Pass-through Cleanup Research Articles

Analysis of 15 bile acids in human plasma based on C18 functionalized magnetic organic polymer nanocomposite coupled with liquid chromatography-tandem mass spectrometry

Because of the “enterohepatic circulation” of bile acid, liver damage can be reflected by monitoring the content of bile acid in the serum of the organism. To monitor the concentration of 15 bile acids in plasma samples, a new technique of PRiME (process, ruggedness, improvement, matrix effect, ease of use) pass-through cleanup procedure combined with high performance liquid chromatography-tandem quadrupole mass spectrometry (HPLC-MS/MS) was developed. The sorbent used in the PRiME pass-through cleanup procedure is a new type of magnetic organic resin composite nano-material modified by C18 (C18-PS-DVB-GMA-Fe3O4), which has high cleanup efficiency of plasma samples. It also shows good performance in the separation and analysis of 15 kinds of bile acids. Under the optimal conditions, the results show higher cleanup efficiency of C18-PS-DVB-GMA-Fe3O4 with recoveries in the range of 82.1–115 %. The limit of quantitative (LOQs) of 15 bile acids were in the range of 0.033 µg/L-0.19 µg/L, and the RSD values of 15 bile acids were in the range of 3.00–11.9 %. Validation results on linearity, specificity, accuracy and precision, as well as on the application to analysis of 15 bile acids in 100 human plasma samples demonstrate the applicability to clinical studies.

Simultaneous determination of 12 illicit drugs in hair sample by the nano-titania-modified covalent organic frameworks (NTM-COFs) on-line pass-through cleanup followed by UPLC-MS/MS

Simultaneous determination of 12 illicit drugs in hair sample by the nano-titania-modified covalent organic frameworks (NTM-COFs) on-line pass-through cleanup followed by UPLC-MS/MS

Simultaneous determination of 12 illicit drugs in human plasma by the PRiME clover-shaped nano-titania functionalized covalent organic frameworks pass-through cleanup procedure followed by ultra-performance liquid chromatography-tandem mass spectrometry

A rapid and sensitive method based on PRiME (process, robustness, improvements, matrix effects, ease of use) pass-through cleanup procedure and ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) for the simultaneous determination of 12 illicit drugs in human plasma was developed and validated. The clover-shaped nano-titania functionalized covalent organic frameworks (CSTF-COFs) has been evaluated in the PRiME pass-through cleanup procedure to remove blood phospholipids from plasma samples. Ion suppression effects can be dramatically reduced by CSTF-COFs based PRiME pass-through cleanup procedure. Under the optimal conditions, the results showed satisfactory recoveries between 85.8% and 109%. Acceptable precision and accuracy were also obtained with RSD values less than 15.0% and RE values below 13.3%. The limits of detections (LODs) of 12 illicit drugs were in the range of 0.018–0.360 µg/L. Furthermore, the PRiME CSTF-COFs cartridge could be conveniently regenerated and reused for 40–50 cycles. The proposed method was applied to real plasma samples from suspected drug abusers, which was proved to be reliable and robust for drug screening in clinical and forensic toxicology.

Nano-titania modified cupric-based metal–organic frameworks for the on-line pass-through cleanup of phenothiazine drugs and metabolites in human plasma

SEM (A) and TEM (B) images of NTMCu-MOFs.

