1
H NMR metabolic profiling of human serum associated with benign
and malignant gallstone diseases
Authors: Kanchan Sonkar1, Anu Behari2, V. K. Kapoor2, Neeraj Sinha1*
Authors affiliations:
1. Centre of Biomedical Magnetic Resonance, Sanjay Gandhi Post Graduate Institute of
Medical Sciences (SGPGIMS) - Campus, Lucknow-226014, Uttar Pradesh, India
2. Department of Surgical Gastroenterology, Sanjay Gandhi Post Graduate Institute of
Medical Sciences, Lucknow-226014, Uttar Pradesh, India
*Correspondence to:
Dr. Neeraj Sinha
Centre of Biomedical Magnetic Resonance, Sanjay Gandhi Postgraduate Institute of
Medical Sciences Campus, Raibarely Road, Lucknow-226014 (INDIA)
E-mail: neerajcbmr@gmail.com, neeraj.sinha@cbmr.res.in
Contact: +91-0522-2495031
Fax: +91-522 – 2668215
Supporting Information Figure 1: Expansion of 1H COSY spectrum (From 0.5 to 6.1 ppm) of
human serum from healthy control along with the assignment of the metabolites (Ile; Isoleucine,
Leu; Leucine, Val; Valine, Lys; Lysine, Glu; Glutamate, Gln; Glutamine, Ala; Alanine, Lac;
Lactate, Thr; Threonine, Asp; Aspartic acid, Asn; Asparagine, Glc; Glucose)
Table 1: Chemical shift assignments of metabolites observed in the 1H NMR CPMG
spectrum of human serum in different pathologies of gallbladder using one dimensional
(1D) chemical shift reported in literature (LIT), two dimensional COSY, HSQC and
comparing standard NMR spectrum (STD) of individual metabolites taken from Biological
Magnetic Resonance Bank (BMRB).
SN
Name of Metabolites
Chem. Shift
Resonances
Methods
1
Acetate
1.92 (s)
CH3
1D, HSQC
2
Alanine
1.48 (d)
β-CH3
1D, COSY
3.78 (q)
α-CH
3
Arginine
1.68 (m)
γ-CH2
1.90 (m)
β-CH2
3.25(m)
δ-CH2
3.77(m)
α-CH
2.87 (dd)
β-CH
2.95 (dd)
β'-CH
4.01 (dd)
α-CH
2.69 (dd)
β-CH
2.82 (dd)
β'-CH
3.90 (dd)
α-CH
3.21 (s)
N(CH3)3
3.53(m)
N-CH2
4.07(m)
O-CH2
2.53 (d)
CH2
2.67 (d)
CH2
3.03( )
N-CH3
3.94( )
N-CH2
0.90/0.96
CH3
1.3( )
.-(CH2)n-
1.59( )
CH2-CH2-CO
2.04/2.07
CH=CH-CH2
2.26( )
CH2-CO
2.81( )
CH=CH-CH2-CH=CH
5.33( )
CH=CH
8.45 (s)
CH
5.24 (d)
C1H
3.24 (dd)
C2H
3.41 ( )
C4H
3.47 ( )
C5H
3.49 (t)
C3H
3.74 ( )
C6H
4
5
6
7
8
9
10
11
Asparagine
Aspartic Acid
Choline
Citric Acid
Creatine
Fatty acids
Formic Acid
β-Glucose
1D, COSY
1D, COSY
1D, COSY
1D, COSY
1D, STD
1D, HSQC
1D, COSY, LIT
1D, STD
1D, COSY
12
13
14
15
Glutamic Acid
Glutamine
Histidine
Isoleucine
16
Lactate
17
LDL/VLDL/HDL
18
Leucine
19
20
21
22
Lysine
Phenylalanine
Threonine
Tyrosine
3.91 ( )
C6'H
4.65 (d)
C1H
2.09 ( )
β-CH2
2.35 (m)
γ-CH2
3.77 ( )
α-CH
2.13 (m)
β-CH2
2.44 (m)
γ-CH2
3.77 ( )
α-CH
3.15 (dd)
β-CH
3.24 (dd)
β'-CH
4.00 (dd)
α-CH
7.13 (s)
C4H-ring
7.98 (s)
C2H-ring
0.94 (t)
δ-CH3
1.01 (d)
γ-CH3
1.26 (m)
γ-CH
1.47 (m)
γ'-CH
1.98 (m)
β-CH
3.68 (d)
α-CH
1.33 (d)
β-CH3
4.12 (q)
α-CH
0.96 (d)
δ-CH3
0.97 (d)
δ'-CH3
1.71 (m)
γ-CH/β-CH2
3.75 ( )
α-CH
1.47 (m)
γ-CH2
1.72 (m)
β-CH2
1.9 (m)
δ-CH2
3.02 ( )
N-CH2
3.74 ( )
α-CH
3.12 ( )
β-CH
3.28 ( )
β'-CH
3.98 ( )
α-CH
7.32 (d)
C2H, C6H-ring
7.37 (m)
C4H-ring
7.41 (m)
C3H, C5H-ring
1.34 (d)
γ-CH3
3.6 (d)
α-CH
4.25 (m)
β-CH
3.06 (dd)
β-CH
3.19 (dd)
β'-CH
3.95 (dd)
α-CH
1D, COSY
1D, COSY
1D, COSY, STD
1D, COSY, STD
1D, STD, COSY
1D, STD, COSY
1D, STD, COSY
1D, STD, COSY
1D, COSY, STD
1D, COSY, STD
23
24
Uracil
Valine
6.89 (d)
C3H, C5H-ring
7.18 (d)
C2H, C6H-ring
5.8 (s)
C5H-ring
7.54 (d)
C6H-ring
0.99 (d)
γ-CH3
1.04 (d)
γ'-CH3
3.62 (d)
β-CH
2.28 ( )
α-CH
1D, COSY, STD
1D, COSY, STD
Table 2: Structure of metabolites identified and assigned by one and two dimensional NMR
experiments:
O
NH2
O
H3C
H3C
H
N
H2N
OH
OH
H2N
Acetic Acid
Alanine

O


HO
OH
NH2
O
Asparagine


Arginine
NH

O
OH
H2N
OH
N
CH3
NH2
Aspartic Acid
O
Creatine
COOH
HO
COOH
N+
HOOC
HO
OH
OH
OH
10
O
6
OH
6
1
4
4
HO
OH
O
5
1
5
3
2
OH
O
Formic Acid
Choline
Citric Acid
COOH
NH
O
H2N


HO
2
OH
3
OH
OH
Beta glucose
Alpha glucose

H2N


COOH


HOOC

1
NH2
NH2
N

2

COOH
HN
O
Glutamic Acid
'
CH3

 COOH

 CH3


H3C
NH2
4
3
Glutamine
Histidine
 COOH


H 2N
CH3
NH2
Isoleucine
COOH
5
NH2

 COOH
NH2
Lysine
Leucine

6
COOH
5
 COOH
NH2
2
4
HO
NH2
3
HO
Lactic Acid
Phenylalanine
OH
4
3
5
2
6
5


COOH
Tyrosine
6
Threonine
 H3C
O
4
NH 3
1
H2N
CH3
1
H3C
COOH

2
N
H
1
Uracil
NH2


COOH
H3C
'
O
Valine