Mass Spectrometric Procedure Research Articles

Comparison of Two Immunoassay Screening Methods and a LC-MS/MS in Detecting Traditional and Designer Benzodiazepines in Urine.

Sensitive and specific immunoassay screening methods for the detection of benzodiazepines in urine represent an important prerequisite for routine analysis in clinical and forensic toxicology. Moreover, emerging designer benzodiazepines force labs to keep their methodologies updated, in order to evaluate the reliability of the immunochemical techniques. This study aimed at evaluating the sensitivity and specificity of two different immunoassay methods for the detection of benzodiazepines in urine, through a comparison with the results obtained by a newly developed liquid chromatographic tandem mass spectrometric (LC-MS/MS) procedure. A cohort of authentic urine samples (N = 501) were processed, before and after a hydrolysis procedure, through two immunoassays and an LC-MS/MS method. The LC-MS/MS target procedure was optimized for monitoring 25 different molecules, among traditional and designer benzodiazepines, including some metabolites. At least one of the monitored substances was detected in 100 out of the 501 samples. A good specificity was observed for the two immunoassays (>0.99), independently of the cut-offs and the sample hydrolysis. The new kit demonstrated a fairly higher sensitivity, always higher than 0.90; in particular, a high cross-reactivity of the new immunoassay was observed for samples that tested positive for lorazepam and 7-aminoclonazepam. The two immunoassays appeared adequate to monitor not only traditional benzodiazepines but also new designer ones.

Analysis of Cannabinoids and Metabolites in Dried Urine Spots (DUS).

Dried urine spots (DUS) represent a potential alternative sample storage for forensic toxicological analysis. The aim of the current study was to develop and validate a liquid chromatographic tandem mass spectrometric procedure for the detection and quantitative determination of cannabinoids and metabolites in DUS. A two-step extraction was performed on DUS and urine samples. An LC-MS/MS system was operated in multiple reaction monitoring and positive polarization mode. The method was checked for sensitivity, specificity, linearity, accuracy, precision, recovery, matrix effects and carryover. The method was applied to 70 urine samples collected from healthy volunteers and drug addicts undergoing withdrawal treatment. The method was successfully developed for DUS. LODs lower than 2.0 ng/mL were obtained for all the monitored substances. All the validation parameters fulfilled the acceptance criteria either for DUS or urine. Among the real samples, 45 cases provided positive results for at least one compound. A good quali-quantitative agreement was obtained between DUS and urine. A good stability of THC, THCCOOH and THCCOOH-gluc was observed after a 24 h storage, in contrast to previously published results. DUS seems to provide a good alternative storage condition for urine that should be checked for the presence of cannabinoids and metabolites.

Hepatoprotective effects of Tagetes lucida root extract in carbon tetrachloride-induced hepatotoxicity in Wistar albino rats through amelioration of oxidative stress

Context The roots of Tagetes lucida Cav. (Asteraceae) have antioxidant and antimicrobial properties. Objective This study aimed to examine the hepatoprotective effects of T. lucida roots ethanol extract (TLRE) using carbon tetrachloride (CCl4)-induced hepatotoxicity in rats. Materials and methods The active ingredients of TLRE were identified by high-performance liquid chromatography, infra-red spectrum, and mass spectrometric procedures. Ninety rats were distributed into four main groups: positive, therapeutic, protective, and negative group. The therapeutic group was implemented using CCl4 (a single dose of 2 mL/kg) before TLRE or silymarin administration. Meanwhile, the protective group was implemented by administering CCl4 (a single dose of 2 mL/kg) after force-feeding TLRE or silymarin. Each therapeutic and protective group was divided into three subgroups: force-fed with saline, TLRE (500 mg/kg), and silymarin (25 mg/kg). The positive group was split into two subgroups that were force-fed TLRE and silymarin. Positive, therapeutic, and protective groups were compared to the negative group (untreated rats). CCl4, TLRE, and silymarin were orally administrated using a gastric tube. Results In the therapeutic and protective groups, TLRE significantly reduced liver enzymes, i.e., aspartate aminotransferase (12.47 and 6.29%), alanine aminotransferase (30.48 and 11.39%), alkaline phosphatase (17.28 and 15.90%), and cytochrome P450-2E1 (39.04 and 48.24%), and tumour necrosis factor-α (53.72 and 53.72%) in comparison with CCl4-induced hepatotoxicity controls. Conclusions TLRE has a potent hepatoprotective effect with a good safety margin. After a repeated study on another type of small experimental animal, their offspring, and an experiment with a large animal, this study may lead to clinical trials.

