatidylinositol (PI)Phosphatidylserine (PS)
Lysophospholipids
Sphingolipids

Other

Lysophosphatidylcholine (LPC)
Lysophosphatidylethanolamine (LPE)
Ceramide (Cer)
Hexosyl ceramide (HexCer)
Acylcarnitines (AC)Coenzyme (Co)
Fatty acid (FA)34:2 (16:0/18:2)
36:2 (18:0/18:2)
34:2 (16:0/18:2)
36:2 (18:0/18:2)
36:5 (18:2/18:3)
34:2 (16:0/18:2)
36:2 (18:0/18:2)
36:5 (18:2/18:3)
34:2 (16:1/18:1)
34:2 (16:0/18:2)

34:3 (16:0/18:3)
36:3 (18:1/18:2)
34:3 (16:0/18:3)
36:3 (18:1/18:2)
36:6 (18:3/18:3)
34:3 (16:0/18:3)
36:3 (18:1/18:2)
36:6 (18:3/18:3)

34:2 (16:0/18:2)
36:2 (18:0/18:2)
18:3

34:3 (16:0/18:3)
36:3 (18:0/18:3)

d36:1 (d18:1/18:0)
t36:2 (t18:1/18:1)

t34:1 (t18:1/16:0)

36:1 (18:0/18:1)

Usage Specifications
Extract TypeNo. of uses
Before reconstitution:
Storage
Recommended retest
Expiration

L-ISO1

L-ISO1-UNL~50*-80°C; protect from lightevery 12 months
3 years from date of manufacture*Conservative estimate, with numbers varying depending on method and application.

Solution Preparation Procedure
1. Reconstitute the lipid yeast extract (L-ISO1 or L-ISO1-UNL) in 1 mL solvent (e.g., isopropanol).
2.	Following a high-speed vortex and brief centrifuge, the resulting clear solution can then be matrix-spiked, diluted (1/10 v/v),
or prepared further for calibrant, QC, and /or sample addition.
Example ResultsA

DG 34:1[M+NH4]+; m/z 649.68028

LPC 18:2[M+H]+; m/z 546.426992

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Cambridge Isotope Laboratories, Inc.

B

PC 34:2[M+HCOO]-; m/z 844.701262

PA 34:2[M-H]-; m/z 710.60551

Figure. Representative XICs of example lipids measured in the U-13C lipid yeast extract by RPLC-MS (ACQUITY HSS T3, Orbitrap ID-X). The ionization mode was ESI+ in A and ESIin B. Data is courtesy of the Patti lab at Washington University.

Table. Example set of identified fatty acids and lipids measured by RPLC-IM-MS (ACQUITY™ Premier CSH™ C18 column, SYNAPT™ XS).
Tabulated is analytical data for unlabeled and U-13C lipids acquired under ESI+ and ESI- (in blue) polarities. Error tolerances on the ion
mobility-derived collision cross-sections (CCS) are ±3%. Data is courtesy of Waters (Manchester, UK).
Lipid (Fatty Acid Level)
Cer d34:1 (d18:1/16:0)
Cer d34:1 (d18:1/16:0)
PC 34:2 (16:0/18:2)
PC 36:4 (18:1/18:3)
PE 34:2 (16:0/18:2)
PE 34:2 (16:0/18:2)
LPC(16:1)
LPC(16:1)

RT (min)
3.31
3.31
2.85
2.53
2.96
2.96
0.82
0.82

Precursor m/z
Unlabeled Lipids
U-13C Lipids
560.5023
594.6124
536.5059
570.6159
758.5700
800.7074
782.5697
826.7160
716.5220755.6510
738.5032
777.6310
494.3223
518.4045
538.3145
562.3950

Adduct[M+Na]+[M-H][M+H]+[M+H]+[M+H]+[M+Na]+[M+H]+[M+HCOO]-CCS (Å2)
Unlabeled Lipids
U-13C Lipids
257.79258.44252.85253.30293.16294.46300.69301.52279.92280.33286.03286.47229.29229.61238.29239.23

Metabolite Yeast Extracts
Compositions
Tabulated are the routinely identified analytes in the metabolite yeast extracts (U-13C, ISO1; unlabeled, ISO1-UNL). Note: Additional
metabolites have been measured with alternate methods and analysis techniques. This includes cofactors (e.g., NMN, ADPR, NADPH)
and coenzymes (e.g., acetyl and malonyl coenzyme A).
Amino Acids and Derivatives (L enantiomer where applicable)
S-Adenosyl-homocysteine (SAH)
Glutamate (Glu)
Alanine (Ala)
Glutamine (Gln)α-Aminoadipate (AAD)
Glycine (Gly)Arginine (Arg)
Guanidineacetate (GAA)
Argininosuccinate (ASA)
Histidine (His)
Asparagine (Asn)
Homoserine (Hse)Aspartate (Asp)Isoleucine (Ile)Betaine (BET)
α-Ketoisovalerate (KIV)
Citrulline (Cit)
Kynurenine (KYN)
Cystathionine (CYS)
Leucine (Leu)
Dihydroxyisovalerate (DIHV)
Lysine (Lys)

Methionine (Met)(±)-3-Methyl-2-oxovalerate (K-IVal)
Ornithine (Orn)
Phenylalanine (Phe)Proline (Pro)Sarcosine (Sar)Serine (Ser)Threonine (Thr)
Tryptophan (Trp)
Tyrosine (Tyr)Valine (Val)

Chemical purity (CP) is 98% or greater, unless otherwise indicated.For research use only. Not for use in diagnostic procedures.

34

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Nucleobases, Nucleosides, and Nucleotides
Adenine (Ade)
Cytidine triphosphate (CTP)
Adenosine (Asin)
Deoxyadenosine monophosphate (dAMP)
Adenosine diphosphate (ADP)
5’-Deoxy-5’-methylthioadenosine (MTAP)
Adenosine monophosphate (AMP)
Guanine (Gnin)
Adenosine triphosphate (ATP)Guanosine (Gsin)
Cyclic adenosine monophosphate (cAMP)
Guanosine diphosphate (GDP)Cyclic guanosine monophosphate (cGMP)
Guanosine monophosphate (GMP)
Cytidine monophosphate (CMP)
Guanosine triphosphate (GTP)
Organic Acids
cis-Aconitate (cis-Ac)
DL-2-Hydroxyglutarate (2-HG)
Citrate (CA)Isocitrate (Iso)Fumarate (Fum)α-Ketoglutarate (α-KG)
Gluconate (GA)
Lactate (Lac)
Sugar and Sugar Phosphates (D enantiomer where applicable)
Dihydroxyacetone phosphate (DHAP)
Glucose (Glc)
Erythritol (Erthrit)
Glucose-6-phosphate (G6P)
Fructose (Fuc)Mannitol (Man-Ol)
Fructose-1,6-bisphosphate (FBP)
Mannose (Man)Fructose-6-phosphate (F6P)
Mannose-6-phosphate (M6P)
Galactose (Gal)
2-Phosphoglycerate (2PG)
Vitamins and Coenzymes
Biotin (B7)*
Nicotinamide (NAM)
Choline (CHOL)

