Fast quantitative determination of methylphenidate levels in rat plasma and brain ex vivo by MALDI-MS/MS.

This study presents a simple and sensitive high-throughput matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry (MALDI-MS/MS) method for ex vivo quantification of methylphenidate (MPH) in rat plasma and brain. The common MALDI matrix alpha-cyano-4-hydroxycinnamic acid was used to obtain an optimal dried droplet preparation. For method validation, standards diluted in plasma and brain homogenate prepared from untreated (control) rats were used. MPH was quantified within a concentration range of 0.1-40 ng/ml in plasma and 0.4-40 ng/ml in brain homogenate with an excellent linearity (R2 ≥ 0.9997) and good precision. The intra-day and inter-day accuracies fulfilled the FDA's ±15/20 critera. The recovery of MPH ranged from 93.8 to 98.5% and 87.2 to 99.8% in plasma and homogenate, respectively. We show that MPH is successfully quantified in plasma and brain homogenate of rats pre-treated with this drug using the internal standard calibration method. By means of this method, a linear correlation between plasma and brain concentration of MPH in rodents pre-treated with MPH was detected. The simple sample preparation based on liquid-liquid extraction and MALDI-MS/MS measurement requires approximately 10 s per sample, and this significantly reduces analysis time compared with other analytical methods. To the best of our knowledge, this is the first MALDI-MS/MS method for quantification of MPH in rat plasma and brain. Copyright © 2015 John Wiley & Sons, Ltd.

Fast quantitative determination of melamine and its derivatives by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

A simple, sensitive and fast method for the determination of melamine and its derivatives in milk powder using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) was developed. Neither time-consuming sample preparation, nor special target plates, or other extra equipment are necessary. The common matrix sinapinic acid (SA) was used with a dried-droplet preparation. Detection limits (signal-to-noise (S/N) ratio = 3) for standard solutions of melamine, ammeline and cyanuric acid were 10, 25 and 10 µg/L, respectively. The limit of quantification (LOQ) for melamine was 25 µg/L and excellent linearity (R(2): 0.9990) was maintained over the range of 10-2000 µg/L. Ammeline and cyanuric acid were analyzed with an LOQ of 50 µg/L and also excellent linearity (R(2): 0.9997 and R(2): 0.9998). Good accuracy and precision were obtained for all concentrations within the range of the standard curve. The developed method was successfully used for the determination of melamine, ammeline and cyanuric acid in milk powder samples with a simple sample preparation. The LOQ of melamine was 0.5 µg/g. Ammeline and cyanuric acid were detectable at 0.5 and 5 µg/g. This method showed excellent accuracy, precision and linearity and significantly reduces the needed analysis time, as only approximately 10 s/sample measuring time is required. To the authors' knowledge, this is the first published method to quantify melamine and derivatives by MALDI-TOF-MS.

Quantitative determination of acetylcholine and choline in microdialysis samples by MALDI-TOF MS.

A simple, sensitive, and fast matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) method was developed for the determination of acetylcholine (ACh) and choline (Ch) in microdialysis samples. An optimized dried droplet preparation with the common matrix alpha-cyano-4-hydroxycinnamic acid (CHCA) was used. Limits of detection [signal-to-noise ratio (S/N) = 3] of 0.3 fmol/microL for ACh and 20 fmol/microL for Ch were found using standards diluted in artificial cerebrospinal fluid (aCSF). The limit of quantification (LOQ) for ACh was 1 fmol/microL, and excellent linearity (R(2) = 0.9996) was maintained over the range of 1-1000 fmol/microL. Choline was quantified over the range of 0.1-50 pmol/microL, also with excellent linearity (R(2) = 0.9995). Good accuracy and precision were obtained for all concentrations within the range of the standard curve. The developed method was successfully used for the determination of ACh and Ch in mouse brain microdialysis samples. The samples were quantified by a calibration curve and also by the method of standard addition. Despite the high salt content of the perfusion fluid (>150 mM), a direct measurement was possible. To the authors' knowledge, this is the first published method to determine acetylcholine and choline in microdialysis samples by MALDI-TOF MS. The method presented significantly reduces the needed analysis time, as only approximately 10 s/sample is required, and it is also possible to improve the temporal resolution, because only approximately 1 microL of sample is needed.

Rapid simultaneous quantitative determination of different small pharmaceutical drugs using a conventional matrix-assisted laser desorption/ionization time-of-flight mass spectrometry system.

The present study establishes a simple, rapid and sensitive method for the simultaneous quantification of different small pharmaceutical drugs using a matrix-assisted laser desorption/ionization source (MALDI) coupled with a time-of-flight (TOF) mass analyzer. Neither time-consuming sample preparation, nor special target plates, isotopically labelled internal standards or other extra equipment are necessary. A simple standard dried-droplet preparation with the common matrix alpha-cyano-4-hydroxycinnamic acid (CHCA) was used. The background signals of CHCA in the low-mass region did not pose the presumed problem, because the sensitivity, resolution and mass accuracy of a modern MALDI-TOF MS system is sufficient to overcome this difficulty. Four experiments were performed in order to verify the quantification method. First, ten different phenothiazines were quantified in the range of 5-2000 nM (1-880 ng/mL). A good precision (relative standard deviation (RSD) 4.4-9.3%), linearity (R2 >0.99) and accuracy (error 4.7-11%) was obtained in all cases. Additionally, simultaneous quantification of these ten phenothiazines was carried out in human plasma without prior chromatographic separation in the range of 2-1750 ng/mL yielding good linearity, precision and accuracy (mean RSD 7.6%; R2 >0.99, mean error 8.0%). Accordingly, a quantitative analysis of ten chemically and pharmaceutically unrelated drugs was performed in the same way. A comparable linearity (R2 >0.99), precision (mean RSD 7.6%) and accuracy (mean error 8.3%) was obtained in the range of 5-2000 nM. Finally, the prazosin content of a commercial tablet was directly determined without further purification steps.

Characterization and quantification of anthocyanins in selected artichoke (Cynara scolymus L.) cultivars by HPLC-DAD-ESI-MSn.

The anthocyanin pattern of artichoke heads (Cynara scolymus L.) has been investigated by high-performance liquid chromatography-electrospray ionization mass spectrometry. For this purpose a suitable extraction and liquid chromatographic method was developed. Besides the main anthocyanins-cyanidin 3,5-diglucoside, cyanidin 3-glucoside, cyanidin 3,5-malonyldiglucoside, cyanidin 3-(3''-malonyl)glucoside, and cyanidin 3-(6''-malonyl)glucoside-several minor compounds were identified. Among these, two peonidin derivatives and one delphinidin derivative were characterized on the basis of their fragmentation patterns. To the best of our knowledge this is the first report on anthocyanins in artichoke heads consisting of aglycones other than those of cyanidin. Quantification of individual compounds was performed by external calibration. Cyanidin 3-(6''-malonyl)glucoside was found to be the major anthocyanin in all the samples analyzed. Total anthocyanin content ranged from 8.4 to 1,705.4 mg kg(-1) dry mass.