Development of a class-specific competitive enzyme-linked immunosorbent assay for the detection of pyrrolizidine alkaloids in vitro

Roseman, David M.; Wu, Xiying; Milco, Larry A.; Bober, Mary A.; Miller, Rebecca; Kurth, Mark J.

doi:10.1021/jf00018a019

Cited by 16 publications

(18 citation statements)

References 29 publications

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“…DMF is usually used as solvent to make active esters of carboxylic acid haptens. This phenomenon has also been observed by other researchers (Tatake et al, 1991;Roseman et al, 1992). Fortunately, this problem was resolved by using the less polar THF.…”

Section: Resultssupporting

confidence: 81%

Development of a Class-Specific Competitive ELISA for the Benzoylphenylurea Insecticides

Wang

Allan

Skerritt

et al. 1998

J. Agric. Food Chem.

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A competitive enzyme-linked immunosorbert assay (ELISA) was developed for the quantitative detection of five benzophenylurea insecticides in soil (90% methanol extraction) and water. No significant matrix effects were observed. Conjugates of diflubenzuron derivatives with proteins were prepared by attachment at either the urea nitrogen of diflubenzuron or the 4-position of the aniline ring, wherein bridging groups of various lengths were used between the hapten and the protein carrier. All combinations of antiserum to each immunogen and enzyme tracer were examined, and an assay was developed using an antibody developed to a hapten conjugate coupled through a succinyl chain at the 4-aniline position. The assay gave 50% inhibition of antibody binding (IC 50 ) at 0.6 ppb for diflubenzuron, 5 ppb for teflubenzuron, 10 ppb for flufenoxuron, 31 ppb for lufenuron, and 45 ppb for chlorfluazuron, with detection limits in the range of 0.05-2.3 ppb for these compounds. The basis for the superior cross-reaction of diflubenzuron was examined by molecular modeling, which suggested that the planarity of the molecules, electron-withdrawing groups, and steric effects of chlorines attached to the phenyl ring may be critical factors affecting antibody binding.

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Section: Resultssupporting

confidence: 81%

Development of a Class-Specific Competitive ELISA for the Benzoylphenylurea Insecticides

Wang

Allan

Skerritt

et al. 1998

J. Agric. Food Chem.

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“…As one might rationally expect, system A (immunogen 6.1 bsa + coating antigen 7.1 ova ) has excellent sensitivity for retrorsine and system B (immunogen 6.2 bsa + coating antigen 7.2 ova ) for monocrotaline (see Table 3). System C (immunogen 7.3 klh + coating antigen 8.3 ova ) showed parts per million level detection for retronecine, sensitivity less than that found with our previously reported retronamine method (Roseman et al, 1992).…”

Section: Scheme 2 Preparation Of Retronecine Antigens Acontrasting

confidence: 67%

“…Indeed, system B holds great potential as a compound-specific detection method. System C detected retronecine at 600-10000 ppb with an I 50 of 3000 ( 600 ppb [less sensitive than that previously reported by our laboratory (Roseman et al, 1992); little cross-reactivity was observed toward other PAs]. All three systems described were capable of detecting retronecine at some level, and none of these systems show cross-reactivity with either swainsonine or lupinine.…”

Section: Discussionmentioning

confidence: 62%

“…The first and more speculative strategy involved direct detection of the retronecine substructure in intact parent PAs (Bober et al, 1990;Kurth et al, 1992), while the second strategy involved a somewhat less direct assay but one which capitalized on the fact that hydrolysis of any retronecinebased macrocyclic diester PA liberates the necine base retronecine (e.g., saponification of monocrotaline yields monocrotalic acid and retronecine). This "class-specific" PA ELISA detects retronecine at an I 50 value of 11 ( 3 ppb, with detection limits of 1.0-100 ppb (Kurth et al, 1989;Roseman et al, 1992).…”

Section: Introductionmentioning

confidence: 99%

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Enzyme-Linked Immunosorbent Assay Detection of Pyrrolizidine Alkaloids: Immunogens Based on Quaternary Pyrrolizidinium Salts

Roseman

Kurth

1996

Bioconjugate Chem.