Nicotinamide adenine dinucleotide,oxidized (NAD+)

Other Small Molecules
Glutamylcysteine (Glu-Cys)
Glutathione, oxidized (GSSG)

Inosine (Isin)
Inosine monophosphate (IMP)5-Methyluridine (m5U)
Pseudouridine (PsU)
Uridine (Uri)
Uridine diphosphate (UDP)
Uridine monophosphate (UMP)
Uridine triphosphate (UTP)
Malate (Mal)Pyruvate (Pyr)Succinate (SA)

6-Phosphogluconate (6PGA)
Ribose (RIB)
Ribose-5-phosphate (R5P)
Sedoheptulose-7-phosphate (S7P)
Trehalose (TRE)

Nicotinamide adenine dinucleotide, reduced(NADH)
Nicotinamide adenine dinucleotide phosphate,oxidized (NADP+)

Glutathione, reduced (GSH)
Mevalonate (MVA)*Identified in ISO1-UNL only.

Usage Specifications
Extract TypeNo. of uses
Before reconstitution:
Storage
Recommended retest
Expiration
Upon reconstitution:
Storage
Recommended retest

ISO1

ISO1-UNL~50*-80°C; protect from lightevery 12 months
4 years from date of manufacture
4ºC
4 weeks*Conservative estimate, with numbers varying depending on method and application.

Solution Preparation Procedure
1. Reconstitute the metabolite yeast extracts (ISO1 or ISO1-UNL) with 2 mL solvent (e.g., water, 50% methanol).
2. Vigorously shake by hand with intermittent high-speed vortexing (2 minute minimum).
3. Centrifuge at 20°C for 5 min at 4000 rcf.
4.	The clear standard solution can then be diluted (1/10 v/v) for direct use or prepared further for calibration and matrix addition.
If particulate matter remains after steps 2) and 3), repeat with extra intense vortexing and shaking until complete dissolution.
 video demonstration of this procedure can be obtained here, as well as in the resources section of the Metabolite Yeast Extracts
A
application page (see isotope.com/applications/metabolic research/metabolomics mixtures and kits).

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Example Results

A

B

Figure. Representative XICs of 13C-labeled cofactors measured by HILIC-MS (positive ESI, Q Exactive HF). Shown in A are the routinely identified metabolites and B the additionallyobserved cofactors. Procedurally, the extract was reconstituted in 2 mL water then diluted 1:10 before analysis. The HILIC separation utilized an Acquity UPLC BEH Amide column(100 × 2.1 mm, 1.7 μm particles).
Example References
Turtoi, E.; Jeudy, J.; Henry, S.; et al. 2023. Analysis of polar primary metabolites in biological samples using targeted metabolomics and LC-MS. STAR Protoc, 4(3),102400-102422.
Apiz Saab, J.J.; Dzierozynski, L.N.; Jonker, P.B.; et al. 2023. Pancreatic tumors exhibit myeloid-driven amino acid stress and upregulate arginine biosynthesis. Elife, 12, e81289. 37254839
Serafimov, K.; Lämmerhofer, M. 2022. Metabolic profiling workflow for cell extracts by targeted hydrophilic interaction liquid chromatography-tandem mass spectrometry.
J Chromatogr A, 1684, 463556-463555.
Li, P.; Su, M.; Chatterjee, M.; et al. 2022. Targeted analysis of sugar phosphates from glycolysis pathway by phosphate methylation with liquid chromatography coupled to tandem
mass spectrometry. Anal Chim Acta, 1221, 340099-34109.
Reiter, A.; Asgari, J.; Wiechert, W.; et al. 2022. Metabolic footprinting of microbial systems based on comprehensive in silico predictions of MS/MS relevant data. Metabolites, 12(3),257-280.
Zhao, X.; Golic, F.T.; Harrison, B.R,; et al. 2022. The metabolome as a biomarker of aging in Drosophila melanogaster. Aging Cell, 21(2), e13548-13561.
Rampler, E.; Hermann, G.; Grabmann, G.; et al. 2021. Benchmarking non-targeted metabolomics using yeast-derived libraries. Metabolites, 11(3), 160-179.
Mairinger, T.; Weiner, T.; Hann, S.; et al. 2020. Selective and accurate quantification of N-acetylglucosamine in biotechnological cell samples via GC-MS/MS and GC-TOFMS.
Anal Chem, 92(7), 4875-4883.
Rusz, M.; Rampler, E.; Keppler, B.K.; et al. 2019. Single spheroid metabolomics: optimizing sample preparation of three-dimensional multicellular tumor spheroids. Metabolites,
9(12), 304-325.
Galvez, L.; Rusz, M.; Schwaiger-Haber, M.; et al. 2019. Preclinical studies on metal based anticancer drugs as enabled by integrated metallomics and metabolomics. Metallomics,
11(10), 1716-1728.
Sullivan, M.R; Danai, L.V.; Lewis, C.A.; et al. 2019. Quantification of microenvironmental metabolites in murine cancers reveals determinants of tumor nutrient availability.
Elife, 8, e44235.
Demarest, T.G.; Truong, G.T.D.; Lovett, J.; et al. 2019. Assessment of NAD+ metabolism in human cell cultures, erythrocytes, cerebrospinal fluid and primate skeletal muscle.
Anal Biochem, 572, 1-8.
 ermann, G.; Schwaiger, M.; Volejnik, P.; et al. 2018. 13C-labelled yeast as internal standard for LC-MS/MS and LC high resolution MS-based amino acid quantification in humanHplasma. J Pharm Biomed Anal, 155, 329-334.
Guijas, C.; Montenegro-Burke, J.R.; Domingo-Almenara, X.; et al. 2018. METLIN: A technology platform for identifying knowns and unknowns. Anal Chem, 90(5), 3156-3164.
Si-Hung, L.; Causon, T.J.; Hann, S. 2017. Comparison of fully wettable RPLC stationary phases for LC-MS-based cellular metabolomics. Electrophoresis, 38(18), 2287-2295.

