Ratiometric fluorescence and colorimetric dual-mode sensing platform based on carbon dots for detecting copper(II) ions and D-penicillamine.

A sensing platform with both ratiometric fluorescence and colorimetric responses towards copper(II) ions (Cu2+) and D-penicillamine (D-pen) was constructed based on carbon dots (CDs). o-Phenylenediamine (OPD) was employed as a chromogenic development reagent for reaction with Cu2+ to generate the oxidation product 2,3-diaminophenazine (oxOPD), which not only emits green fluorescence at 555 nm, but also quenches the blue fluorescence of CDs at 443 nm via the inner filter effect (IFE) and Förster resonance energy transfer (FRET). Additionally, oxOPD exhibits obvious absorption at 420 nm. Since the intense chelation affinity of D-pen to Cu2+ greatly inhibits the oxidation of OPD, the intensity ratio of fluorescence at 443 nm to that at 555 nm (F443/F555) and the absorbance at 420 nm (A420) were conveniently employed as spectral response signals to represent the amount of D-pen introduced into the testing system. This dual-signal sensing platform exhibits excellent selectivity and sensitivity towards both Cu2+ and D-pen, with low detection limits of 0.019 µM and 0.092 µM, respectively. In addition, the low cytotoxicity of the testing reagents involved in the proposed sensing platform facilitates its application for live cell imaging.

Development of carbon dot-thiochrome-based sensing system for ratiometric fluorescence detection of D-penicillamine.

A simple and rapid ratiometric fluorescent sensing system for D-penicillamine (D-PA) determination is developed based on yellow carbon dots (Y-CDs) combined with thiochrome (oxVB1) for the first time. The oxidization of thiamine (VB1) can be catalyzed by Alkaline-hydrolyzed artemisinin (a-ART) to form oxVB1, which leads to the occurrence of fluorescence emission peak at 466 nm. Furthermore, the oxidation reaction between a-ART and VB1 could be inhibited by D-PA, and accompanied with the decrease of fluorescence at 466 nm. However, the fluorescence peak of Y-CDs as an internal reference at 566 nm was almost unchanged. The ratiometric signal changes contributed to a robust and sensitive D-PA sensing. Under the optimal condition, a good linear response for the D-PA detection was obtained in the ranges of 0.5-50 µM with a detection limit of 0.33 µM. In addition, Y-CDs and thiochrome-based sensing system was applied to D-PA determination in real samples and obtained acceptable results. We developed a new carbon dots/thiochrome fluorescent nanoprobe for ratiometric fluorescence sensing of D-penicillamine.

A graphene quantum dots-Pb2+ based fluorescent switch for selective and sensitive determination of D-penicillamine.

Taking consideration of metal-induced fluorescence quenching and excellent coordination effect of D-penicillamine (D-PA), a graphene quantum dots (GQDs)-based fluorescent switch for D-PA detection was designed and established firstly with the help of lead ions. GQDs obtained from citric acids made them rich in carboxyl and hydroxyl groups, giving GQDs the ability to combine with lead ions. As anticipated, the fluorescence intensity was quenched by Pb2+ through electron transfer process. Further, the addition of D-PA effectively recovered the fluorescence due to the departure of Pb2+ from GQDs aroused by the strong coordination between D-PA and Pb2+. Thus, a fluorescent switch was activated for D-PA detection. The fluorescence recovery efficiencies were found to be proportional to the concentration of D-PA in the range of 0.6-50 µmol L-1 and the detection limit was 0.47 µmol L-1. The real sample detection was performed in human urea sample and satisfactory recoveries of 96.84%-102.13% were obtained. The GQDs-Pb2+ based fluorescent switch sensing method was firstly established with low detection limit and wide linear range, making it a supplement and improvement for D-PA detection.

Copper nanocluster-based fluorescence enhanced determination of d-penicillamine.

Taking advantage of the compelling properties of d-penicillamine (d-PA) combined with copper, a method for the sensitive and selective determination of d-PA was established using copper nanocluster (Cu NC)-based fluorescence enhancement. d-PA molecules containing a thiol compound showed a strong tendency to combine with the surface of Cu NCs, causing the re-dispersion of nanoclusters and therefore fluorescence intensity was enhanced. Fluorescence enhancement efficiency of Cu NCs induced by d-PA was linear, with the d-PA concentration varying from 0.6-30 µg ml-1 (R2  = 0.9952) and with a detection limit of 0.54 µg ml-1 . d-PA content in human urine samples was detected with recoveries of 104.8-112.99%. Fluorescence-enhanced determination of d-PA using Cu NCs was established for the first time and this rapid, easy and sensitive method should attract much attention for this application.

