Yongliang Cui

Design, Synthesis, and Insecticidal Activities of Phthalamides Containing a Hydrazone Substructure

Fluobendiamide is the first commercialized artificial synthetic insecticide acting on the ryanodine receptor. This new molecule possesses a combination of excellent insecticidal activity and eco-friendly characteristics with a skeleton structure of phthalamide. In this study, we incorporated hydrazone, present in many pesticidal compounds reported during the last two decades, into phthalamide derivatives via Schiff-base condensation and aminolysis to obtain 21 new compounds; these compounds were characterized by proton nuclear magnetic resonance ((1)H NMR), infrared spectroscopy (IR), and high-resolution mass spectrometry (HRMS) or elemental analysis. A preliminary bioassay against peach aphids ( Myzus persicae ) revealed that the title compounds exhibited good stomach toxicity at 600 mg/L. Twelve new compounds were found to display higher activity than postive control flubendiamide (LC(50) = 184.099 mg/L), however, LC(50) was less than 100 mg/L only for compounds 4e, 4o, 4s, 4t (59-77 mg/L). That is, combinations of a p-fluorophenyl or (methyl)thienyl group at the Ar position with an isopropyl or cyclohexyl group at the R position might improve the lethality of the designed phthalamide derivative. Preliminary results of a bioassay at 600 mg/L against diamondback moth ( Plutella xylostella , Linnaeus) showed that only the title compound 4e possessed good larvicidal activity. On comparison of the bioassay results of stomach toxicity and larvicidal activity, it is noteworthy that the compound incorporating phenylpyrazolyl exhibited good larvicidal activity and poor stomach activity.

Development of a Specific Monoclonal Antibody-Based ELISA to Measure the Artemether Content of Antimalarial Drugs

Artemether is one of the artemisinin derivatives that are active ingredients in antimalarial drugs. Counterfeit and substandard antimalarial drugs have become a serious problem, which demands reliable analytical tools and implementation of strict regulation of drug quality. Structural similarity among artemisinin analogs is a challenge to develop immunoassays that are specific to artemisinin derivatives. To produce specific antibodies to artemether, we used microbial fermentation of artemether to obtain 9-hydroxyartemether, which was subsequently used to prepare a 9-O-succinylartemether hapten for conjugation with ovalbumin as the immunogen. A monoclonal antibody (mAb), designated as 2G12E1, was produced with high specificity to artemether. 2G12E1 showed low cross reactivities to dihydroartemisinin, artemisinin, artesunate and other major antimalarial drugs. An indirect competitive enzyme linked immunosorbent assay (icELISA) developed showed a concentration causing 50% of inhibition for artemether as 3.7 ng mL−1 and a working range of 0.7–19 ng mL−1. The icELISA was applied for determination of artemether content in different commercial drugs and the results were comparable to those determined by high-performance liquid chromatography analysis. In comparison with reported broad cross activity of anti-artemisinin mAbs, the most notable advantage of the 2G12E1-based ELISA is its high specificity to artemether only.

Validation of ELISA for Quantitation of Artemisinin-Based Antimalarial Drugs

The circulation of counterfeit or substandard artemisinins (ARTs) in malaria-endemic areas poses a serious threat to the long-term use of these drugs. Here, we validated an indirect competitive enzyme-linked immunosorbent assay (icELISA) for quantification of ARTs and found that 50% of inhibitory concentrations of dihydroartemisinin, artemether, and artesunate were 8.1, 207.0, and 4.7 ng/mL, respectively. We compared the icELISA with high-performance liquid chromatography (HPLC) for quantifying ART and its derivatives in 22 convenience samples of commercial antimalarial drugs. Paired t tests showed a borderline significant difference between the two methods (mean = 0.03, 95% confidence interval [CI] 0.00-0.07, P = 0.074) and the icELISA results were more variable than those of the HPLC analysis (P < 0.001), suggesting that further improvement is needed to enhance the performance of the icELISA. Our results showed that the icELISA has the potential to be improved for quality assurance of ARTs at the point of care in endemic settings.