Application Notes
Percy, A.J.; Munjoma, N.; Heywood, D.; et al. 2022. Targeted MRM screening of U-13C lipid yeast extracts for robust lipidomics applications. (CIL application note #54)
Mohsin, S.B.; Batoon, P. 2022. Absolute quantitation of fragile metabolites by isotope dilution mass spectrometry on the Agilent 6495 Triple Quadrupole LC/MS.
(Agilent application no. 5994-4439EN)
Chemical purity (CP) is 98% or greater, unless otherwise indicated.

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For research use only. Not for use in diagnostic procedures.

| Stable Isotope-Labeled Mixtures, Sets, and Kits

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E. coli Cell Extracts
Compositions
Condensed table of example metabolites detected in the E. coli extract samples (see PMID: 29256075 for application details). The spreadof logPs (source: ALOGPS) highlights the breadth of physicochemical diversity.
Compound (Abbrev. or Alt. Name)
Adenine (Ade)
Adenosine monophosphate (AMP)
Biotin (vitamin B7)
Coenzyme Q10 (CoQ10; ubiquinone)
Diacylglycerol (DG) 17:0/17:0
Diacylglycerol (DG) 18:0/18:2
Diacylglyercol (DG) 18:1/18:1
Elaidic acid (EA; trans 18:1)
Glutathione (GSH)
Oleic acid (OA; 18:1)
Palmitic acid (PAL; 16:0)
Palmitoyl CoA (PAL-CoA)
Phosphatidylcholine (PC) 18:2/18:2
Phosphatidylethanolamine (PE) 16:1/16:1
Phosphatidylethanolamine (PE) 18:1/18:1
Phenethylamine (PEA)
Proline (Pro)
Phosphatidylserine (PS) 18:1/18:1
Retinol (vitamin A)
Stearic acid (STE; 18:0)
Thymidine (Thd)
Tryptophan (Trp)
Uracil (U)Uridine (Uri)

LogP
-0.38
-2.30
0.30
9.94
10.16
10.28
10.26
7.68
-2.70
7.68
7.23
2.35
5.68
7.89
8.81
1.41
-2.70
4.96
6.38
8.02
-1.30
1.04
-1.20
-1.80

Metabolite Class
Nucleobase
Nucleotide
VitaminCoenzyme
Lipid
Lipid
Lipid
Fatty acid
PeptideFatty acid
Fatty acid
Coenzyme
Lipid
Lipid
Lipid
Neurotransmitter
Amino acid
Lipid
VitaminFatty acid
Nucleoside
Amino acid
Nucleobase
Nucleoside

Usage Specifications
Criteria
No of sample runs
Storage
Recommended retest

Recommendation≤20 (with 5 µL LC-MS injections)-80ºC; protect from light, air, and moisture
2 years

Solution Preparation Procedure
1.	Reconstitute the dried-down extracts in 100 μL of ACN:water (e.g., 1:1 v/v). Note: Alternate solvent ratios are possible without impact
to the credentialing approach. Changing the ratio to increase ACN or water will simply increase the coverage of organic or watersoluble metabolites, respectively.
2.	Sonicate briefly the solutions (equates to 40 μL/mg of E. coli extract) then centrifuge (e.g., for 15 min at 13,000 RPM and 4°C) andincubate overnight at 4°C. This results in a clear solution.
3.	Mix aliquots of the unlabeled and 13C-labeled extracts (at 99% isotopic enrichment) into autosampler vials at distinct ratios (e.g., 1:2
and 1:1 v/v). Vortex briefly.
4. Load the vials onto an autosampler rack for the benchmarking optimizations.

Chemical purity (CP) is 98% or greater, unless otherwise indicated.For research use only. Not for use in diagnostic procedures.

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Example Results

A

B

Figure. Credentialing benchmark approach for untargeted metabolomics. A Schematic of the credentialing workflow (abridged from PMID: 25160088) used to discriminate
features of biological origin from contaminants and artifacts. The filtering criteria is based on feature ratios, with I being the intensity. B Credentialed metabolite measured on
a Q Exactive Plus. Shown are the LC-MS details and spectra for glutathione (C10H17N3O6S). The M+0 in the MS survey scan is at m/z 308.0914, while the M+U is at 318.1249.
Example References
Li, S.; Zheng, S. 2023. Generalized tree structure to annotate untargeted metabolomics and stable isotope tracing data. Anal Chem, 95(15), 6212-6217.
Dodds, J.N.; Wang, L.; Patti, G.J.; et al. 2022. Combining isotopologue workflows and simultaneous multidimensional separations to detect, identify, and validate metabolites in
untargeted analyses. Anal Chem, 94(5), 2527-2535.
Giné, R.; Capellades, J.; Badia, J.M.; et al. 2021. HERMES: a molecular-formula-oriented method to target the metabolome. Nat Methods, 18(11), 1370-1376.
Cho, K.; Schwaiger-Haber, M.; Naser, F.J.; et al. 2021. Targeting unique biological signals on the fly to improve MS/MS coverage and identification efficiency in metabolomics.
Anal Chim Acta, 1149, 338210-338227.
Sindelar, M.; Patti, G.J. 2020. Chemical discovery in the era of metabolomics. J Am Chem Soc, 142(20), 9097-9105.
Wang, L.; Naser, F.J.; Spalding, J.L.; et al. 2019. A protocol to compare methods for untargeted metabolomics. Methods Mol Biol, 1862, 1-15.
Spalding, J.L.; Naser, F.J.; Mahieu, N.G.; et al. 2018. Trace phosphate improves ZIC-pHILIC peak shape, sensitivity, and coverage for untargeted metabolomics. J Proteome Res,
17(10), 3537-3546.
Naser, F.J.; Mahieu, N.G.; Wang, L.; et al. 2018. Two complementary reversed-phase separations for comprehensive coverage of the semipolar and nonpolar metabolome.
Anal Bioanal Chem, 410(4), 1287-1297.
Mahieu, N.G.; Patti, G.J. 2017. Systems-level annotation of a metabolomics data set reduces 25,000 features to fewer than 1,000 unique metabolites. Anal Chem, 89(19),10397-10406.
Benton, H.P.; Ivanisevic, J.; Mahieu, N.G.; et al. 2015. Autonomous metabolomics for rapid metabolite identification in global profiling. Anal Chem, 87(2), 884-891.
Mahieu, N.G.; Huang, X.; Chen, Y.; et al. 2014. Credentialing features: a platform to benchmark and optimize untargeted metabolomic methods. Anal Chem, 86(19), 9583-9589.