Smartphone-based Ellman's colourimetric methods for the analysis of d-penicillamine formulation and thiolated polymer.

A new, smartphone-based colourimetric method for the assay of d-penicillamine formulations relying on the Ellman's reaction was developed by performing the colourimetric reaction in a microplate. Subsequently, the plate was positioned on a white illuminating screen of an iPad placed in a dark box in order to capture a top-view image using an iPhone 5s back camera. The intensity of yellow colour was converted to Red-Green-Blue pixels using a free mobile application. Under the optimal conditions for the reaction and photography, the intensity of blue colour, which was logarithmically transformed, showed an excellent linearity over the drug concentration range of 5-40 µg/mL. The assay was validated and successfully applied to the assay of drug content and the determination of drug amount released in the dissolution test in the capsule dosage forms. Apart from that, a smartphone was employed for the colour measurement as an alternative to a spectrophotometer in the currently used method for the quantitation of free sulfhydryl groups in polymers. Using cysteine-conjugated chitosan as a sample and l-cysteine as a standard, the smartphone method gave the results in agreement with those obtained from the absorbance measurement on a microplate reader. In conclusion, smartphone-based colourimetry has been proved to be a reliable, fast, simple and affordable alternative means for the analysis of d-penicillamine and cysteine-conjugated polymer and can be potentially applied to other thiol-containing drugs and excipients.

Sensitive and Selective Detection of Antibiotic D-Penicillamine Based on a Dual-Mode Probe of Fluorescent Carbon Dots and Gold Nanoparticles.

This paper reported a dual-mode probe for D-penicillamine on the basis of pH-mediated gold nanoparticles aggregation and fluorescence resonance energy transfer (FRET) from carbon dots. D-penicillamine is a zwitterionic compound and has different forms depending on specific pH ranges. The thiol group of D-penicillamine has high affinity towards the surface of gold nanoparticles and can replace other surface ligands. When pH values were close to its isoelectrical point (pH(I)), the D-penicillamine capped gold nanoparticles aggregated through hydrogen bonding or electrostatic interactions, resulting in the releasing of carbon dots from gold nanoparticles. The dual-mode probe consisted of fluorescent carbon dots and gold nanoparticles, and the fluorescence of carbon dots was quenched by the attached gold nanoparticles due to the FRET. Then, the fluorescence can be recovered in presence of D-penicillamine due to the gold nanoparticles aggregation in specific pH range. Under the optimum conditions, the probe has linear response for D-penicillamine in the 0.25-1.5 µM concentration range with a detection limit of 0.085 µM. This method provides a potential application in sensitive detection of D-penicillamine.

A water-stable homochiral luminescent MOF constructed from an achiral acylamide-containing dicarboxylate ligand for enantioselective sensing of penicillamine.

A water-stable homochiral luminescent metal-organic framework (HLMOF), {[Cd(l)(4,4'-bipy)]·DMA·5H2O}n (1), was obtained by a spontaneous symmetry-breaking crystallization from achiral precursors without any enantiopure auxiliary. The homochirality of 1 was confirmed by single-crystal X-ray diffraction and CD spectroscopy. Complex 1 exhibits an interesting enantioselective sensing of d/l-penicillamine in water.

Nanoparticle-based Chemiluminescence for Chiral Discrimination of Thiol-Containing Amino Acids.

The ability to recognize the molecular chirality of enantiomers is extremely important owing to their critical role in drug development and biochemistry. Convenient discrimination of enantiomers has remained a challenge due to lack of unsophisticated methods. In this work, we have reported a simple strategy for chiral recognition of thiol-containing amino acids including penicillamine (PA), and cysteine (Cys). We have successfully designed a nanoparticle-based chemiluminescence (CL) system based on the reaction between cadmium telluride quantum dots (CdTe QDs) and the enantiomers. The different interactions of CdTe QDs with PA enantiomers or Cys enantiomers led to different CL intensities, resulting in the chiral recognition of these enantiomers. The developed method showed the ability for determination of enantiomeric excess of PA and Cys. It has also obtained an enantioselective concentration range from 1.15 to 9.2 mM for PA. To demonstrate the potential application of this method, the designed platform was applied for the quantification of PA in urine and tablet samples. For the first time, we presented a novel practical application of nanoparticle-based CL system for chiral discrimination.

Fluorescent MUA-stabilized Au nanoclusters for sensitive and selective detection of penicillamine.

