Identification and characterization of impurities in an insecticide, commercial chlorantraniliprole using liquid chromatography-tandem mass spectrometry.

RATIONALE: Ensuring the global safety and effectiveness of agrochemicals has become imperative. An in-depth understanding of impurity profiles of products is crucial, especially for high-demand agrochemicals, where impurities may be more toxic and persistent than original agrochemicals. This study focuses on the detection and identification of impurities in a commercial chlorantraniliprole (CAP), an anthranilic diamide class broad-spectrum insecticide. METHODS: Commercial CAP was collected from an agrochemical supplier in India and was analyzed using a high-performance liquid chromatography-photodiode array (HPLC-PDA) (Agilent 1260; wavelength, 220 nm) with a Zorbax RP SB-C18 (250 × 4.6 mm, 5 µm) column and liquid chromatography-mass spectrometry (LC-MS) (Agilent 6545 quadrupole time of flight (Q-TOF)) techniques to identify the impurities. The impurities were isolated by preparative HPLC using a Zorbax-DB C18 (250 × 9.4 mm, 5 µm) column. liquid chromatography- tandem mass spectrometry (LC-MS/MS) experiments (Q-TOF) were performed on CAP and its impurities to obtain their structural data. RESULTS: HPLC-PDA analysis of CAP showed four major impurities (IM-1 to IM-4) ranging from 0.76% to 4.1%. The positive ion electrospray ionization (ESI) mass spectra of CAP and its impurities showed dominant [M + H]+ ions in addition to [M + Na]+ , [M + K]+ , and [2M + Na]+ ions. High-resolution mass spectrometry (HRMS) data provided the elemental composition of the compounds, and isotopic distribution patterns revealed the number of Cl and/or Br atoms present in them. The structures of impurities were proposed based on the LC-MS/MS) data and further confirmed by nuclear magnetic resonance (NMR) data on isolated impurities/synthesis. CONCLUSION: The quality and impurities of CAP, a popular insecticide, must be assessed and described for its efficacy and safety. In this study, four impurities of CAP were detected using HPLC and successfully characterized using LC-HRMS, LC-MS/MS, and NMR data. The method is useful for verifying the purity of CAP as well as helping in the identification of its possible impurities.

LC-MS/MS Analysis of Reaction Products of Arginine/Methylarginines with Methylglyoxal/Glyoxal.

Methylglyoxal (MGO) and glyoxal (GO) are toxic α-dicarbonyl compounds that undergo reactions with amine containing molecules such as proteins and amino acids and result in the formation of advanced glycation end products (AGEs). This study aimed at investigating the reactivity of arginine (Arg) or dimethylarginine (SDMA or ADMA) with MGO or GO. The solutions of arginine and MGO or GO were prepared in PBS buffer (pH 7.4) and incubated at 37 °C. Direct electrospray ionization-high-resolution mass spectrometry (ESI-HRMS) analysis of the reaction mixture of Arg and MGO revealed the formation of Arg-MGO (1:1) and Arg-2MGO (1:2) products and their corresponding dehydrated products. Further liquid chromatography (LC)-MS analyses revealed the presence of isomeric products in each 1:1 and 1:2 product. The [M + H]+ of each isomeric product was subjected to MS/MS experiments for structural elucidation. The MS/MS spectra of some of the products showed a distinct structure indicative fragment ions, while others showed similar data. The types of products formed by the arginines with GO were also found to be similar to that of MGO. The importance of the guanidine group in the formation of the AGEs was reflected in similar incubation experiments with ADMA and SDMA. The structures of the products were proposed based on the comparison of the retention times and HRMS and MS/MS data interpretation, and some of them were confirmed by drawing analogy to the data reported in the literature.

Identification and Characterization of the Isomeric Impurity of the Fungicide "Cyazofamid".

Identification and characterization of biproducts/ impurities present in agrochemicals are critical in view of their efficacy and safety towards public health. We herein present our study on identification and characterization of an impurity, 5-chloro-2-cyano-N,N-dimethyl-4-p-tolylimidazole-1-sulfonamide (2) present in the fungicide, "cyazofamid". Intermittent HPLC analysis of the reaction of substituted imidazole (1) with N,N-dimethylsulfamoyl chloride suggested that 2 is formed during the reaction. Isolation by preparative HPLC and characterization by NMR, LC/HRMS, MS/MS and single crystal XRD analysis confirmed 2 as an isomer of cyazofamid, wherein the N,N-dimethyl sulfonamide group was positioned on the other nitrogen of imidazole in close proximity to chloride group. Computational studies further supported the formation of 2 and ruled out the other possible isomeric structures.

Dietary Lactobacillus acidophilus and Mannan-Oligosaccharides Alter the Lipid Metabolism and Health Indices in Broiler Chickens.

The effects of dietary Lactobacillus acidophilus (LBA) and mannan-oligosaccharide (MOS) supplementation on lipid metabolism and consequent lipid profile and health indices in broiler chicken were investigated in this study. Supplementation of 0.2% MOS along with either 106 or 107 LBA/g feed in broiler chicken downregulated hepatic expression of genes involved in lipogenesis, and upregulated expression of lipolytic genes. It caused decline of lipogenesis and increase of lipid oxidation which resulted in lower carcass fat content. None of the genes studied influenced fatty acid profile of chicken meat except the expression of stearoyl CoA (Δ9) desaturase-1 (SCD-1) whose upregulation increased monounsaturated fatty acid (MUFA) content at the cost of saturated fatty acid (SFA) content. The lipid metabolism indices of chicken meat such as ∆9 desaturase index (DI) increased in birds supplemented with 0.2% MOS along with either 106 or 107 CFU LBA/g feed, whereas no effect was observed on ∆5 + ∆6 DI. The supplementation of 0.2% MOS along with either 106 or 107 CFU LBA/g feed in birds improved the health indices of chicken meat due to upregulation of SCD-1 expression. The supplementation of 0.2% MOS along with either 106 or 107 CFU LBA/g feed in broiler chicken produced hypocholesterolemic and hypolipidemic effects with improved serum cardio-protective indices.

