Chemical Analysis of Gallstones of Nepali Patients

Page: [57 - 66] Pages: 10

  • * (Excluding Mailing and Handling)

Abstract

Background: The bile concretions formed inside the gallbladder are called gallstones. Gallstone disease is one of the major causes of morbidity, mortality, and economic burden throughout the world. Approximately 10% to 20% of the world population and 4.87% in Nepal are suffering from a gallstone.

Objective: The aim is to classify the gallstone samples based on morphological and cross-sectional evidences, and a comparative study of their chemical composition to understand the mechanism of formation.

Methods: Two types of gallstones were studied; combined cholesterol gallstone and black pigment gallstone. SEM, UV-Vis, and EDS analyzed the gallstones for morphological and elemental study.

Results: The UV-Vis spectrum showed characteristic peaks for cholesterol, calcium bilirubinate, and copper bilirubinate. SEM image of combined cholesterol gallstone reveals that cholesterol is the major phase in the shell, whereas bilirubin is the major phase in the core. EDS detection of C and O as the major elements confirmed cholesterol, whereas the detection of C, O, N, and Ca as the major elements confirmed bilirubin. In the shell of the black pigment gallstone, cholesterol, calcium carbonate, calcium phosphate, and bilirubinate were detected, whereas, in the core, bilirubinate of calcium and copper, cholesterol, protein, and calcium carbonate were detected.

Conclusion: Cholesterol and bilirubin are the major components of the gallstones. Metals like Al, Ca and Cu play a crucial role in the initial formation of the gallstones. The combined cholesterol gallstone has a yellow pigment center composed mainly of unconjugated bilirubin and an outer shell primarily composed of cholesterol and calcium carbonate. Precipitation of bilirubinate of calcium and copper is important during the formation of black PGS.

