Fiber enhanced Raman spectroscopic analysis as a novel method for diagnosis and monitoring of diseases related to hyperbilirubinemia and hyperbiliverdinemia.

Fiber enhanced resonance Raman spectroscopy (FERS) is introduced for chemically selective and ultrasensitive analysis of the biomolecules hematin, hemoglobin, biliverdin, and bilirubin. The abilities for analyzing whole intact, oxygenated erythrocytes are proven, demonstrating the potential for the diagnosis of red blood cell related diseases, such as different types of anemia and hemolytic disorders. The optical fiber enables an efficient light-guiding within a miniaturized sample volume of only a few micro-liters and provides a tremendously improved analytical sensitivity (LODs of 0.5 μM for bilirubin and 0.13 μM for biliverdin with proposed improvements down to the pico-molar range). FERS is a less invasive method than the standard ones and could be a new analytical method for monitoring neonatal jaundice, allowing a precise control of the unconjugated serum bilirubin levels, and therefore, providing a better prognosis for newborns. The potential for sensing very low concentrations of the bile pigments may also open up new opportunities for cancer research. The abilities of FERS as a diagnostic tool are explored for the elucidation of jaundice with different etiologies including the rare, not yet well understood diseases manifested in green jaundice. This is demonstrated by quantifying clinically relevant concentrations of bilirubin and biliverdin simultaneously in the micro-molar range: for the case of hyperbilirubinemia due to malignancy, infectious hepatitis, cirrhosis or stenosis of the common bile duct (1 μM biliverdin together with 50 μM bilirubin) and for hyperbiliverdinemia (25 μM biliverdin and 75 μM bilirubin). FERS has high potential as an ultrasensitive analytical technique for a wide range of biomolecules and in various life-science applications.

[1]  Zuo-wei Li,et al.  Raman sensitivity enhancement for aqueous absorbing sample using Teflon-AF 2400 liquid core optical fibre cell. , 2007, Analytica chimica acta.

[2]  Don McNaughton,et al.  Resonance Raman spectroscopy reveals new insight into the electronic structure of beta-hematin and malaria pigment. , 2004, Journal of the American Chemical Society.

[3]  M. Perutz,et al.  Structure of Hæmoglobin: A Three-Dimensional Fourier Synthesis at 5.5-Å. Resolution, Obtained by X-Ray Analysis , 1960, Nature.

[4]  L. Ståhle,et al.  A novel mutation in the biliverdin reductase‐A gene combined with liver cirrhosis results in hyperbiliverdinaemia (green jaundice) , 2009, Liver international : official journal of the International Association for the Study of the Liver.

[5]  D. Houlihan,et al.  Investigation of jaundice , 2011, Medicine.

[6]  M. Pelletier,et al.  Raman sensitivity enhancement for aqueous protein samples using a liquid-core optical-fiber cell. , 2001, Analytical chemistry.

[7]  Jürgen Popp,et al.  Rapid monitoring of intermediate states and mass balance of nitrogen during denitrification by means of cavity enhanced Raman multi-gas sensing. , 2015, Analytica chimica acta.

[8]  N. Nemoto,et al.  Inhibition of benzo[a]pyrene-induced mutagenesis in Chinese hamster V79 cells by hemin and related compounds. , 1983, Mutation research.

[9]  Waveguide Capillary Cell for Low-Refractive-Index Liquids , 1997 .

[10]  B. Davis,et al.  Hyperbiliverdinemia in the bronze baby syndrome. , 1987, Journal of the American Academy of Dermatology.

[11]  G. T. Evans,et al.  A study of the serum biliverdin concentration in various types of jaundice. , 1947, The Journal of laboratory and clinical medicine.

[12]  Stefan W. Ryter,et al.  Bile Pigments in Pulmonary and Vascular Disease , 2012, Front. Pharmacol..

[13]  M. Perutz,et al.  Structure of haemoglobin: a three-dimensional Fourier synthesis at 5.5-A. resolution, obtained by X-ray analysis. , 1960, Nature.

[14]  R. Suzuki,et al.  Bilirubin exhibits a novel anti-cancer effect on human adenocarcinoma. , 2006, Biochemical and biophysical research communications.

[15]  J. Pople,et al.  Self‐Consistent Molecular‐Orbital Methods. I. Use of Gaussian Expansions of Slater‐Type Atomic Orbitals , 1969 .

[16]  C. Hammerman,et al.  Understanding severe hyperbilirubinemia and preventing kernicterus: adjuncts in the interpretation of neonatal serum bilirubin. , 2005, Clinica chimica acta; international journal of clinical chemistry.

[17]  Mi-Sook Won,et al.  An amperometric bilirubin biosensor based on a conductive poly-terthiophene-Mn(II) complex. , 2008, Biosensors & bioelectronics.