In this study, we have developed a facile method for preparation of highly fluorescent Au nanoclusters (AuNCs) using 11-mercaptoundecanoic acid (MUA) as both the reducing and stabilizing agent. The as-prepared MUA functionalized AuNCs (MUA-AuNCs) have good water solubility, excellent photostability, and strong fluorescence emission at 610 nm with a quantum yield of 7.28% in water. The fluorescence of MUA-AuNCs was first quenched by copper ions through electron transfer, subsequently caused obvious restoration by competitive effect after adding penicillamine, making it a potential fluorescent sensor for penicillamine with a detection limit of 0.08 µM. Furthermore, the newly designed fluorescence "on-off-on" assay was explored for the measurement of penicillamine in complex real water and urine samples with satisfactory results.

First Report for Determination of d-Penicillamine in the Presence of Tryptophan Using a 2-Chlorobenzoyl Ferrocene/Ag-Supported ZnO Nanoplate-Modified Carbon Paste Electrode.

In this work, a carbon paste electrode modified with Ag-ZnO nanoplates and 2-chlorobenzoyl ferrocene was prepared and applied for the determination of d-penicillamine in the presence of tryptophan. The morphologies of Ag-ZnO nanoplates were examined by scanning electron microscopy. It was found that under an optimum condition (pH 7.0), the oxidation of d-penicillamine at the surface of such an electrode occurs at a potential about 165 mV less positive than that of an unmodified carbon paste electrode. The diffusion coefficient (D = 7.6 × 10-6 cm2/s) and the electron transfer coefficient (α = 0.54) for d-penicillamine oxidation were also determined. The proposed method exhibited a wide linear dynamic range of 0.03-250.0 µM, with an LOD (S/N = 3) of 0.015 µM. Moreover, the modified electrode exhibited good reproducibility and high selectivity, demonstrating its feasibility for the analytical purpose. Lastly, this new sensor was used for the determination of d-penicillamine and tryptophan in real samples.

Theoretical spectroscopic insights of tautomers and enantiomers of penicillamine.

B3LYP and MP2 calculations have been carried out to investigate tautomers and enantiomers of penicillamine (Pen). Their infrared (IR), ultraviolet (UV), circular dichroism (CD) and nuclear magnetic resonance (NMR) spectra were obtained at linear-response, time-dependent DFT (TD-DFT). IR, UV and NMR spectra cannot be used to identify Pen enantiomers, showing nearly equal spectral profiles. CD spectra, however, give rise to completely symmetric signals, forming a perfect specular image to each other. Distinct CD profiles were also obtained for Pen tautomers. Important IR differences were found in positions and intensities of the vibrational stretching bands involving acid and amine groups of Pen tautomers. The highest electron transitions involving HOMO-LUMO orbitals show to be of major importance in the computed UV spectra, showing a large red-shift around 30nm as the zwitterionic and neutral Pen spectra are compared. NMR results show to be quite useful for identification of Pen tautomers since clear differences are found by means of the computed shielding tensors as well as spin-spin coupling constants 1J(N,H) data.

Recognition of D-Penicillamine Using Schiff Base Centered Fluorescent Organic Nanoparticles and Application to Medicine Analysis.

Schiff base centered fluorescent organic compound 1,1'-[(1E,2E)-hydrazine-1,2-diylidenedi(E)methylylidene]- dinaphthalen-2-ol (HN) was synthesized followed by spectral characterization viz., NMR, IR and Mass spectroscopy. The fluorescent nanoparticles of HN prepared using reprecipitation method shows red shifted aggregation induced enhanced emission (AIEE) with respect to HN solution in acetone. The average particle size of nanoparticles (HNNPs) is of 67.2 nm shows sphere shape morphology. The surfactant cetyltrimethyl ammonium bromide (CTAB) used to stabilize HNNPs induces positive charge surface with zeta potential of 11.6 mV. The positive charge of HNNPs responsible to adsorb oppositely charged analyte on its surface with binding interactions. The fluorescence experiments performed with and without addition of different analytes to the aqueous suspension of HNNPs shows selective fluorescence quenching of HNNPs by D-Penicillamine (D-PA). The effect of other coexisting analytes does not affect the selective sensing behavior of D-PA. The mechanism of binding between HNNPs and D-PA was discussed on the basis of electrostatic interaction and adsorption phenomenon. The results interpreted by using DLS-Zeta sizer, Fluorescence lifetime measurements, conductometric titration supports the electrostatic adsorption between HNNPs and D-PA. The method has extremely low limit of detection (LOD) value 0.021 ppm is of significant as compared to reported methods. The proposed fluorescence quenching method was effectively used for quantitative estimation of D-PA from pharmaceutical medicine. Graphical Abstract The fluorescence quenching based selective recognition of D-Penicillamine (D-PA) by using Schiff base centered fluorescent organic nanoparticles was developed and successfully applied to quantitative determination of D-PA from pharmaceutical samples viz. capsule and tablet.