A rapid method for the simultaneous stereoselective determination of the triazole fungicides in tobacco by supercritical fluid chromatography-tandem mass spectrometry combined with pass-through cleanup

This work presents a simple, rapid and green chiral analysis method for five triazole fungicides (penconazole, tebuconazole, triadimefon, myclobutanil, and triadimenol) in tobacco, by which the samples were cleaned up by the novel pass-through solid phase extraction and subsequently the stereoisomers were separated and determined by the supercritical fluid chromatography-tandem mass spectrometry (SFC-MS/MS). Optimized separation of the stereoisomers was achieved on an ACQUITY UPC2 Trefoil AMY 1 column within 6 min. Under fortified concentration levels of 0.1, 0.5 and 2.0 mg/kg, the mean recoveries were 82.8-106.6%, the intra-day relative standard deviations (RSDs) were 1.1-6.6%, and the inter-day RSDs were 2.5-5.6%. The correlation coefficient was greater than 0.9926 for all studied analytes within the range of 10-500 ng/mL. The limits of detection (LODs) for all stereoisomers ranged from 0.26 μg/kg to 3.24 μg/kg. The established method was subsequently successfully applied to analyze authentic samples, confirming that this method is a novel, rapid and environmentally friendly method for the stereoselective separation of triazole fungicides in tobacco.

Development of a Pass-through SPE Cartridge for the Rapid Determination of Fipronil and Its Metabolites in Chicken Eggs by LC-MS/MS

In this work, a novel pass-through cleanup cartridge was developed for the rapid determination of fipronil and its metabolites in chicken eggs by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The systemic elimination experiments on pigments, lipids, proteins, and so on were performed, and then a multi-function SPE cartridge containing three sorbents (PestiCarb, Alumina-N, and COOH) was designed. Using this cartridge, a simple cleanup procedure with the ideal elimination effects was achieved by gravity in less than 1 min. The involved LC-MS/MS method provided satisfactory performance. The tested recoveries were in the range of 78.6–87.1% with the relative standard deviations (RSDs) less than 8.3%. The limits of quantitation (LOQs) of fipronil, fipronil-desulfinyl, fipronil-sulfone, and fipronil-sulfide were 1, 0.5, 1, and 0.5 μg/kg, respectively. This method was successfully applied to detect some chicken eggs from local markets and farms. A positive sample of 5.3 μg/kg fipronil and 31.8 μg/kg fipronil-sulfone was found and the potential risk root was traced. In the future, a specific device equipped with a lot of the above-developed cartridges could be designed for the rapid and high-throughput cleanup. It would be helpful to reduce the manual labor and save the time of sample pretreatment.

Simultaneous determination of ten aminoglycoside antibiotics in aquatic feeds by high-performance liquid chromatography quadrupole-orbitrap mass spectrometry with pass-through cleanup.

A simple and sensitive method has been established based on pass-through cleanup and high-performance liquid chromatography quadrupole-orbitrap mass spectrometry (HPLC-Q/Orbitrap MS) for the simultaneous determination of ten aminoglycosides (AGs) in aquatic feeds. The extraction solution and cleanup procedure had been optimized, and good sensitivity, accuracy, and precision were obtained. The calibration curves of AGs were linearity (R2 > 0.99) in the range of 2.0 to 200 μg/L (or 5.0 to 500 μg/L). The limits of detection of AGs were between 10 and 25 μg/kg. The recoveries of AGs ranged from 74.9% to 94.3%, and the intraday and interday relative standard deviations were less than 15%. Finally, this method was successfully applied to determine ten AGs in 30 aquatic feed samples. It might be the first time to use pass-through cleanup approach combined with HPLC-Q/Orbitrap MS method for AGs determination in aquatic feed samples.

Matrix effect-free on-line pass-through cleanup procedure for the fast determination of local anesthetic drug by LC-MS/MS

