Renato Haddad

Easy ambient sonic-spray ionization-membrane interface mass spectrometry for direct analysis of solution constituents.

Using a cellulose dialysis membrane and aqueous solutions of common drugs as a proof-of-principle example, we demonstrate that solid but permeable and flexible membranes can be used as interfaces for the direct analysis of solution constituents via easy ambient sonic-spray ionization mass spectrometry. This new combination of MS techniques, herein termed EASI-MIMS, promotes droplet pick up of the analyte from the external surface of the membrane from where the analyte has selectively permeated for proper mass spectrometry characterization and quantitation. Possible application of EASI-MIMS such as the environmental analyses of effluents, on-line monitoring of fermentation and biotransformations and on-line pharmacokinetic blood analysis are discussed.

Easy Ambient Sonic-Spray Ionization Mass Spectrometry Combined with Thin-Layer Chromatography

On-spot detection and analyte characterization on thin-layer chromatography (TLC) plates is performed via ambient desorption/ionization and (tandem) mass spectrometry detection, that is, via easy ambient sonic spray ionization mass spectrometry (EASI-MS). As proof-of-principle cases, mixtures of semipolar nitrogenated compounds as well as pharmaceutical drugs and vegetable oils have been tested. The technique has also been applied to monitor a chemical reaction of synthetic importance. EASI is the simplest and gentlest ambient ionization technique currently available, assisted solely by N2 (or air). It uses no voltages, no electrical discharges; no UV or laser beams, and no high temperature and is most easily implemented in all API mass spectrometers. TLC is also the simplest, fastest, and most easily performed chromatographic technique. TLC plus EASI-MS therefore provide a simple and advantageous combination of chromatographic separation and sensitive detection of the TLC spots as well as on-spot MS or MS/MS characterization. The favorable characteristics of TLC-EASI-MS indicate advantageous applications in several areas such as drug and oil analysis, phytochemistry and synthetic chemistry, forensics via reliable counterfeit detection, and quality control.

Oil Wastes as Unconventional Substrates for Rhamnolipid Biosurfactant Production by Pseudomonas aeruginosa LBI

Oil wastes were evaluated as alternative low‐cost substrates for the production of rhamnolipids by Pseudomonas aeruginosa LBI strain. Wastes obtained from soybean, cottonseed, babassu, palm, and corn oil refinery were tested. The soybean soapstock waste was the best substrate, generating 11.7 g/L of rhamnolipids with a surface tension of 26.9 mN/m, a critical micelle concentration of 51.5 mg/L, and a production yield of 75%. The monorhamnolipid RhaC10C10 predominates when P. aeruginosa LBI was cultivated on hydrophobic substrates, whereas hydrophilic carbon sources form the dirhamnolipid Rha2C10C10 predominantly.

Perfume fingerprinting by easy ambient sonic‐spray ionization mass spectrometry: nearly instantaneous typification and counterfeit detection

Perfume counterfeiting is an illegal worldwide practice that involves huge economic losses and potential consumer risk. EASI is a simple, easily performed and rapidly implemented desorption/ionization technique for ambient mass spectrometry (MS). Herein we demonstrate that EASI-MS allows nearly instantaneous perfume typification and counterfeit detection. Samples are simply sprayed onto a glass rod or paper surface and, after a few seconds of ambient drying, a profile of the most polar components of the perfume is acquired. These components provide unique and reproducible chemical signatures for authentic perfume samples. Counterfeiting is readily recognized since the exact set and relative proportions of the more polar chemicals, sometimes at low concentrations, are unknown or hard to reproduce by the counterfeiters and hence very distinct and variable EASI-MS profiles are observed for the counterfeit samples.

New bioactive metabolites produced by Phomopsis cassiae, an endophytic fungus in Cassia spectabilis

Two new metabolites, ethyl 2,4-dihydroxy-5,6-dimethylbenzoate (1) and phomopsilactone (2) were isolated from Phomopsis cassiae, an endophytic fungus in Cassia spectabilis. Their structures were elucidated by 1D and 2D NMR, MS and IR spectral data. Compounds 1 and 2 displayed strong antifungal activity against the phytopatogenic fungi Cladosporium cladosporioides and C. sphaerospermum, as well as cytotoxicity against human cervical tumor cell line (HeLa), in in vitro assays.

