3D spheroid HepaRG and fluorescent biphasic tracer for CYP3A4-mediated antibiotic interaction monitoring in sepsis.

Cytochrome P450 3A4 (CYP3A4) is a crucial enzyme in the metabolism of xenobiotics, particularly in drug metabolism interactions (DDIs), making it a significant factor in clinical drug use. However, current assay techniques are both laborious and costly, making it difficult to construct a high-throughput monitoring method that can be used in conjunction with the clinic. This poses certain safety hazards for drug combination. Therefore, it is crucial to develop a synchronized monitoring method for the inhibition and induction of CYP3A4. In this study, we utilized 3D culture technology to develop a HepaRG cells spheroid model. The CYP450 and transporter expression, the albumin secretion, and urea synthesis capacity characteristics were analyzed. The NEN probe was utilized as a tracer molecule for CYP3A4. The fluorescence intensity of metabolites was characterized by laser confocal technique to determine the inhibition and expression of CYP3A4 in the HepaRG cell spheroid model by the antibiotics for sepsis. The results indicate that the HepaRG sphere model successfully possessed the physiological phenotype of the liver, which could be used for drug interaction monitoring. Through positive drug testing, NEN probe was able to achieve bidirectional characterization of CYP3A4 induction and inhibition. The monitoring method described in this paper was successfully applied to drug interaction monitoring of commonly used antibiotics in sepsis patients, which is a convenient and rapid monitoring method. The proposed method offers a new strategy for monitoring CYP3A4-mediated drug-drug interactions with a high-throughput assay, which will help to improve the safety of clinical drug combination.

Antibiotic combinations prediction based on machine learning to multicentre clinical data and drug interaction correlation.

BACKGROUND: With increasing antibiotic resistance and regulation, the issue of antibiotic combination has been emphasised. However, antibiotic combination prescribing lacks a rapid identification of feasibility, while its risk of drug interactions is unclear. METHODS: We conducted statistical descriptions on 16 101 antibiotic coprescriptions for inpatients with bacterial infections from 2015 to 2023. By integrating the frequency and effectiveness of prescriptions, we formulated recommendations for the feasibility of antibiotic combinations. Initially, a machine learning algorithm was utilised to optimise grading thresholds and habits for antibiotic combinations. A feedforward neural network (FNN) algorithm was employed to develop antibiotic combination recommendation model (ACRM). To enhance interpretability, we combined sequential methods and DrugBank to explore the correlation between antibiotic combinations and drug interactions. RESULTS: A total of 55 antibiotics, covering 657 empirical clinical antibiotic combinations were used for ACRM construction. Model performance on the test dataset showed AUROCs of 0.589-0.895 for various antibiotic recommendation classes. The ACRM showed satisfactory clinical relevance with 61.54-73.33% prediction accuracy in a new independent retrospective cohort. Antibiotic interaction detection showed that the risk of drug interactions was 29.2% for strongly recommended and 43.5% for not recommended. A positive correlation was identified between the level of clinical recommendation and the risk of drug interactions. CONCLUSIONS: Machine learning modelling of retrospective antibiotic prescriptions habits has the potential to predict antibiotic combination recommendations. The ACRM plays a supporting role in reducing the incidence of drug interactions. Clinicians are encouraged to adopt such systems to improve the management of antibiotic usage and medication safety.

Preparation of a broad-specificity antibody against zearalenone and its primary analogues and development of immunoassay of Coicis Semen and related products.

The purpose of this study was to prepare a highly sensitive and specific zearalenone (ZEN) monoclonal antibody, which was then used to develop an indirect enzyme-linked immunosorbent assay (ic-ELISA) and a colloidal gold immunochromatographic assay (GICA). These techniques were used for the detection of Coicis Semen and related products (Coicis Semen flour, Yimigao, and Yishigao). Immunogens were synthesized by oxime active ester techniques and characterized via ultraviolet spectrophotometry. Immunogens were injected subcutaneously into the abdominal cavities and backs of mice. Using the prepared antibodies, we developed ic-ELISA and GICA rapid detection methods, which were then applied for the rapid detection of ZEN and its analogues from Coicis Semen and related products. For ic-ELISA, the half maximal inhibitory concentration (IC50 ) values for ZEN, α-zearalenol (α-ZEL), ß-zearalenol (ß-ZEL), zearalanone (ZAN), α-zearalanol (α-ZAL), and ß-zearalanol (ß-ZAL) were determined to be 1.13, 1.69, 2.06, 0.66, 1.20, and 0.94 ng•mL-1 , respectively. For GICA, the cutoff values of ZEN, α-ZEL, ß-ZEL, α-ZAL, and ß-ZAL on test strips were 0.5 ng•mL-1 in phosphate buffer saline (0.01 M, pH 7.4), while ZAN was found to be 0.25 ng•mL-1 . Furthermore, the cutoff values of test strips were between 10 and 20 µg∙kg-1 in Coicis Semen and related products. The results of these two detection methods were in good agreement with results from liquid chromatography-tandem mass spectrometry. This study provides technical support for the preparation of broad-specificity monoclonal antibodies against ZEN and lays the foundation for the simultaneous detection of multiple mycotoxins from food and herbal medicines.