A novel nano-titania modified covalent organic frameworks (NTM-COFs) has been synthesized and characterized by transmission electron microscopy (TEM) and scanning electron micrographs (SEM). Besides, NTM-COFs, as efficient sorbent, has also been evaluated in the on-line pass-through cleanup procedure prior to the analysis of local anesthetic drugs (lidocaine, bupivacaine and tetracaine) in human plasma by liquid chromatography-tandem quadrupole mass spectrometry (LC-MS/MS). Under optimum conditions, the level of matrix effects can be dramatically reduced by the NTM-COFs based on-line pass-through cleanup procedure, with acceptable recoveries ranging from 88.8% to 103%. Satisfactory trueness and precision of the proposed method were also obtained, with the within- and between-run RSDs less than 7.0% and relative error (REs) less than 12%. The limits of detections (LODs) of lidocaine, bupivacaine and tetracaine were 0.029 μg·L−1, 0.027 μg·L−1 and 0.016 μg·L−1, respectively. The analytical method has been successfully applied for the determination of the plasma concentrations of bupivacaine in five parturients who received an epidural administration of bupivacaine hydrochloride during vaginal delivery. Results demonstrate the applicability of the developed NTM-COFs on-line pass-through cleanup procedure coupled with LC-MS/MS method to clinical studies.

Simultaneous determination of eight microcystins in fish by PRiME pass-through cleanup and online solid phase extraction coupled to ultra high performance liquid chromatography-tandem mass spectrometry

A rapid and sensitive method was developed based on online solid phase extraction coupled to ultra high performance liquid chromatography-tandem mass spectrometry (online SPE-UHPLC-MS/MS) for the simultaneous determination of eight common microcystins (MCs) in fish. Analytes, including MC-LR, MC-RR, MC-LY, MC-LW, MC-YR, MC-WR, MC-LF and MC-LA, were extracted by the solvent of MeOH: water (80:20, v/v) with 0.1% FA, and the extracted solution was cleaned up by one-step PRiME (process, robustness, improvements, matrix effects, ease of use) procedure before online SPE-UHPLC-MS/MS analysis. After the optimization of sample pretreatment conditions and instrumental parameters, method validation experiments were carried out to evaluate the performance of the method Good linearities were obtained in the range of 0.5–20 μg/L with r2 larger than 0.99. The average recoveries of eight MCs spiked at 2, 10 and 20 μg/kg in fish samples were from 70.5% to 98.9% with relative standard deviations (RSDs) 1.3%–3.5% for repeatability and 2.2%–7.3% for reproducibility. Limits of detections (LODs) were ranged from 0.1 to 0.5 μg/kg and Limits of quantifications (LOQs) were in the range of 0.3–1.5 μg/kg. The developed method was further applied to the analysis of fish samples collected from Lake Taihu, China. MC-LR, MC-YR and MC-RR were detected in the range of 0.5–3.1 μg/kg. The experimental results proved the feasibility of the developed method for the identification and quantification of various MCs in fish products.

Multiply-functionalized graphene oxide on-line pass-through cleanup for the fast determination of aconitum alkaloids in human plasma by LC-MS/MS

A novel ionic liquid coupled with nano-titania coated modified magnetic GO composites (IL-Ti-Mag-GO) has been synthesized.

PRiME pass-through purification of lignans in Silybum marianum and UPLC−MS/MS analysis

A PRiME (process, robustness, improvements, matrix effects, ease of use) pass-through cleanup procedure was developed for the extraction and purification of silychristins A and B, silybins A and B, isosilybins A and B, and silydianin in Silybum marianum. After optimizing the extracting solvent types and the sample loading volume, the crude extract was diluted to 3 mL with 95% acetonitrile and then loaded on the PRiME cartridge. The eluate was analyzed by ultra-performance liquid chromatography and tandem mass spectrometry (UPLC-MS/MS). All the target analytes were deprotonated as [M-H]− at m/z 481 by conducting collision-induced dissociation (CID), and the major fragment ions were m/z 463 ([M-H2O-H]−), 453 ([M-CO-H]−), 355 ([M-C6H6O3-H]−), 301 ([M355-CO2-H]−), and 179 ([C10H11O3]−). Afterwards, this method was validated in terms of linearity (R2 > 0.9990), intra-day precision (1.02%–3.79%), inter-day precision (1.59%–4.87%), sensitivity (LOD ≤ 0.45 μg·kg−1 and LOQ ≤ 1.50 μg·kg−1), and recovery (76.9–103.4%, RSD < 8.90%). Finally, the proposed protocol was successfully applied to eight batches of S. marianum samples. The total content of the seven active compounds varied amongst the batches from different places of origin.