Detection of clostebol in sports: Accidental doping?

The detection of clostebol misuse in sports has been growing recently, especially in Italy, due to the ample availability of pharmaceutical formulations containing clostebol acetate (Trofodermin®) and the use of more sensitive instrumentation by the antidoping laboratories. Most of these cases have been claimed to be related to a nonconscious use of the drug or through contact with relatives or teammates using it. We have investigated, through the application of the well-known and currently used gas chromatographic mass spectrometric procedures, the likelihood of these allegations and have demonstrated that after a single transdermal administration of 5 mg of clostebol acetate and a transient contact with the application area, it is possible to generate adverse analytical findings in antidoping controls. We have reviewed the Phase I and Phase II clostebol metabolism in order to generate evidences that may help the sport authorities reviewing these cases. The main clostebol metabolite (4-chloro-androst-4-en-3α-ol-17-one, M1) generally used at the screening level as well as other three metabolites (M2-M4) are mainly excreted as glucuronides, whereas M5 (4ζ-chloro-5ζ-androstan-3β-ol-17-one) is predominantly excreted as sulfate. Neither the 5α-reductases activity (impaired by the presence of the chlorine in C4) nor specific sulfotransferases present in the skin allowed a clear distinction of the administration route. Studies with a larger number of volunteers and probably investigating another physiological fluid allowed in antidoping such as blood are needed for a deeper investigation. It is not unreasonable to establish a reporting level for M1, maybe creating some false negatives but excluding nonintentional doping scenarios.

Linking a rapid throughput plate-assay with high-sensitivity stable-isotope label LCMS quantification permits the identification and characterisation of low \u03b2-L-ODAP grass pea lines

BackgroundGrass pea (Lathyrus sativus) is an underutilised crop with high tolerance to drought and flooding stress and potential for maintaining food and nutritional security in the face of climate change. The presence of the neurotoxin β-L-oxalyl-2,3-diaminopropionic acid (β-L-ODAP) in tissues of the plant has limited its adoption as a staple crop. To assist in the detection of material with very low neurotoxin toxin levels, we have developed two novel methods to assay ODAP. The first, a version of a widely used spectrophotometric assay, modified for increased throughput, permits rapid screening of large populations of germplasm for low toxin lines and the second is a novel, mass spectrometric procedure to detect very small quantities of ODAP for research purposes and characterisation of new varieties.ResultsA plate assay, based on an established spectrophotometric method enabling high-throughput ODAP measurements, is described. In addition, we describe a novel liquid chromatography mass spectrometry (LCMS)-based method for β-L-ODAP-quantification. This method utilises an internal standard (di-13C-labelled β-L-ODAP) allowing accurate quantification of β-L-ODAP in grass pea tissue samples. The synthesis of this standard is also described. The two methods are compared; the spectrophotometric assay lacked sensitivity and detected ODAP-like absorbance in chickpea and pea whereas the LCMS method did not detect any β-L-ODAP in these species. The LCMS method was also used to quantify β-L-ODAP accurately in different tissues of grass pea.ConclusionsThe plate-based spectrophotometric assay allows quantification of total ODAP in large numbers of samples, but its low sensitivity and inability to differentiate α- and β-L-ODAP limit its usefulness for accurate quantification in low-ODAP samples. Coupled to the use of a stable isotope internal standard with LCMS that allows accurate quantification of β-L-ODAP in grass pea samples with high sensitivity, these methods permit the identification and characterisation of grass pea lines with a very low ODAP content. The LCMS method is offered as a new ‘gold standard’ for β-L-ODAP quantification, especially for the validation of existing and novel low- and/or zero-β-L-ODAP genotypes.