Development of a Specific Monoclonal Antibody for the Quantification of Artemisinin in Artemisia annua and Rat Serum

Artemisinin, extracted from Artemisia annua, and its derivatives are important frontline antimalarials. To produce specific antibodies for the detection and quantification of artemisinin, artemisinin was transformed to 9-hydroxyartemisinin by microbial fermentation, which was used to prepare a 9-succinate artemisinin hapten for conjugation with ovalbumin. A monoclonal antibody (mAb), designated as 3H7A10, was selected from hybridoma cell lines which showed high specificity to artemisinin. No competitive inhibition was observed with artesunate, dihydroartemisinin, and artemether for up to 20,000 ng mL(-1). An indirect competitive enzyme-linked immunosorbent assay (icELISA) was developed, which showed a concentration causing 50% of inhibition (IC50) for artemisinin as 2.6 ng mL(-1) and a working range of 0.6-11.5 ng mL(-1). The icELISA was applied for the quantification of artemisinin in crude extracts of wild A. annua and the study of pharmacokinetics of artemisinin in rat serum after intraperitoneal injection. The results were highly correlated with those determined by HPLC-UV analysis (R(2) = 0.9919). In comparison with reported antiartemisinin mAbs which have broad cross-reactivity with other artemisinin derivatives, the high specificity of 3H7A10 for artemisinin will enable development of methods for quantification of artemisinin in Artemisia plants and antimalarial drugs such as Arco and for pharmacokinetic studies.

Production of Monoclonal Antibody to Herbicide Fenoxaprop-ethyl

Fenoxaprop-ethyl is a selective aryloxyphenoxypropionate herbicide used widely to control annual and perennial grasses. A monoclonal antibody (MAb) against fenoxaprop-ethyl (FE), designated as 3E6B9C, was produced and had very low cross-reactivity with some of its structural analogs, such as clodinafop-propargyl, diclofop-methyl, lactofen, and quizalofop-p-ethyl. An indirect competitive enzyme-linked immunosorbent assay (icELISA) was developed. The concentration of R-(+)-fenoxaprop-ethyl (R-FE) producing 50% of inhibition (IC(50)) and the working range of icELISA were 3.1 ng/mL and 0.6-29 ng/mL, respectively. This assay is also sensitive to R-fenoxaprop, S-(-)-fenoxaprop-ethyl, and metamifop with IC(50) of 3.4, 2.7, and 3.5 ng/mL, respectively. The recoveries of R-FE in soil samples with the icELISA were 86-102%.

Development of a monoclonal antibody-based ELISA for the detection of the novel insecticide cyantraniliprole

Cyantraniliprole is the newest anthranilic diamide insecticide acting on ryanodine receptors with higher efficacy, and broader spectrum than chlorantraniliprole in pest control. To study the effects of its residues on the environment and food samples, an enzyme-linked immunosorbent assay (ELISA) has been developed. The hapten was synthesized and conjugated with bovine serum albumin (BSA) and ovalbumin (OVA) as the immunogen and coating antigen, respectively. A sensitive and specific monoclonal antibody (mAb) was generated and was designated as mAb3B2. mAb3B2 was used to develop an indirect competitive ELISA (icELISA). The concentration of cyantraniliprole producing 50% inhibition (IC50) was 1.57 μg L−1 and the effective range of icELISA was 0.43–6.15 μg L−1. The icELISA showed low cross-reactivity with chlorantraniliprole, and very low or no cross-reactivity with the other cyantraniliprole analogues. Average recoveries from pakchoi (Brassica rapa) and tap water samples were 93.7–101.0% and 94.4–100.6%, respectively. Meanwhile, to confirm the results of icELISA, high-performance liquid chromatography was applied for the determination of cyantraniliprole residues in pakchoi samples. The comparable results suggest that the ELISA is suitable for rapid detection of cyantraniliprole residues in environmental and agricultural samples.

Development of a Sensitive Monoclonal Antibody-Based Enzyme-Linked Immunosorbent Assay for the Analysis of Paclobutrazol Residue in Wheat Kernel

An indirect competitive enzyme-linked immunosorbent assay (icELISA) was developed with monoclonal antibody (mAb) mAb6H73C9 recognizing the plant growth regulator paclobutrazol (PBZ). The icELISA had a half-maximum inhibition concentration (IC50) and working range of approximately 8.7 and 2.0-50.4 ng/mL, respectively. Average recoveries of PBZ in the wheat (Triticum aestivum) kernel samples were between 84.3 and 118.9% with relative standard deviations between 3.9 and 14.2%. As determined by the icELISA and further confirmed by liquid chromatography-electrospray ionization quadrupole Orbitrap mass spectrometry (LC-ESI-MS) analysis, the maximum residue concentration was about 0.07 mg/kg in the kernel samples, which indicated that PBZ could transfer from PBZ- treated seedlings to the kernel samples. The correlation coefficient (R(2)) between icELISA and LC-ESI-MS results was 0.979, which manifested that the developed icELISA was sensitive enough for monitoring PBZ residues in wheat kernels.