RP‐HPLC method with fluorescence detection for amino acids <scp>D</scp>/<scp>L</scp> ratio determination in fossil bones as a marker of DNA preservation

Iuliani, Piera; Federico, Lucia Di; Fontecchio, G; Carlucci, Giuseppe

doi:10.1002/jssc.201000151

Cited by 6 publications

(7 citation statements)

References 35 publications

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“…Thus, it has been possible to identify situations that increase the variability between samples as this may affect the results independent of the accuracy of the technique. In another study conducted using this information, the contamination of the fossil DNA was checked . Armstrong et al.…”

Section: Discussionmentioning

confidence: 99%

Estimation of Chronological Age from Postmortem Tissues Based on Amino Acid Racemization

Eyüboğlu

Gören

Yurtsever

2018

Journal of Forensic Sciences

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Skin and cartilage tissue specimens from 32 male and 13 female corpses aged between 17 and 50 years were collected within 24 h after the death. Each specimen was analyzed for the composition of dextro (D) and levo (L) forms of aspartate, glutamate, and alanine. Linear regression models were constructed using ln [(1 + D/L)/(1 - D/L] equation to define the relationship between the extent of racemization and the chronological age. Aspartate D/L rates from cartilage showed high correlation (r = 0.779, p < 0.001, n = 45). Aspartate D/L rates from skin showed very low correlation (r = 0.356, p < 0.002, n = 44). The multilinear regression model of both aspartate D/L rates of cartilage and skin tissues in 44 cases yielded a coefficient of r = 0.828 (p < 0.001). In conclusion, only racemization rate of Aspartate both in the skin and the cartilage tissues correlated with the chronological age. Our results may imply that the age can be estimated more precisely if two different tissue specimens are obtained from one corpse.

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Section: Discussionmentioning

confidence: 99%

Estimation of Chronological Age from Postmortem Tissues Based on Amino Acid Racemization

Eyüboğlu

Gören

Yurtsever

2018

Journal of Forensic Sciences

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“…149 mg of the Cardinal's bone fragment were processed as previously discussed (11,13). In short, this RP-HPLC method is carried out by on-line pre-column derivatization with o-phthalaldehyde-N-acetyl-L-cysteine system and fluorescence detection.…”

Section: Hplc Analysis Ofd/lasp and D/lala Racemizationmentioning

confidence: 99%

“…This information is obtained on the aDNA preservation and its contamination by measuring the OIL enantiomeric ratio of aspartic acid (Asp) and alanine (Ala) using HPLC methods before performing molecular studies on ancient human remains (10). To avoid any introduction of modern DNA, we subjected the Cardinal's bone to our own RP-HPLC-FL analysis (11). We used a compact bone, considered the most suited aDNA source for genetic assays thanks to bony lacunae osteocytes protecting against the introduction of exogenous DNA (12).…”

mentioning

confidence: 99%

Immunogenetics and Hplc Analyses Contribute to Understanding the Etiopathology of Rheumatoid Arthritis through Studies on Ancient Human Remains

Poma

Carlucci

Fontecchio

2012

Int J Immunopathol Pharmacol

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Genetic investigations on ancient human remains affected by rheumatological pathologies are a research field of particular interest for identifying origins and the etiopathology of diseases, especially those having an autoimmune background such as rheumatoid arthritis (RA). We wish to demonstrate how reliable studies concerning this topic require collaboration between multiple disciplines, usually starting from paleopathologic observations up to immunogenetic screening, even involving analytical chemistry. Here, we focused our investigation on the skeleton of Cardinal Carlo de'Medici (1595-1666) for whom RA and psoriatic arthritis (PsA) were postulated after paleopathologic examination. RA susceptibility is linked to specific HLA alleles belonging to DRB1 04 locus, such as DRB1 0401, while Cw 0602 and DRB1 07 predispose to PsA. Thus, we genotyped the Cardinal?s remains to search for RA or PsA risk genes. Ancient DNA is often subjected to hydrolysis followed by fragmentation. For this reason, all immunogenetic tests were preceded by an original RP-HPLC-FL method able to inform on the ancient DNA preservation and the extent of contamination, with the purpose of avoiding the risk of false positive results. After DNA isolation from a piece of bone from the Cardinal, PCR-SSP and reverse-SSO hybridization assays were applied to perform genomic HLA-typing. RP-HPLC-FL analysis revealed a good preservation of DNA without contamination by exogenous genomes. Molecular tests assigned to the Cardinal the genotype DRB1 0401/1102 for HLA-DRB locus and Cw 04/ 12 for HLA-C locus, data that support a genetic predisposition for RA but not for PsA. This multidisciplinary study has allowed us: (i) to ascertain that the remains undoubtedly belonged to the specific subject, Cardinal Carlo de?Medici; (ii) to sustain that the subject suffered from RA rather then that PsA, and (iii) to state that RA was already widespread in Europe at the Renaissance age, despite some authors claiming that the disease was introduced to the Old Continent from America after colonization during the 18th century.

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“…Traditionally, AAs were separated and analyzed by the means of HPLC and GC . Compared with HPLC and GC, CE has some significant advantages, such as high efficiency, less sample consumption, and shorter analysis time .…”

Section: Introductionmentioning

confidence: 99%

Study of the effect of culture mediums on the amino acid metabolites for Corynebacterium glutamicum using high‐speed micellar electrokinetic chromatography

et al. 2019

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Corynebacterium glutamicum (C. glutamicum) is a well‐known workhorse for the industrial production of amino acids. Different carbon, nitrogen, and sulfur source may force the bacterium to produce specific metabolites. In this work, a method of high‐speed MEKC with LIF detection was developed to rapidly analyze the amino acid metabolites released by C. glutamicum, which is fed with different culture mediums. Corynebacterium glutamicum was cultured in microbial fuel cells to monitor its metabolism process and collect its metabolites. In the CE system, a microliter‐scale sample reservoir was designed and applied to perform tiny volume sample injection. With the assistance of microwave, the derivatization time for amino acids with FITC was greatly shortened to 6 min. Under the optimized condition, the eight candidate amino acids of metabolites could be rapidly separated within 2 min. The whole analysis process for real samples, from sampling to determination, could be shortened to less than 10 min. The results showed that C. glutamicum could produce additional l‐lysine and l‐valine as the metabolites when fed with glucose and l‐methionine, respectively. The method proved that culture mediums used to feed C. glutamicum had great effect on the bacterium's metabolites.

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RP‐HPLC method with fluorescence detection for amino acids D/L ratio determination in fossil bones as a marker of DNA preservation

Cited by 6 publications

References 35 publications

Estimation of Chronological Age from Postmortem Tissues Based on Amino Acid Racemization

Estimation of Chronological Age from Postmortem Tissues Based on Amino Acid Racemization

Immunogenetics and Hplc Analyses Contribute to Understanding the Etiopathology of Rheumatoid Arthritis through Studies on Ancient Human Remains

Study of the effect of culture mediums on the amino acid metabolites for Corynebacterium glutamicum using high‐speed micellar electrokinetic chromatography

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