Comparison and correlation study of polar biomarkers of CKD patients in saliva and serum by UPLC-ESI-MS

Jielian yan (Research Center of Analytical Instrumentation, Analytical & Testing Center, Sichuan University)
Li Zhou ()
Xiaohong Tang ()
Yong Liu ()
Xiaohong Tang (Department of Nephrology, West China Hospital, Sichuan University)
Yong Liu (Research Center of Analytical Instrumentation, Analytical & Testing Center, Sichuan University)
Yixiang Duan (Research Center of Analytical Instrumentation, Analytical & Testing Center, Sichuan University Research Center of Analytical Instrumentation, Key Laboratory of Bio-resource and Eco-environment, Ministry of Education, College of Life Science, Sichuan University)

Article ID: 640



There are abundant reports on the use of aromatic amino acids and creatinineas biomarkers in serum and urine forCKD(chronic kidney disease). However, investigationsinto these bio-species in bio-fluids, such as saliva and sweat, are rarely reported. Increasing interest in non-invasive methods for medical diagnosis advocates for the testing of these bio-fluidstoidentify potential biomarkers for prompt clinical andpreliminary screening using advanced analytical equipment.

Unstimulated whole saliva samples were obtained from twenty-seven CKD patients and an equivalent number of healthy individuals. Saliva was assayed with ultra-performance liquid chromatography coupled with electro-spray ionization tandem mass spectrometry (UPLC-ESI-MS) in hydrophilic interaction chromatography mode. The data were analyzed using a student’s t test and receiver operatingcharacteristic(ROC) to evaluate the predictive power of several potential biomarkers (P<0.01) in saliva for preliminary CKD screening. Through testing of salivary samples between CKD patients and healthy individuals, we found three possible salivary biomarkers that demonstrated significant differences(P<0.01) from the nine reported species in serum and/or urine. The area under the curve (AUC) values for control vs CKDpatientsfor on L-phenylalanine, L- tryptophan, and creatinine were 0.863, 0.834, and 0.916, respectively. This is the first report to compare serum and urine biomarkers in saliva between CKD patients and healthy people. This study explores the potential of CKD diagnosis by saliva, and demonstrates a positive correlation between salivary and serum creatinine.

Full Text:



[1] Reference

[2] VivekanandJha, Guillermo Garcia-Garcia, Kunitoshi Iseki, et al. Chronic kidney disease: global dimension and perspectives. Lancet.2013, 382: 260-272.

[3] Linden Heflin, Sarah Walsh, Miguel Bagajewicz. Design of medical diagnostics products: A case-study of a saliva diagnostics kit. Computers and Chemical Engineering. 2009, 33(5): 1067-1076.

[4] Yerkey MW, Kernis SJ, Franklin BA, et al. Renal dysfunction and acceleration of coronary disease. Heart.2004, 90: 961-966.

[5] Mohamad A. El Sawy, Mona M. Zaki, Ihab Z. EL-Hakim, et al. Serum amino acid abnormalities in pediatric patients with chronic renal failure with and without history of thromboembolic manifestations. Egyptian Journal of Medical Human Genetics. 2012, 13(1): 73-80.

[6] V. Faure, L. Dou, F. Sabatier, et al. Elevation of circulating endothelial microparticles in patients with chronic renal failure. Journal of Thrombosis and Haemostasis.2006, 4: 566-573.

[7] GiacomoGaribotto. Muscle amino acid metabolism and the control of muscle protein turnover in patients with chronic renal failure. Nutrition. 1999, 15(2): 145-155.

[8] Masahiro Sugimoto, Juri Saruta, ChisaMatsuki, et al. Physiological and environmental arameters associated with mass spectrometry-based salivary metabolomic profiles. Metabolomics. 2013, 9(2): 454-463.

[9] Yoshiaki Nomura, Yoh Tamaki, Akiko Eto, et al. Screening for periodontal diseases using salivary lactate dehydrogenase, hemoglobin level, and statistical modeling. Journal of Dental Sciences. 2012, 7(4): 379-383.

[10] Akira Katakura, Nobuharu Yamamoto, Takumi Sakuma, et al. A screening test for oral cancer using saliva samples: Proteomic analysis ofbiomarkers in whole saliva. Journal of Oral and Maxillofacial Surgery, Medicine, and Pathology. 2015, 27(1): 1-5.