Combined cysteine and homocysteine quantitation in plasma by trap and release membrane introduction mass spectrometry

Recently, a new and efficient method for total homocysteine (tHcy) quantitation in plasma using trap and release membrane introduction mass spectrometry (T&R-MIMS) with a versatile removable direct introduction membrane probe (DIMP) was described [R. Haddad, M. A. Mendes, N. F. Hoehr and M. N. Eberlin, Analyst, 2001, 126, 1212]. Herein we report on the use of the DIMP-T&R-MIMS technique for total cysteine (tCys) quantitation; hence combined tCys and tHcy quantitation in plasma or serum can be achieved. The method employs Cys and Hcy derivatization with ethyl chloroformate (after disulfide bond reduction with dithiothreitol and protein precipitation with trichloroacetic acid), preconcentration in a capillary silicone membrane, and their thermal desorption to the gas phase inside the ion source region of a mass spectrometer, at a point exactly between the two ionization filaments. Thermal desorption uses the uniform heat radiation provided by the two ionization filaments. The analytes are then ionized by electron ionization and both Cys and Hcy are quantitated by mass spectrometry using selected ion monitoring. For tCys quantitation, good linearity and reproducibility was observed for concentrations ranging from 5 to 350 microM, recovery was near 95%, and the limit of detection (LOD) was of 2 microM. This LOD is well below the mean Cys concentration in plasma, and serum samples from a large group of healthy people showed a mean tCys concentration of 132 +/- 45 microM.

Amino acid quantitation in aqueous matrices via trap and release membrane introduction mass spectrometry: homocysteine in human plasma

Trap and release membrane introduction mass spectrometry (T&R-MIMS) using a removable direct insertion membrane probe (DIMP) is employed to determine the total homocysteine concentration (tHcy) directly from human plasma after derivatization with ethyl chloroformate. The method uses no chromatographic separation, is linear, reproducible, and displays limit of quantitation (2 pM) sufficiently below the threshold concentration of tHcy in plasma. It also combines chemical, membrane, and mass spectrometric discrimination, and can be used to determine selected amino acids in human plasma simultaneously. After derivatization with ethyl chloroformate, many amino acids in aqueous solution are observed to be efficiently detected; hence T&R-MIMS is promising as a simple and sensitive technique for simultaneous quantitation of selected amino acids in plasma and urine, and in other aqueous matrices.

Increased endothelin-1 reactivity and endothelial dysfunction in carotid arteries from rats with hyperhomocysteinemia

Background and purpose:  There are interactions between endothelin‐1 (ET‐1) and endothelial vascular injury in hyperhomocysteinemia (HHcy), but the underlying mechanisms are poorly understood. Here we evaluated the effects of HHcy on the endothelin system in rat carotid arteries.

Chronic Methionine Load-Induced Hyperhomocysteinemia Enhances Rat Carotid Responsiveness for Angiotensin II

The purpose of the present study was to investigate the effects of chronic methionine treatment on vascular smooth muscle contractility for angiotensin II (Ang II). Methionine at 0.1, 1 and 2 g/kg body weight was administered daily in the drinking water for 2, 4, 8 and 16 weeks. Rat carotid rings from control and treated animals were placed in an organ chamber containing Krebs solution. Concentration-response curves for Ang II and potassium chloride (KCl) were determined. Methionine-rich diet enhanced the plasmatic homocysteine concentration, and the magnitude of the contractile response to Ang II was increased in carotid rings from treated animals after 8 and 16 weeks. However, the treatment did not alter KCl-induced contraction. In another set of experiments, the rings were incubated with indomethacin and curves for Ang II were obtained. Exposure of the rings to indomethacin inhibited the enhancement in the contractile response to Ang II. The present findings show that chronic methionine treatment enhances homocysteine plasmatic concentration leading to an enhanced Ang-II-induced contraction, which appears to be related to the release of vasoconstrictor prostanoid(s).

Easy Ambient Sonic-Spray Ionization Mass Spectrometric of Olive Oils: Quality Control and Certification of Geographical Origin

Herein, we show that easy ambient sonic spray ionization mass spectrometry in the negative ion mode [EASI(-)-MS] of water:methanol extracts of olive oil samples from 5 different countries (Portugal, Italy, Spain, Lebanon, and Greece) provides very characteristic profiles of chemotaxonomic markers, that is, free fatty acids and phenols. These EASI(-)-MS fingerprints, acquired with great speed and simplicity after minimal sample preparation, permits secure identification of the samples as olive oils via their unique profiles of fatty acids plus phenolic constituents as well the certification of geographical regions via characteristic features of the profiles of phenolic constituents.