Chemical purity (CP) is 98% or greater, unless otherwise indicated.

38

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| Stable Isotope-Labeled Mixtures, Sets, and Kits

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PeptiQuant™ Plus Assay Kits
Researchers in academia and life science industries continue to implement a bottom-up MS-based workflow for protein biomarker screening.
Biomarker verification/validation requires absolute quantification of surrogate peptides in a sample matrix, a requirement that is best
achieved using well-characterized stable isotope-labeled standards. To ensure robust quantitative measurements, QC checks should beroutinely performed. CIL offers a collection of PeptiQuant Assay Kits (from MRM Proteomics Inc.) for QC and biomarker assessment using
bottom-up LC-MS/MS methodologies. The QC kits are designed to evaluate the performance of an LC-MS platform, either alone or in
combination with a human plasma proteomics workflow (see corresponding flyer for panel details). The biomarker assessment kits (BAKs)
are intended to help researchers screen target panels of candidate protein disease biomarkers in humanor mouse plasma samples (see corresponding flyer for panel details). The current platform-specific offeringsfor each kit type are listed in the overview below.
Overview
QC Kits
Catalog No.
LCMSP-QC

WFPKCambridge Isotope
Laboratories, Inc.
isotope.com

RESEARCH PRODUCTS

Description

Kit Contents

PeptiQuant Plus Human
Plasma Daily QC Kit• C/ N-labeled peptide mix• USB (e.g., user manual,acquisition and analysis files)• 13C/ 15N-labeled peptide mix• unlabeled peptide mix• trypsin• BSA• human plasma• USB (e.g., user manual,acquisition and analysis files)

PeptiQuant Plus Human
Plasma Workflow QC Kit

Biomarker Assessment Kits (BAKs)
Catalog No. Description

Human
BAK-125

BAK-270

13

15

No. ofPeptides35

35

Unit Size
10, 20, or 50
injections
1 or 2 runs

Optimized
Instrument• 6490/6495 QqQ• QTRAP® 6500
• Q Exactive™ Plus
• 6490/6495 QqQ• QTRAP 6500
• Q Exactive Plus

PeptiQuant Plus
Quality Control Kits
For Targeted and Untargeted
Proteomics

Cambridge Isotope Laboratories, Inc. is pleased to offer
PeptiQuant™ Plus quality control (QC) kits for performanceassessment of targeted (or untargeted) plasma proteomic
workflows. The daily QC kit is intended for routine LC-MS/MS
performance checks, while the workflow QC kit is designed tomeasure the quality of one type of bottom-up LC-MS/MS method.
It should be noted that the utilized workflow is the same as that
employed in the PeptiQuant Plus Biomarker Assessment Kits
(BAKs),1,2 which could enable their parallel usage. As with theBAKs, these QC kits are optimized for platform-dependent
operation (see table below for current offerings), with a condensed
panel of 35 peptides (one peptide/protein) monitored per kit for
rapid QC analysis. All assays have been validated according to the
CPTAC (Clinical Proteomic Tumor Analysis Consortium) guidelines,
with their characterization results uploaded to the CPTAC Assay
Portal3 for reference.
Platforms Available
Kit Type

Agilent
6490/6495

Sciex QTRAP®
6500

Thermo Scientific™
Q Exactive™ Plus

Daily QCnnn

Workflow QC

n

n

n

Note: Please inquire if an alternate LC-MS/MS platform or alternate kit number(i.e., for daily injection or workflow run) is desired.

1. Percy, A.J.; Borchers, C.H. 2021. Detailed method for performing the ExSTA approach in quantitative bottom-up plasma proteomics. Methods Mol Biol, doi: 10.1007/978-10716-1024-4.
2. Gaither, C.; Popp, R.; Mohammed, Y.; et al. 2020. Determination of the concentration range for 267 proteins from 21 lots of commercial human plasma using highly multiplexed
multiple reaction monitoring mass spectrometry. Analyst, 145(10), 3634-3644.
3. Whiteaker, J.R.; Halusa, G.N.; Hoofnagle, A.N.; et al. 2014. CPTAC Assay Portal: a repository of targeted proteomic assays. Nat Methods, 11(7), 703-704.

Continued ➤Cambridge Isotope Laboratories, Inc.
North America: 1.800.322.1174 cilsales@isotope.com | International: +1.978.749.8000 intlsales@isotope.com | fax: 1.978.749.2768 | isotope.com

Cambridge Isotope
Laboratories, Inc.
isotope.com

RESEARCH PRODUCTSRESEARCH PRODUCTS

PeptiQuant™ Plus
Biomarker
Assessment Kits
For Targeted Quantitative
Proteomics
Cambridge Isotope Laboratories, Inc. is pleased to offer PeptiQuant™ Plus biomarker assessment kits (BAKs) for the targeted
and precise quantitation of ≥125 proteins in human and mouse plasma samples.1-9 These PeptiQuant kits utilize a well-defined,bottom-up LC-MS/MS methodology together with heavy (stable isotope-labeled standards, SIS) and light (natural, NAT) peptide standards
for the quantitation of moderate-to-high abundance proteins. The peptides have been carefully selected, formulated, and characterized
to serve as effective, molecular surrogates for their corresponding proteins. The proteins are candidate disease markers (e.g., cancer,
cardiovascular) that are suitable for biomarker discovery and verification studies. All assays have been validated according to the complete
set of CPTAC (Clinical Proteomic Tumor Analysis Consortium) guidelines and are available for viewing on the CPTAC Assay Portal.10

Kit Contents

Optimized Platforms Available

Kit Contents

No. ofQuant.
Proteins

Unit Size

Optimized
Instrument

Kit Type

6490/6495
QqQ

Human BAK-125

nMouse BAK-125

nMouse BAK-125-2

nMouse BAK-125-3nHuman BAK-270

n

Human Cancer BAK-50

n

6545
Q-TOF

QTRAP®
6500nnn

Q Exactive™
Plus

Xevo
TQ-XS

n

n

n
n
n

n

n
n

Most kits are available for 20, 50, and 100 samples.
Note: The -2 and -3 demarcations in the catalog number reflect an alternate setof 125 target proteins. Please inquire if an alternate LC-MS/MS platform is desiredfrom the current panel offerings.
Example References
1. Gaither, C.; Popp, R.; Zahedi, R.P.; et al. 2022. Multiple reaction monitoring-mass spectrometry
enables robust quantitation of plasma proteins regardless of whole blood processing delays that
may occur in the clinic. Mol Cell Proteomics, 21(5), 100212-100223.
2. Brzhozovskiy, A.; Kononikhin, A.; Bugrova, A.E.; 2022. The parallel reaction monitoring-parallel
accumulation-serial fragmentation (prm-PASEF) approach for multiplexed absolute quantitationof proteins in human plasma. Anal Chem, 94(4), 2016-2022.

