Innovative approaches to biologic development on the trail of CT-P13: biosimilars, value-added medicines, and biobetters.

The biosimilar concept is now well established. Clinical data accumulated pre- and post-approval have supported biosimilar uptake, in turn stimulating competition in the biologics market and increasing patient access to biologics. Following technological advances, other innovative biologics, such as "biobetters" or "value-added medicines," are now reaching the market. These innovative biologics differ from the reference product by offering additional clinical or non-clinical benefits. We discuss these innovative biologics with reference to CT-P13, initially available as an intravenous (IV) biosimilar of reference infliximab. A subcutaneous (SC) formulation, CT-P13 SC, has now been developed. Relative to CT-P13 IV, CT-P13 SC offers clinical benefits in terms of pharmacokinetics, with comparable efficacy, safety, and immunogenicity, as well as increased convenience for patients and reduced demands on healthcare system resources. As was once the case for biosimilars, nomenclature and regulatory pathways for innovative biologics require clarification to support their uptake and ultimately benefit patients.

Method development, matrix effect, and risk assessment of 49 multiclass pesticides in kiwifruit using liquid chromatography coupled to tandem mass spectrometry.

In the present study, a liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) method with a minimal matrix effect (ME) was developed and validated for simultaneous determination of a diverse range of pesticides (49) in kiwifruit. Samples extracted by the quick, easy, cheap, effective, rugged, and safe (QuEChERS) citrate-buffered method were analyzed either without purification or following purification (with primary secondary amine (PSA) or PSA + graphitized carbon black (GCB)). With the addition of a clean-up step, the suppression of the ME decreased, with a higher number of pesticides determined by the application of PSA + GCB. The method exhibited good linearity with coefficients of determination (R2) ≥ 0.9972 and satisfactory recoveries (70-120%) with a relative standard deviations (RSDs) <10%. The limits of quantification (LOQ) were lower than the maximum residue limits (MRLs) set by the Korean Ministry of Food and Drug Safety (MFDS) and the CODEX Alimentarius. The developed method was applied to the real samples and the results indicated that the quantitated levels of all pesticides, except for pyraclostrobin and carbendazim, are lower than the MRLs set by the regulatory authorities. The percentage of the acceptable daily intake was <20%, suggesting that there is no risk associated with the intake of residual pesticides through kiwifruit.

Trichloroethylene and Parkinson's Disease: Risk Assessment.

This study was conducted to investigate the mechanism of action and extent of selective dopaminergic neurodegeneration caused by exposure to trichloroethylene (TCE) leading to the endogenous formation of the neurotoxin 1-trichloromethyl-1,2,3,4-tetrahydro-ß-carboline (TaClo) in rodents. Beginning at 3 months of age, male C57BL/6 mice received oral TCE dissolved in vehicle for 8 months. Dopaminergic neuronal loss was assessed by nigral tyrosine hydroxylase (TH) immunoreactivity. Selective dopaminergic neurodegeneration was determined based on histological analysis of non-dopaminergic neurons in the brain. Behavioral assays were evaluated using open field activity and rotarod tests. Mitochondrial complex I activity, oxidative stress markers, and microglial activation were also examined in the substantia nigra. The level of TaClo was detected using HPLC-electrospray ionization tandem mass spectrometry. Dopaminergic neurotoxicity of TaClo was determined in midbrain organotypic cultures from rat pups. Following 8 months of TCE treatment, there was a progressive and selective loss of 50% of the dopaminergic neurons in mouse substantia nigra (SN) and about 50% loss of dopamine and 72% loss of 3,4-dihydroxyphenylacetic acid in the striatum, respectively. In addition, motor deficits, mitochondrial impairment, oxidative stress, and inflammation were measured. TaClo content was quantified in the brain after TCE treatment. In organotypic cultures, TaClo rather than TCE induced dopaminergic neuronal loss, similar to MPP+. TCE exposure may stimulate the endogenous formation of TaClo, which is responsible for dopaminergic neurodegeneration. However, even prolonged administration of TCE was insufficient for producing a greater than 50% loss of nigral dopamine neurons, indicating that additional co-morbid factors would be needed for mimicking the profound loss of dopamine neurons seen in Parkinson's disease.

Dissipation kinetics, pre-harvest residue limits, and dietary risk assessment of the systemic fungicide metalaxyl in Swiss chard grown under greenhouse conditions.

The residual behavior of the systemic fungicide, metalaxyl, in Swiss chard cultivated at two different locations under greenhouse conditions was investigated using high-performance liquid chromatography coupled with an ultraviolet detector (HPLC-UVD). Samples were randomly collected over 14 days and extracted using acetonitrile, partitioned using solid sodium chloride, and a solid-phase extraction (SPE) NH2 cartridge was used for cleanup. The linearity over a concentration range 0.05-50 mg/L was excellent with a coefficient of determination (R2) of 0.9997. The recovery rate ranged from 77.05 to 88.92% with relative standard deviations (RSDs) ≤ 10.74, and the limits of detection (LOD) and quantification (LOQ) were 0.0033 and 0.01 mg/kg, respectively. The initial (2 h after application) deposits were 4.69 and 5.90 mg/kg for sites 1 and 2, respectively, which increased to 4.95 and 6.57 mg/kg, respectively, one day post-application, owing to the systemic properties of the fungicide. The dissipation half-life was 5.3 and 6.0 days for sites 1 and 2, respectively. The pre-harvest residue limit (PHRL) suggested that if 55.38 and 47.23 mg/kg was applied 10 days before harvest or 33.28 and 30.73 mg/kg was applied 5 days before harvest (for sites 1 and 2, respectively) then the concentration will fall below the maximum residue limit (MRL = 20.0 mg/kg) at the time of harvest. The dietary risk assessment, estimated as hazard quotient (RQ%), indicate that metalaxyl can be safely used in/on Swiss chard, with no hazardous effects expected for consumers.

