UHPLC-MS analysis of juvenile hormone II in Mediterranean corn borer (Sesamia nonagrioides) hemolymph using various ionization techniques

The juvenile hormones (JHs) have been considered the most versatile hormones in the animal kingdom. JH-II is the most abundant JH in Sesamia nonagrioides, important maize pests in the Mediterranean basin. This study compared the sensitivities and matrix effects of four ionization modes on analyzing JH-II in S. nonagrioides hemolymph using ultrahigh-performance liquid chromatography-mass spectrometry (UHPLC-MS) in single ion monitoring (SIM) mode. The ionization techniques tested were electrospray ionization (ESI), atmospheric pressure chemical ionization (APCI), atmospheric pressure photoionization (APPI), and APPI with the lamp turned off, which corresponds to atmospheric pressure thermospray ionization (APTSI). ESI was discarded because of the high matrix effect. APPI was discarded because the correlation responses between solvent and matrix on the instrumental quality parameters were worse than those for APTSI and APCI. In our analytical conditions, APCI has shown the best validation parameter values. APCI ionization is widely available in instrumental laboratories.     

Isoflavonoids isolated from Cuban propolis

Chemical investigation of a red-type Cuban propolis sample has led to the isolation of 11 isoflavonoids (2 isoflavones, 3 isoflavans, and 6 pterocarpans), together with gallic acid, isoliquiritigenin, and (-)-liquiritigenin. Structural determination, including the absolute stereochemistry, was accomplished by spectroscopic analysis, particularly CD and 2D NMR techniques. The fragmentation behavior of pterocarpans was studied by electrospray ionization (ESI) tandem mass spectrometry (MS/MS) using an ion-trap analyzer, and a generalized fragmentation pathway, useful in the identification and structural characterization of pterocarpans, is proposed. Isoflavonoids are reported for the first time from propolis samples.     

LC/MS/MS characterization of phenolic constituents in dried plums

Dried plums are known as a healthy food in the West and used as medicine in India. They have been characterized by high concentrations of phenolic compounds, which are believed to play a crucial role in protection against various age-related diseases. Liquid chromatography/mass spectrometry/mass spectrometry (LC/MS/MS) with four different conditions was used to analyze the phytochemicals in commercial dried plums. The major components were neochlorogenic acid and cryptochlorogenic acid. Forty minor components were characterized by their MS/MS spectra and LC retention time. Six of them are novel ester isomers formed by two caffeic acids and one quinic acid. The diagnostic fragmentation patterns of different phenolics are presented on the basis of electrospray ionization (ESI) MS/MS data of components in dried plums and fourteen authentic standards.     

Identification of benzalkonium chloride in commercial grapefruit seed extracts

Commercial grapefruit seed extracts (GSE) were extracted with chloroform. The solvent was evaporated, and the resulting solid was subsequently analyzed by high-performance liquid chromatography (HPLC), electrospray ionization mass spectrometry (ESI/MS), tandem mass spectrometry (ESI/MS/MS), and elemental analysis (by proton-induced X-ray emission analysis). Three major constituents were observed by HPLC and were identified as benzyldimethyldodecylammonium chloride, benzyldimethyltetradecylammonium chloride, and benzyldimethylhexadecylammonium chloride. This mixture of homologues is commonly known as benzalkonium chloride, a widely used synthetic antimicrobial ingredient used in cleaning and disinfection agents.     

Quantitation of total folate in whole blood using LC-MS/MS

An accurate method for measuring whole blood total folate using liquid chromatography with tandem mass spectrometry is described and compared to GC/MS and a chemiluminescence assay. Whole blood from normal adults (n = 15) was fortified with a [(13)C(6)]para-aminobenzoic acid (pABA) internal standard and treated with 12.1 N hydrochloric acid at 110 degrees C for 4 h to hydrolyze all folates to pABA. Contaminants in the hydrolysate were adsorbed onto a C18 SPE cartridge. The eluate containing the folate catabolite pABA was partitioned into ethyl acetate and methylesterified with trimethylsilyldiazomethane. The methyl-pABA derivatives were quantified by positive-ion atmospheric pressure chemical ionization (APCI)LC-MS/MS. An isocratic mobile phase of acetonitrile-water (70:30) (v/v) on a C18 analytical column was used with a postcolumn reagent of 0.025% formic acid. The limit of quantitation for folate was 56.6 nmol/L RBC, and the limit of detection was 22.6 nmol/L RBC. Folate levels as determined by LC-MS/MS correlated well with the chemiluminescence assay and a GC/MS method. This new LC-MS/MS method provides enhanced sample throughput (n = 36 per day) as compared to GC/MS methods. LC-MS/MS will enable accurate measurements of red blood cell (RBC) folate in nutrition surveys and clinical trials.     