[11] C. Streckfus, L. Bigler, W. Dubinsky. Using LC-MS/MS mass spectrometry to identify breast cancer biomarkers in saliva.Oral Surgery, Oral Medicine, Oral Pathology, Oral Radiology, and Endodontology. 2006, 102(3): 337-338.

[12] Qihui Wang, Qiaoling Yu, Qingyu Lin, et al. Emerging salivary biomarkers by mass spectrometry. ClinicaChimicaActa.2015, 438: 214-221.

[13] Jie Li, YulanPeng, YixiangDuan. Diagnosis of breast cancer based on breath analysis: An emerging method. Critical Reviews in Oncology/Hematology.2013, 87: 28-40.

[14] Makio Mogi, Toshitaka Kage, Takehiro Chino, et al. Increased β2-microglobulin in both parotid and submandibular/sublingual saliva from patients with Sjögren's syndrome. Archives of Oral Biology. 1994, 39(10): 913-915.

[15] Satoru Fukiya , Miki Arata , Hiroko Kawashima, et al. Conversion of cholic acid and chenodeoxycholic acid intotheir 7-oxo derivatives by Bacteroidesintestinalis AM-1 isolatedfrom human feces. FEMS Microbiology Letters.2009, 293: 263-270.

[16] Yingyong Zhao, Xianlong Cheng, Feng Wei, et al. Application of faecalmetabonomics on an experimental

[17] model of tubulointerstitial fibrosis by ultra performance liquid chromatography/high-sensitivity mass spectrometry with MSE data collection technique. Biomarkers. 2012, 17(8): 721-729.

[18] David O Mcgregor, Warwick J Dellow, Richard A Robson, et al. Betaine supplementation decreases post-methionine hyperhomocysteinemia in chronic renal failure. Kidney International.2002, 61: 1040-1046.

[19] Henricus A. M. Mutsaers, Udo F. H. Engelke, Martijn J. G. Wilmer, et al. Optimized Metabolomic Approach to Identify Uremic Solutes in Plasma of Stage 3–4 Chronic Kidney Disease Patients. Plos one. 2013, 8(8): e71199.

[20] LewenJia, Jing Chen, Peiyuan Yin et al. Serum metabonomics study of chronic renal failure by ultra performance liquid chromatography coupled with Q-TOF mass spectrometry. Metabolomics. 2008, 4(2): 183-189.

[21] Jonas Bergström, Anders Alvestrand and Peter Fürst. Plasma and muscle free amino acid in maintenance hemodialysis patients without protein malnutrition. Kidney International. 1990, 38(1): 108-114.

[22] Ewa M. Slominska, MarekSzolkiewicz, Ryszard T. Smolenski, et al. High Plasma Adenine Concentration in

[23] Chronic Renal Failure and Its Relation to Erythrocyte ATP.Nephron.2002, 91: 286-291.

[24] Kaori Hayashi, Hiroyuki Sasamura, TakakoHishiki, et al. Use of serum and urine metabolome analysis for the detection of metabolic changes in patients with stage 1-2 chronic kidney disease. Nephro-Urology Monthly. 2011, 3(3): 164-171.

[25] Marian Wang, Irma, Vyhmeister, Marian E. Swendseid, et al. Phenylalanine hydroxylase and tyrosine aminotransferase activities in chronically uremic rats. The Journal of Nutrition. 1975, 105(1): 122-127.

[26] Anna Tankiewicz, DariuszPawlak, Joanna Topczewska-Bruns, et al. Kidney and liver kynurenine pathway enzymes in chronic renal failure. Advances in experimental medicine and biology.2003, 527: 409-14.

[27] Vishal Diwan, David Small, Kate Kauter, et al. Gender differences in adenine-induced chronic kidney disease and cardiovascular complications in rats. American Journal of Physiology. 2014, 307(11): 1169-1178.

[28] Jennifer A. Larsen, Elizabeth M. Parks, Cailin R. Heinze, et al. Evaluation of recipes for home-​prepared diets for dogs and cats with chronic kidney disease. Journal of the American Veterinary Medical Association. 2012, 240(5): 532-538.