A Multi-Class, Multi-Residue Method for Detection of Veterinary Drugs in Multiple Meat Using a Pass-Through Cleanup SPE Technique and UPLC-MS/MS Analysis

A quantitative method using ultra-performance liquid chromatography coupled with tandem mass spectrometry (UPLC-MS/MS) was developed for the determination of 60 compounds, belonging to a variety of veterinary drug (VD) classes, in meat. The included analytes belong to the following VD classes: β-agonists, sulfanilamides, quinolones, macrolides, tetracyclines, β-lactams, nitroimidazoles, glucocorticoids, sex hormones, chloromycetins, sedatives, and olaquindox metabolite. The effective PRiME pass-through cleanup procedure was used to ensure high extraction efficiency and good sample cleanup after a simple liquid extraction of the meat samples with acetonitrile/water. The developed method was validated successfully. Mean recoveries for all analytes ranged from 80 to 116%, with the relative standard deviations (RSDs) ≤ 7.8%. Limits of quantification were in the range 0.05–3.0 μg kg−1 and limits of detection were in the range 0.1–10 μg kg−1. The matrix effect was evaluated for the different meat matrices and was found to be markedly different in different matrices. The validated method was used in a pilot study to analyze real samples of pork, beef, mutton, chicken, and pork liver, lambs’ liver, and chicken liver. Trace amounts of β-agonists, oxytetracycline, quinolones, chloromycetin sulfadimidine, and 3-methyl-quinoxaline-2-carboxylicacid were detected in these samples. In conclusion, this workflow can provide a simpler and more cost-effective alternative to conventional analytical methods and is compatible with processing large sample numbers in a short time period.

Hydrophilicity nano-titania coating modified magnetic graphene oxide for pass-through cleanup of fipronil and its metabolites in human blood

A novel hydrophilicity nano-titania coating modified magnetic graphene oxide (HTC-Mag-GO) has been synthesized. It has been evaluated in PRiME (process, robustness, improvements, matrix effects, ease of use) pass-through cleanup procedure for human blood prior to analysis of fipronil and its metabolites, i.e., fipronil sulphone, fipronil sulphide and fipronil desulfinyl by liquid chromatography-tandem quadrupole mass spectrometry (LC–MS/MS). Compared with the Oasis PRiME HLB cartridge, HTC-Mag-GO is much more effective for the removal of matrix effect. Furthermore, it is beneficial to protect the chromatographic column and ESI source by the HTC-Mag-GO PRiME pass-through cleanup procedure, resulting from the much cleaner blood samples. Under the optimum conditions, the results show higher cleanup efficiency of HTC-Mag-GO with recoveries in the range of 92.4%–108%. Especially, the HTC-Mag-GO is also evaluated for reuse (20 times) without much sacrifice of the cleanup efficiency. The limits of quantification (LOQs) for fipronil, fipronil sulphone, fipronil sulphide and fipronil desulfinyl are 8.9 ng/L, 7.2 ng/L, 8.0 ng/L and 42 ng/L, respectively. The developed method has also been successfully applied to monitoring fipronil and its metabolites in 120 blood samples, and fipronil sulphone is detected in six samples with concentrations in the range of 12.1 ng/L–106 ng/L. Further, the well validation results and the application to analysis of fipronil and its metabolites in real samples demonstrate the applicability to toxico kinetic studies and clinical studies.

