Preparation of monodisperse curcumin-imprinted polymer by precipitation polymerization and its application for the extraction of curcuminoids from Curcuma longa L.

A monodisperse molecularly imprinted polymer (MIP) for curcumin was first prepared by precipitation polymerization using methacrylamide (MAM) and 4-vinylpyridine as functional co-monomers, divinylbenzene as a crosslinker, and a mixture of acetonitrile and toluene as a porogen. The use of MAM as the co-monomer resulted in the formation of a monodisperse MIP and non-imprinted polymer (NIP). MIP and NIP, respectively, were monodispersed with a narrow particle size distribution (3.3?±?0.09 and 3.5?±?0.10 μm). In addition to shape recognition, hydrophobic and hydrogen-bonding interactions affected the retention and molecular-recognition of curcumin on the MIP. The MIP for curcumin could extract curcuminoids (curcumin, demethoxycurcumin, and bisdemethoxycurcumin) in Curcuma longa L.

Identification of 3-chloro-1,2-propandiol using molecularly imprinted composite solid-phase extraction materials

A novel molecularly imprinted material based on silica microparticles was synthesized by surface polymerization with 3-chloro-1,2-propandiol (3-MCPD) as a template molecule. The molecularly imprinted polymer (MIP) was characterized by infrared spectroscopy and scanning electron microscopy. The adsorption of 3-MCPD by MIP was measured by gas chromatography with electron capture detection (GC-ECD) and an equilibrium binding experiment. Scatchard analysis revealed that the maximum apparent binding capacities of the MIP and non-imprinted polymer (NIP) were 67.64 and 23.31 μmol/g, respectively. The new adsorbent was successfully used in solid-phase extraction (SPE) to selectively enrich and determine 3-MCPD in soy sauce samples. The MIP-SPE column achieves recoveries higher than 92.7 % with a relative standard deviation of less than 1.83 %. The MIP-SPE-GC protocol improved the selectivity and eliminated the effects of template leakage on quantitative analysis and could be used for the determination of 3-MCPD in other complex food samples. Graphical Abstract

The MIP-SPE column developed by us achieves recoveries higher than 92.7 % with a relative standard deviation of less than 1.83 % for determining the 3-MCPD in the soy sauce matrix (mixed with 3-MCPD, 2-MPCD and 1,3-DCP).      

Hydrophilic molecularly imprinted polymers for bisphenol A prepared in aqueous solution

We have prepared a hydrophilic molecularly imprinted polymer (MIP) for the hydrophobic compound bisphenol A (BPA) in aqueous solution using 3-acrylamido-N,N,N-trimethylpropan-1-aminium chloride (AMTC) as the functional monomer. Under redox-polymerization conditions, BPA forms an ion-pair with AMTC, which was confirmed by ¹H-NMR titration. The imprinting effect in aqueous solution was evaluated by comparison of this material with the corresponding non-imprinted polymer (NIP) and with a control polymer (CP) bearing no AMTC. The MIP showed the highest activity among the three polymers, and the imprinting factors as calculated from the amount of BPA bound to the MIP divided by the amounts bound to NIP and CP, respectively, are 1.8 and 6.0. The MIP was selective for BPA in aqueous solution, while structurally related compounds are not recognized. Such a selectivity for a hydrophobic compound is rarely observed in aqueous medium because non-specific binding of BPA inevitably leads to hydrophobic interaction.

Determination of t,t-muconic acid in urine samples using a molecular imprinted polymer combined with simultaneous ethyl chloroformate derivatization and pre-concentration by dispersive liquid–liquid microextraction

