Pressure Chemical Ionization Mass Research Articles

Rapid Identification for Authenticity of Mutton by DAPCI-MS

Following the method of surface desorption atmospheric pressure chemical ionization mass spectrometry(DAPCI-MS),without any pretreatment to the samples,the mass spectra of a series of meat,including pure samples(duck,beef,pork,and mutton)and mutton-flavored meat samples of duck,beef and pork,were obtained directly.The ionization part relied on a self-made equipment of DAPCI.The DAPCI-MS raw data were further analyzed by principle component analysis(PCA).The result shows that the mass spectra of duck,beef,pork,mutton and mutton-flavored fake meat samples are distinguishing,indicating that the PCA model can discriminate the different meat samples successfully.This method can achieve the rapid identification as well as rapid analysis of meat samples at high throughput without any pretreatment.This study provides a method that achieves rapid detection of fake mutton and identification of meat samples for food security.

Separation of enantiomeric triacylglycerols by chiral-phase HPLC.

Liquid chromatography-atmospheric pressure chemical ionization mass spectrometry (LC-MS/APCI) with a chiral phase was used for separation of triacylglycerols (TAG) obtained either by organic synthesis or isolated from different algal species. We present chromatographic characteristics and tandem mass spectra of enantiomers and positional isomers (regioisomers) of C16, C18 and C20 polyunsaturated fatty acids. The retention time was found to depend on the structure of the given TAG, increasing with increasing number of double bonds and decreasing with increasing number of the carbons in TAG, with the exception of dieicosapentaenoyl-palmitoyl-glycerols.

Enhanced detection of the algal toxin PbTx-2 in marine waters by atmospheric pressure chemical ionization mass spectrometry.

Karenia brevis, a marine dinoflagellate, biosynthesizes a unique class of polyether toxins called brevetoxins that produce significant health, environmental and economic impacts in and along coastal waters. Previous application of liquid chromatography/mass spectrometry for detection of the most common brevetoxin, PbTx-2, has relied almost exclusively upon electrospray ionization (ESI). A different ionization source is proposed in this study with improved sensitivity ultimately leading to lower limit of detection compared to (+) ESI. Brevetoxin standards and samples (PbTx-2) were analyzed by liquid chromatography/mass spectrometry using both (+) atmospheric pressure chemical ionization and (+) electrospray ionization sources. LC/MS with (+) APCI exhibited an order of magnitude improvement in the limit of detection (7.7 × 10(-4) pg mass on-column) compared to the same method using (+) ESI (7.5 × 10(-3) pg mass on-column). The calibration sensitivity of (+) APCI (1.3 × 10(3)) was also five times higher than positive mode (+) ESI (0.26 × 10(3)). Positive mode APCI represents a significant improvement in detection and quantification of PbTx-2 by LC/MS allowing for smaller sample sizes compared to previous studies using (+) ESI. This in turn leads to higher throughput of samples during and after bloom events giving stakeholders detailed information on the fate of this potent marine toxin.

Profiling of triacylglycerols in plant oils by high-performance liquid chromatography-atmosphere pressure chemical ionization mass spectrometry using a novel mixed-mode column.

In this investigation, a rapid and high-throughput method for profiling of TAGs in plant oils by liquid chromatography using a single column coupled with atmospheric pressure chemical ionization (APCI) mass spectrometry was reported. A novel mixed-mode phenyl-hexyl chromatographic column was employed in this separation system. The phenyl-hexyl column could provide hydrophobic interactions as well as π-π interactions. Compared with two traditionally columns used in TAG separation - the C18 column and silver-ion column, this column exhibited much higher selectivity for the separation of TAGs with great efficiency and rapid speed. By comparison with a novel mix-mode column (Ag-HiSep OTS column), which can also provide both hydrophobic interactions as well as π-π interactions for the separation of TAGs, phenyl-hexyl column exhibited excellent stability. LC method using phenyl-hexyl column coupled with APCI-MS was successfully applied for the profiling of TAGs in soybean oils, peanut oils, corn oils, and sesame oils. 29 TAGs in peanut oils, 22 TAGs in soybean oils, 19 TAGs in corn oils, and 19 TAGs in sesame oils were determined and quantified. The LC-MS data was analyzed by barcodes and principal component analysis (PCA). The resulting barcodes constitute a simple tool to display differences between different plant oils. Results of PCA also enabled a clear identification of different plant oils. This method provided an efficient and convenient chromatographic technology for the fast characterization and quantification of complex TAGs in plant oils at high selectivity. It has great potential as a routine analytical method for analysis of edible oil quality and authenticity control.