Systemic availability of lipophilic organic UV filters through dermal sunscreen exposure

BackgroundChemical UV filters are common components in sunscreens and cosmetic products and used to protect the skin against harmful effects of sunlight like sunburn. However, the effectiveness of sunscreens in the prevention of skin cancer is in some parts still controversial. Meanwhile, questions about negative effects of the chemical UV filters on human health arise and request an effective risk assessment. Real-life exposure data in humans after application of these products are still rare. Thus, we explored whether and to what extent UV filters are absorbed through the skin into the human body. Material and methodsPlasma and urine samples from 20 healthy volunteers were collected before, during and after a real-life exposure scenario (1st application: 2 mg/cm2; 2nd and 3rd (after 2 and 4 h): 1 mg/cm2 each) using a commercial sunscreen formulation for one day. These samples were analyzed for their content of the currently prominent UV filters octocrylene and avobenzone as well as 2-cyano-3,3-diphenylacrylic acid (CDAA) as the main octocrylene metabolite by using different liquid chromatography electrospray-ionization tandem mass spectrometric procedures. ResultsFollowing dermal sunscreen exposure, avobenzone, octocrylene and CDAA reached concentrations up to 11 μg/L, 25 μg/L and 1352 μg/L in plasma. In urine detection rates of avobenzone and octocrylene were low while CDAA showed a high detection rate and reached up to 5207 μg/g creatinine. Kinetic models could be fitted for octocrylene and CDAA in plasma and CDAA in urine. Concentration peaks were reached between 10 and 16 h after first application and half-life periods were in the range of 1.5 to 2 days. The lipophilic UV filter octocrylene and its metabolite CDAA showed a much slower elimination than other more hydrophilic UV filters. Concordantly, the metabolite CDAA in particular showed a markedly increased renal excretion over the whole sampling period and indicated high internal exposure to OC. DiscussionReal-life sunscreen usage leads to considerable bioavailability of organic UV filters and their metabolites which is rarely seen for other environmental exposures. A combined monitoring of the parent compound and its metabolites is important to fully address internal exposure to the UV filter in humans. Considering the kinetic profiles a prolonged systemic release due to depot formation in skin and a potential accumulation through multi-day exposure is presumed. High in-vivo loads call for a critical toxicological assessment of the UV filters and their metabolites.

Toxicokinetics of urinary 2-ethylhexyl salicylate and its metabolite 2-ethyl-hydroxyhexyl salicylate in humans after simulating real-life dermal sunscreen exposure.

Chemical UV filters are common components in sunscreens and cosmetic products. The question of adverse health risks is not completely resolved, partly owing to lacking human data from dermal exposure, which are essential for sound risk assessment. Therefore, we investigated the urinary toxicokinetics of 2-ethylhexyl salicylate (EHS) after a 1-day dermal real-life sunscreen application scenario. Twenty human volunteers were dermally exposed to a commercial sunscreen for 9h under real-life conditions (2mg/cm2 body surface area; double re-application; corresponding to 3.8g EHS). Urine samples were analyzed for EHS and one of its specific metabolites 2-ethyl-5-hydroxyhexyl salicylate (5OH-EHS) using a two-dimensional liquid chromatographic electrospray-ionization tandem mass spectrometric procedure. EHS and 5OH-EHS were excreted after sunscreen application and reached up to 525µg/g and 213µg/g creatinine, respectively. The toxicokinetic models showed concentration peaks between 7 and 8h after first application. First-phase terminal half-lives were 8-9h. For 5OH-EHS, a second-phase terminal half-life could be determined (87h). EHS and 5OH-EHS showed a faster elimination with 70-80% of the overall excretion occurring within 24h after application compared to more lipophilic UV filters. Cumulative excreted amounts over 24h reached up to 334µg EHS and 124µg of 5OH-EHS. Simulated real-life sunscreen use for 1day leads to the bioavailability of the UV filter EHS in humans. The kinetic profiles with a prolonged systemic availability indicate a skin depot and make accumulation during consecutive multi-day exposure likely.