The present communication describes the preparation and evaluation of a molecularly imprinted polymer (MIP) as a solid-phase extraction (SPE) sorbent and simultaneous ethyl chloroformate (ECF) derivatization and pre-concentration by dispersive liquid–liquid microextraction (DLLME) for the analysis of t,t-muconic acid (t,t-MA) in urine samples using gas chromatography–mass spectrometry. The imprinting polymer was prepared using methacrylic acid as a functional monomer, ethylene glycol dimethacrylate as a cross-linker, 2,2-azobisisobutyronitrile as the initiator and t,t-MA as a template molecule. The imprinted polymer was evaluated for its use as a SPE sorbent by comparing both imprinted and non-imprinted polymers in terms of the recovery of t,t-MA from urine samples. Molecular modelling studies were performed in order to estimate the binding energy and efficiency of the MIP complex formed between the monomer and the t,t-MA. Various factors that can affect the extraction efficiency of MIP, such as the loading, washing and eluting conditions, were optimized; other factors that can affect the derivatization and DLLME pre-concentration were also optimized. MIP in combination with ECF derivatization and DLLME pre-concentration for t,t-MA exhibits good linearity, ranging from 0.125 to 2 μg?mL^?1 (R ²?=?0.9971), with limit of detection of 0.037 μg?mL^?1 and limit of quantification of 0.109 μg?mL^?1. Intra- and inter-day precision was found to be <6 %. The proposed method has been proven to be effective and sensitive for the selective pre-concentration and determination of t,t-MA in urine samples of cigarette smokers.

Optimization of the polar organic chemical integrative sampler for the sampling of acidic and polar herbicides

This paper presents an optimization of the pharmaceutical Polar Organic Chemical Integrative Sampler (POCIS-200) under controlled laboratory conditions for the sampling of acidic (2,4-dichlorophenoxyacetic acid (2,4-D), acetochlor ethanesulfonic acid (ESA), acetochlor oxanilic acid, bentazon, dicamba, mesotrione, and metsulfuron) and polar (atrazine, diuron, and desisopropylatrazine) herbicides in water. Indeed, the conventional configuration of the POCIS-200 (46 cm² exposure window, 200 mg of Oasis® hydrophilic lipophilic balance (HLB) receiving phase) is not appropriate for the sampling of very polar and acidic compounds because they rapidly reach a thermodynamic equilibrium with the Oasis HLB receiving phase. Thus, we investigated several ways to extend the initial linear accumulation. On the one hand, increasing the mass of sorbent to 600 mg resulted in sampling rates (R _s s) twice as high as those observed with 200 mg (e.g., 287 vs. 157 mL day^?1 for acetochlor ESA). Although detection limits could thereby be reduced, most acidic analytes followed a biphasic uptake, proscribing the use of the conventional first-order model and preventing us from estimating time-weighted average concentrations. On the other hand, reducing the exposure window (3.1 vs. 46 cm²) allowed linear accumulations of all analytes over 35 days, but R _s s were dramatically reduced (e.g., 157 vs. 11 mL day^?1 for acetochlor ESA). Otherwise, the observation of biphasic releases of performance reference compounds (PRC), though mirroring acidic herbicide biphasic uptake, might complicate the implementation of the PRC approach to correct for environmental exposure conditions.

Solvent extraction of europium trifluoromethanesulfonate into nitrobenzene by using three electroneutral receptors

From extraction experiments and $ \gamma $ -activity measurements, the extraction constants corresponding to the general equilibrium Eu³⁺(aq) + 3 A^?(aq) + L(nb) $ \Leftrightarrow $ EuL³⁺(nb) + 3A^?(nb) taking place in the two-phase water–nitrobenzene system ( $ {\text{A}}^{ - } = {\text{CF}}_{ 3} {\text{SO}}_{3}^{ - } $ ; L = electroneutral receptors denoted by 1, 2, and 3 – see Scheme 1; aq = aqueous phase, nb = nitrobenzene phase) were evaluated. Further, the stability constants of the EuL³⁺ complexes in nitrobenzene saturated with water were calculated; they were found to increase in the series of 3 < 2 < 1.

Method for determination of microcystin-leucine-arginine in water samples based on the quenching of the fluorescence of bioconjugates between CdSe/CdS quantum dots and microcystin-leucine-arginine antibody

We have developed a method for the determination of microcystin-leucine-arginine (MC-LR) in water samples that is based on the quenching of the fluorescence of bioconjugates between CdSe/CdS quantum dots (QDs) and the respective antibody after binding of MC-LR. The core-shell CdSe/CdS QDs were modified with 2-mercaptoacetic acid to improve water solubility while their high quantum yields were preserved. Monoclonal MC-LR antibody was then covalently bioconjugated to the QDs. It was found that the fluorescence intensity of the bioconjugates was quenched in the presence of MC-LR. A linear relationship exists between the extent of quenching and the concentration of MC-LR. Parameters affecting the quenching were investigated and optimized. The limit of detection is 6.9?×?10^?11 mol L^?1 (3σ). The method was successfully applied to the determination of MC-LR in water samples.