CHR-insertion (R=H, CH3) into cyclohexyl-substituted silsesquioxanes: reactivity and decomposition studies.

Amorphous silica plays an important role in heterogeneous catalysis as a support and is frequently presumed to be "inert". The structure of the supported catalyst is key to understanding the stability and reactivity of catalytic systems. To provide vital insights into the surface reactivity of silica, Polyhedral oligomeric silsesquioxanes (POSSs) can act as realistic homogeneous molecular models for silica surfaces. Here, we report novel reactivities associated with the silica surface, derived from our insights obtained by means of such model systems with potentially significant implications in catalysis when employing silica-supported catalysts. In this work, the gas-phase reactivities of two cyclohexyl-substituted POSSs, namely the completely condensed triganol prism [Si6cy6O9] (a6b0), and the incompletely-condensed partial cube [Si7cy7O9(OH)3] (a7b3), with cy = c-C6H11, were studied by using atmospheric pressure chemical ionisation (APCI) and collision-induced decomposition (CID) spectroscopies. Silsesquioxane a6b0, containing three-membered rings, was found to be much more reactive, undergoing novel CH2-insertion on reaction with gas phase molecules-a reaction not observed for a7b3, containing only four-membered rings. Both silsesquioxanes displayed the ability to trap ammonia formed in situ within the mass spectrometer from N2 in the instrument. This work also demonstrates the applicability of APCI and the role of CID in elucidating reactive POSS structures, highlighting novel gas-phase reactivities of POSS.

Inter-individual retronasal aroma release variability during cheese consumption: Role of food oral processing

The aim of our study was to explain inter-individual differences on in vivo aroma release during cheese consumption by oral physiological parameters.To reach this objective, 34 subjects were recruited. Their salivary flow, oral volume and velum opening were determined. Six cheddar-based melted cheeses with different fat levels and firmness were flavoured with nonan-2-one (NO) and ethyl propanoate (EP). During their consumption (free protocol), in vivo retro nasal aroma release was followed by Atmospheric Pressure Chemical Ionisation–Mass Spectrometry (APCI–MS). Chewing activity was evaluated by electromyography recordings. Bolus saliva content, mouth-coating, and bolus rheology were also determined.Based on the quantity of aroma released before and after swallowing, subjects can be clustered into three groups: the first one (HRG) is characterized by a large quantity of aroma release whatever the aroma compound; the second one (MRG) showed a large release for EP and a lower one for NO; the third group (LRG) was characterized by a low quantity of aroma release whatever the compound.Whatever the group of subjects, fat and firmness effects differed according to the aroma compound. EP release increased with firmness and fat content, whereas NO release was not affected by firmness and decreased when fat content increased. Physiological parameters which better differentiated the three groups of subjects according to their release behaviour were chewing activity, mouth coating and frequency of velum opening.Subjects from HRG were differentiated from LRG subjects by a higher chewing activity, and more frequent velum opening. Subjects from MRG presented a lower mouth coating explaining their lower release of NO, the more hydrophobic compound.This study shows that the total amount of aroma released in the nasal cavity during food consumption depends not only on the characteristics of the product but also on the oral physiology of the subjects and on their food oral processing.

Determination of Fatty Acids in Three Nitraria Species by Precolumn Fluorescence Labeling for High-Performance Liquid Chromatography and Atmospheric Pressure Chemical Ionization–Mass Spectrometry

A novel fluorescence method using 2-(7-methyl-1H-pyrazolo-[3,4-b] quinoline-1-yl) ethyl-4-methyl benzenesulfonate as the labeling reagent was established for high-performance liquid chromatography determination of fatty acids. The conditions were optimized, including the identity of the organic solvent, identity, and amount of catalyst, amount of derivatization reagent, derivatization temperature, and derivatization time. The results indicated that quantitative yields of derivative were obtained with a five-fold molar excess of reagent at 90°C for 30 min using 25 mg of potassium carbonate as the catalyst. Atmospheric pressure chemical ionization mass spectrometry results indicated that collision-induced dissociation of protonated fatty acid derivatives produced fragments at m/z 228.2, m/z 210.2, and m/z 183.8. The method was validated in terms of linearity, limits of detection, limits of quantification, precision, and accuracy, and the results showed that the method exhibited excellent sensitivity, selectivity, and reproducibility. Limits of detection and limits of quantification were in the ranges of 0.52–2.34 ng mL−1 and 1.23–6.63 ng mL−1, respectively. This method was successfully applied to the determination of fatty acids in sarcocarps, seeds, and leaves of Nitraria tangutorum Bobr., Nitraria sibirica Pall., and Nitraria roborowskii Kom. The results indicated that the main components were oleic, linoleic, linolenic, and hexadecanoic acids. However, the composition of fatty acids in the tissues varied considerably.

