**Biomedicine and Environment: The Future is Now?**

334 Stoichiometry and Research – The Importance of Quantity in Biomedicine

Kellner, R. A. (1998). UV-Vis Spectrophotometry, Emission and Luminescence, In: *Analytical* 

Martell, A. E. & Motekaitis, R. J. (1992). *Determination and Use of Stability Constants*, John

McBride, L. (1980). *The Iron Reagents*, 3rd edition, G. Frederick Smith Chemical Co.,

McBryde, W. A. E. (1964). A Spectrophotometric Reexamination of the Spectra and

Miller, J. C. & Miller, J. N. (1993). *Statistics for Analytical Chemistry*, 3rd edition, Ellis Horwood Limited, ISBN 0-13-030990-7, Chichester, West Sussex, United Kingdom Riemer, J.; Hoepken, H. H.; Czerwinska, H.; Robinson, S. R. & Dringen, R. (2004).

Skoog, D. A.; Holler, F. J. & Crouch, S. R. (2007). *Principles of Instrumental Analysis*, 6th edition, Cengage Learning, ISBN 9780495012016, Florence, Kentucky, USA Smith, F. E.; Herbert, J.; Gaudin, J.; Hennessy, D. J & Reid, G. R. (1984). Serum Iron

Stauffer, M. T. (2007). Undergraduate Analytical Chemistry: To Use and Evaluate Organic

Stauffer, M. T.; Hunter, L. J. & Troncone, S. K. (2007). Determination of Iron in Abandoned

Stookey, L. L. (1970). Ferrozine – A New Spectrophotometric Reagent for Iron. *Analytical* 

Thomas, M. (1996). *Ultraviolet and Visible Spectroscopy*, 2nd edition, John Wiley & Sons, Ltd.,

Thompsen, J. C. & Mottola, H. A. (1984). Kinetics of the Complexation of Iron(II) with

Viollier, E.; Inglett, P. W.; Hunter, K.; Roychoudhury, A. N. & Van Cappellen, P. (2000). The

Yamamoto, M.; Nishida, A.; Otsuka, K.; Komai, T. & Fukushima, M. (2010). Evaluation of

*Geochemistry*, Vol. 15, No. 6, (July 2000), pp. 785-790, ISSN 0883-2927 Vosburgh, W. C. & Cooper, G. R. (1941). Complex Ions I. The Identification of Complex Ions

*Society*, Vol. 63, No. 2, (February 1941), pp. 437-442, ISSN 0002-7863

2010), pp. 4456-4460, ISSN 0960-8524

Ferrozine. *Analytical Chemistry*, Vol. 56, No. 4, (April 1984), pp. 755-757, ISSN 0003-

ferrozine method revisited: Fe(II)/Fe(III) determination in natural waters. *Applied* 

in Solution by Spectrophotometric Measurements. *Journal of the American Chemical* 

the binding of iron(II) to humic substances derived from a compost sample by a colorimetric method using ferrozine. *Bioresource Technology*, Vol. 101, No. 12, (June

*Chemistry*, Vol. 42, No. 7, (June 1970), pp. 779-781, ISSN 0003-2700

ISBN 0-471-96743-2, West Sussex, England, United Kingdom

Wiley & Sons, ISBN 978-0-471-18817-9, New York, New York, USA

540, Wiley-VCH, ISBN 3-527-28610-1, Weinheim, Germany

No. 8, (August 1964), pp. 1917-1927, ISSN 0008-4042

40, No. 3, (May-June 2007), pp. 439-452, ISSN 0038-7010

Columbus, Ohio, USA

New York, New York, USA

pp. 306-310, ISSN 0009-9120

437, ISSN 0038-7010

2700

*Chemistry*, Kellner, R. A.; Mermet, J.-M.; Otto, M. & Widmer, H. M., Eds., pp. 527-

Stabilities of the Iron(III)-Tiron Complexes. *Canadian Journal of Chemistry*, Vol. 42,

Colorimetric ferrozine-based assay for the quantitation of iron in cultured cells. *Analytical Biochemistry*, Vol. 331, No. 2, (August 2004), pp. 370-375, ISSN 0003-2697 Rossotti, F. C. & Rossotti, H. (1961). *The Determination of Stability Constants*, McGraw-Hill,

Determination Using Ferene Triazine. *Clinical Biochemistry*, Vol. 17, (October 1984),

Chelators for Spectrophotometric Determination of Iron. *Spectroscopy Letters*, Vol.

Mine Drainage by UV-Vis Spectrophotometry and Flame Atomic Absorption Spectrophotometry. *Spectroscopy Letters*, Vol. 40, No. 3, (May-June 2007), pp. 429-

**15** 

*Brazil* 

**Recent New Characterizations** 

Jose F.R. Bachega2 and Patrícia S. Santiago1,3

*3Campus Experimental de Registro, UNESP, SP,* 

**on the Giant Extracellular Hemoglobin** 

**of** *Glossoscolex paulistus* **and Some Other** 

*1Instituto de Química de São Carlos, Universidade de São Paulo, São Carlos, SP, 2Instituto de Física de São Carlos, Universidade de São Paulo, São Carlos, SP,* 

Giant extracellular hemoglobins, also known as erythrocruorins, have been investigated as a model of extreme complexity in oxygen-binding heme proteins [1,2]. They are characterized by a very high molecular mass (*MM*) of several megadalton (MDa), and their oligomeric structure together with the crowded and protected heme environment are two of the main factors responsible for the high redox stability [3,4]. Superoxide dimustase (SOD)-like intrinsic activity, observed for hemoglobins of *Lumbricus terrestris* (HbLt) and of *Arenicola marina* (HbAm), is another important factor [5,6]. These hemoglobins present a highly cooperative oxygen binding and a peculiar behavior associated to their oligomeric dissociation into smaller subunits and possible rearrangement back into the native

Moreover, a strong motivation to study these giant hemoglobins is related to their potential use in medicine as blood substitutes. Studies have been performed in the past for HbLt [9], and are presently underway to test and validate the use of HbAm in this direction [6,10]. They seem to be very promising due to the lack of undesirable immunological reactions in tests with animals, explained by the absence of cell membranes as occurs with human hemoglobin in red blood cells [6,10]. Besides, the resistance to oxidation of extracellular hemoglobins, as noticed by their redox stability, is also an advantage as compared to the use

The giant extracellular hemoglobin of *Glossoscolex paulistus* (HbGp) is similar to several homologous proteins described in the literature [11-14]. These extracellular hemoglobins are constituted by a large number of globin-like subunits containing heme groups with *MM* in the range 15–19 kDa. These globin subunits form monomers of 16 kDa (*d*) and disulphide bound hetero-trimers of 51-52 kDa (*abc*), linked by non-heme structures (24–32 kDa), named linkers (*L*) [12,13]. Recent partial characterization of *MM* of HbGp by matrix assisted laser

**1. Introduction** 

oligomeric structure [7,8].

of human hemoglobin in this medical application.

**Giant Hemoglobins from Different Worms** 

Marcel Tabak1, Francisco A.O. Carvalho1, José W.P. Carvalho1,