Identification and quantification of astaxanthin esters in shrimp (Pandalus borealis) and in a microalga (Haematococcus pluvialis) by liquid chromatography-mass spectrometry using negative ion atmospheric pressure chemical ionization

Negative ion liquid chromatography-atmospheric pressure chemical ionization mass spectrometry [negative ion LC-(APCI)MS] was used for the identification of astaxanthin esters in extracts of commercial shrimp (Pandalus borealis) and dried microalga (Haematococcus pluvialis) samples. A cleanup step using a normal phase solid phase extraction (SPE) cartridge was applied prior to analysis. Recovery experiments with astaxanthin oleate as model compound proved the applicability of this step (98.5 +/- 7.6%; n = 4). The assignment of astaxanthin esters in negative ion LC-(APCI)MS was based on the detection of the molecular ion (M*-) and the formation of characteristic fragment ions, resulting from the loss of one or two fatty acids. Quantification of individual astaxanthin esters was performed using an astaxanthin calibration curve, which was found to be linear over the required range (1-51 micromol/L; r2 = 0.9996). Detection limits, based on the intensity of M*-, a signal-to-noise ratio of 3:1, and an injection volume of 20 microL, were estimated to be 0.05 microg/mL (free astaxanthin), 0.28 microg/mL (astaxanthin-C16:0), and 0.78 microg/mL (astaxanthin-C16:0/C16:0), respectively. This LC-(APCI)MS method allows for the first time the characterization of native astaxanthin esters in P. borealis and H. pluvialis without using time-consuming isolation steps with subsequent gas chromatographic analyses of fatty acid methyl esters. The results suggest that the pattern of astaxanthin-bound polyunsaturated fatty acids of P. borealis does not reflect the respective fatty acid pattern found in triacylglycerides. Application of the presented LC-(APCI)MS technique in common astaxanthin ester analysis will forestall erroneous xanthophyll ester assignment in natural sources.     

Atmospheric pressure ionization LC-MS-MS determination of urushiol congeners

This paper describes atmospheric pressure ionization (API) LC-MS-MS determination of urushiols, 3-n-alkenyl- and -alkyl-substituted catechols responsible for poison oak dermatitis. Urushiol was isolated from Western poison oak according to the method of Elsohly et al. (1) (J. Nat. Prod. 1982, 45, 532-538)-the purified preparation contained C(17)- and C(15)-substituted urushiols with zero, one, two, and three double bonds as determined from GC-MS analysis of trimethylsilyl derivatives. Urushiol mixtures were separated on a C(18) reversed phase HPLC column with a methanol-water gradient with urushiols eluting in 100% methanol. Atmospheric pressure chemical ionization (APCI) produced primarily [M - H](-) and MH(+) molecule ions. Electrospray ionization (ESI) yielded [M - H](-) and adduct ions including [M + Cl](-). Daughter ions of [M - H](-) included quinoid radical anions ([M - H - H(2)](-) and m/z 122(-)) and a benzofuran phenate (m/z 135(-)). A suite of hydrocarbon fragments were produced by collision-induced dissociation of MH(+) directly or via an intermediate [MH - H(2)O](+) daughter ion. Six urushiol congeners, one not previously reported in poison oak, were determined by negative ion API-LC-MS-MS with detection limits of approximately 8 pg/microL (ESI) and approximately 800 pg/microL (APCI). API-LC-MS-MS was used to determine urushiol in surface wipes, air samples, and plant materials.     

Separation and characterization of phenolic compounds in fennel (Foeniculum vulgare) using liquid chromatography-negative electrospray ionization tandem mass spectrometry

Liquid chromatography (LC) diode array detection (DAD) coupled to negative electrospray ionization (ESI) tandem mass spectrometry (MS/MS) was used for the rapid and sensitive identification of water-soluble phenolic compounds in fennel waste. The plant material was first extracted and then chromatographed on Sephadex LH-20 to afford seven fractions, each of them being subjected to LC-MS analysis. Identification of the compounds was carried out by interpretation of UV, MS, and MS/MS spectra. Forty-two phenolic substances were identified, 27 of which had not previously been reported in fennel, including hydroxycinnamic acid derivatives, flavonoid glycosides, and flavonoid aglycons.     