Enhanced cleanup efficiency hydroxy functionalized-magnetic graphene oxide and its comparison with magnetic carboxyl-graphene for PRiME pass-through cleanup of strychnine and brucine in human plasma samples

An enhanced cleanup efficiency hydroxy functionalized-magnetic graphene oxide (EH-Mag-GO) fully covered porous nano-titania as coating has been designed and synthesized. It has been evaluated in PRiME (process, robustness, improvements, matrix effects, ease of use) pass-through cleanup procedure for human plasma prior to analysis of strychnine and brucine by liquid chromatography-tandem quadrupole mass spectrometry (LC-MS/MS). Comparing with the magnetic carboxyl-graphene (Mag-CG), EH-Mag-GO is much more effective for the removal of matrix effect resulted from blood phospholipids. Under optimal conditions, the results show higher cleanup efficiency of EH-Mag-GO with recoveries in the range of 89.4%–118%. The limits of quantification (LOQs) for strychnine and brucine are 0.088 μg/L and 0.092 μg/L, respectively. Especially, the EH-Mag-GO is also evaluated for reuse (20 times) without much sacrifice of the cleanup efficiency. Validation results on linearity, specificity, accuracy and precision, as well as on the application to analysis of strychnine and brucine in six cases of suspected semen strychni poisoning demonstrate the applicability to clinical studies.

Nano-titania-coated modified magnetic graphene oxide for pass-through cleanup of chloramphenicol, thiamphenicol, and florfenicol in human blood

To develop a rapid, effective, and accurate method for the simultaneous determination of chloramphenicol, thiamphenicol, and florfenicol in human blood by pass-through cleanup solid-phase extraction combined with liquid chromatography-tandem mass spectrometry (LC-MS/MS), novel nano-titania-coated modified magnetic graphene oxide (TiO2-Mag-GO) is used as the PRiME pass-through cleanup solid-phase extraction sorbent for the cleanup of blood phospholipids. The chromatographic separation was performed on an Eclipse Plus C18 column (100 mm×2.1 mm, 1.8 μm) using 0.08% (v/v) aqueous ammonia solution and acetonitrile having aqueous ammonia (0.08%, v/v) as mobile phases. The tandem mass spectrometer was operated using electrospray ion source (ESI) in the multiple reaction monitoring (MRM) mode. The results showed that the linearities were in the range of 0.1-10.0 μg/L with the determination coefficients (r2) greater than 0.9990 for chloramphenicol, thiamphenicol, and florfenicol. The limits of quantification (LOQs) (S/N>10) in the blood samples were between 0.056 and 0.082 μg/L and the recoveries ranged from 90.0% to 105% at three spiked levels. Precision values expressed as relative standard deviations (RSDs) were in the range of 1.2%-6.6%. The developed method can be used for routine analyses to determine chloramphenicol, thiamphenicol, and florfenicol in clinical studies.

Fast determination of fipronil and its metabolites in seafood using PRiME pass-through cleanup followed by isotope dilution UHPLC-MS/MS

A simple and effective method based on the PRiME pass-through cleanup procedure followed by UHPLC-MS/MS was developed for analyzing fipronil and its metabolites in seafoods.

Determination of Nitrite in Milk- and Soy-Based Nutritional Ingredients by Derivatization with 2,3-Diaminonaphthalene and Fluorescence Spectrometry.

Nitrite (NO2-) is an inorganic anion that can be found in various powdered milk- and soy-based nutritional ingredients as an incidental contaminant. Reliable determination of NO2- in nutritional ingredients is of paramount importance to ensure the safety of finished products. The derivatization reaction of NO2- with 2,3-diaminonaphthalene with the formation of fluorescent 2,3-naphtotriazole has been adapted to milk- and soy-based nutritional ingredients. The sample preparation consisted of protein precipitation with Carrez solution, simple pass-through cleanup of extracts utilizing a carbon black-based cartridge and derivatization, followed by batch fluorometry. The method was validated in six representative ingredient matrixes-i.e., whole-milk powder, nonfat dry milk, milk protein concentrate, whey protein concentrate, sodium caseinate, and soy protein isolate. Recovery values were 82-109%, whereas within-day and intermediate precision were 0.6-5.2 and 3.6-11% (RSDs), respectively. The method LOQ was 0.1 or 0.2 µg/g sodium nitrite (NaNO2), depending on the ingredient matrix. Surveyed NO2- concentration levels in 25 lots of 10 types of nutritional ingredients ranged from between less than 0.1 to 29 µg/g NaNO2. This method is proposed as a more sensitive and rugged alternative to the widely used ion chromatographic and colorimetric approaches.