Effect of processing conditions on the content of cis/trans carotene isomers as provitamin A carotenoids in Korean sweet potato varieties

The present investigation intends to evaluate the changes in the content of cis/trans carotene isomers as provitamin A carotenoids by steaming and roasting processes in the roots of four Korean sweet potato varieties viz. Shinzami, Younwhangmi, Chuwhangmi and Jinhongmi using a liquid chromatography with diode array detection and the negative ion atmospheric pressure chemical ionization mass spectrometric (LC-DAD-APCI/MS) method and UV spectral pattern library created from several reference data. Except Shinzami, the content of all trans β-carotenes was found to slightly decreased or remained constant when steamed or roasted. The content of cis α-/β-carotenes was potentially increased about 2-fold or greater when raw or steamed and the content was slightly decreased while roasted. In Chuwhangmi, the content of 13-cis α-carotene and all trans α-carotenes were rapidly increased when steamed and slightly decreased when roasted. Chuwhangmi exhibited 27.2 mg/100 g DW content of all trans β-carotenes when roasted and thus, it was considered as a relatively superior cultivar.

Extract-filter-shoot liquid chromatography with mass spectrometry for the analysis of vitamin D2 in a powdered supplement capsule and standard reference material 3280.

An "extract-filter-shoot" method for the analysis of vitamin D2, ergocalciferol, in a dry powdered dietary supplement capsule containing rice flour excipient and in a National Institute of Standards and Technology standard reference material 3280 is reported. Quantification of vitamin D2 was done by atmospheric pressure chemical ionization mass spectrometry using selected ion monitoring, two transitions of selected reaction monitoring, and extracted ion chromatograms from full scans. UV detection was used for the quantification of Vitamin D2 in the dry powder capsule, whereas interfering species rendered UV detection unreliable for standard reference material 3280. Average values for standard reference material 3280 ranged from 8.27 ± 0.58 to 8.33 ± 0.57 μg/g using internal standard calibration and response factor approaches, compared to the previous National Institute of Standards and Technology internal value for vitamin D2 of 8.78 ± 0.11 μg/g, and the recently updated reference value of 8.6 ± 2.6 μg/g. The powdered supplement capsule was found to contain 28.19 ± 0.35 to 28.67 ± 0.90 μg/capsule for a capsule labeled to contain 25.00 μg. The triacylglycerol composition of the rice flour excipient in the powdered supplement capsule determined by atmospheric pressure chemical ionization mass spectrometry is also reported.

A new flavonol glycoside from the medicinal halophyte Suaeda fruticosa

A new flavonol glycoside, namely 3-(α-rhamnopyranosyl-(1 → 2)-[β-xylopyranosyl-(1 → 6)]-β-glucopyranosyloxy) isorhamnetin was reported from methanol extracts of aerial parts of Suaeda fruticosa for the first time. In this work, liquid chromatography coupled to atmospheric pressure chemical ionisation mass spectrometry, high-resolution mass spectrometry and nuclear magnetic resonance spectroscopy were used to identify this new compound. Structure was elucidated on the basis of extensive spectroscopic analysis, including HSQC, HMBC and 1H–1H COSY. Antioxidant potentialities of a pure compound were evaluated. The estimation of antioxidant capacities using oxygen radical absorbance capacity (ORAC method) and a cell based-assay (WS1) indicated that this new flavonol exhibited the highest antioxidant activities with an ORAC value of 5.0 ± 0.3 μmol Trolox/μmol and inhibited the tBH-induced oxidation of 2′,7′-dichlorofluorescin with an IC50 value of 4.9 ± 0.6 μM.

Quantification of levornidazole and its metabolites in human plasma and urine by ultra-performance liquid chromatography-mass spectrometry.