Dietary synbiotic supplementation improves the growth performance, body antioxidant pool, serum biochemistry, meat quality, and lipid oxidative stability in broiler chickens.

The present study investigated the effects of Lactobacillus acidophilus (LBA) and mannan-oligosaccharides (MOS) supplementation on the production performance, serum biochemistry, antioxidant profile, health indices, meat quality, and lipid oxidative stability of broiler chicken. A total of 252 commercial broiler chickens at 1 d old of uniform body weight were randomly allocated to 6 maize-soybean-based dietary treatments: T1 (control diet), T2 ( antibiotic bacitracin methylene di-salicylate [BMD] at 20 mg/kg diet), T3 (MOS at 0.1% + LBA at 106 CFU/g feed), T4 (MOS at 0.1% + LBA at 107 CFU/g feed), T5 (MOS at 0.2% + LBA at 106 CFU/g feed), and T6 (MOS at 0.2% + LBA at 107 CFU/g feed). Each treatment was assigned to 6 replicates of 7 birds. The samples for meat quality and serum biochemistry analysis were taken from 12 birds per treatment (2 birds/replicate). The results revealed better (P < 0.01) growth performance and production efficiency of birds fed either T5 or T6 diet compared to control or BMD supplemented diet and BMD-supplemented birds superseded the control birds. Higher (P < 0.01) serum and liver antioxidant enzyme activities, meat antioxidant capacity (2, 2-azinobis-3-ethylbenzothiazoline-6-sulfonic acid [ABTS] and 1, 1-diphenyl-2-picrylhydrazyl [DPPH] assays], serum total protein, high-density lipoproteins (HDL) cholesterol (P < 0.05), and globulin levels (P < 0.01) were observed in birds fed either T5 or T6 diet compared to control or BMD supplemented birds, whereas, lower lipid oxidation (P < 0.01), cardiac risk ratio, atherogenic coefficient, atherogenic index of plasma, serum glucose, triglyceride, total cholesterol levels (P < 0.01), and serum albumin-to-globulin ratio (P < 0.05) were observed in the chickens. The pH of meat from birds fed T4, T5 or T6 diet was lower (P < 0.01) compared to control and other treatments. The extract release volume (ERV), water holding capacity (WHC), and protein content of meat were higher (P < 0.05) in birds fed either T5 or T6 diet compared to control or BMD supplemented birds. Thus, it was concluded that the supplementation of 0.2% MOS along with LBA at 106 CFU/g is optimum for better growth performance, serum biochemistry, antioxidant profile, health indices, meat quality, and lipid oxidative stability of broiler chickens.

Identification and characterization of impurities in an insecticide, bifenthrin technical.

The HPLC-DAD and GC/MS methods were successfully used for the identification and characterization of the impurities in an agrochemical insecticide, bifenthrin technical. Three impurities ranging from 0.175%-0.541% were detected by the HPLC-DAD method. The LC/MS technique with ESI or APCI source failed to detect the impurities detected by HPLC-DAD, due to lack of ionization in ESI or APCI. The three impurities were enriched by prep-HPLC, and then their structures were elucidated based on the GC/EIMS and CIMS data. The EI mass spectra of bifenthrin and its impurities displayed molecular ion and provided structure indicative fragment ions; the CIMS data further confirmed their molecular weight. The identity of the impurity 1 was further confirmed by the synthesis of the authentic sample followed by NMR and GC/MS data.

Dietary Mannan-oligosaccharides potentiate the beneficial effects of Bifidobacterium bifidum in broiler chicken.

This study investigated the effects of dietary Bifidobacterium bifidum (BFD) and mannan-oligosaccharide (MOS), as a synbiotic, on the production performance, gut microbiology, serum biochemistry, antioxidant profile and health indices of broiler chicken. Six dietary treatments were T1 (negative control), T2 (positive control-20 mg antibiotic BMD kg-1 diet; BMD: bacitracin methylene disalicylate), T3 (0·1% MOS + 106  CFU BFD per g feed), T4 (0·1% MOS + 107  CFU BFD per g feed), T5 (0·2% MOS + 106  CFU BFD per g feed) and T6 (0·2% MOS + 107  CFU BFD per g feed). Significantly (P < 0·01) better growth performance and efficiency was observed in birds supplemented with 0·2% MOS along with 106  CFU BFD per g of feed compared to BMD and control birds. Supplementation with 0·2% MOS along with either 106 or 107  CFU BFD per g feed reduced (P < 0·01) the gut coliform, Escherichia coli, total plate count, and Clostridium perfringens count and increased the Lactobacillus and Bifidobacterium count. Significantly (P < 0·01) higher serum and liver antioxidant enzyme pool, serum HDL cholesterol and lower serum glucose, triglyceride, total cholesterol, cardiac risk ratio, atherogenic coefficient and atherogenic index of plasma were observed in birds supplemented with 0·2% MOS along with 106  CFU BFD per g of feed compared to control or BMD supplemented birds. Better production performance, gut microbial composition, serum biochemistry, antioxidant profile and health indices were depicted by broiler chicken supplemented with 0·2% MOS and 106  CFU BFD per g of feed.