Development of multiclass methods for drug residues in eggs: hydrophilic solid-phase extraction cleanup and liquid chromatography/tandem mass spectrometry analysis of tetracycline, fluoroquinolone, sulfonamide, and beta-lactam residues

A method was developed for detection of a variety of polar drug residues in eggs via liquid chromatography/tandem mass spectrometry (LC/MS/MS) with electrospray ionization (ESI). A total of twenty-nine target analytes from four drug classes-sulfonamides, tetracyclines, fluoroquinolones, and beta-lactams-were extracted from eggs using a hydrophilic-lipophilic balance polymer solid-phase extraction (SPE) cartridge. The extraction technique was developed for use at a target concentration of 100 ng/mL (ppb), and it was applied to eggs containing incurred residues from dosed laying hens. The ESI source was tuned using a single, generic set of tuning parameters, and analytes were separated with a phenyl-bonded silica cartridge column using an LC gradient. In a related study, residues of beta-lactam drugs were not found by LC/MS/MS in eggs from hens dosed orally with beta-lactam drugs. LC/MS/MS performance was evaluated on two generations of ion trap mass spectrometers, and key operational parameters were identified for each instrument. The ion trap acquisition methods could be set up for screening (a single product ion) or confirmation (multiple product ions). The lower limit of detection for screening purposes was 10-50 ppb (sulfonamides), 10-20 ppb (fluoroquinolones), and 10-50 ppb (tetracyclines), depending on the drug, instrument, and acquisition method. Development of this method demonstrates the feasibility of generic SPE, LC, and MS conditions for multiclass LC/MS residue screening.     

Comparison of proanthocyanidins in commercial antioxidants: grape seed and pine bark extracts

The major constituents in grape seed and pine bark extracts are proanthocyanidins. To evaluate material available to consumers, select lots were analyzed using high-performance liquid chromatography, gas chromatography/mass spectrometry (GC/MS), liquid chromatography/mass spectrometry (LC/MS), gel permeation chromatography (GPC), and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Atmospheric pressure chemical ionization (APCI) LC/MS was used to identify monomers, dimers, and trimers present. GC/MS analyses led to the identification of ethyl esters of hexadecanoic acid, linoleic acid, and oleic acid, as well as smaller phenolic and terpene components. The GPC molecular weight (MW) distribution indicated components ranging from approximately 162 to approximately 5500 MW (pine bark less than 1180 MW and grape seed approximately 1180 to approximately 5000 MW). MALDI-TOF MS analyses showed that pine bark did not contain oligomers with odd numbers of gallate units and grape seed contained oligomers with both odd and even numbers of gallate. Reflectron MALDI-TOF MS identified oligomers up to a pentamer and heptamer, and linear MALDI-TOF MS showed a mass range nearly double that of reflectron analyses.     

Characterization of soluble non-covalent complexes between bovine serum albumin and beta-1,2,3,4,6-penta-O-galloyl-D-glucopyranose by MALDI-TOF MS

Beta-1,2,3,4,6-penta-O-galloyl-D-glucopyranose (PGG) and soluble complexes of PGG with bovine serum albumin (BSA) were characterized by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). PGG was also characterized by electrospray ionization mass spectrometry (ESI-MS). Similar fragmentation patterns of PGG were found in ESI-MS and MALDI-TOF MS. The apparent stoichiometries of non-covalent BSA-PGG complexes were determined by MALDI-TOF MS.     

Mulberrofuran G and Isomulberrofuran G from Morus alba L.: Anti-hepatitis B Virus Activity and Mass Spectrometric Fragmentation

Mulberrofuran G (1) and isomulberrofuran G (2), a pair of isomeric Diels-Alder-type adducts, were isolated from the root bark of Morus alba L. Isomulberrofuran G (2) as a new IIB-type Diels-Alder-type adduct, was elucidated by extensive (1)H, (13)C, and two-dimensional (2D) nuclear magnetic resonance (NMR) and mass spectrometry (MS) spectroscopic analyses. A fragmentation study on compounds 1 and 2 was performed by high-resolution electrospray ionization (ESI) multistage tandem mass spectrometry linked with ion-trap (IT) and time-of-flight (TOF) mass analyzers (ESI-MS(n)/IT-TOF) in negative mode, which resulted in obviously different fragmentations. In the MS(2) experiments, the characteristic ions at m/z 451 and 439 could be revealed as their respective diagnostic ions. Mulberrofuran G (1) showed moderate activity, inhibiting hepatitis B virus (HBV) DNA replication with the IC(50) value of 3.99 μM, according to the anti-HBV assay on the HepG 2.2.15 cell line in vitro.     