Cambridge Isotope Laboratories, Inc.

PeptiQuant Plus Mouse
Plasma Proteomics Kit• C/ N-labeled peptide mix• unlabeled peptide mix• trypsin• BSA• USB (e.g., user manual,acquisition and analysis files)• 13C/ 15N-labeled peptide mix• unlabeled peptide mix• trypsin• BSA• USB (e.g., user manual,acquisition and analysis files)• 13C/ 15N-labeled peptide mix• unlabeled peptide mix• trypsin• BSA• USB (e.g., user manual,acquisition and analysis files)

125

20, 50, or
100 samples• 6490/6495 QqQ• QTRAP 6500
• Q Exactive Plus
• Xevo TQ-XS

270

20, 50, or
100 samples• 6490/6495 QqQ• QTRAP 6500
• Q Exactive Plus

50

50 or 100
samples• 6490/6495 QqQ• Q Exactive Plus

• 13C/ 15N-labeled peptide mix• unlabeled peptide mix• trypsin• BSA• USB (e.g., user manual,acquisition and analysis files)

125

20, 50, or
100 samples• 6490/6495 QqQ• QTRAP 6500
• Q Exactive Plus
• 6545 Q-TOF

13

15• 4 vials of starting material (lyophilized)
Vial 1 (“S” on cap): SIS peptide mix (for spiking into standard
curve, QC, and experimental sample digests)
Vial 2 (clear cap): NAT peptide mix (for spiking into standard
curve and QC sample digests)
Vial 3 (purple cap): TPCK-treated trypsin
Vial 4 (green cap): BSA• USB containing user manual, target details (with specifications),
vendor-specific acquisition method, and Skyline analysis file

Features and Benefits• CPTAC-certified assays• SIS and NAT peptides for each target protein• Robust and diverse protein quantitation• Reduces development time and cost

3. Percy, A.J.; Borchers, C.H. 2021. Detailed method for performing the ExSTA approach inquantitative bottom-up plasma proteomics. Methods Mol Biol, 2228, 353-384.
4. Mohammed, Y.; Bhowmick, P.; Michaud, S.A.; et al. 2021. Mouse Quantitative Proteomics
Knowledgebase: reference protein concentration ranges in 20 mouse tissues using 5000
quantitative proteomics assays. Bioinformatics, in press.

7. Gaither, C.; Popp, R.; Mohammed, Y.; et al. 2020. Determination of the concentration range for267 proteins from 21 lots of commercial human plasma using highly multiplexed multiple reaction
monitoring mass spectrometry. Analyst, 10, 3449-3760.

Expanded PeptiQuant Plus
Human Plasma Proteomics
Kit• CPTAC-certified assays• SIS and NAT peptides for each target protein• Rapid performance evaluation of LC-MS/MS platform
and /or analytical workflow• Reduces development time and cost

• Starting materials (lyophilized), with all five vials utilized in theworkflow QC kit (for one analytical run) and vial 1 only in thedaily QC kit (enables five injections):
Vial 1 (“S” on cap): SIS peptide mix for spiking into standard
curve, QC, and experimental sample digests
Vial 2 (clear cap): NAT peptide mix for spiking into standard
curve and QC sample digests
Vial 3 (purple cap): TPCK-treated trypsin (1 mg/mL after
100 mM Tris pH 8.0 addition)
Vial 4 (green cap): BSA (10 mg/mL after PBS addition)
Vial 5 (blue cap)
Human plasma

References

6. Bhardwaj, M.; Weigl, K.; Tikk, K.; et al. 2020. Multiplex quantitation of 270 plasma protein
markers to identify a signature for early detection of colorectal cancer. Eur J Cancer, 127, 30-40.

PeptiQuant Plus Human
Plasma Proteomics Kit

Features and Benefits

Kit Contents• USB containing user manual, guideline performance values,
acquisition method (for given instrument platform), andSkyline analysis file

The daily QC kits are available for 10, 20, and 50 injections.
The workflow QC kits are available for 1 and 2 runs.

5. Ayton, S.; Janelidze, S.; Roberts, B.; et al. 2021. Acute phase markers in CSF reveal inflammatorychanges in Alzheimer’s disease that intersect with pathology, APOE ε4, sex and age. Prog
Neurobiol, 101904-101915.

BAK-CNCR50 DiseaseQuant Human
Tissue Cancer Pathway
Proteomics Kit

Mouse
M-BAK-125*

RESEARCH PRODUCTS™

8. Michaud, S.A.; Sinclair, N.J.; Pětrošová, H.; et al. 2018. Molecular phenotyping of laboratory
mouse strains using 500 multiple reaction monitoring mass spectrometry plasma assays.
Commun Biol, 1, 78.
9. Mohammed, Y.; Goodlett, D.; Borchers, C.H. 2023. Absolute quantitative targeted proteomics
assays for plasma proteins. Methods Mol Biol, 628, 439-473.
10. Whiteaker, J.R.; Halusa, G.N.; Hoofnagle, A.N.; et al. 2014. CPTAC Assay Portal: a repositoryof targeted proteomic assays. Nat Methods, 11(7), 703-704.
Application Note
Percy, A.J.; Trouvé, R.; Lehmann, S.; Hirtz, C. Vialaret, J. 2021. Translation and implementation of thePeptiQuant™ Plus Human Plasma BAK-270. (CIL application note #50)

Continued ➤North America: 1.800.322.1174 cilsales@isotope.com | International: +1.978.749.8000 intlsales@isotope.com | fax: 1.978.749.2768 | isotope.com

Click on the thumbnails or visit
isotope.com for more information.

*Alternate sets of 125 target proteins are available (see PeptiQuant Plus BAK flyer for details).
PeptiQuant is a trademark of MRM Proteomics Inc.