Analysis of fragmented oxidized phosphatidylcholines in human plasma using mass spectrometry: Comparison with immune assays

Circulating oxidized phospholipids are increasingly recognized as biomarkers of atherosclerosis. Clinical association studies have been mainly performed using an immune assay based on monoclonal antibody E06, which recognizes a variety of molecular species of oxidized phosphatidylcholine (OxPC) in lipoproteins, cell membranes or covalently bound to plasma proteins. Accumulating evidence shows that individual molecular species of OxPC demonstrate different biological activities and have different half-life times. Therefore, it is likely that certain molecular species can be associated with pathology more strongly than others. This hypothesis can only be tested using LC-MS/MS allowing quantification of individual molecular species of OxPCs. In order to ensure that laborious LC-MS/MS methods do not simply replicate the results of a technically simpler E06-OxPCs assay, we have performed relative quantification of 8 truncated molecular species of OxPCs in plasma of 132 probands and compared the data with the results of the E06-OxPCs and OxLDL assays. We have found a strong correlation between individual molecular species of OxPCs but only a weak correlation of LC-MS/MS-OxPCs data with the E06-OxPCs assay and no correlation with the OxLDL assay. Furthermore, in contrast to the results of E06-OxPCs or OxLDL assays, 7 out of 8 OxPC species were associated with hypertension. The data suggest that the results of the LC-MS/MS-OxPCs assay do not replicate the results of two ELISA-based lipid oxidation tests and therefore may produce additional diagnostic information. These findings necessitate development of simplified mass spectrometric procedures for high-throughput and affordable analysis of selected molecular species of OxPCs.

Improving the diagnosis, treatment, and prevention of chronic kidney disease through accurate and reliable parathyroid hormone measurements by establishing a mass spectrometric reference measurement procedure

Improving the diagnosis, treatment, and prevention of chronic kidney disease through accurate and reliable parathyroid hormone measurements by establishing a mass spectrometric reference measurement procedure

Incorporation of zolpidem and methoxyphenamine into white hair strands after single administrations: Influence of hair pigmentation on drug incorporation

In order to investigate the influence of pigmentation on the incorporation of drugs into hair, time-course changes in drug distribution along non-pigmented (white) hairs as well as pigmented (black) hairs plucked from the same subject was observed following single administrations of two basic drugs with different properties, zolpidem and methoxyphenamine. These drugs in 1-mm sections of single hair specimens were each determined by a liquid chromatography–tandem mass spectrometric procedure. During the early stage (12–36 h) after intake, for black hairs, both drugs were detected over the entire area of hair root (4–5 mm in length), in which notable concentration of these drugs in the hair bulb (0–1-mm segment from the bottom of hair root, Region 1) and lower concentrations in the upper dermis zone (1–2-mm to 3–4-mm or to 4–5-mm segments, Region 2) were commonly observed. Meanwhile, for white hairs, high drug concentrations in Region 1 as detected in black hairs were not observed although only small amounts of these drugs were detected over Region 2. Subsequent time-course changes in the concentration of drugs in hair demonstrated that the drugs once incorporated into white hair via Region 2 decreased gradually over the period from 24 h to 35 days after intake, but those of black hairs remained almost unchanged. These findings revealed here suggest that hair pigments have two important roles in the distribution of drugs: (1) incorporation of drugs into hair via Region 1, and (2) retention of already incorporated drugs in the hair tissue. These findings would be useful for discussing individual drug-use history based on hair analysis in the forensic fields.

Certification of ERM‐EB400, the First Matrix Reference Material for Lead Isotope Amount Ratios, and ERM‐AE142, a Lead Solution Providing a Lead Isotopic Composition at the Edge of Natural Variation