Characterization of vegetable oils: detailed compositional fingerprints derived from electrospray ionization fourier transform ion cyclotron resonance mass spectrometry

Adulteration of vegetable oil is of concern for both commercial and health reasons. Compositional based fingerprints can potentially reveal both the oil source and its possible adulteration. Here, electrospray ionization (ESI) Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) resolves and identifies literally thousands of distinct chemical components of commercial canola, olive, and soybean oils, without extraction or other wet chemical separation pretreatment. In negative-ion ESI FT-ICR MS, the acidic components of soybean oil are easily distinguished from those of canola and olive oil based on relative abundances of C(18) fatty acids, whereas olive oil differs from canola and soybean oil based on relative abundances of tocopherols. In positive-ion ESI FT-ICR MS, the three oils are readily distinguished according to the relative abundances of di- and triacylglycerols with various numbers of double bonds in the fatty acid chains. We demonstrate the detection of soybean oil as an adulterant of olive oil, based on relative abundances of members of each of several chemical families. We suggest that the detailed chemical compositions of vegetable oils can be used to characterize them and to detect and identify adulterants.     

Hop as an interesting source of resveratrol for brewers: optimization of the extraction and quantitative study by liquid chromatography/atmospheric pressure chemical ionization tandem mass spectrometry

Nowadays, hop is used almost exclusively by brewers for bitterness and flavor. Although hop polyphenols have been widely studied in the past decade for their antioxidant activity in the boiling kettle, very little is known about their real impact on health. The discovery of resveratrol in hop pellets highlights the potential health-promoting effect of moderate beer consumption. Here, we have optimized a quantitative extraction procedure for resveratrol in hop pellets. Preliminary removal of hydrophobic bitter compounds with toluene and cyclohexane at room temperature allows 99% trans-resveratrol recovery by ethanol:water (75:25, v/v) solid/liquid extraction at 60 degrees C. Reverse phase liquid chromatography proves an excellent means of separating isomers. In addition, we have compared two mass spectrometry ionization methods-atmospheric pressure chemical ionization (APCI) and electrospray ionization (ESI)-in both the positive and the negative modes. On the basis of standard additions applied with the optimized extraction procedure and reverse phase high-performance liquid chromatography-APCI(+)-tandem mass spectrometry, it appears that Tomahawk hop pellets (T90, harvest 2002) contain 0.5 ppm trans-resveratrol, 2 ppm trans-piceid, no cis-resveratrol, and 0.9 ppm cis-piceid.     

Liquid chromatography analysis of erythromycin A in salmon tissue by electrochemical detection with confirmation by electrospray ionization mass spectrometry

A rapid and sensitive method is described for the quantitation of erythromycin A (EA) in edible salmon tissue by liquid chromatography (LC) analysis using either electrochemical detection (ED) or electrospray ionization mass spectrometric (ESI/MS) detection. The salmon tissue is extracted with 10 mM ammonium formate. The extract is then purified by solid phase extraction using a hydrophilic-lipophilic balanced (HLB) polymeric-based C18 packing, followed by partitioning of EA into methylene chloride at alkaline pH, evaporation, and final dilution. The mean recoveries of EA at 50, 100, 200, and 400 ppb levels in fortified salmon tissue were 63.8 +/- 6.0 and 75.5 +/- 5.4% by LC-ED and LC-ESI/MS, respectively. There was no evidence of formation of the anhydro-EA (m/z 716) decomposition product of EA (m/z 734) that was reported to occur by other published methods.     

Mass spectrometry and identification of sterols in vegetable oils as butyryl esters and relative quantitation by gas chromatography with flame ionization detection

Electron ionization mass spectrometry (MS) of sterol butyrates is described. Fragmentation of common sterol butyrates is related to structure and is discussed in relation to the fragmentation of free sterols and of commonly used sterol derivatives. Derivatized samples of vegetable oils are introduced using a 10 m capillary gas chromatographic (GC) column for complete separation of the sterol butyrates. Quantitation of sterol butyrates in vegetable oils by packed column GC/flame ionization detection is based on percent relative area of peaks identified by MS. Results of analyses of sunflower, castor, rapeseed, and virgin olive oils, and other oils are presented. These techniques have been applied to the rapid screening of marketed olive oils for possible adulteration.     