Determination of nitrotyrosine in Arabidopsis thaliana cell cultures with a mixed-mode solid-phase extraction cleanup followed by liquid chromatography time-of-flight mass spectrometry

In this work, a method for the determination of trace nitrotyrosine (NO₂Tyr) and tyrosine (Tyr) in Arabidopsis thaliana cell cultures is proposed. Due to the complexity of the resulting extracts after protein precipitation and enzymatic digestion and the strong electrospray signal suppression displayed in the detection of both Tyr and NO₂Tyr from raw A. thaliana cell culture extracts, a straightforward sample cleanup step was proposed. It was based on the use of mixed-mode solid-phase extraction (SPE) using MCX-type cartridges (Strata?-X-C), prior to identification and quantitation using fast liquid chromatography–electrospray time-of-flight mass spectrometry. Unambiguous confirmation of both amino acids was accomplished with accurate mass measurements (with errors lower than 2 ppm) of each protonated molecule along with a characteristic fragment ion for each species. Recovery studies were accomplished to evaluate the performance of the SPE sample preparation step obtaining average recoveries in the range 92–101 %. Limit of quantitation obtained for NO₂Tyr in A. thaliana extracts was 3 nmol L^?1. Finally, the proposed method was applied to evaluate stress conditions of the plant upon different concentrations of peroxynitrite, a protein-nitrating compound, which induces the nitration of Tyr at the nanomolar range. Detection and confirmation of the compounds demonstrated the usefulness of the proposed approach.

Analysis of selected pharmaceuticals in fish and the fresh water bodies directly affected by reclaimed water using liquid chromatography-tandem mass spectrometry

A comprehensive method for the analysis of 11 target pharmaceuticals representing multiple commonly used therapeutic classes was developed for biological tissues (fish), reclaimed water, and the surface water directly affected by irrigation with reclaimed water. One gram of fish tissue homogenate was extracted by accelerated solvent extraction with methylene chloride followed by mixed-mode cation exchange solid phase extraction (SPE) cleanup and analyzed by liquid chromatography-tandem mass spectrometry. Compared to previously reported methods, the protocol produces cleaner extracts resulting in lower method detection limits. Similarly, an SPE method based on Oasis HLB cartridges was used to concentrate and cleanup reclaimed and surface water samples. Among the 11 target compounds analyzed, trimethoprim, caffeine, sulfamethoxazole, diphenhydramine, diltiazem, carbamazepine, erythromycin, and fluoxetine were consistently detected in reclaimed water. Caffeine, diphenhydramine, and carbamazepine were consistently detected in fish and surface water samples. Bioaccumulation factors for caffeine, diphenhydramine, and carbamazepine in mosquito fish (Gambusia holbrooki) were calculated at 29?±?26, 821?±?422, and 108?±?144, respectively. This is the first report of potential accumulation of caffeine in fish from a water body directly influenced by reclaimed water.

A microvolume molecularly imprinted polymer modified fiber-optic evanescent wave sensor for bisphenol A determination

A fiber-optic evanescent wave sensor for bisphenol A (BPA) determination based on a molecularly imprinted polymer (MIP)-modified fiber column was developed. MIP film immobilized with BPA was synthesized on the fiber column, and the sensor was then constructed by inserting the optical fiber prepared into a transparent capillary. A microchannel (about 2.0 μL) formed between the fiber and the capillary acted as a flow cell. BPA can be selectively adsorbed online by the MIP film and excited to produce fluorescence by the evanescent wave produced on the fiber core surface. The conditions for BPA enrichment, elution, and fluorescence detection are discussed in detail. The analytical measurements were made at 276 nm/306 nm (λ _ex/λ _em), and linearity of 3?×?10^?9–5?×?10^?6 g mL^?1 BPA, a limit of detection of 1.7?×?10^?9 g mL^?1 BPA (3σ), and a relative standard deviation of 2.4 % (n?=?5) were obtained. The sensor selectivity and MIP binding measurement were also evaluated. The results indicated that the selectivity and sensitivity of the proposed fiber-optic sensor could be greatly improved by using MIP as a recognition and enrichment element. Further, by modification of the sensing and detection elements on the optical fiber, the proposed sensor showed the advantages of easy fabrication and low cost. The novel sensor configuration provided a platform for monitoring other species by simply changing the light source and sensing elements. The sensor presented has been successfully applied to determine BPA released from plastic products treated at different temperatures.