Lead isotope amount ratios are commonly used in diverse fields such as archaeometry, geochemistry and forensic science. Currently, five reference materials with certified lead isotope amount ratios are available, namely NIST SRM 981, 982 and 983, GBW‐04442 and NMIJ 3681‐a. Only NIST SRM 981 and NMIJ 3681‐a have approximately natural isotopic compositions, and NIST SRM 981 is predominantly used for correcting mass discrimination/mass fractionation in the applied mass spectrometric procedures. Consequently, there is no other certified reference material available to be used for validation and/or quality control of the analytical procedures applied to lead isotope amount ratio measurements. To fill this gap, two new reference materials have been produced and certified for their lead isotope amount ratios. For both certified reference materials, complete uncertainty budgets have been calculated and SI traceability has been established. This provides the users with independent means for validating and verifying their analytical procedures and for conducting quality control measures. ERM‐EB400 is a bronze material with a nominal lead mass fraction of 45 mg kg−1 and certified lead isotope amount ratios of n(206Pb)/n(204Pb) = 18.072(17) mol mol−1, n(207Pb)/n(204Pb) = 15.578(18) mol mol−1 and n(208Pb)/n(204Pb) = 38.075(46) mol mol−1 with the associated expanded uncertainties (k = 2) given in brackets. ERM‐AE142 is a high‐purity solution of lead in 2% nitric acid with a nominal mass fraction of 100 mg kg−1 and certified Pb isotope amount ratios of n(206Pb)/n(204Pb) = 21.114(17) mol mol−1, n(207Pb)/n(204Pb) = 15.944(17) mol mol−1 and n(208Pb)/n(204Pb) = 39.850(44) mol mol−1 with the associated expanded uncertainties (k = 2) given in brackets. Both materials are specifically designed to fall within the natural lead isotopic variation and to assist users with the validation and verification of their analytical procedures. Note that while one of these reference materials requires the chemical separation of Pb from its matrix (ERM‐EB400), the other does not (ERM‐AE142). As additional information, δ208/206PbNIST SRM981 values are provided for both materials. For ERM‐AE142, a delta value of δ208/206PbNIST SRM981 = −28.21(30)‰ was obtained, and for ERM‐EB400, a delta value of δ208/206PbNIST SRM981= −129.47(38)‰ was obtained, with the associated expanded uncertainties (k = 2) given in brackets.

Determination of Calcium in Blood

Human blood is tested in clinical diagnostic laboratories for numerous indicators. One of these indicators is the concentration of calcium. This review presents a chronological development of procedures for determining the concentration of calcium in plasma, serum, and whole blood, which are based on various analytical methods such as titrimetry, manometry, X-ray fluorescence, flame photometry, spectrophotometry, spectrofluorimetry, potentiometry, flame atomic absorption, inductively coupled plasma atomic emission spectrometry, chromatography, and mass spectrometry. A brief overview of these methods and the results of comparative tests are given. Currently, potentiometric procedures for determining the concentration of calcium in plasma, serum, and whole blood are the most widespread in small, medium, and large clinical diagnostic laboratories. Flame photometric procedures often act as expert methods. Mass spectrometric procedures are used to validate reference sample solutions, while inductively coupled plasma atomic emission spectrometric procedures and chromatographic procedures are used in research organizations.

Characterization of the artist's palette from the polychrome decorations of the El Bahia Palace doors (Marrakesh, Morocco)

Abstract We investigated the cedar wood doors of the Bahia Palace in Marrakesh (Morocco), which are richly decorated with geometric motifs. A gas chromatographic mass spectrometric analytical procedure (GC/MS) was used to characterize the organic binders in micro samples collected from one of the doors. In order to examine the artist's palette, X-ray fluorescence (XRF), Raman and Fourier transform infrared spectroscopy (FTIR) and high performance liquid chromatography (HPLC-DAD), were applied to samples in fragments or on cross-sections of the embedded samples. This multi analytical approach effectively identified the polychrome decoration technique, mainly based on the use of animal glue as a proteinaceous binder, and characterized the superficial varnishes. Lastly many of the different pigments used for the creation of these works of art were identified. We believe that this study contributes significantly to the knowledge of Moroccan art from a material and technological point of view.