Chemical purity (CP) is 98% or greater, unless otherwise indicated.For research use only. Not for use in diagnostic procedures.

cilsales@isotope.com | intlsales@isotope.com | isotope.com 39

Cambridge Isotope Laboratories, Inc.

| Stable Isotope-Labeled Mixtures, Sets, and Kits

Usage Specifications
Before reconstitution (for peptide mixes):
Storage-80°C
Recommended retest
6 months from date of manufacture

Note: The dried-down trypsin and BSA materials supplied with the BAKs are not stable at room temperature and must be stored at 4°C.
Trypsin is to be prepared immediately prior to use and stored on ice until then.
Example Results

A

B

Figure. Bottom-up LC-MRM/MS data for two representative, low abundance targets from a PeptiQuant Plus BAK-270 analysis. Illustrated are Skyline screenshots for peptide
DVIVHPLPLK (from Adhesion G protein-coupled receptor F5, gene ADGRF5, UniProtKB Q8IZF2) in A and peptide LLEYDTVTR (from Adipocyte plasma membrane-associated protein,gene APMAP, UniProtKB Q9HDC9) in B. The tabs refer to the monitored panel, two example XICs (for curve calibrant level F and a pooled human plasma sample analysis), thestandard curve (with calibrant levels marked with gray squares, QC samples with green diamonds, experimental sample with red arrow), and the results grid. The protein
concentration in the pooled human plasma sample was determined to be 321.2 ng/mL for ADGRF5 and 439.9 ng/mL for APMAP. Results courtesy of MRM Proteomics.

Chemical purity (CP) is 98% or greater, unless otherwise indicated.

40

tel: +1.978.749.8000 | 1.800.322.1174 (North America) | fax: +1.978.749.2768

For research use only. Not for use in diagnostic procedures.

| Stable Isotope-Labeled Mixtures, Sets, and Kits

isotope.com

Example References
Mohammed, Y.; Goodlett, D.; Borchers, C.H. 2023. Absolute quantitative targeted proteomics assays for plasma proteins. Methods Mol Biol, 628, 439-473.
Stakhneva, E.M.; Kashtanova, E.V.; Polonskaya, Y.V.; et al. 2022. The search for associations of serum proteins with the presence of unstable atherosclerotic plaque in coronary
atherosclerosis. Int J Mol Sci, 23(21), 12795-12805.
Richard, V.R.; Gaither, C.; Popp, R.; et al. 2022. Early prediction of COVID-19 patient survival by targeted plasma multi-omics and machine learning. Mol Cell Proteomic, 21(10),100277-100290.
Pastushkova, L.K.; Goncharov, I.N.; Koloteva, M.I.; et al. 2022. Characteristics of blood plasma proteome changes associated with the hemorrhagic purpura of cosmonauts on thefirst day after long-term space missions. Life Sci Space Res, 33, 7-12.
Gaither, C.; Popp, R.; Zahedi, R.P.; et al. 2022. Multiple reaction monitoring-mass spectrometry enables robust quantitation of plasma proteins regardless of whole blood processing
delays that may occur in the clinic. Mol Cell Proteomics, 21(5), 100212-100223.
Brzhozovskiy, A.; Kononikhin, A.; Bugrova, A.E.; et al. 2022. The parallel reaction monitoring-parallel accumulation-serial fragmentation (prm-PASEF) approach for multiplexed
absolute quantitation of proteins in human plasma. Anal Chem, 94(4), 2016-2022.
Percy, A.J.; Borchers, C.H. 2021. Detailed method for performing the ExSTA approach in quantitative bottom-up plasma proteomics. Methods Mol Biol, 2228, 353-384.
Mohammed, Y.; Bhowmick, P.; Michaud, S.A.; et al. 2021. Mouse Quantitative Proteomics Knowledgebase: reference protein concentration ranges in 20 mouse tissues using 5000
quantitative proteomics assays. Bioinformatics, in press.
Ayton, S.; Janelidze, S.; Roberts, B.; et al. 2021. Acute phase markers in CSF reveal inflammatory changes in Alzheimer’s disease that intersect with pathology, APOE ε4, sex and age.
Prog Neurobiol, 101904.
Gaither, C.; Popp, R.; Mohammed, Y.; et al. 2020. Determination of the concentration range for 267 proteins from 21 lots of commercial human plasma using highly multiplexed
multiple reaction monitoring mass spectrometry. Analyst, 145(10), 3634-3644.
Bhardwaj, M.; Weigl, K.; Tikk, K.; et al. 2020. Multiplex quantitation of 270 plasma protein markers to identify a signature for early detection of colorectal cancer. J Cancer, 127,
30-40.
Michaud, S.A.; Sinclair, N.J.; Petrošová, H.; et al. 2018. Molecular phenotyping of laboratory mouse strains using 500 multiple reaction monitoring mass spectrometry plasma assays
Commun Biol, 1, 78.
Orti, V.; Mertens, B.; Vialaret, J.; et al. 2018. Data from a targeted proteomics approach to discover biomarkers in saliva for the clinical diagnosis of periodontitis. Data Brief, 18,294-299.
Mohammed, Y.; Pan, J.; Zhang, S.; et al. 2018. ExSTA: External standard addition method for accurate high-throughput quantitation in targeted proteomics experiments.
Proteomics Clin Appl, 12(2), 1600180.
Application Note
Percy, A.J.; Trouvé, R.; Lehmann, S.; Hirtz, C. Vialaret, J. 2021. Translation and implementation of the PeptiQuant™ Plus Human Plasma BAK-270. (CIL application note #50)

PeptiQuant Plus Platform Performance Kit has proven to be a vital component of our everyday quality assurance that enables us to deliver“ Thehigh-quality
targeted proteomics data in an accurate and timely manner. This kit has a ‘dilute and shoot’ operation and comes with vendorspecific LC-MRM/MS parameters and a Skyline analysis file for quick input and results output. Altogether, the performance kit is an excellent
means to rapidly assess LC-MS performance that should become a routine staple in a proteomic user’s toolbox.
– Tasso Miliotis, PhD
Associate Principal Scientist at AstraZeneca Gothenburg (Sweden)”

Kits contain all the essential materials, including the standards and methods, for performing absolute protein“ PeptiQuantquantificationPlusbyAssay
LC-MRM/MS in a standardized way. The standard protocol helped us reduce the assay development time, while improve thereproducibility and precision of multiplex protein quantification. In addition to the biomarker assessment kits, the quality control kits enable
the instrument performance and assay reproducibility to be monitored and assessed, which ultimately provided us confidence in the reliabilityof the quantification results.
– Elaine Wong, PhD
Scientific Officer at Queen Mary Hospital, Fu Lam (Hong Kong)”

Chemical purity (CP) is 98% or greater, unless otherwise indicated.For research use only. Not for use in diagnostic procedures.

cilsales@isotope.com | intlsales@isotope.com | isotope.com 41

| Stable Isotope-Labeled Mixtures, Sets, and Kits

Cambridge Isotope Laboratories, Inc.