Chiral recognition of penicillamine enantiomers using hemoglobin and gold nanoparticles functionalized graphite-like carbon nitride nanosheets via electrochemiluminescence.

A new stable and stereo-selective electrochemiluminescence (ECL) interface has been designed for specific recognition of penicillamine (Pen) enantiomers by using hemoglobin (Hb) and gold nanoparticles functionalized graphite-like carbon nitride nanosheets composite (Au-g-C3N4 NHs) modified glassy carbon electrodes (Hb/Au-g-C3N4/GCE). The advantages of Hb as chiral selector and Au-g-C3N4 NHs as luminophore were perfectly displayed in this novel interface. The obviously different ECL intensity was exhibited after l-Pen and d-Pen adsorbed on Hb/Au-g-C3N4/GCE, and a larger response was observed on d-Pen/Hb/Au-g-C3N4/GCE. Under the optimum conditions, the developed ECL chiral sensor showed excellent analytical property for detection of Pen enantiomers in a linear range of 1.0×10-4M to 5.0×10-3M, and the detection limits of l-Pen and d-Pen were 3.1×10-5M and 3.3×10-5M (S/N=3) respectively. This work with high selectivity, stability and reproducibility may open a new door based on ECL to discriminate Pen enantiomers.

Circular dichroism sensor based on cadmium sulfide quantum dots for chiral identification and detection of penicillamine.

A new chemical sensor based on the measuring of circular dichroism signal (CD) was fabricated from cysteamine capped cadmium sulfide quantum dots (Cys-CdS QDs). The chiral-thiol molecules, d-penicillamine (DPA) and l-penicillamine (LPA), were used to evaluate potentials of this sensor. Basically, DPA and LPA provide very low CD signals. However, the CD signals of DPA and LPA can be enhanced in the presence of Cys-CdS QDs. The CD spectra of DPA and LPA exhibited a mirror image profile. Parameters affecting the determination of DPA and LPA were thoroughly investigated in details. Under the optimized condition, the CD signals of DPA and LPA displayed a linear relationship with the concentrations of both enantiomers, ranging from 1 to 35 µM. Detection limits of this sensor were 0.49 and 0.74 µM for DPA and LPA, respectively. To demonstrate a potential application of this sensor, the proposed sensor was used to determine DPA and LPA in real urine samples. It was confirmed that the proposed detection technique was reliable and could be utilized in a broad range of applications.

Spectrophotometric sequential injection determination of D-penicillamine based on a complexation reaction with nickel ion.

A simple and sensitive spectrophotometric method, based on reaction between Ni(II) ion and D-penicillamine (PEN), was developed. The proposed SIA system enhanced the analytical applicability of the reaction of complexation, and allowed the determination of PEN in the concentration range of 3.0 × 10(-6) - 2.0 × 10(-4) mol L(-1) with a sampling rate of 200 h(-1). With the proposed SIA system, PEN could be accurately determinated up to 0.9 nmol quantity. The method was successfully applied to the determination of PEN in laboratory samples and pharmaceuticals.

An electrochemical sensor based on 1-benzyl-4-ferrocenyl-1H-[1,2,3]-triazole/carbon nanotube; detection of D-penicillamine in the presence of tryptophan.

A glassy carbon electrode modified with 1-benzyl-4-ferrocenyl-1H-[1,2,3]-triazole (BFT) and carbon nanotubes have been applied to the electrocatalytic oxidation of D-penicillamine (D-PA) which reduced the overpotential by about 470 mV with obviously increase the current response. Due to its strong electrocatalytic activity towards D-PA, the modified electrode can resolve the overlapped voltammetric waves of D-PA and tryptophan (TRP) into two well-defined voltammetric peaks with peak-to-peak separation in potentials of about 270 mV. This property allows to selective determination of D-PA in the presence of TRP. The transfer coefficient (a) for the electrocatalytic oxidation of D-PA and diffusion coefficient of this substance under the experimental conditions were also investigated. In phosphate buffer solution (PBS) of pH8.0, the oxidation current increased linearly with two concentration intervals of D-PA, one is 1.0 to 10.0 µM and, the other is 10.0 to 800.0 µM. The detection limit (3σ) obtained by square wave voltammetry (SWV) was 0.1 µM. The proposed method was successfully applied to the determination of D-PA, and TRP in real samples.

