Identification and Quantitation of Source from Hemoglobin of Blood and Blood Mixtures by High Performance Liquid Chromatography

Abstract

The described technique offers a sensitive and reproducible method for inferring the source of over 50 different animal species from bloodstains and blood mixtures. Hemoglobins from each of the species were examined using reversed-phase high performance liquid chromatography (HPLC) in chromatographic times of less than 25 mins. The HPLC method complements and furthers current methodology for identification of species of origin. HPLC analysis is particularly well suited for the quantitative analysis of blood and blood mixtures and is applicable to species for which antisera are unavailable. The sensitivity of the method (hemoglobin amounts down to 1.2 µg) lends itself to the analysis of blood mixtures in which only a small percentage of the mixture represents blood from a given species. Such resolution and quantitation is applicable to wildlife forensic casework.

Meat species identification based on the loop mediated isothermal amplification and electrochemical DNA sensor

“…High-performance liquid chromatography (Espinoza, Kirms, & Filipek, 1996;Toorop, Murch, & Ball, 1997), electrophoretic techniques (Ozgen Arun & Ugur, 2000;Renon, Bernardi, Scocca, Cantoni, & Gridavilla, 2003), and enzyme-linked immunosorbent assays (Hajmeer, Cliver, & Provost, 2003;Macedo-Silva et al, 2000) have also been reported. Although most of these protein based methods identified the species-origin of raw meats, they were significantly less sensitive in heat-treated processed meats because of the thermal alteration of the specific epitopes in meat proteins (Hird, Goodier, & Hill, 2003).…”

A TaqMan real-time PCR system for the identification and quantification of bovine DNA in meats, milks and cheeses

“…Lipid analysis is only applicable for gross measurement of animal-derived fats (Lumley, 1996;Saeed, Ali, Abdul Rahman, & Sawaya, 1989). Protein-based methods such as high performance liquid chromatography (HPLC) (Espinoza, Kirms, & Filipek, 1996), enzyme-linked immunosorbent assays (ELISA) (Chen & Hsieh, 2000) or isoelectric focusing protein profiles (Skarpeid, Kvaal, & Hildrum, 1998) are effective mainly for unprocessed food and are unable to differentiate species such as lamb and goat or chicken and turkey. Both require complicated procedures and it has proved difficult to accurately quantify the analytes in a short time (Mayer, 2005).…”

Electrospray Ionization Mass Spectrometric Analysis of Blood for Differentiation of Species

“…The ␣/␤-pair centered at 15112/ 16009 is the coincidental ␣/␤-pair marker for all of these animals. However, HPLC analysis of blood samples from bear, elk, and horse would indicate a difference between the primary structures of the genera's ␣-chains and those of the genera's ␤-chains based upon differing retention times (9). Consequently, HPLC analysis of blood samples from overlapping bear, elk, or horse samples prior to the mass spectrometric analysis should differentiate these species.…”