LC–MS/MS procedure for the simultaneous determination of N-acetyl-S-(1-naphthyl)cysteine and N-acetyl-S-(2-napthyl)cysteine in human urine

A two-dimensional liquid chromatographic electrospray-ionization tandem mass spectrometric (LC/LC–ESI–MS/MS) procedure for the simultaneous determination of the expected mercapturic acids of naphthalene (1- and 2-naphthylmercapturic acids; 1- and 2-NpMA) and of the well-established parameter for benzene biomonitoring (S-phenylmercapturic acid; PhMA) in human urine was developed, validated and applied to human urine samples. Apart from sample acidification, the enrichment of analytes and sample clean-up as well as the separation of all analytes were completely automated using both a restricted access material column (RAM C18) and a core-shell biphenyl material. Sensitive, specific and reliable detection of all target substances, with limits of detection ranging from 0.03 to 0.04 μg/L, was achieved using structurally well matching isotope-labelled internal standard substances for each analyte. Intraday and interday precision were determined, ranging from 2.2 to 4.3%, and mean accuracy from 98.4 to 100.8%. Due to the low limits of detection, the good precision and accuracy of the developed procedure, it is well suited for application in biomonitoring studies to evaluate the validity of mercapturic acids as biomarkers after naphthalene exposure.

Supercharging Synthetic Polymers: Mass Spectrometric Access to Nonpolar Synthetic Polymers

We report a mass spectrometric access route to analyze nonpolar poorly ionizing synthetic polymers exploiting supercharging technology by chloride attachment via high-resolution electrospray ionization mass spectrometry (HR ESI MS). The novel mass spectrometric procedure allows for the characterization of polyhydrocarbons, which include hardly ionizable polymers such as poly(styrene) (PS) (ranging from 1700 to 18 000 g mol–1) and—for the first time reported using ESI as ionization method—poly(1,4-butadiene) (PBD) (ranging from 1000 to 10 000 g mol–1). The method is also applied to additional synthetic polymers including poly(2-vinylpyridine) (P2VP) and poly(acrylamide) (PAAm). The powerful chloride attachment enables the detection of multiply charged polyhydrocarbons (up to quadruply charged). A systematic assessment of the manipulation of these charge states using supercharging agents (sulfolane, propylene carbonate, and m-nitrobenzyl alcohol) is carried out. Our investigations include an assessment of t...

Accurate quantification of sphingosine-1-phosphate in normal and Fabry disease plasma, cells and tissues by LC-MS/MS with 13C-encoded natural S1P as internal standard

We developed a mass spectrometric procedure to quantify sphingosine-1-phosphate (S1P) in biological materials. The use of newly synthesized 13C5 C18-S1P and commercial C17-S1P as internal standards rendered very similar results with respect to linearity, limit of detection and limit of quantitation. Caution is warranted with determination of plasma S1P levels. Earlier it was reported that S1P is elevated in plasma of Fabry disease patients. We investigated this with the improved quantification. No clear conclusion could be drawn for patient plasma samples given the lack of uniformity of blood collection and plasma preparation. To still obtain insight, plasma and tissues were identically collected from α-galactosidase A deficient Fabry mice and matched control animals. No significant difference was observed in plasma S1P levels. A significant 2.3 fold increase was observed in kidney of Fabry mice, but not in liver and heart. Comparative analysis of S1P in cultured fibroblasts from normal subjects and classically affected Fabry disease males revealed no significant difference. In conclusion, accurate quantification of S1P in biological materials is feasible by mass spectrometry using the internal standards 13C5 C18-S1P or C17-S1P. Significant local increases of S1P in the kidney might occur in Fabry disease as suggested by the mouse model.

Determination of N-nitrosamines by automated dispersive liquid-liquid microextraction integrated with gas chromatography and mass spectrometry.