Determination of a broad spectrum of endocrine-disrupting pesticides in fish samples by UHPLC-MS/MS using the pass-through cleanup approach.

We describe a new methodology for the simultaneous determination of the endocrine-disrupting herbicides (acetochlor, alachlor, amitrole, and atrazine), fungicides (carbendazim, triadimefon, penconazole, and propiconazole), and insecticides (carbaryl and carbofuran) in fish samples followed by high-performance liquid chromatography with tandem mass spectrometry. Samples were extracted and purified using the pass-through cleanup approach. The recoveries of the pesticides were in the range 71.8-116.5%, with relative standard deviations lower than 15.28%. Limits of quantitation were in the range of 0.03-2.50μg/kg. Validation results on linearity, accuracy, and precision, as well as on application to the analysis of the endocrine-disrupting pesticides in 20 fish samples, demonstrated the applicability to screen the presence of pesticides in fish.

Application of carbon nanosorbent for PRiME pass-through cleanup of 10 selected local anesthetic drugs in human plasma samples

A novel PRiME (process, robustness, improvements, matrix effects, ease of use) pass-through cleanup procedure has been developed to improve the existing commercially available designs. Carbon nanosorbents, i.e., magnetic modified carboxyl-graphene (Mag-CG) and magnetic modified carboxyl-carbon nanotubes (Mag-CCNTs), have been synthesised and evaluated in PRiME pass-through cleanup procedure for human plasma prior to analysis of 10 selected local anesthetic drugs by liquid chromatography-tandem quadrupole mass spectrometry (LC-MS/MS). The matrix effect, an interesting phenomenon of ion suppression for local anesthetic drugs containing ester group and ion enhancement for other drugs containing acylamino group, has been minimized using carbon nanosorbents PRiME pass-through cleanup procedure. Under the optimal conditions, the obtained results show higher cleanup efficiency of the carbon nanosorbents with recoveries between 70.2% and 126%. Furthermore, the carbon nanosorbents are also evaluated for reuse up to 80–100 times. The limits of quantification (LOQs) for local anesthetic drugs are in the range of 0.024–0.15 μg/L. Validation results on linearity, specificity, accuracy, and precision, as well as the application to the analysis of lidocaine in five patients recruited from the lung cancer demonstrate the applicability to clinical studies.

PRiME pass-through cleanup for the fast determination of aflatoxins in human serum by using LC-MS/MS

A novel, simple and sensitive method based on the use of a PRiME (process, robustness, improvements, matrix effects, ease of use) pass-through cleanup procedure combined with ultra-fast liquid chromatography-tandem quadrupole mass spectrometry (UFLC-MS/MS) for the determination of 4 AFs, i.e., AFB1, AFB2, AFG1 and AFG2 in human serum, was developed and validated. The effective PRiME pass-through cleanup procedure using the Oasis PRiME HLB cartridge was optimized for the extraction of AFB1, AFB2, AFG1 and AFG2 in human serum. The obtained results demonstrated the higher extraction capacity of the PRiME pass-through cleanup procedure with recoveries between 92.1% and 102%. The limits of detection (LODs) for AFB1, AFB2, AFG1 and AFG2 were 0.048 μg L−1, 0.069 μg L−1, 0.043 μg L−1 and 0.14 μg L−1, respectively. Validation results on linearity, specificity, accuracy and precision, as well as on application to the analysis of AFs in 10 liver cancer volunteers demonstrated the applicability to clinical studies.