[18]  Bayden R. Wood,et al.  Raman excitation wavelength investigation of single red blood cells in vivo , 2002 .

[19]  J. Ostrow,et al.  Bilirubin chemistry and metabolism; harmful and protective aspects. , 2009, Current pharmaceutical design.

[20]  Robert I. Altkorn,et al.  Raman Performance Characteristics of Teflon®-AF 2400 Liquid-Core Optical-Fiber Sample Cells , 1999 .

[21]  Jens Kobelke,et al.  Origins of modal loss of antiresonant hollow-core optical fibers in the ultraviolet. , 2015, Optics express.

[22]  S. Asher.  UV resonance Raman studies of molecular structure and dynamics: applications in physical and biophysical chemistry. , 1988, Annual review of physical chemistry.

[23]  L. Burgess,et al.  A Raman waveguide detector for liquid chromatography. , 1999, Analytical chemistry.

[24]  Jürgen Popp,et al.  In situ localization and structural analysis of the malaria pigment hemozoin. , 2007, The journal of physical chemistry. B.

[25]  Andrew J Berger,et al.  Quantitative concentration measurements of creatinine dissolved in water and urine using Raman spectroscopy and a liquid core optical fiber. , 2005, Journal of biomedical optics.

[26]  M. Schmitt,et al.  Device for Raman difference spectroscopy. , 2007, Analytical chemistry.

[27]  J. Prichard.  Biliverdin Appearing in a Case of Malnutrition , 1972, The British journal of clinical practice.

[28]  Richard P. Van Duyne,et al.  Intensity Considerations in Liquid Core Optical Fiber Raman Spectroscopy , 2001 .

[29]  J. J. Lauff,et al.  Separation of bilirubin species in serum and bile by high-performance reversed-phase liquid chromatography. , 1981, Journal of chromatography.

[30]  T. Dunning,et al.  A Road Map for the Calculation of Molecular Binding Energies , 2000 .

[31]  Jürgen Popp,et al.  Fiber array based hyperspectral Raman imaging for chemical selective analysis of malaria-infected red blood cells. , 2015, Analytica chimica acta.

[32]  Robert F. Hout,et al.  Molecular orbital studies of vibrational frequencies , 2009 .

[33]  Jürgen Popp,et al.  Enhanced Raman multigas sensing - a novel tool for control and analysis of (13)CO(2) labeling experiments in environmental research. , 2014, The Analyst.

[34]  T. Spiro.  Resonance Raman spectroscopy. New structure probe for biological chromophores , 1974 .

[35]  A. Becke.  Density-functional thermochemistry. II: The effect of the Perdew-Wang generalized-gradient correlation correction , 1992 .

[36]  G. Babcock,et al.  Unconjugated bilirubin induces apoptosis in colon cancer cells by triggering mitochondrial depolarization , 2004, International journal of cancer.

[37]  M. Frisch,et al.  Ab Initio Calculation of Vibrational Absorption and Circular Dichroism Spectra Using Density Functional Force Fields , 1994 .

[38]  Tse-Chuan Chou,et al.  A portable potentiostat for the bilirubin-specific sensor prepared from molecular imprinting. , 2007, Biosensors & bioelectronics.

[39]  Jens Kobelke,et al.  Double antiresonant hollow core fiber--guidance in the deep ultraviolet by modified tunneling leaky modes. , 2014, Optics express.

[40]  Jürgen Popp,et al.  Structural analysis of the antimalarial drug halofantrine by means of Raman spectroscopy and density functional theory calculations. , 2010, Journal of biomedical optics.

[41]  E. Sasse,et al.  Standardization in bilirubin assays: evaluation of selected methods and stability of bilirubin solutions. , 1973, Clinical chemistry.

[42]  A. Bulmer,et al.  The anti-mutagenic properties of bile pigments. , 2008, Mutation research.

[43]  Alison G. Smith,et al.  Heme, Chlorophyll, and Bilins , 2001, Humana Press.

[44]  M. Schmitt,et al.  Raman spectroscopic investigation of the antimalarial agent mefloquine , 2007, Analytical and bioanalytical chemistry.

[45]  K. Walters,et al.  Serum bilirubin and risk of respiratory disease and death. , 2011, JAMA.

[46]  Jürgen Popp,et al.  Morphology-sensitive Raman modes of the malaria pigment hemozoin. , 2009, The Analyst.

[47]  S. Strom,et al.  Treatment of the Crigler-Najjar syndrome type I with hepatocyte transplantation. , 1998, The New England journal of medicine.

[48]  M. Schmitt,et al.  In vitro polarization‐resolved resonance Raman studies of the interaction of hematin with the antimalarial drug chloroquine , 2004 .