We developed and validated an ultra-performance liquid chromatographic (UPLC) method coupled with atmospheric pressure chemical ionization (APCI) mass spectrometry for simultaneous determination of levornidazole and its first-pass metabolites, l-chloro-3-(2-hydroxymethyl-5-nitro-l-imidazolyl)-2-propanol (Ml), 2-methyl-5-nitroimidazole (M2) and 3-(2-methyl-5-nitro-1-imidazolyl)-1,2-propanediol (M4), in human plasma and urine. The biological samples were pretreated by protein precipitation and liquid-liquid extraction and analyzed using an ACQUITY UPLC CSH C18 column (2.1×50 mm, 1.7 μm) and a QTRAP mass spectrometer in multiple reaction monitoring mode via APCI. Acetonitrile and 0.1% formic acid in water was used as the mobile phase in gradient elution at a flow rate of 0.6 mL/min. The lower limit of quantification of this method was 0.0100, 0.00500, 0.0200 and 0.00250 μg/mL for levornidazole, M1, M2 and M4, respectively. The linear calibration curves were obtained for levornidazole, M1, M2, and M4 over the range of 0.0100-5.00, 0.00500-2.50, 0.0200-10.0 and 0.00250-1.25 μg/mL, respectively. The intra- and inter-batch precision was less than 12.2% in plasma and less than 10.8% in urine. The intra- and inter-batch accuracy was 87.8-105.7% in plasma and 92.8-109.2% in urine. The mean recovery of levornidazole, M1, M2 and M4 was 91.1-105.1%, 95.8-103.8%, 87.8-96.8%, 96.8-100.6% from plasma and 96.0-100.9%, 96.9-107.9%, 95.1-102.7%, 103.7-105.9% from urine respectively. This method was validated under various conditions, including room temperature, freeze-thaw cycles, long-term storage at -40 ± 5°C, after pretreatment in the autosampler (at 10°C), and 10- and 100-fold dilution. This newly established analytical method was successfully applied in a pharmacokinetic study following single intravenous infusion of levornidazole in 24 healthy Chinese subjects.

A method for simultaneous analysis of phytosterols and phytosterol esters in tobacco leaves using non aqueous reversed phase chromatography and atmospheric pressure chemical ionization mass spectrometry detector

While numerous analytical methods for phytosterols have been reported, the similar polarity and large molecules of phytosterol esters have made the methods lengthy and complicated. For this reason, an analytical method that could completely separate phytosterol esters including the higher fatty acids such as palmitic acid, stearic acid, oleic acid, linoleic acid and linolenic acid in addition to phytosterols without preliminary separation was developed. The separation was accomplished by non-aqueous reversed phase chromatography technique using only acetone and acetonitrile. An atmospheric pressure chemical ionization/mass spectrometry detector configured at selected ion monitoring mode was hyphenated with the separation system to detect phytosterols and phytosterol esters. Twenty-four types of these were consequently separated and then identified with their authentic components. The calibration curve was drawn in the range of about 5 to 25,000ng/mL with a regression coefficient over 0.999. The limit of detection and limit of quantification, respectively, ranged from 0.9 to 3.0ng/mL and from 3.0 to 11.0ng/mL. Recovery rates ranged from 80 to 120%. The quantification results were subjected to statistical analysis and hierarchical clustering analysis, and were used to determine the differences in the amounts of phytosterols and phytosterol esters across tobacco leaves. The newly developed method succeeded in clarifying the whole composition of phytosterols and phytosterol esters in tobacco leaves and in explaining compositional differences across the variety of tobacco leaves.

Chaos Entropy Index Combined with Desorption Atmospheric Pressure Chemical Ionization Mass Spectrometry for Rapid Analysis of Complex Viscous Sample, Edible Oil

Abstract By means of surface desorption atmospheric pressure chemical ionization mass spectrometry (DAPCI-MS), mass-spectra fingerprints of various oil samples were rapidly recorded in the mass range of m/z 50–1000 without any sample pretreatment in negative ion mode. A new spectrogram analysis method (Chaos Entropy Index) for identifying complex mixtures was established and applied to the authentication of expired oil. Multivariate statistics analysis showed that expired edible oils can be successfully separated from fresh edible oils with 100% accuracy on the basis of Global CEIs and the most significant mass ranges presenting major feature peaks of expired oil were determined based on Local CEIs. These experimental findings demonstrate that as a rapid, simple, and accurate analytical method, CEI approach in conjunction with DAPCI-MS has an extensive prospect for differential analysis of complex viscous mixtures.