Analytical approach, dissipation pattern and risk assessment of pesticide residue in green leafy vegetables: A comprehensive review.

The category of 'leafy vegetables' comprises a wide range of plants, including cabbage, lettuce, leeks, spinach, Swiss chard and kale, and it forms a significant component of the human diet. Typically, leafy vegetables are low in calories and fat, are great sources of vitamins, protein, dietary fibre and minerals (including iron, calcium, and nitrates), and are rich in phytochemicals. To counter the impact of pests on vegetables, a broad variety of pesticides are used. Because of their large surface areas, leafy vegetables are expected to have high residual pesticide levels. As such, a sound analytical approach is needed to detect and quantify residue levels that are equal to or lower than the maximum residue limits, thus rendering the products safe for consumption. Overall, leafy vegetables consumed raw (after a tap water wash only), boiled or steamed contribute 2% of total vegetable consumption globally, and they might have a comparatively greater influence on health than cereal ingestion. Consequently, in this review paper, we highlight the importance of leafy vegetables, the pesticides that are commonly used on them and various analytical techniques, including sample preparation, extraction, clean-up and final detection. The effects on dissipation patterns, pre-harvest residue limits and safety/risks imposed by various pesticides are also reviewed and discussed. In conclusion, environmentally friendly extraction methods coupled with high-throughput techniques with greater reproducibility and lower uncertainty are needed for quantifying residues in leafy vegetables at very low concentrations. Commercial and household food preparation, such as washing, peeling, blanching and cooking are effective in removing most of the pesticide residues that are loosely attached on vegetables.

Decline patterns and risk assessment of 10 multi-class pesticides in young sprout amaranth (Amaranthus mangostanus) under greenhouse growing conditions.

The present study was designed to investigate the residual decline pattern and the risk assessment of 10 different class pesticides, namely azoxystrobin, boscalid, diazinon, diethofencarb, difenoconazole, etofenprox, flubendiamide, paclobutrazol, and pyraclostrobin in young vegetative amaranth (Amaranthus mangostanus) sprayed once or twice under greenhouse growing conditions. Field-incurred samples, collected at 3, 7, or 10 days after application of both treatments, were extracted and purified with the quick, easy, cheap, effective, rugged, and safe "QuEChERS" citrate-buffered method and analyzed with liquid chromatography-electrospray ionization tandem mass spectrometry (LC-MS/MS) in positive ion mode. The linearity was satisfactory with determination coefficients (R 2) falling between 0.9817 and 0.9999 and limits of detection (LOD) and quantification (LOQ) values of 0.0007 and 0.002 mg/kg, respectively. The mean recovery rate at four spiking levels (equivalent to 5, 10, 50, and 100 × LOQ) ranged from 78.1 to 131.6% with a relative standard deviation (RSD) of < 11%. Substantial differences in the initial deposit between the tested analytes were observed and clearly indicated that the structure, as well as the initial concentration of applied products, greatly affected the residue deposit. From the obtained residual data, the provisional marginal maximum residue limits (MRLs) and the pre-harvest intervals (PHI) were proposed. Risk assessment was evaluated by comparing the theoretical maximum daily intake (TMDI) with the acceptable daily intake (ADI). Herein, the TMDI was lower than the ADI (TMDI/ADI ratio ≤ 80% set by the Korean Ministry of Food and Drug Safety) except for difenoconazole (80.92%, marginally higher), indicating that the vegetative amaranth is not hazardous and can be consumed safely by Korean consumers.

UPLC-MS/MS method for determination of bepotastine in human plasma.

A sensitive and rapid method for quantitation of bepotastine in human plasma has been established using ultra performance liquid chromatography-electrospray ionization tandem mass spectrometry (UPLC-ESI-MS/MS). Valsartan was used as an internal standard. Bepotastine and internal standard in plasma sample were extracted using ethylacetate (liquid-liquid extraction). A centrifuged upper layer was then evaporated and reconstituted with the mobile phase of acetonitrile--5 mM ammonium formate (pH 3.5) (85:15, v/v). The reconstituted samples were injected into a phenyl column. Using MS/MS in the multiple reaction monitoring mode, bepotastine and valsartan were detected without severe interference from human plasma matrix. Bepotastine produced a protonated precursor ion ([M+H](+)) at m/z 389 and a corresponding product ion at m/z 167. And the internal standard produced a protonated precursor ion ([M+H](+)) at m/z 436 and a corresponding product ion at m/z 291. Detection of bepotastine in human plasma by the UPLC-ESI-MS/MS method was accurate and precise with a quantitation limit of 0.2 ng/mL. The validation, reproducibility, stability and recovery of the method were evaluated. The method has been successfully applied to pharmacokinetic studies of bepotastine in human plasma.