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Polyclonal antibody-based enzyme-linked immunosorbent assays (ELISAs) were developed for the detection of retrorsine (1, 351 g/mol), monocrotaline (2, 325 g/mol), and retronecine (3, 155 g/mol) in the parts per billion (ppb) range. A set of three bifunctional linking arms (6-8) was synthesized. By N-alkylation of pyrrolizidine alkaloids (PAs) retrorsine, monocrotaline, and retronecine acetonide (9), six haptens (6.1, 6.2, 7.1, 7.2, 7.9, and 8.9) were synthesized and used to generate rabbit antisera. The resulting anti-retrorsine antiserum gave a 50% inhibition (I50) value of 0.9 +/- 0.2 ppb for retrorsine with detection limits of 0.5-10 ppb. The same ELISA system also detected isatidine (4, retrorsine N-oxide) dihydrate (403 g/mol) with an I50 of 1 ppb and senecionine (5,352 g/mol) with an I50 of 100 ppb. A second monocrotaline-based ELISA detected monocrotaline with an I50 of 36 +/- 9 ppb 2 with detection limits of 5-500 ppb and shows no cross-reactivity with 1 or 5; this ELISA demonstrates the potential for the substrate-specific detection method. A third retronecine-based ELISA detects 3 with an I50 of 3000 +/- 600 ppb (3 +/- 0.6 ppm) and detection limits of 600-10,000 ppb. None of these ELISAs cross-react with the structurally similar swainsonine (10) or lupinine (11) alkaloids. PAs were detected in extracts of Senecio vulgaris and Crotalaria retusa, but not in Lupinus spp., as a demonstration of the ELISA's usefulness.

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“…Some earlier studies related to the detection of alkaloids is presented in Table 2. In comparison with earlier reports on SEN detection protocols [27][28][29][30][31][32][33][34], rapid and sensitive voltammetric detection of SEN was performed in the present study by an easy-to-use and low-cost disposable sensor in combination with an electrochemical measurement protocol. Our electrochemical assay was also successfully applied for SEN detection in food products, such as, flour and herbal tea.…”

Section: Discussionmentioning

confidence: 95%

Detection of Senecionine in Dietary Sources by Single-Use Electrochemical Sensor

et al. 2021

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Pyrrolizidine alkaloids (PAs) are produced by plants as secondary compounds that are the most widely distributed natural toxins. There have been many cases of human toxicity caused by consumption of toxic plant species, as herbal teas and grain or grain products contaminated with PA-containing seeds have been reported. Companies that produce dried spices and tea leaves should examine the PA level in their products. For the first time in the literature, a simple and inexpensive electrochemical assay based on a single-use sensor was introduced for quantitative determination of senecionine (SEN) in the most frequently contaminated food sources. SEN was immobilized on a pencil graphite electrode surface by the passive adsorption technique. Differential pulse voltammetry (DPV) was used to evaluate the oxidation signal of SEN, which was observed to be around +0.95 V. The oxidation signal was specific to the SEN in the sample, and the current value was proportional to its concentration. The selectivity of our assay was also tested in the presence of other similar PAs such as intermedine, lycopsamine, and heliotrine. The detection limit is calculated by developed assay and found to be 5.45 µg/mL, which is an acceptable concentration value of SEN occurring at toxic levels for consumers. As an application of the developed sensor in food products, the electrochemical detection of SEN was successfully performed in flour and herbal tea products.

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Development of a class-specific competitive enzyme-linked immunosorbent assay for the detection of pyrrolizidine alkaloids in vitro

Cited by 16 publications

References 29 publications

Development of a Class-Specific Competitive ELISA for the Benzoylphenylurea Insecticides

Development of a Class-Specific Competitive ELISA for the Benzoylphenylurea Insecticides

Enzyme-Linked Immunosorbent Assay Detection of Pyrrolizidine Alkaloids: Immunogens Based on Quaternary Pyrrolizidinium Salts

Detection of Senecionine in Dietary Sources by Single-Use Electrochemical Sensor

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