Accurate quantitation of pentaerythritol tetranitrate and its degradation products using liquid chromatography–atmospheric pressure chemical ionization–mass spectrometry

After an explosion of pentaerythritol tetranitrate (PETN), its degradation products pentaerythritol trinitrate (PETriN), dinitrate (PEDiN) and mononitrate (PEMN) were detected using liquid chromatography–atmospheric-pressure chemical-ionization–mass spectrometry (LC–APCI–MS). Discrimination between post-explosion and naturally degraded PETN could be achieved based on the relative amounts of the degradation products. This information can be used as evidence when investigating a possible relationship between a suspect and a post-explosion crime scene. The present work focuses on accurate quantitation of PETN and its degradation products, using PETriN, PEDiN and PEMN standards specifically synthesized for this purpose. With the use of these standards, the ionization behavior of these compounds was studied, and a quantitative method was developed. Quantitation of PETN and trace levels of its degradation products was shown to be possible with accuracy between 85.7% and 103.7% and a precision ranging from 1.3% to 11.5%. The custom-made standards resulted in a more robust and reliable method to discriminate between post-explosion and naturally-degraded PETN.

Formation of deaminated dimer species of amino acids by atmospheric pressure chemical ionization

Interactions of biological molecules to form cluster species play a key role in biological processes and investigation of non-covalent complexes is one of the research fields using mass spectrometry. Atmospheric pressure chemical ionization mass spectrometry (APCI-MS) is a useful method for the investigation of cluster formation of amino acids (AAs) by ion-molecule reactions. A mixture of 20 protein AAs was ionized by APCI and the product ions were analyzed. The ionization was performed in the positive and negative ion modes. Formation of the homo- and heterocluster ions of AAs was investigated. Mechanism for the formation of AA homo- and heterocluster ions was examined using hydrogen/deuterium (H/D)-exchange experiments. In the positive ion mode, of the dimer species only the [2Pro+H](+) ion was detected. In the negative ion mode, the [2M - H](-) ions of His, Val, Ser, and Gln were observed. The deaminated dimers such as the [2Gln - H - NH3](-) and [His + Gln - H - NH3](-) ions were also observed. In the negative ion mass spectra of the His/Arg, His/Asn, and His/Lys binary mixture solutions, the [His + AA - H - NH3](-) ions of Asn, Arg, and Lys were also detected. The number and abundances of the negative product ions were much greater than those of the positive ones. Mechanism for the formation of [2Gln - H - NH3](-) and [His+AA - H - NH3](-) was examined by deuterium replacement of the amine and hydroxide groups to distinguish the deamination and dehydration reactions with a single quadrupole mass spectrometer. The [His + AA - H - NH3](-) ion is formed by ion-molecule reaction between the [His-H](-) ion and a neutral AA.

A novel derivatization method for the determination of Fosfomycin in human plasma by liquid chromatography coupled with atmospheric pressure chemical ionization mass spectrometric detection via phase transfer catalyzed derivatization

An analytical method employing novel sample preparation and liquid chromatography coupled with atmospheric pressure chemical ionization mass spectrometric detection (LC–APCI/MS) was developed for the determination of fosfomycin in human plasma. Sample preparation involves derivatization through phase transfer catalysis (PTC) which offers multiple advantages due to the simultaneous extraction, preconcentration and derivatization of the analyte. Using a PT catalyst, fosfomycin was extracted from plasma in an organic phase and, then converted to a pentafluorobenzyl ester with the use of pentafluorobenzyl bromide (PFBBr) derivatization reagent. The method was fully optimized by taking into account both PTC and derivatization parameters. Several catalysts, in a wide range of concentrations, with different counter ions and polarities were tested along with different extraction solvents and pH values. Thereafter, the derivatization procedure was optimized by altering the amount of the derivatization reagent, the temperature of the reaction and finally, the derivatization duration. As internal standard (I.S.) ethylphosphonic acid was chosen and underwent the same pretreatment. The derivatives were separated on a pentafluorophenyl (PFP)-C18 analytical column, which provides unique selectivity, using an isocratic elution with acetonitrile–water (70–30, v/v). The method was validated according to US Food and Drug Administration (FDA) guidelines and can be used for a bioequivalence study of fosfomycin in human plasma. The correlation coefficient (r2) of the calibration curve of spiked plasma solutions in the range of 50–12000ng/mL was found greater than 0.999 with a limit of quantitation (LOQ) equal to 50ng/ml (for 500μL plasma sample).