Graphical Abstract

[1]
Johnston DE, Kaplan MM. Pathogenesis and treatment of gallstones. N Engl J Med 1993; 328(6): 412-21.
[http://dx.doi.org/10.1056/NEJM199302113280608] [PMID: 8421460]
[2]
Bateson MC. Fortnightly review: Gallbladder disease. BMJ 1999; 318(7200): 1745-8.
[http://dx.doi.org/10.1136/bmj.318.7200.1745] [PMID: 10381713]
[3]
Yoo EH, Lee SY. The prevalence and risk factors for gallstone disease. Clin Chem Lab Med 2009; 47(7): 795-807.
[http://dx.doi.org/10.1515/CCLM.2009.194] [PMID: 19499973]
[4]
Harish B. A cross sectional study on causes and risk factors of gallstone disease among patients with symptomatic cholilithiasis. Int J Nurs Res Pract 2014; 1(1): 20.
[5]
Jaisawal RK, Mishra C, Panthee MR, Pathak YR, Acharya AP. Prevalence of gall stone disease in Nepal: Multi center ultrasonographic study. Post-Grad Med J NAMS 2007; 7(2)
[6]
Kameda A. The history of gall stone. Nippon Naika Gakkai Zasshi 2002; 91: 105-9.
[7]
Takagi I, Toda G. Definition, classification and clinical symptoms of cholelithiasis. Jpn J Clin Med 1993; 51(7): 1705-10.
[PMID: 8366583]
[8]
Jarrar BM, Al-Rowaili MA. Chemical composition of gallstones from Al-jouf province of Saudi Arabia. Malays J Med Sci 2011; 18(2): 47-52.
[PMID: 22135586]
[9]
Peter A, Cozmuta LM, Nicula C, et al. Multi-analyses of gallstones and correlation between their properties with the laboratory results. Anal Biochem 2020; 593113587
[http://dx.doi.org/10.1016/j.ab.2020.113587] [PMID: 31978457]
[10]
Mohan H. Textbook of Pathology. 5th ed. New Delhi: Jaypee Brothers Medical Publishers (P) Ltd. 2005.
[http://dx.doi.org/10.5005/jp/books/10939]
[11]
Kim IS, Myung SJ, Lee SS, Lee SK, Kim MH. Classification and nomenclature of gallstones revisited. Yonsei Med J 2003; 44(4): 561-70.
[http://dx.doi.org/10.3349/ymj.2003.44.4.561] [PMID: 12950109]
[12]
Gupta U, Singh V, Kumar V, Khajuria Y. Spectroscopic studies of cholesterol: Fourier transform infra-red and vibrational frequency analysis. Materials Focus 2014; 3(3): 211-7.
[http://dx.doi.org/10.1166/mat.2014.1161]
[13]
Jaswal BBS, Kumar V, Swart HC, Sharma J, Rai PK, Singh VK. Multi-spectroscopic analysis of cholesterol gallstone using TOF-SIMS, FTIR and UV-Vis spectroscopy. Appl Phys B 2015; 121(1): 49-56.
[http://dx.doi.org/10.1007/s00340-015-6200-3]
[14]
Chekroune M, Benamara S. Gallstones-dissolving capacity of lemon (Citrus limon) juice, Herniaria hirsuta L. extract and lemon juice-based natural vinaigrette in vitro. Indian J Tradition Knowledge 2017; 16(2): 197-202.
[15]
Das I, Singh Y, Ansari SA, Agrawal NR. In vitro dissolution of gallbladder stone by edible leaves, fruits and homoeopathic medicines. J Cryst Growth 2005; 283(3-4): 508-13.
[http://dx.doi.org/10.1016/j.jcrysgro.2005.06.006]
[16]
Rai AK, Rai SB, Rai DK, Singh VB. Spectroscopic studies and normal coordinate analysis of bilirubin. Spectrochim Acta A Mol Biomol Spectrosc 2002; 58(10): 2145-52.
[http://dx.doi.org/10.1016/S1386-1425(01)00685-0] [PMID: 12212739]
[17]
Suzuki N. On bilirubin-metal complex compounds in relation to black pigments of gallstones. Tohoku J Exp Med 1966; 90(2): 195-205.
[http://dx.doi.org/10.1620/tjem.90.195] [PMID: 5971087]
[18]
Cavalu S, Popa A, Bratu I, Borodi G, Maghiar A. New evidences of key factors involved in “silent stones” etiopathogenesis and trace elements: Microscopic, spectroscopic, and biochemical approach. Biol Trace Elem Res 2015; 168(2): 311-20.
[http://dx.doi.org/10.1007/s12011-015-0361-0] [PMID: 26006095]
[19]
Malet PF, Williamson CE, Trotman BW, Soloway RD. Composition of pigmented centers of cholesterol gallstones. Hepatology 1986; 6(3): 477-81.
[http://dx.doi.org/10.1002/hep.1840060326] [PMID: 3710436]
[20]
Qiao T, Ma RH, Luo XB, Luo ZL, Zheng PM, Yang LQ. A microstructural study of gallbladder stones using scanning electron microscopy. Microsc Res Tech 2013; 76(5): 443-52.
[http://dx.doi.org/10.1002/jemt.22185] [PMID: 23401308]
[21]
Sharma R, Soy S, Kumar C, Sachan SG, Sharma SR. Analysis of gallstone composition and structure in Jharkhand region. Indian J Gastroenterol 2015; 34(1): 29-37.
[http://dx.doi.org/10.1007/s12664-014-0523-6] [PMID: 25586075]
[22]
Weerakoon H, Navaratne A, Ranasinghe S, Sivakanesan R, Galketiya KB, Rosairo S. Chemical characterization of gallstones: An approach to explore the aetiopathogenesis of gallstone disease in Sri Lanka. PLoS One 2015; 10(4)e0121537
[http://dx.doi.org/10.1371/journal.pone.0121537] [PMID: 25853583]
[23]
Jayasoma K, Koralegedara NH, Dharmapala A, Chandrajith R. Microstructural and geochemical characterization of gallstones: Implication for biomineralization. Biol Trace Elem Res 2022; 200(12): 4891-902.
[http://dx.doi.org/10.1007/s12011-021-03076-4] [PMID: 35089502]
[24]
Qiao T, Ma R, Luo X, Yang L, Luo Z, Zheng P. The systematic classification of gallbladder stones. PLoS One 2013; 8(10)e74887
[http://dx.doi.org/10.1371/journal.pone.0074887] [PMID: 24124459]
[25]
Li WH, Shen GR, Soloway RD, et al. Copper bilirubinate and black pigment gallstone. Biospectroscopy 1995; 1(2): 149-56.
[http://dx.doi.org/10.1002/bspy.350010209] [PMID: 7696561]
[26]
Suzuki N, Nakamura Y, Kobayashi N, Sato T. On metal elements in pure pigment gallstones. Tohoku J Exp Med 1975; 116(3): 233-40.
[http://dx.doi.org/10.1620/tjem.116.233] [PMID: 1179420]
[27]
Mondal B, Maulik D, Mandal M, Sarkar GN, Sengupta S, Ghosh D. Analysis of carcinogenic heavy metals in gallstones and its role in gallbladder carcinogenesis. J Gastrointest Cancer 2017; 48(4): 361-8.
[http://dx.doi.org/10.1007/s12029-016-9898-1] [PMID: 27888389]
[28]
Athanasiadou D, Godelitsas A, Sokaras D, et al. New insights into the chemical and isotopic composition of human-body biominerals. I: Cholesterol gallstones from England and Greece. J Trace Elem Med Biol 2013; 27(2): 79-84.
[http://dx.doi.org/10.1016/j.jtemb.2012.08.004] [PMID: 23117070]
[29]
Shen Y, Xie A, Huang L, Huang F, Chen Z, Ma D. Biomimetic synthesis of calcium bilirubinate in different inverse microemulsions. Synth React Inorg Met-Org Nano-Met Chem 2005; 35(5): 359-64.
[http://dx.doi.org/10.1081/SIM-200059199]