[29] Yingyongzhao. Metabolomics in chronic kidney disease.Clinica Chimica Acta.2013, 422: 59-69.

[30] Tessari P, Deferrari G, Robaudo C, et al. Phenylalanine hydroxylation across the kidney in humans Rapid Communication. Kidney International. 1999, 56(6): 2168-2172.

[31] Niels Moller, Shon Meek, Maureen Bigelow, et al. The kidney is an important site for in vivo phenylalanine-to-tyrosine conversion in adult humans:a metabolic role of the kidney. Proceedings of the National Academy of Sciences. 2000, 97(3): 1242-1246.

[32] GiacomoGaribotto, GiacomoDeferrari, Cristina Robaudo, et al. Effects of a protein meal on blood amino acid profile in patients with chronic renal failure. Nephron.1993, 64: 216-225.

[33] Mackenzie Walser, Sylvia B Hill. Free and protein-bound tryptophan in serum of untreated patients with chronic renal failure.Kidney International.1993, 44: 1366-1371.

[34] D. Pawlak, A. Tankiewicz, P. Myslwiec, et al. Tryptophan metabolism via the kynurenine pathway in experi-mental chronic renal failure. Nephron.2002, 90: 328-335.

[35] Hirschberg P, Kopple JD. Protein calories and fat in the predialysis patients. In: Mitch WE, Klahr S, eds. Nutrition and the kidney. Boston: Little, Brown. 1988: 131.

[36] Mohamed E Suliman, Björn Anderstam, Jonas Bergström. Evidence of taurinedeple-tion and accumulation of cysteinesulfinic acid in chronic dialysis patients.Kidney International.1996, 50: 1713-1717.

[37] William M. Pardridge, Elliot M. Landaw. Tracer kinetic model of blood-brain barrier transport of plasma protein-boundligands.The Journal of Clinical Investigation. 1984, 74(3): 745-752.

[38] Tizianello A, Deferrari G, Garibotto G, et al. Abnormalities in amino acid metabolism in chronic renal failure. Contribtuion to Nephrol.1990, 81: 169-180.

[39] Manuel Hidalgo. Pancreatic Cancer.New England Journal of Medicine.2010, 362: 1605-1617.

[40] Jie Wei, GuoxiangXie, Zengtong Zhou, et al. Salivary metabolite signatures of oral cancer and leukoplakia .International Journal of Cancer. 2011, 129(9): 2207-2217.

[41] W.A. HuubWaterval, Jean L.J.M. Scheijen, Marjon M.J.C. Ortmans-Ploemen, et al. Quantitative UPLC-MS/MS analysis of underivatised amino acids in body fluids is a reliable tool for the diagnosis and follow-up of patients with inborn errors of metabolism. ClinicaChimicaActa.2009, 407: 36-42.

[42] Keitaro Yokoyama, Masamich Tajima, Hiraku, et al. Plasma pteridine concentrations in patients with chronic renal failure. Nephrology Dialysis Transplantion. 2002, 17(6): 1032-1036.

[43] Ramesh Venkatapathy, VasupradhaGovindarajan, NirimaOza, et al. Salivary creatinine estimation as an alternative to serum creatinine in chronic kidney disease patients. International Journal of Nephrology. 2014, 2014: 1-6.

[44] A. C. Guyton and J. E. Hall, “The body fluids and kidneys,” in Textbook of Medical Physiology. Elsevier Saunders. 2006, 11: 291-415.

[45] Ivanovski K, Naumovski V, Kostadinova M, et al. Xerostomia and salivary levels of glucose and urea in patients with diabetes. Prilozi. 2012, 33(2): 219-229.

[46] T. Nakahari,H. Yoshida, Y. Imai. Transepithelial fluid shift generated by osmolarity gradients in unstimulated perfused rat submandibular glands.Experimental Physiology. 1996, 81(5): 767-779.

[47] Yuncheng Xia, CanhuiPeng, Zhifang Zhou, et al. Clinical significance of saliva urea, creatinine, and uric acid levels in patients with chronic kidney disease. Journal of Central South University.Medical sciences. 2012, 37(11): 1171-1176.



  • There are currently no refbacks.
Copyright © 2019 Jielian yan, Yixiang Duan, Li Zhou, Xiaohong Tang, Yong Liu

Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.