Mitochondrial lineage A2ah found in a pre-Hispanic individual from the Andean region.

OBJECTIVES: The aim of this study was to contribute to the knowledge of pre-Hispanic Andean mitochondrial diversity by analyzing an individual from the archaeological site Pukara de La Cueva (North-western Argentina). The date of the discovery context (540 ± 60 BP) corresponds to the Regional Developments II period. METHODS: Two separate DNA extractions were performed from dentin powder of one tooth. HVR I was amplified by PCR from each extract in three overlapping fragments and the haplotype was determined by consensus among all obtained sequences. The procedures were carried out under strict protocols developed for working with ancient DNA. RESULTS: The individual belonged to the A2ah lineage due to the presence of the 16097C and 16098G transitions, which constitute its distinctive motif. This lineage is very rare in Native American populations and was described in four individuals from current groups inhabiting the Bolivian Llanos, two from South-eastern Brazil, and one from the Gran Chaco region. In addition, two other mutations (16260T and 16286T) were shared with one of the individuals from the Bolivian Llanos region. CONCLUSIONS: Considering that the origin of this lineage was postulated for the South American lowlands, the present pre-Hispanic discovery in the Andean area could be taken as a new evidence of gene flow between these regions. Also, it allows the questioning of the geographical origin of this mitochondrial lineage.

Biofluid sampler: A new gateway for mail-in-analysis of whole blood samples.

The current study reports the development of a novel biofluid sampler (BFS) which is capable of sampling and sample preparation of whole blood without converting it into plasma or serum. The sampler can retain a whole blood sample from 10 to 1000 µL. Although the device shares the same working principle of dried blood spot (DBS) cards, it eliminates most of the technological shortcomings of DBS cards such as low maximum sample volume (~50 µL), sample inhomogeneity due to haematocrit, and poor physical adsorption driven analyte retention by incorporating sol-gel derived high efficiency, multi-functional sorbents on cellulose fabric substrate. The performance of BFS was tested via "Mail-in-Analysis" using three non-steroidal anti-inflammatory drugs (NSAIDs, ketoprofen, carprofen and diclofenac) as the test compounds. Human whole blood samples were fortified with the test compounds and sampled on conventional DBS cards and biofluid samplers (BFSs) in the USA. After drying the blood samples at room temperature, the samples were shipped to Italy for chromatographic analysis. The analytes were back-extracted from the DBS cards and BFSs using methanol and subsequently analysed using a short Symmetry C18 column (75 × 4.6 mm, 3.5 µm). Acetonitrile (ACN) and PBS (30 mM; pH = 2.5) were used as the mobile phases and the elution was performed under isocratic conditions. Compared to the classical dried blood spot cards (DBS), BFSs offer better performance in retaining the selected NSAIDs under conventional postal shipment. By substantially expanding the sampling capacity, eliminating most of the shortcomings of classical DBS cards and exploiting the better materials properties of sol-gel based functional sorbents, BFSs offer a new and profoundly simplified approach for whole blood sampling and analysis and is expected to change the current practice of blood analysis, allowing accurate quantitative analyses either in a local laboratory (on site) or using mail-in-analysis (off site) without compromising the quality of bioanalytical data.

Fast off-line FPSE-HPLC-PDA determination of six NSAIDs in saliva samples.

A fast off-line FPSE-HPLC-PDA method has been reported that allows simultaneous clean up and determination of six non-steroidal anti-inflammatory drugs (NSAIDs) in saliva samples from healthy volunteers. Particularly, furprofen, indoprofen, ketoprofen, fenbufen, flurbiprofen, and ibuprofen were chromatographically resolved. Benzyl paraben was chosen as the internal standard (BzPB, IS). These target compounds were successfully extracted from human saliva using fabric phase sorptive extraction (FPSE) and then analysed in the liquid chromatographic system by means of a short analytical column (Symmetry C18, 75 × 4.6 mm, 3.5 µm) using acetonitrile (AcN) and phosphate buffer (PBS, 30 mM; pH = 2.5) as the mobile phases. The method, validated through the calculation of all analytical parameters in accordance of International Guidelines, was applied to real saliva sample analysis collected from informed volunteers. The proposed approach that included the use of sol-gel polytetrahydrofuran (sol-gel PTHF) sorbent immobilized on cellulose support and C18 stationary phase used in HPLC, showed high potential as a fast tool for future clinical and forensic applications. The herein reported results encourage potential future application of FPSE in the forensic field. Furthermore, the FPSE membrane was tested in dried saliva spot mode (DSS) in order to check its potential use as a sampling device, also for forensic applications.