Quantification of Carbonic Anhydrase Inhibitors and Metabolites in Urine and Hair of Patients and Their Relatives

Faro, Alfredo Fabrizio Lo; Tini, Anastasio; Bambagiotti, Giulia; Pirani, Filippo; Faragalli, Andrea; Carle, Flavia; Pacella, Elena; Ceka, Artan; Moretti, Marco; Gottardi, Massimo; Lassandro, Nicola Vito; Nicolai, Michele; Lupidi, Marco; Mariotti, Cesare; Busardò, Francesco Paolo; Carlier, Jérémy

doi:10.3390/biology11101379

Cited by 2 publications

(2 citation statements)

References 16 publications

(21 reference statements)

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“…The analyses of these three sulfonamide molecules indicate that our “Signal-ON” ASLFA approach shows comparable analytical sensitivity to that of existing small molecule “Signal-OFF” LFA detection methods. The median urine concentration in human was about 72.8 ppb for acetazolamide . Therefore, our ASLFA can probably be used to detect acetazolamide in human urine samples.…”

Section: Resultsmentioning

confidence: 88%

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Affinity-Switchable Interaction of Biotin and Streptavidin for the Signal-ON Detection of Small Molecules

Chung,

Liu,

Jian

et al. 2023

ACS Sens.

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Lateral flow assay (LFA) based on gold nanoparticles (AuNPs) is a widely used analytical device for the rapid analysis of environmental hazards and biomarkers. Typically, a sandwich-type format is used for macromolecule detection, in which the appearance of a red test line indicates a positive result (Signal-ON). In contrast, small molecule detection usually relies on a competitive assay, where the absence of a test line indicates positive testing (Signal-OFF). However, such a “Signal-OFF” reading is usually detected within a narrower dynamic range and tends to generate false-negative signals at a low concentration. Moreover, inconsistent readings between macromolecule and small molecule testing might lead to misinterpretation when used by nonskilled individuals. Herein, we report a “Signal-ON” small molecule competitive assay based on the sterically modulated affinity-switchable interaction of biotin and streptavidin. In the absence of a small molecule target, a large steric hindrance can be imposed on the biotin to prevent interaction with streptavidin. However, in the presence of the small molecule target, this steric effect is removed, allowing the biotin to bind to streptavidin and generate the desired test line. In this article, we demonstrate the selective detection of two small molecule drugs, sulfonamides and trimethoprim, using this simple and modular affinity-switchable lateral flow assay (ASLFA). We believe that this affinity-switchable approach can also be adapted in drug discovery and clinical diagnosis, where the competitive assay format is always used for the rapid analysis of small molecules.

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Section: Resultsmentioning

confidence: 88%

“…The median urine concentration in human was about 72.8 ppb for acetazolamide. 47 Therefore, our ASLFA can probably be used to detect acetazolamide in human urine samples. Furthermore, we also studied the long-term stability of Au@ ASB-SFM in the absence or presence of hCAII.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

Affinity-Switchable Interaction of Biotin and Streptavidin for the Signal-ON Detection of Small Molecules

Chung,

Liu,

Jian

et al. 2023

ACS Sens.

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Elimination profile of low‐dose chlortalidone and its detection in hair for doping analysis—Implication for unintentional non‐therapeutic exposure

Thieme,

Weigel,

Anielski

et al. 2024

Drug Testing and Analysis

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Chlortalidone (CLT) is a thiazide‐type diuretic with high affinity for the erythrocyte carbonic anhydrase. Therapeutically, it is mostly used to treat edema and hypertension due to liver cirrhosis, heart insufficiency, or renal dysfunction. Although diuretics and masking agents are prohibited by the World Anti‐Doping Agency (WADA) at all times in sports, substances belonging to this category are constantly detected in athlete samples, according to WADA's annual testing figures. Within this group of structurally diverse compounds, a threshold of 20 ng/mL has been introduced for six substances solely due to their presence as contaminants in other permitted drugs because of pharmaceutical production processes. In a recent presumptive doping case with a low urinary CLT concentration, the question of unintentional doping, for example, by contaminated non‐steroidal anti‐inflammatory drug intake, arose. To examine this potential scenario, a co‐elimination of low‐dose CLT and hydrochlorothiazide (HCTA; 20 × 50 μg, 0.2 mg/day each) was conducted on five consecutive days in two volunteers. Urine samples were subjected to liquid chromatography‐tandem mass spectrometry (LC–MS/MS). Moreover, we examined the incorporation of CLT in scalp hair. HCTA is rapidly excreted renally in comparatively high concentrations. In contrast, the elimination of CLT is considerably slower (terminal elimination half‐life extended by a factor of 12) and, consequently, much less concentrated in corresponding urine samples (45 and 53 ng/mL, respectively). Conversely, a higher hair incorporation of chlorthalidone was observed with simultaneous dosing of both. The results suggest that an unintentional intake of sub‐therapeutic CLT doses due to contamination might result in an adverse analytical finding.

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Quantification of Carbonic Anhydrase Inhibitors and Metabolites in Urine and Hair of Patients and Their Relatives

Cited by 2 publications

References 16 publications

Affinity-Switchable Interaction of Biotin and Streptavidin for the Signal-ON Detection of Small Molecules

Affinity-Switchable Interaction of Biotin and Streptavidin for the Signal-ON Detection of Small Molecules

Elimination profile of low‐dose chlortalidone and its detection in hair for doping analysis—Implication for unintentional non‐therapeutic exposure

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