Fluorescent turn-on detection of cysteine using a molecularly imprinted polyacrylate linked to allylthiol-capped CdTe quantum dots

CdTe quantum dots capped with thioglycolic acid (TGA) display a strong turn-on fluorescence response if exposed to solutions of cysteine (Cys). In order to exploit this effect, a molecularly imprinted polymer (MIP) for Cys was covalently linked to the QDs via allyl mercaptan. The resulting nanomaterials (QDs, MIP-coated QDs, and nonimprint-coated QDs) were characterized by FTIR and scanning electron microscopy. The adsorption of Cys was studied in phosphate buffer (pH 7.4) with respect to equilibration times (5, 15, and 40 min, respectively), binding constants [2.98, 2.42, and 0.96 (×10⁴ M^?1)], and Langmuir isotherms (R²?=?0.9995, 0.9999, and 0.9983) in the Cys concentration range between 3.33 μM to 500 μM. The method has a detection limit of 0.85 μM (3σ, blank, for n?=?10). The selectivity of the MIP-coated QDs for Cys over 19 other amino acids is similar to that of bare QDs, but MIP-QDs afford better recoveries of Cys from solutions also containing bovine serum albumin (90 %) and fetal bovine serum (97 %), respectively, when compared to the recoveries that are obtained with bare (non-imprinted) QDs (135 % and 120 %). This is probably due to the fact that the outer MIP shell largely reduces protein wrapping, dot aggregation, and matrix inclusion.

Development of a molecularly imprinted polymer-matrix solid-phase dispersion method for selective determination of β-estradiol as anabolic growth promoter in goat milk

A simple, fast, and sensitive method for determination of 17 β-estradiol (E2) in goat milk samples has been developed by combining selective molecularly imprinted matrix solid-phase dispersion (MIP–MSPD) and liquid chromatography with diode-array detection (DAD). The molecularly imprinted polymer was synthesized by use of 17β-estradiol as template molecule, methacrylic acid as functional monomer, ethylene glycol dimethacrylate as crosslinker monomer, azobisisobutyronitrile as initiator, and acetonitrile as porogen, and was used as selective solid support for matrix solid-phase dispersion. The selected dispersant had high affinity for E2 in the goat milk matrix and the extract obtained was sufficiently clean for direct injection for HPLC analysis without any interferences from the matrix. The proposed MIP–MSPD method was validated for linearity, precision, accuracy, decision limit (CCα) and detection capability (CCβ), in accordance with European Commission Decision 2002/657/EC criteria. Linearity ranged from 0.3–10 μg g^?1 (correlation coefficient r ²?>?0.999). Mean recovery of E2 from goat milk samples at different spiked levels was between 89.5 and 92.2%, with RSD values within 1.3–2%. CCα and CCβ values were 0.36 and 0.39 μg g^?1, respectively. The developed MIP–MSPD method was successfully applied to direct determination of E2 in goat milk samples.

Evaluation of human plasma sample preparation protocols for untargeted metabolic profiles analyzed by UHPLC-ESI-TOF-MS

Eight human plasma preparation protocols were evaluated for their suitability for metabolomic studies by ultra-high-performance liquid chromatography coupled with electrospray ionization time-of-flight mass spectrometry: organic solvent protein precipitation (PPT) with either methanol or acetonitrile in 2:1 and 3:1 (v/v) ratios with plasma; solid-phase extraction (SPE) using C₁₈ or HybridSPE cartridges; and a combination of PPT and SPE C₁₈ cartridges and microextraction by packed sorbent. A study design in which the order of injection of the samples was not randomized is presented. The analyses were conducted in a BEH C₁₈ column (1.7 μm, 2.1 mm?×?100 mm) using a linear gradient from 100 % water to 100 % methanol, both with 0.1 % formic acid, in 21 min. The most reproducible protocol considering both the univariate and the multivariate analysis results was PPT with acetonitrile in a 2:1 (v/v) ratio with plasma, offering a mean coefficient of variation of the area of all the detected features of 0.15 and one of the best clusterings in the principal component analysis plots. On the other hand, the highest number of extracted features was achieved using methanol in a 2:1 (v/v) ratio with plasma as the PPT solvent, closely followed by the same protocol with acetonitrile in a 2:1 (v/v) ratio with plasma, which offered only 1.2 % fewer repeatable features. In terms of concentration of remaining protein, protocols based on PPT with acetonitrile provided cleaner extracts than protocols based on PPT with methanol. Finally, pairwise comparison showed that the use of PPT- and SPE-based protocols offers a different coverage of the metabolome. Graphical Abstract

