Supplementary Information
Cholesterol and its esters as serum biomarkers in malignant Obstructive Jaundice: A single step 1H NMR metabolomic approach
Authors: Shatakshi Srivastava1 and Raja Roy1
Santosh Kumar2, Hari Om Gupta2, Jitendra Kumar Kushwaha2, Abhijit Chandra3, Abhinav Arun Sonkar2
Authors affiliations:
- Centre of Biomedical Magnetic Resonance, SGPGIMS Campus, Raibarely Road, Lucknow, Uttar Pradesh, India
- Departments of General Surgery, King George’s Medical University, Lucknow, India.
- Departments of Surgical Gastroentology, King George’s Medical University, Lucknow, India.
*Correspondence to:
1:Professor Raja Roy
Centre of Biomedical Magnetic Resonance
Sanjay Gandhi Postgraduate Institute of Medical Sciences Campus
Raibarely Road, Lucknow-226014 (INDIA)
Email:
Contact: +91-522-2668215 Ext: 5032
Fax: +91-522-2668215
2:Prof. Abhinav A. Sonkar
Department of General Surgery
King George’s Medical University
Lucknow 226001, Uttar Pradesh, India
E-mail:
Phone No.: +91-9839017609
Running title:Proton NMR Spectroscopy of Malignant Obstructive Jaundice
Supplementary FigureS1:Comparison of serum spectra obtained after Folch method of lipid extraction and after DMSO method. The expansions of region 0.6 to 1.0 ppm are provided for depicting the changes in Cholesterol and Cholesterol ester peaks under control and benign conditions. (a) and (b) are lipid extract and DMSO dissolved serum spectra of same benign case and ( c) lipid extract (* denotes EtOH as impurity) and (d) DMSO dissolved serum spectra of same control case.
Supplementary FigureS2: The assignments of the metabolites in bile dissolved in DMSO are labeled according to the previous available literature.
Supplementary Figure S3:Expansion and overlay of 1H- 13C HSQC NMR spectrum (From 0.6 to 2.4 ppm) of control DMSO dissolved serum on spectrum of standard cholesterol dissolved in DMSO. Two dimensional 1H- 13C HSQC spectrum with the assignment of the cholesterol and cholesterol esters. The red spectra = standard cholesterol spectrum and black spectrum = control serum dissolved in DMSO.
Supplementary Figure S4: Expansion and overlay of 1H- 13C HSQC NMR spectrum (From 0.6 to 2.4 ppm) of control DMSO dissolved serum on spectrum of standard cholesterol dissolved in DMSO. Two dimensional 1H- 13C HSQC spectrum with the assignment of the cholesterol and cholesterol esters. The red spectra = control serum dissolved in DMSO and black spectrum = malignant serum dissolved in DMSO.
Supplementary Table S1: Comparisons of NMR parameters among three groups viz. healthy control, benign OBJ and malignant OBJ, where * =p<0.05, ** = p<0.005, *** = p<0.001.
S. No. / Metabolites / Control (n=57) vs. Benign (n=28) / Control (n=57) vs. Malignant (n=36) / Benign (n=28) vs. Malignant (n=36)1. / Isobutyrate / p = 0.03* / p = 0.004** / p = 0.08
2. / Alanine / p = 0.004** / p = 0.000*** / p = 0.08
3. / Acetone / p = 0.004** / p = 0.000*** / p = 0.06
4. / Glutamine / p = 0.005* / p = 0.000*** / p = 0.04*
5. / Creatine / p = 0.06 / p = 0.06 / p = 0.07
6. / Lactate / p = 0.06 / p = 0.06 / p = 0.06
7. / Threonine / p = 0.002** / p = 0.006* / p = 0.05
8. / 1-methylhistidine / p = 0.04* / p = 0.004** / p = 0.06
Supplementary Table S2: The comparisons of ratios of serum Chol/tChol, CE/tChol and Chol/CE between different groups of benign, malignant and healthy control, where, * = p<0.05, ** = p<0.005, ***= p<0.001.
S. No. / Metabolic ratios / Control (n=57) vs. Benign (n=28) / Control (n=57) vs. Malignant (n=36) / Benign (n=28) vs. Malignant (n=36)1. / Chol/tChol / p = 0.07 / p = 0.004** / p = 0.04*
2. / CE/tChol / p = 0.06 / p = 0.004** / p = 0.04*
3. / Chol/CE / p = 0.06 / p = 0.003** / p = 0.003**
Supplementary Table S3: Comparison of NMR parameters (concentration of chief biliary constituents) in patients of benign OBJ and malignant OBJ. * = p<0.05, ** = p<0.005, *** = p<0.001.
S. No. / Metabolites / Malignant (n=36) vs. Benign (n=28)1. / Total bile acids (BA) / p = 0.004**
2. / Cholesterol / p = 0.003**
3. / Phosphatidylcholine / p = 0.00*
4. / Glycerophosphatidylcholine / p = 0.01*