INLIGHT® Glycan Tagging Kit

RESEARCH PRODUCTS

Cambridge Isotope Laboratories, Inc.
isotope.com

Glycans participate in a large number of cellular, molecular, and biological processes and are implicated in a numberof diseases (e.g., Alzheimer’s, cancer). To aid the identification and relative quantification of glycans by LC-MS,
CIL offers the innovative INLIGHT® (Individuality Normalization when Labeling with Isotopic Glycan Hydrazide Tag)
glycan-tagging kit. This kit employs both natural (NAT – 12C6) and stable isotope-labeled (SIL – 13C6) phenyl 2-GPN
reagents in the hydrazide derivatization of free N-glycans. While this tagging strategy was developed for N-glycans,it has also been adapted to sample analysis of O-glycan and heparin oligomer profiles. The INLIGHT kit contains fivevials of NAT reagent and five vials of SIL reagent, which in total provides sufficient tagging for approximately 125
relative quantification experiments. The user manual provides step-by-step instructions for executing the modified
INLIGHT strategy using maltoheptaose and fetuin A as examples. Data processing and analysis of derivatized glycans
can be facilitated in GlycoHunter or Skyline.

INLIGHT Glycan
Tagging Kit®

New KitProtocolCatalog No. GTK-1000
The INLIGHT® glycan-tagging kit, developed by the David Muddiman group in collaboration
with synthetic chemist Daniel Comins at North Carolina State University,1,2 represents a commendable
tool for the relative quantification of glycans in binary samples using MS-based techniques.
CIL is pleased to offer the INLIGHT kit with an updated procedure enabling faster preparationand enhanced MS analysis / interpretation. The kit contents include phenyl 2-GPN hydrazide
tagging reagents (unlabeled and 13C6), an unlabeled maltoheptaose oligosaccharide, and auser manual. The manual provides detailed instructions on N-glycan release (with PNGase F),tagging, and LC-MS analysis using fetuin A and maltoheptaose as examples. The enhanced
protocol and software solutions have been described previously,3-5 while examples of INLIGHT
applications have been further demonstrated.6-9 Overall, the INLIGHT kit can be applied tocomplex glycome samples (e.g., human plasma) enabling ~125 relative quantification
experiments of N-glycans in binary biosamples (i.e., case and control).

1. PNGase
2. INLIGHT light

Kit Benefits• Dried-down hydrazide reagents forflexible preparation• Detailed instruction manual for simpler,
faster, and more efficient analysis• Amenable to RPLC- and HILIC-MS• Software solutions for expediting
analysis• Relative quantification of N-linked
glycomes

Figure. Hydrazone formation mechanismof N-glycans with the INLIGHT reagents.
Following derivatization, the glycome
samples are mixed 1:1 (v/v) then analyzed
by LC-MS. Ratios of each individual glycan
pair will reveal differences between thetwo samples in a quantitative fashion.

Asn – Peptide
1. PNGase
2. INLIGHT heavy

Example References
1. Walker, S.H.; Lilley, L.M.; Enamorado, M.F.; et al. 2011. Hydrophobic derivatization of N-linked glycans forincreased ion abundance in electrospray ionization mass spectrometry. J Am Soc Mass Spectrom, 22(8),1309-1317.
2. Walker, S.H.; Taylor, A.D.; Muddiman, D.C. 2013. Individuality Normalization when Labeling with Isotopic
Glycan Hydrazide Tags (INLIGHT): A novel glycan-relative quantification strategy. J Am Soc Mass Spectrom,24(9), 1376-1384.
3. Kalmar, J.G.; Garrard, K.P.; Muddiman, D.C. 2021. GlycoHunter: An open-source software for the detection
and relative quantification of INLIGHT®-labeled N-linked glycans. J Proteome Res, 20(4), 1855-1863.
4. Kalmar, J.G.; Butler, K.E; Baker, E.S.; et al. 2020. Enhanced protocol for quantitative N-linked glycomics
analysis using Individuality Normalization when Labeling with Isotopic Glycan Hydrazide Tags (INLIGHT)®.
Anal Bioanal Chem, 412(27), 7569-7579.
5. De Leoz, M.L.A.; Duewer, D.L.; Fung, A.; et al. 2020. NIST interlaboratory study on glycosylation analysis ofmonoclonal antibodies: comparison of results from diverse analytical methods. Mol Cell Proteomics, 19(1),11-30.

6. King, S.R.; Hecht, E.S.; Muddiman, D.C. 2018. Demonstration of hydrazide tagging for O-glycans and acentral composite design of experiments optimization using the INLIGHT® reagent. Anal Bioanal Chem,
410(5), 1409-1415.
7. Mangrum, J.B.; Mehta, A.Y.; Alabbas, A.B.; et al. 2017. Comparative analysis of INLIGHT®-labeled
enzymatically depolymerized heparin by reverse-phase chromatography and high-performance mass
spectrometry. Anal Bioanal Chem, 409(2), 499-509.
8. Loziuk, P.L.; Hecht, E.S.; Muddiman, D.C. 2017. N-linked glycosite profiling and use of Skyline as a platform
for characterization and relative quantification of glycans in differentiating xylem of Populus trichocarpa.
Anal Bioanal Chem, 409(2), 487-497.
9. Hecht, E.S.; Scholl, E.H.; Walker, S.H.; et al. 2015. Relative quantification and higher-order modeling of theplasma glycan cancer burden ratio in ovarian cancer case-control samples. J Proteome Res, 14(10),4394-4401.

Cambridge Isotope Laboratories, Inc.
North America: 1.800.322.1174 cilsales@isotope.com | International: +1.978.749.8000 intlsales@isotope.com | fax: 1.978.749.2768 | isotope.com
GLY_INLIGHT_KIT (10/4/22)
Supersedes all previously published literature

Click on the thumbnail or visit
isotope.com for more information.

