[Rapid detection of zearalenone in Coicis Semen based on ELISA].

Zearalenone(ZEN) is a toxic metabolite produced by Fusarium culmorum, F. graminearum, F. tricinctum, and other fungi, with estrogenic characteristics. Exposure to or ingestion of ZEN during pregnancy can cause reproductive dysfunction, miscarriage, stillbirth, and malformation, and seriously endanger human life and health. The detection methods for ZEN in the Chinese Pharmacopoeia(2020 edition) are liquid chromatography(LC) and liquid chromatography-mass spectrometry(LC-MS), and it is stipulated that ZEN should not exceed 500 µg in 1 000 g of Coicis Semen. Although these detection methods by instruments can achieve the qualitative and quantitative analysis of ZEN in Coicis Semen, their high detection cost and long periods hinder the rapid screening of a large number of samples in the field. In this study, the synthesized ZEN hapten was conjugated with bovine serum albumin(BSA) and ovalbumin(OVA) to obtain the complete ZEN antigen. By virtue of antibody preparation techniques, ZEN monoclonal antibody 4F6 was prepared, which showed 177.5%, 137.1%, and 109.7% cross-reactivity with ZEN structural analogs zearalanol, zearalenone, and α-zearalenol, respectively, and no cross-reactivity with other fungal toxins such as aflatoxin. Direct competitive enzyme-linked immunosorbent assay(dcELISA) based on ZEN monoclonal antibody 4F6 was developed for the determination of ZEN in Coicis Semen with an IC_(50) of 1.3 µg·L~(-1) and a detection range of 0.22-21.92 µg·L~(-1). The recoveries were 83.91%-105.3% and the RSD was 4.4%-8.0%. The established dcELISA method was used to determine the ZEN residuals in nine batches of Coicis Semen samples, and the results were validated by LC-MS. The correlation between the two detection methods was found to be 0.993 9, indicating that the established dcELISA could be used for the rapid qualitative and quantitative detection of ZEN residuals in Coicis Semen.

[Development of a monoclonal antibody-based enzyme-linked immunosorbent assay for determination of vomitoxin (DON) in Coicis Semen].

Coicis Semen is a common bulk medicinal material used for both medicine and food, which has the effect of promoting diuresis, draining dampness, invigorating the spleen and checking diarrhea. It is derived from Coix lacryma-jobi var. ma-yuen of the family Poaceae, and is easily contaminated by fungi such as Fusarium graminearum and F. flavum due to climate reasons to produce vomitoxin. The guiding principles for determination of vomitoxin in Chinese medicinal materials in Chinese Pharmacopoeia are mainly HPLC and LC-MS, which have long detection period and are time-consuming and laborious, and thus cannot meet the requirements of on-site quality inspection of drugs. The complete antigen of vomitoxin-protein was obtained by chemical derivatization of vomitoxin. The monoclonal antibody against vomitoxin was prepared by classic monoclonal antibody preparation technology, and enzyme-linked immunosorbent assay(ELISA) method for detection of vomitoxin in Coicis Semen was established through methodological investigation. The IC_(50) based on the ELISA for vomitoxin in Coicis Semen was 3.88 µg·L~(-1), and the average recoveries and the RSD were 77.32%-93.73% and 4.6%-9.7%, respectively. With the established ELISA method, the vomitoxin residue in 14 batches of Coicis Semen samples were determined and validated by LC-MS, and the correlation between the two assays was found to be 0.997 8, indicating that the established ELISA method could be used for quantitative determination of vomitoxin residue in Coicis Semen and could achieve the rapid quantitative determination of the vomitoxin residue.

[Effect of Gansui Banxia Tang plus-minus Gansui and Gancao anti-drug combination that preferred dose close to clinical application on diuretic effect in malignant ascites rats].

OBJECTIVE: To observe the effect of Gansui Banxia Tang plus-minus Gansui and Gancao anti-drug combination on hepatic and renal functions in malignant ascites rats to explore whether the efficacy or toxicity associated with the anti-drug combination. METHOD: The male wistar rats were randomly divided into a blank group, model group, furosemide group, Gansui Banxia Tang group, Gansui Banxia Tang removed Zhigancao group, Gansui Banxia Tang removed Cugansui group, Gansui Banxia Tang removed Zhigancao and Cugansui group. In addition to normal feeding, every morning except for the blank group and model group, the rest of the group was given drugs, the control group and the model group was given distilled water, the volume is 10 mL x kg(-1). Administered five days, all rats were fasted but except water for 24 hours to collect urine. Administered nine days all rats were fasted but except water for 12 hours, we need to weigh weight of rats. When we remove the ascites, we also need to weigh weight of rats. We use the weight before removing ascites minus weight after removing ascites to indirectly measure the amount of ascites. When we remove the ascites, we need to abdominal aortic blood, centrifuge testing renin, angiotensin II, aldosterone, antidiuretic hormone and other indicators. RESULT: The effect of Gansui Banixa Tang on increasing the net weight, lowering abdominal circumference and body weight ratio, lowering renin, angiotensin, aldosterone, antidiuretic hormone is better than the other treatment group. CONCLUSION: In diuresis party, the group of Gansui Banxia Tang is better than the group of Gansui Banxia Tang remove Zhigancao or Cugansui or Zhigancao and Cugansui, renin-angiotensin-aldosterone system may play a diuretic effect of its one way.