Enantioseparations by high-performance liquid chromatography based on chiral ligand-exchange.

Chiral ligand-exchange chromatography (CLEC) first described in the late 1960s to early 1970s by Davankov and Rogozhin can be still considered as an elective choice for the direct enantioseparation of compounds endowed with chelating moieties. Among the numerous chelating species that have been evaluated as chiral selectors in ligand-exchange (LE) chromatography, a special role is played by a group of amino acids including proline, hydroxyproline, cysteine, phenylalanine, and penicillamine. More to the point, relevant chromatographic performances are also provided by amino alcohol-based chiral selectors, among which, those carrying a leucinol residue as the basic scaffold are worth to be mentioned. Among the various enantiomer chromatographic separation techniques, CLEC has been exploited in all the main techniques including a chiral mobile phase (CMP), a covalently bound chiral stationary phase (B-CSP), and a coated chiral stationary phase (C-CSP). It is the objective of this chapter to describe selected CLEC applications dealing with the above three distinct approaches.

A novel HPLC-electrochemical detection approach for the determination of D-penicillamine in skin specimens.

D-penicillamine is a thiol drug mainly used for Wilson's disease, rheumatoid arthritis and cystinuria. Adverse effects during normal use of the drug are frequent and may include skin lesions. To evaluate its toxic effects in clinical cases an original method based on high performance liquid chromatography coupled to amperometric detection in a specific biological matrix such as skin has been developed. The chromatographic analysis of D-penicillamine was carried out on a C18 column using a mixture of acid phosphate buffer and methanol as the mobile phase. Satisfactory sensitivity was obtained by oxidizing the molecule at +0.95 V with respect to an Ag/AgCl reference electrode. A chemical reduction of D-penicillamine-protein disulphide bonds using dithioerythritol combined with microwaves was necessary for the determination of the total amount of D-penicillamine in skin specimens. A further solid-phase extraction procedure on C18 cartridges was implemented for the sample clean-up. The whole analytical procedure was validated: high extraction yield (>91.0%) and satisfactory precision (RSD<6.8%) values were obtained. It was successfully applied to skin samples from a patient who was previously under a long-term, high-dose treatment with the drug and presented serious D-penicillamine-related dermatoses. Thus, the method seems to be suitable for the analysis of D-penicillamine in skin tissues.

Automated tagging of pharmaceutically active thiols under flow conditions using monobromobimane.

The thiol-specific derivatization reagent monobromobimane (MBB) is applied--for the first time--under flow conditions. Sequential injection analysis allows the handling of precise volumes of the reagent in the micro-liter range. The effect of the main chemical and instrumental variables was investigated using captopril (CAP), N-acetylcysteine (NAC) and penicillamine (PEN) as representative pharmaceutically active thiols. Previously reported hydrolysis of MBB due to interaction with nucleophilic components of the buffers was avoided kinetically under flow conditions. The proposed analytical scheme is suitable for the fluorimetric determination of thiols at a sampling rate of 36 h(-1).

p-Aminophenol-multiwall carbon nanotubes-TiO2 electrode as a sensor for simultaneous determination of penicillamine and uric acid.

In this work, p-aminophenol-multiwall carbon nanotubes-TiO(2) is proposed as a sensor for the rapid, sensitive, and highly selective voltammetric determination of penicillamine (PA) in the presence of uric acid (UA). The electrochemical behavior of the compounds at this modified electrode was studied by cyclic voltammetry (CV), differential pulse voltammetry (DPV), and electrochemical impedance spectroscopy (EIS). The results indicated that the chemically modified electrode exhibits efficient electrocatalytic activity in the oxidation of PA which occurs at a potential of about 530 mV, less positive than that for the unmodified carbon nanotubes paste electrode at pH 6.0. Peak potentials of PA and UA were separated with a difference of 215 mV using DPV. These conditions were sufficient to allow for the determination of PA and UA, both individually and simultaneously. At pH 6.0, the catalytic peak currents were linearly dependent on PA and UA concentrations in the ranges 0.4-200 micromol L(-1) PA and 3.0-1000 micromol L(-1) UA. Detection limits for PA and UA were 0.1 and 1.1 micromol L(-1), respectively. The RSD% for 1.2 and 1.5 micromol L(-1) PA were 1.6% and 2.1%, respectively, whereas they were 1.5% and 1.1% for 15.0 and 30.0 micromol L(-1) UA, respectively. Finally, the sensor was examined as a selective, simple, and precise new electrochemical sensor for the determination of PA in real samples in the presence of UA in drugs and urine.