An automated dispersive liquid-liquid microextraction integrated with gas chromatography and mass spectrometric procedure was developed for the determination of three N-nitrosamines (N-nitroso-di-n-propylamine, N-nitrosopiperidine, and N-nitroso di-n-butylamine) in water samples. Response surface methodology was employed to optimize relevant extraction parameters including extraction time, dispersive solvent volume, water sample pH, sodium chloride concentration, and agitation (stirring) speed. The optimal dispersive liquid-liquid microextraction conditions were 28 min of extraction time, 33 μL of methanol as dispersive solvent, 722 rotations per minute of agitation speed, 23% w/v sodium chloride concentration, and pH of 10.5. Under these conditions, good linearity for the analytes in the range from 0.1 to 100 μg/L with coefficients of determination (r(2) ) from 0.988 to 0.998 were obtained. The limits of detection based on a signal-to-noise ratio of 3 were between 5.7 and 124 ng/L with corresponding relative standard deviations from 3.4 to 5.9% (n = 4). The relative recoveries of N-nitroso-di-n-propylamine, N-nitrosopiperidine, and N-nitroso di-n-butylamine from spiked groundwater and tap water samples at concentrations of 2 μg/L of each analyte (mean ± standard deviation, n = 3) were (93.9 ± 8.7), (90.6 ± 10.7), and (103.7 ± 8.0)%, respectively. The method was applied to determine the N-nitrosamines in water samples of different complexities, such as tap water, and groundwater, before and after treatment, in a local water treatment plant.

From Plant Extracts to Historical Textiles: Characterization of Dyestuffs by GC–MS

A gas chromatographic–mass spectrometric procedure for the analysis of dyes from plant extracts was optimized and applied for the detection of components in extracts from 12 dyeing plants native from all over the world. The main marker molecules in each of the dyestuffs, as well as accessory compounds, were successfully separated and identified by means of their electron impact mass spectra, thus demonstrating that a single GC–MS procedure can be conveniently applied to the detection of natural dyes such as flavonoids, neoflavonoids, anthraquinones and tannins. Other constituents of plant extracts, such as organic acids, oils and sugars, that hydrolyse during the extraction process, were also detected and recognized in the same chromatographic run. The GC–MS method was tested on woollen references dyed according to traditional recipes, and on historical wool samples taken from a tapestry of the sixteenth century, and the main dyestuffs used for colouring could be traced even with the low amounts usually available with the archaeometric samples. Besides some limitations, that are addressed, the results show that the GC analysis is a useful tool for quick assessment and control of natural extracts and the application of the technique for the characterization of dyes in historical textiles, in addition or in place of the more widely used procedures that employ liquid chromatography, is encouraged.

Clinical and forensic examinations of glycaemic marker methylglyoxal by means of high performance liquid chromatography–tandem mass spectrometry

The postmortem determination of hyperglycaemic coma is quite difficult because of the lack of morphological findings and the difficult interpretation of biochemical parameters. Methylglyoxal (MG) is a reactive oxoaldehyde, which is mainly derived from glycolysis. An electrospray ionisation liquid chromatography-tandem mass spectrometric procedure for the determination of methylglyoxal in human serum and postmortem blood was developed. It involves protein precipitation with perchloric acid and a derivatisation step with 2,3-diaminonaphthalene. The assay was validated according to international guidelines. Serum samples from diabetics obtained at a diabetes clinic and from non-diabetics were used to assess data about reference concentrations in human serum. The assay showed linearity within the physiological concentrations in serum (5-500ng/ml). Intraday imprecision at three concentrations was 10.3, 9.2 and 8.3%, and interday imprecision was 15.3, 14.2 and 9.4%; the limit of detection was 1.3ng/ml, and limit of quantification, 3.2ng/ml. One hundred and eighteen clinical (100 diabetics, 18 non-diabetics) and 98 forensic samples (84 non-diabetics, 14 in a status of hyperglycaemic coma) were measured. During life, diabetics showed significantly (p < 0.001) higher serum concentrations of MG than non-diabetics. After death, concentrations of MG increased significantly (p < 0.001). However, there was no correlation between the sum formula of Traub in vitreous humour and MG femoral blood concentrations (R = 0.237). This indicates that MG concentrations in the deceased cannot distinguish deaths due to a hyperglycaemic coma from other causes of death.

Abstract 4784: Rapid mass spectrometric profiling of blood sera detects cancer with high accuracy

Abstract Cancer diagnosis is problematic because the disease is often asymptomatic at early stages of progression and/or recurrence. We report here the integration of a mass spectrometric metabolic profiling procedure coupled with a support vector machine computational method for use in early cancer diagnostics, and describe successful application of the method to ovarian, pancreatic and prostate cancers. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4784. doi:1538-7445.AM2012-4784