Overview
Catalog No.
GTK-1000

Description
INLIGHT® Glycan Tagging Kit

Kit Contents• Light phenyl 2-GPN INLIGHT reagent (5 × 0.25 mg)• Heavy phenyl 2-GPN INLIGHT reagent (13C6; 5 × 0.25 mg)• Unlabeled maltoheptaose (5 × 10 μg)

Usage Specifications
Criteria
Recommendation
No. of Uses
25 per vial
Before reconstitution:
Storageambient temperature; protect from light and moisture
Recommended retest
5 years from date of manufacture

Figure. LC-MS examples of INLIGHT derivatized maltoheptaose andN-linked glycans from bovine fetuin.
A. Extracted ion chromatogram of [NAT + H]+ maltoheptaose.
B. Mass spectrum at 10 minutes illustrating NAT and SIL derivatized
maltoheptaose, maltohexaose, maltopentaose, and maltotetraose.
C. Extracted ion chromatogram of a fetuin-based glycan at m/z
1012.3980 corresponding to [NAT + 2H + (Fuc)1(Gal)2(GlcNAc)4(Man)3]2+.
D. Average mass spectrum of a fetuin analysis showing the NAT and SIL
derivatized N-linked glycan pairs in the LC window 8-11 min.

Example References
Butler, K.E.; Kalmar, J.G.; Muddiman, D.C.; et al. 2022. Utilizing liquid chromatography, ion mobility spectrometry, and mass spectrometry to assess INLIGHT™ derivatized N-linked
glycans in biological samples. Anal Bioanal Chem, 414(1), 623-637.
Kalmar, J.G.; Garrard, K.P.; Muddiman, D.C. 2021. GlycoHunter: An open-source software for the detection and relative quantification of INLIGHT®-labeled N-linked glycans.
J Proteome Res, 20(4), 1855-1863.
Kalmar, J.G.; Butler, K.E; Baker, E.S.; et al. 2020. Enhanced protocol for quantitative N-linked glycomics analysis using Individuality Normalization when Labeling with Isotopic Glycan
Hydrazide Tags (INLIGHT).® Anal Bioanal Chem, 412(27), 7569-7579.
King, S.R.; Hecht, E.S.; Muddiman, D.C. 2018. Demonstration of hydrazide tagging for O-glycans and a central composite design of experiments optimization using the INLIGHT®
reagent. Anal Bioanal Chem, 410(5), 1409-1415.
Loziuk, P.L.; Hecht, E.S.; Muddiman, D.C. 2017. N-linked glycosite profiling and use of Skyline as a platform for characterization and relative quantification of glycans indifferentiating xylem of Populus trichocarpa. Anal Bioanal Chem, 409(2), 487-497.
Hecht, E.S.; Scholl, E.H.; Walker, S.H.; et al. 2015. Relative quantification and higher-order modeling of the plasma glycan cancer burden ratio in ovarian cancer case-control samples.
J Proteome Res, 14(10), 4394-4401.

Chemical purity (CP) is 98% or greater, unless otherwise indicated.

42

tel: +1.978.749.8000 | 1.800.322.1174 (North America) | fax: +1.978.749.2768

For research use only. Not for use in diagnostic procedures.

isotope.com

| Stable Isotope-Labeled Mixtures, Sets, and Kits

Q&As
Listed below are a series of general Q&As for CIL’s mixes and kits. Product-specific FAQs can be located on isotope.com’s FAQs page andon their corresponding product application page (e.g., Life Sciences → Metabolomics Mixtures and Kits → QC Mixes and Kits).
What are the advantages of selecting an off-the-shelf mix vs.
a collection of individual isotope standards for self-mixing?
The MSK/NSK products are formulated to exacting standards
following detailed batch records developed from over 20 years offormulation experience. Following production, randomly selectedvials are analyzed to ensure both accuracy and consistency. The
astute attention and process control generates exceptional vial-tovial and lot-to-lot reproducibility. This high reproducibility returnis one of the merits of utilizing CIL prepared mixes. Additional
advantages are:• reduced development time and cost;• enhanced data quality;• ease of user implementation; and• improved confidence in analytical results.
The majority of the mixes are 13C- and/or 15N-labeled.
What are the advantages of 13C/15N-labeling vs. D-labeledin MS measurements?
The nature of the stable isotope can potentially impact thepreanalytical (e.g., storage and handling) and analytical (e.g.,sample preparation and processing) phases of an experiment.
In comparison to D labels, 13C/15N labels can have:• improved isotope stability;• negligible isotope scrambling issues;•	conserved chromatographic elution (relative to its unlabeledstandard); and• heightened analytical reliability.
In cases where D-labeled compounds were selected, the D-label
is located at nonexchangeable positions and was stability tested
for preservation as well as product application. For further
background, please refer to our technical note that describes
the benefits of 13C vs. D standards in MS-based studies.

How often should QC measurements be performed,
and what mixes are most suitable?
Prior to first use, QCs should be conducted several times
in succession prior to sample analysis to establish baseline
performance and intervention limits. Once established, QCs
should be performed routinely (i.e., before, during, and after
sample analysis) to monitor the effectiveness of the analyticalmethod and instrument platform over time. While all of CIL’s
mixes could be used in these types of operations, the ones ideallysuited for these type of measurements are the metabolomic QC kits(see MSK-QC-KIT and MSK-QReSS-KIT) and the proteomic QC kits(see PeptiQuant Plus daily and workflow QC kits).
In a targeted quantitative application, how many MRM
transitions should be monitored per analyte?
Ideally, a minimum of two MRM transitions per compound (be it
metabolite or peptide) should be targeted. This will allow for aquantifier and qualifier(s) assignment, with the ion ratios serving
as an additional metric for performance qualification. Nonetheless,this may not be possible for all analytes given the compound’s
fragmentation chemistry, the employed MS/MS parameters,
and the mode of operation utilized. Empirical experiments withisotope-labeled standards should therefore be first performed
to optimize the MS/MS parameters prior to conducting thequantitative study with precious experimental samples.
Can custom mixes or add-on vials be formulated?
Yes, we have the ability to customize. We would first review
feasibility and then provide a quotation on your specific mix.
To start this process, please provide the necessary details on
this Custom Mix Request form or contact your local sales
representative.

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Cambridge Isotope Laboratories, Inc.

| Stable Isotope-Labeled Mixtures, Sets, and Kits

Please visit isotope.com for a complete list of isotope-labeled compounds.
Research products are distributed and sold worldwide via our extensive network.
CIL’s distributor listing is available at isotope.com.

To request a quotation or place an order:
North America: 1.978.749.8000 | 1.800.322.1174 | cilsales@isotope.com
International: +1.978.749.8000 | intlsales@isotope.com
Fax: 1.978.749.2768 | isotope.com
For research use only. Not for use in diagnostic procedures.

Cambridge Isotope Laboratories, Inc. 3 Highwood Drive, Tewksbury, MA 01876 USA
North America: 1.800.322.1174 | International: +1.978.749.8000 | fax: +1.978.749.2768 | isotope.com

MIXES_KITS_CATALOG (8/28/24)
Supersedes all previously published literature