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Lipidomic analysis of polyunsaturated fatty acids and their oxygenated metabolites in plasma by solid-phase extraction followed by LC-MS

The present work describes the development of a robust and sensitive targeted analysis platform for the simultaneous quantification in blood plasma of lipid oxygenated mediators and fatty acids using solid-phase extraction (SPE) and high-performance liquid chromatography coupled to tandem mass spectrometry (HPLC-MS/MS). The concurrent analysis of these lipid mediators is challenging because of their instability, differences in solubility, and the frequent occurrence of isobaric forms with similar fragmentation patterns. Results demonstrated that the reduction of SPE temperature to 4 °C is a critical parameter for preserving the hydroperoxy derivatives. Polymeric HLB cartridges increased 40–50 % ARA, EPA, and DHA sensitivity compared to C18 sorbent and also provided higher global performance for most hydroxides and other oxidation products. The proposed method for the two tested mass analyzers yields high sensitivity, good linearity, and reproducibility, with detection limits ranging 0.002–7 ng/mL and global recoveries as high as 85–112 %. However, the additional advantage of the linear ion trap (LIT) mass analyzer working in full scan product ion mode, compared to the triple quadrupole (QqQ) operating in multiple reaction monitoring (MRM), should be noted: the full scan product ion mode provides the full fragmentation spectra of compounds that allowed the discrimination of coeluting isomers and false positive identifications without additional chromatography development. The proposed lipidomic procedure demonstrates a confident, simple, and sensitive method to profile in plasma a wide range of lipid eicosanoid and docosanoid mediators, including innovatively the analysis of hydroperoxy congeners and nonoxidized PUFA precursors.

The calculation of the detection efficiency in the calibration of gross alpha–beta systems

This paper presents a method for efficiency calibration of a measuring alpha–beta system PROTEAN ORTEC, MPC-2000-DP, using standard radioactive sources. The system is used to measure gross alpha–beta activity concentrations in environmental samples. The calculated efficiencies of detection were subsequently introduced in the system for two working geometries: measuring geometry—gross alpha–beta $ \varepsilon_{\alpha }^{g} $  = 31,37 ± 0.25 (%)—the alpha efficiency and $ \varepsilon_{\beta }^{g} $  = 44.94 ± 0.69 (%)—the beta efficiency, where the spillover factor is $ X_{\text{talk}}^{g} $  = 25.59 ± 0.50 (%) and measuring geometry up alpha–beta $ \varepsilon_{\alpha }^{u} $  = 36.23 ± 0.29 (%)—the alpha efficiency and $ \varepsilon_{\beta }^{u} $  = 48.53 ± 0.74 (%)—the beta efficiency, where the spillover factor is $ X_{\text{talk}}^{u} $  = 31.08 ± 0.60 (%).     

Excess Molar Volumes, Excess Molar Enthalpies and Excess Isentropic Compressibilities of 1-Methyl Pyrrolidin-2-one with Water and Propanols

Excess molar volumes V ^E, excess molar enthalpies H ^E, and speeds of sound u for 1-methyl pyrrolidin-2-one (1) + water or propan-1-ol or propan-2-ol (2) binary mixtures have been measured over the entire composition range (at 308.15 K) using a dilatometer, calorimeter and interferometer. Speeds of sound data, u, of (1 + 2) binary mixtures have been utilized to determine excess isentropic compressibilities, $ \kappa_{S}^{\text{E}} $ . The observed V ^E, H ^E and $ \kappa_{S}^{\text{E}} $ data have been analyzed in terms of (1) Graph theory (which involves the topology of the constituents of mixture), and (2) the Prigogine–Flory–Patterson theory. Analysis of V ^E data in terms of Graph theory suggests that 1-methyl pyrrolidin-2-one, water, propan-1-ol, and propan-2-ol exist as associated molecular entities. IR studies lend additional support to the proposed molecular entities in (1 + 2) mixtures. It has been observed that V ^E, H ^E and $ \kappa_{S}^{\text{E}} $ values predicted by Graph theory compare well with their corresponding experimental values.     