CHEMOMETRIC APPROACHES FOR THE CHARACTERIZATION OF TURKISH RHODODENDRON AND HONEYDEW HONEYS DEPENDING ON AMINO ACID COMPOSITION

The amino acid composition of twenty-five honey samples obtained from different regions of Turkey (rhododendron and honeydew honey) was determined by a liquid chromatography–atmospheric pressure chemical ionization–mass spectrometry (LC–APCI–MS). Principal component analysis (PCA) and hierarchical cluster analysis (HCA) were used to characterize and classify the honey samples. In rhododendron honeys, major amino acids were detected as aspartic acid, lysine, and arginine, for honeydew honeys, lysine, arginine, and histidine were found, following the proline. Five principal components (PCs) and four main clusters were obtained from the chemometric analysis that classifies the samples. Five PCs were detected to be explanatory of more than 79% of the total variance in the data matrix. It can be concluded that free amino acids are good indicators for determination of botanical origins and chemometric studies are highly useful for honey characterization.

Rapid differentiation of microbial cultures based on the analysis of headspace volatiles by atmospheric pressure chemical ionization mass spectrometry

APCI-MS fingerprinting of headspace volatiles was used to differentiate microbial cultures without any sample pretreatment, chromatographic separation and wet chemistry.

Development and validation of a shipboard system for measuring high-resolution vertical profiles of aqueous dimethylsulfide concentrations using chemical ionisation mass spectrometry

Environmental context Dimethylsulfide, a trace gas produced by oceanic plankton, is a key chemical species in the global cycles of sulfur and aerosols, with implications that span marine ecology to climate regulation. Knowledge of what governs dimethylsulfide production in the surface ocean depends on our ability to measure concentration changes over time and depth. We describe a sampling and analytical system that provides continuous shipboard measurements of dimethylsulfide concentrations in high-resolution vertical profiles. Abstract A sampling and analytical system has been developed for shipboard measurements of high-resolution vertical profiles of the marine trace gas dimethylsulfide (DMS). The system consists of a tube attached to a conductivity–temperature–depth (CTD) probe with a peristaltic pump on deck that delivers seawater to a membrane equilibrator and atmospheric pressure chemical ionisation mass spectrometer (Eq-APCIMS). This allows profiling of DMS concentrations to a depth of 50m, with a depth resolution of 1.3–2m and a detection limit of nearly 0.1nmolL–1. The seawater is also plumbed to allow parallel operation of additional continuous instruments, and simultaneous collection of discrete samples for complementary analyses. A valve alternates delivery of seawater from the vertical profiler and the ship’s underway intake, thereby providing high-resolution measurements in both the vertical and horizontal dimensions. Tests conducted on various cruises in the Mediterranean Sea, Atlantic, Indian, and Pacific Oceans show good agreement between the Eq-APCIMS measurements and purge and trap gas chromatography with flame photometric detection (GC-FPD) and demonstrate that the delivery of seawater from the underway pump did not significantly affect endogenous DMS concentrations. Combining the continuous flow DMS analysis with high-frequency hydrographic, optical, biological and meteorological measurements will greatly improve the spatial–temporal resolution of seagoing measurements and improve our understanding of DMS cycling.

Characterization of potential degradation products in a PEGylating reagent 20 kDa monomethoxy polyethylene glycol propionaldehyde by RP-HPLC, APCI-MS and NMR

Ensuring quality of PEGylating reagents is essential for the successful development and manufacturing of PEGylated biopharmaceuticals. However, little is known about how to maintain and verify the quality of PEG raw materials for PEGylated protein manufacturing. In this study, monomethoxy polyethylene glycol propionaldehyde (mPEG-aldehyde) was subjected to conditions that mimic accelerated stability conditions. Separation of trace-level degradation products in the presence of mPEG-aldehyde was achieved by derivatization with 2,4-dinitrophenylhydrazine (DNPH), followed by reversed phase high performance liquid chromatography with ultraviolet detection (RP-HPLC-UV) at 355nm. Structural characterization by atmospheric pressure chemical ionization mass spectrometry (APCI-MS) identified formaldehyde, acetaldehyde, crotonaldehyde, acrolein, benzaldehyde, and tolualdehyde as major degradation products or process-related impurities. The presence of formaldehyde and acrolein was confirmed by 1H NMR in the forced degraded mPEG-aldehyde samples without derivatization of mPEG-aldehyde. Findings from this study imply that reactive impurities could form as a result of inappropriate mPEG-aldehyde handling or storage. Further, a rapid screening method based on reversed phase HPLC was shown to be an effective screening assay used for routine screening of mPEG-aldehyde to ensure consistent PEGylated protein product quality.