Solid phase extraction as sample pretreatment method for top-down quantitative analysis of low molecular weight proteins from biological samples using liquid chromatography – triple quadrupole mass spectrometry

Maráková, Katarína; Renner, Beatriz J.; Thomas, Shannon; Opetová, Martina; Tomašovský, Radovan; Alex, J.; Schug, Kevin A.

doi:10.1016/j.aca.2023.340801

Cited by 8 publications

(6 citation statements)

References 58 publications

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“…A previous study reported the use of liquid chromatography with mass spectrophotometry to determine glabridin in human serum 25 . Sample extraction from human serum by solid‐phase extraction needs to be implemented before analysis to reduce impurity disturbance 26 . In the current study, the developed ELISA method is specific and sensitive enough to analyse glabridin without requiring any sample pretreatment.…”

Section: Resultsmentioning

confidence: 91%

“…extraction from human serum by solid-phase extraction needs to be implemented before analysis to reduce impurity disturbance. 26 In the current study, the developed ELISA method is specific and sensitive enough to analyse glabridin without requiring any sample pretreatment. Therefore, this method may facilitate the analysis of clinical samples, including pharmacokinetic studies.…”

Section: Application For Glabridin Determination In Human Serummentioning

confidence: 85%

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Construction of a monoclonal antibody against glabridin (2G4) and development of an enzyme‐linked immunosorbent assay

Krittanai,

Choonong,

Butdapheng

et al. 2023

Phytochemical Analysis

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Introduction Glabridin is a unique isoflavonoid found only in Glycyrrhiza glabra L. The pharmacological effects of glabridin are well established, especially for beauty‐ and wellness‐related uses, such as antioxidant, anti‐inflammatory, ultraviolet (UV) protection, and skin‐lightening effects. Therefore, glabridin is often found in commercial products such as creams, lotions, and dietary supplements. Objective This study aimed to develop an enzyme‐linked immunosorbent assay (ELISA) using a glabridin‐specific antibody. Method Immunogen conjugation of glabridin‐bovine serum albumin was performed via the Mannich reaction, and the resulting conjugates were injected into BALB/c mice. Subsequently, hybridomas were produced. An ELISA method for glabridin determination was developed and validated. Result A highly specific antibody against glabridin was produced using clone 2G4. The assay range for the determination of glabridin was 0.28–7.02 μg/ml, with a detection limit of 0.16 μg/ml. The validation parameters in terms of accuracy and precision met the acceptable criteria. Standard curves of glabridin in various matrices were compared to evaluate the matrix effect on human serum using ELISA. Standard curves of the human serum and water matrix were obtained in the same manner, and the measurement range was 0.41–10.57 μg/ml. Conclusion The developed ELISA method was used to quantify glabridin in plant materials and products with high sensitivity and specificity, and has potential applications in quantifying compounds in plant‐derived products and human serum samples.

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Section: Resultsmentioning

confidence: 91%

Section: Application For Glabridin Determination In Human Serummentioning

confidence: 85%

Construction of a monoclonal antibody against glabridin (2G4) and development of an enzyme‐linked immunosorbent assay

Krittanai,

Choonong,

Butdapheng

et al. 2023

Phytochemical Analysis

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“…Standard and sample preparation, including the molecular weight cut‐off (MWCO) and SPE processes [29], are listed in the Supporting Information section.…”

Section: Methodsmentioning

confidence: 99%

Capillary electrophoresis on‐line hyphenated with mass spectrometry for analysis of insulin‐like growth factor‐1 in pharmaceutical preparations

et al. 2023

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Insulin‐like growth factor‐1 (IGF‐1) is a 70‐amino acid single‐chain polypeptide, which has found application in diagnostics as a biomarker of growth hormone disorders and as a therapy for growth failure in children and adolescents. Due to its strong anabolic effects, it is often abused by athletes for doping purposes. Here, we developed an on‐line hyphenated method based on capillary zone electrophoresis (CZE) and triple quadrupole mass spectrometry (MS) detection with electrospray ionization (CZE–electrospray ionization source–MS [CZE–ESI–MS]) for the determination of IGF‐1 in pharmaceutical matrices. We achieved a highly efficient, accurate, repeatable, sensitive, and selective analysis of IGF‐1 with favorable migration times (<15 min). Optimized and validated CZE–ESI–MS method was successfully applied for the determination of IGF‐1 in injectable solutions (Increlex®), and its presence was also confirmed in nutritional preparations (tablets and liquid colostrum). This is the first validated CZE–ESI–MS method for the determination of IGF‐1 in pharmaceutical matrices revealing the potential of capillary electrophoresis for its use in drug quality control laboratories with benefits, such as high separation efficiency, high‐speed analysis, low sample consumption, as well as environmental and cost aspects.

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“…Proteins can be extracted from biological matrices employing various offline and on-line techniques such as homogenization, centrifugation, precipitation, liquid-liquid extraction, SPE, SPME, (micro)dialysis, and ultrafiltration [86,87]. Even though, there is an effort to develop non-immunoaffinity sample preparation methods for the extraction of multiple intact proteins from biological matrices [88], sample enrichment methods based on immunoaffinity are still predominantly applied for intact protein analysis. Sample preparation techniques for extraction and fractionation of intact proteins from biological matrices for MS analysis were extensively reviewed in our recent paper [89].…”

Section: Peptide-and Protein-based Biomarkersmentioning

confidence: 99%

Capillary electrophoresis‐mass spectrometry for intact protein analysis: Pharmaceutical and biomedical applications (2018–March 2023)

Maráková

Opetová

Tomašovský

2023

J of Separation Science

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Capillary electrophoresis is recognized as a valued separation technique for its high separation efficiency, low sample consumption, good economic and ecological aspects, reproducibility, and complementarity to traditional liquid chromatography techniques. Capillary electrophoresis experiments are generally performed utilizing optical detection, such as ultraviolet or fluorescence detectors. However, in order to provide structural information, capillary electrophoresis hyphenated to highly sensitive and selective mass spectrometry has been developed to overcome the limitations of optical detections. Capillary electrophoresis‐mass spectrometry is increasingly popular in protein analysis, including biopharmaceutical and biomedical research. It is frequently applied for the determination of physicochemical and biochemical parameters of proteins, offers excellent performance for in‐depth characterizations of biopharmaceuticals at various levels of analysis, and has been also already proven as a promising tool in biomarker discovery. In this review, we focus on the possibilities and limitations of capillary electrophoresis‐mass spectrometry for protein analysis at their intact level. Various capillary electrophoresis modes and capillary electrophoresis‐mass spectrometry interfaces, as well as approaches to prevent protein adsorption and to enhance sample loading capacity, are discussed and the recent (2018–March 2023) developments and applications in the field of biopharmaceutical and biomedical analysis are summarized.

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Solid phase extraction as sample pretreatment method for top-down quantitative analysis of low molecular weight proteins from biological samples using liquid chromatography – triple quadrupole mass spectrometry

Cited by 8 publications

References 58 publications

Construction of a monoclonal antibody against glabridin (2G4) and development of an enzyme‐linked immunosorbent assay

Construction of a monoclonal antibody against glabridin (2G4) and development of an enzyme‐linked immunosorbent assay

Capillary electrophoresis on‐line hyphenated with mass spectrometry for analysis of insulin‐like growth factor‐1 in pharmaceutical preparations

Capillary electrophoresis‐mass spectrometry for intact protein analysis: Pharmaceutical and biomedical applications (2018–March 2023)

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