Capacitive sensing of microcystin variants of Microcystis aeruginosa using a gold immunoelectrode modified with antibodies,gold nanoparticles and polytyramine

We report on the application of an automated and easy-to-use device to directly measure the immunoreactions between adda-specific monoclonal antibodies and microcystins. The antibodies were immobilized on a gold electrode whose surface was modified first with polytyramine and then with gold nanoparticles. The immunoreaction leads to a change in the capacitance of the system. Under optimum conditions, the sensor is capable of performing stable regeneration-assay cycles and has a low detection limit at a concentration of 0.01 pM level of microcystin-leucine-arginine (MC-LR). The surface of the biosensor can be regenerated with pH 2.5 glycine buffer which dissociates the antibody-antigen complex. The biosensor was used to monitor the production of microcystins during batch cultivation of Microcystis aeruginosa (isolated from ponds in Botswana). Liquid chromatography coupled to MS/MS detection was used to identify three variants, viz. MC-LR (995.6 Da), DmMC-LR (981.2 Da) and MC-LA (910.5 Da).

An Improved and Validated LC Method for Resolution of Panthenol

An improved LC method was developed and validated for determination of enantiomeric purity of panthenol in bulk drugs. The method is based on derivatization of panthenol with 3,5-dinitrobenzoyl chloride. Baseline separation with resolution >2.7 was achieved within 20 min on Kromasil CHI-DMB (250 × 4.6 mm) column using n-hexane:ethanol (95:5 v/v) as mobile phase at a flow rate of 1.5 mL min^?1. The analytes were detected by their UV absorbance at 265 nm. The effects of ethanol, 2-propanol and temperature on enantioselectivity and resolution of enantiomers were evaluated. The method was extensively validated and proved to be robust. The recoveries were between 98.3 and 101.4% with <1.6% relative standard deviation. The regression equations for the derivatives of d-panthenol and l-panthenol were y ₁ = 18.01x ₁ ? 32.56 (r ₁ ²  = 0.9984) and y ₂ = 17.855x ₂ ? 28.16 (r ₂ ²  = 0.9990), respectively. The LOD and LOQ for the derivative of d-panthenol were 10.6 and 37.4 μg mL^?1 and for the derivative of l-panthenol were 12.1 and 40 μg mL^?1, respectively. The improved method was found to be simple, rapid, and sensitive for the determination of enantiomeric purity of panthenol in bulk drugs.     

Preparation of MIP grafts for quercetin by tandem aryl diazonium surface chemistry and photopolymerization

The food antioxidant quercetin was used as a template in an ultrathin molecularly imprinted polymer (MIP) film prepared by photopolymerization. Indium tin oxide (ITO) plates were electrografted with aryl layers via a diazonium salt precursor bearing two terminal hydroxyethyl groups. The latter act as hydrogen donors for the photosensitizer isopropylthioxanthone and enabled the preparation of MIP grafts through radical photopolymerization of methacrylic acid (the functional monomer) and ethylene glycol dimethacrylate (the crosslinker) in the presence of quercetin (the template) on the ITO. The template was extracted, and the remaining ITO electrode used for the amperometric determination of quercetin at a working potential of 0.26 V (vs. SCE). The analytical range is from 5.10^?8 to 10^?4 mol L^?1, and the detection limit is 5.10^?8 mol L^?1.

Complexation of some univalent organic cations with benzo-18-crown-6 in nitrobenzene saturated with water

From extraction experiments and γ-activity measurements, the exchange extraction constants corresponding to the general equilibrium C⁺(aq) + 1·Na⁺(nb) ? 1·C⁺(nb) + Na⁺(aq) taking place in the two-phase water–nitrobenzene system (C⁺ = univalent organic cation, 1 = benzo-18-crown-6; aq = aqueous phase, nb = nitrobenzene phase) were evaluated. Moreover, the stability constants of the 1·C⁺ complex species in nitrobenzene saturated with water were calculated; they were found to increase in the following cation order: protonated tyramine, protonated l-valine methyl ester < protonated dopamine < protonated serotonin < methylammonium < protonated hexamethylenetetramine < ethanolammonium < protonated dl-noradrenaline.