[49]  Li Zhang.  Heme Biology: The Secret Life of Heme in Regulating Diverse Biological Processes , 2011 .

[50]  G. Kikuchi,et al.  Heme oxygenase and heme degradation. , 2003, Biochemical and biophysical research communications.

[51]  I. Kuntz,et al.  Bile pigments as HIV-1 protease inhibitors and their effects on HIV-1 viral maturation and infectivity in vitro. , 1996, The Biochemical journal.

[52]  J. Kirk.  Neonatal jaundice: a critical review of the role and practice of bilirubin analysis , 2008, Annals of clinical biochemistry.

[53]  T. H. Dunning.  Gaussian basis sets for use in correlated molecular calculations. I. The atoms boron through neon and hydrogen , 1989 .

[54]  J. Popp,et al.  Relationship between molecular structure and Raman spectra of quinolines , 2009 .

[55]  M. Schmitt,et al.  Ultrasensitive in situ tracing of the alkaloid dioncophylline A in the tropical liana Triphyophyllum peltatum by applying deep-UV resonance Raman microscopy. , 2007, Analytical chemistry.

[56]  Don McNaughton,et al.  Raman microspectroscopy and imaging provides insights into heme aggregation and denaturation within human erythrocytes. , 2005, Journal of biomedical optics.

[57]  Jin‐Ming Lin,et al.  Determination of total bilirubin in human serum by chemiluminescence from the reaction of bilirubin and peroxynitrite. , 2004, Talanta.

[58]  M. Schmitt,et al.  FT‐Raman and NIR‐SERS characterization of the antimalarial drugs chloroquine and mefloquine and their interaction with hematin , 2006 .

[59]  Jürgen Popp,et al.  Ultrasensitive fiber enhanced UV resonance Raman sensing of drugs. , 2013, Analytical chemistry.

[60]  Takashi Ohrui,et al.  Transient relief of asthma symptoms during jaundice: a possible beneficial role of bilirubin. , 2003, The Tohoku journal of experimental medicine.

[61]  Parr,et al.  Development of the Colle-Salvetti correlation-energy formula into a functional of the electron density. , 1988, Physical review. B, Condensed matter.

[62]  M. Prezelj.  Enzymic determination of total bilirubin in serum with the BA-1000. , 1988, Clinical chemistry.

[63]  T. W. Wu,et al.  Isolation and preliminary characterization of a fraction of bilirubin in serum that is firmly bound to protein. , 1982, Clinical chemistry.

[64]  R. Tukey,et al.  Human UDP-glucuronosyltransferases: metabolism, expression, and disease. , 2000, Annual review of pharmacology and toxicology.

[65]  Prichard Js.  Biliverdin appearing in a case of malnutrition. , 1972 .

[66]  Jürgen Popp,et al.  In vivo localization and identification of the antiplasmodial alkaloid dioncophylline A in the tropical liana Triphyophyllum peltatum by a combination of fluorescence, near infrared Fourier transform Raman microscopy, and density functional theory calculations. , 2006, Biopolymers.

[67]  Jürgen Popp,et al.  Investigation of gas exchange processes in peat bog ecosystems by means of innovative Raman gas spectroscopy. , 2013, Analytical chemistry.

[68]  Andrew J Berger,et al.  Quantitative Analysis of Raman Signal Enhancement from Aqueous Samples in Liquid Core Optical Fibers , 2004, Applied spectroscopy.

[69]  H. Kesteloot,et al.  Serum bilirubin and 10-year mortality risk in a Belgian population , 2001, Cancer Causes & Control.

[70]  Zeev Rosenzweig,et al.  A fiber optic sensor for rapid analysis of bilirubin in serum , 1997 .

[71]  A. Wolkoff.  Inheritable Disorders Manifested by Conjugated Hyperbilirubinemia , 1983, Seminars in liver disease.

[72]  M. Litorja,et al.  Low-loss liquid-core optical fiber for low-refractive-index liquids: fabrication, characterization, and application in Raman spectroscopy. , 1997, Applied optics.

[73]  Tadashi Yoshida.  [Heme oxygenase and heme degradation]. , 2003, Seikagaku. The Journal of Japanese Biochemical Society.

[74]  A. Greenberg,et al.  Green jaundice , 1971, The American Journal of Digestive Diseases.

[75]  Kevin R Ward,et al.  Measurement of hemoglobin oxygen saturation using Raman microspectroscopy and 532-nm excitation. , 2008, Journal of applied physiology.

[76]  D Lindhout,et al.  The genetic basis of the reduced expression of bilirubin UDP-glucuronosyltransferase 1 in Gilbert's syndrome. , 1995, The New England journal of medicine.