Quantitative confirmation of dimetridazole and ipronidazole in swine feed by capillary gas chromatography/mass spectrometry with multiple ion detection

Extracts from 4 types of swine feed containing 0.11 ppm each of dimetridazole (DMZ) and ipronidazole (IPR) were analyzed by capillary gas chromatography/mass spectrometry (GC/MS) using multiple ion detection (MID) techniques. We demonstrate in this paper that the quantitative results obtained by capillary GC/MS with MID are comparable for both compounds to results obtained by liquid chromatography and have a lower coefficient of variation for DMZ. Moreover, consistency in the ion ratios (5 ions in DMZ and 6 ions in IPR) permits identification of these compounds by electron ionization MS.     

Creatinine measurements in 24 h urine by liquid chromatography--tandem Mass Spectrometry

A simple, sensitive, and specific liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for determining urinary creatinine was developed and used to evaluate 24 h urine samples collected during an exposure study. Urine (1 microL) was diluted with methanol and then directly applied to LC-MS/MS. Under electrospray ionization (ESI) conditions, the transition molecules of creatinine and creatinine- d3 were observed at m/ z 114 > 44 and m/ z 117 > 47, respectively. The retention time of creatinine was 0.59 min. The linear range was 1-2000 ng/mL, with a detection limit in urine of 1 ng/mL. LC-MS/MS and colorimetric end-point methods were significantly associated ( R2 = 0.8785, p < 0.0001). The LC-MS/MS method to determine creatinine in 24 h urine samples had shorter retention times, was more sensitive, reliable, reproducible, simple, selective, and used a smaller sample size than other LC-MS/MS or commercial methods.     

Determination of ascorbic acid and carotenoids in food commodities by liquid chromatography with mass spectrometry detection

Two methods, one to determine ascorbic acid and one to determine lycopene and beta-carotene, in vegetables and fruits by liquid chromatography coupled with mass spectrometry (LC-MS) have been established. The chromatographic separation of the studied compounds and their MS parameters were optimized to improve selectivity and sensitivity. In both methods, separation was carried out with two coupled columns, first a C(18) and then a dC(18), using as mobile phase 70% methanol (0.005% acetic acid) and 30% acetic acid 0.05% for ascorbic acid determination and a mixture of methanol, tetrahydrofuran, and acetonitrile (60:30:10 v/v/v) for carotenoid analysis in isocratic mode. The molecular ion was selected for the quantification in selective ion monitoring (SIM) mode. Ascorbic acid was detected with electrospray ionization probe (ESI) in negative mode, while chemical ionization atmospheric pressure (APCI) in positive mode was used for the target carotenoids. The methodology for ascorbic acid analysis is based on an extraction with polytron using methanol and a mixture of methaphosphoric acid and acetic acid. Extraction of the carotenoids was carried out with tetrahydrofuran/methanol (1:1) (v/v). The proposed methods were applied, after their corresponding validations, to the analysis of four varieties of tomatoes, tomato in tin enriched and dried tomato, and to the analysis of mango and kiwi fruits, to compare the content in these compounds. Moreover, the influence of the process of freezing and the effect that the manipulation/preservation has in the content of ascorbic acid in tomato have also been studied.     

Electrospray ionization mass spectrometry fingerprinting of Brazilian artisan cachaça aged in different wood casks

We have investigated the capability of direct infusion electrospray ionization mass spectrometry in the negative ion mode, ESI(-)-MS, to differentiate representative samples of artisan cacha?a, a Brazilian sugar cane distillate of large production, aged in four different types of wood casks: amburana (Amburana cearensis), jequitibA (Cariniana estrellensis), balm (Myroxylon peruiferum), and oak (Quercus rubra). The ESI(-)-MS were found to be very characteristic, showing sets of diagnostic ions for each of the four types of samples: amburana (m/z 271, 313, 377), jequitibA (m/z 143, 171, 255), balm (m/z 137, 269, 283, 297), and oak (m/z 197, 301, 307). Furthermore, principal component (PCA) and hierarchical cluster analysis (HCA), applied to the ESI(-)-MS data, divided these samples into four definite categories. The influence of the aging time on the ESI(-)-MS fingerprints of the cacha?a samples stored in oak casks was also established. An inversion in the relative intensity of the diagnostic ions of m/z 307 and 301 is detected in the ESI(-)-MS as the aging time increased from 1 to 2 years. The chemical structures of the major cacha?a components were proposed on the basis of the following: (a) the comparison of the ESI(-)-MS/MS of the diagnostic anions with those of the authentic anions or (b) the interpretation of the fragmentation patterns of the previously unknown diagnostic anions. Hence, direct infusion ESI(-)-MS allows not only a rapid, simple, and accurate way to distinguish among cacha?a samples stored in different wood casks but also monitoring changes in their chemical composition according to the aging time.