Efficient removal of phytochrome using rice straw-derived biochar: Adsorption performance, mechanisms, and practical applications.

Rice straw derived biochar was fabricated and applied as a purification agent. The adsorption kinetics, isotherms, and thermodynamics for adsorbates were determined using the biochar. Adsorption kinetics and isotherms were best fitted by the pseudo-second order and Langmuir models. Biochar could effectively remove chlorophyll in 9 different solutions. Biochar was employed as a clean-up reagent for 149 pesticides detection, which revealed that biochar had a higher phytochrome removal capacity than graphitized carbon black and 123 pesticides had satisfactory recovery values. The biochar was prepared into a sample pad by electrospinning and was then used for online sample clean-up in a test strip, and it showed high ability of removing phytochrome and improving detection sensitivity. Thus, biochar could be applied as a purification agent to remove pigmentation, making it a promising candidate not only for sample pretreatment but also in the fields of food, agriculture and environment.

Residue detection and correlation analysis of multiple neonicotinoid insecticides and their metabolites in edible herbs.

In this work, a green analytical method was established for the simultaneous extraction and detection of 20 analytes-10 neonicotinoid insecticides and their 10 major toxic metabolites in edible herbs. QuEChERS and LC-MS/MS were used to analyze the 20 analytes in five edible herbs. The residues of the 20 neonicotinoid insecticides and their metabolites in 109 herbal samples were detected, of which 90 samples were positive, and the residue of total neonicotinoid insecticides ranged from 0.26 to 139.28 µg/kg. Acetamiprid (77.06 %, ≤85.95 µg/kg), imidacloprid (67.89 %, ≤32.49 µg/kg) and their metabolites (N-desmethyl-acetamiprid (44.04 %, ≤18.42 µg/kg) and desnitro imidacloprid (48.62 %, ≤16.55 µg/kg) were most frequently detected in herbs. Significant positive correlations were found between imidacloprid/acetamiprid and their metabolites in Lycii fructus and Citri reticulatae pericarpium. Therefore, more attention may be given to the neonicotinoid insecticide residues in edible herbs in the future.

DeepIDC: A Prediction Framework of Injectable Drug Combination Based on Heterogeneous Information and Deep Learning.

BACKGROUND AND OBJECTIVE: In clinical practice, injectable drug combination (IDC) usually provides good therapeutic effects for patients. Numerous clinical studies have directly indicated that inappropriate IDC generates adverse drug events (ADEs). The clinical application of injections is increasing, and many injections lack relevant combination information. It is still a significant need for experienced clinical pharmacists to participate in evidence-based drug decision making, monitor medication safety, and manage drug interactions. Meanwhile, a large number of injection pairs and dosage combinations limit exhaustive screening. Here, we present a prediction framework, called DeepIDC, that can expediently screen the feasibility of IDCs using heterogeneous information with deep learning. This is the first specific prediction framework to identify IDCs. METHODS: Since the interaction between the injected drugs may occur in the direct physical and chemical reactions at the time of mixing or may be the indirect interaction of their drug targets and pathways, we used molecular fingerprints, drug-target associations, and drug-pathway associations to convert injections into a string of digital vectors. Then, based on these injection vectors, we combined a bidirectional long short-term memory and a feed-forward neural network to build a prediction model for accurate and instructive prediction of IDC. RESULTS: In three realistic evaluation scenarios, DeepIDC has achieved ideal prediction results. Furthermore, compared with the other five machine-learning methods, the proposed predictor is more efficient and robust. Among the top 30 potential IDCs of each IDC class predicted by DeepIDC, we found that 9 cases were experimentally verified in the literature or available on Drug.com. CONCLUSION: The information we extracted in vivo and in vitro can effectively characterize injectable drugs. DeepIDC developed based on deep learning algorithm provides a valuable unified framework for new IDC discovery, which can make up for the lack of IDC information and predict potential IDC events.

Hepatotoxicity of food-borne mycotoxins: molecular mechanism, anti-hepatotoxic medicines and target prediction.

Mycotoxins are metabolites produced by fungi. The widespread contamination of food and feed by mycotoxins is a global food safety problem and a serious threat to people's health. Most food-borne mycotoxins have strong hepatotoxicity. However, no effective methods have been found to prevent or treat Mycotoxin- Induced Liver Injury (MILI) in clinical and animal husbandry. In this paper, the molecular mechanisms and potential anti-MILI medicines of six food-borne MILI are reviewed, and their targets are predicted by network toxicology, which provides a theoretical basis for further study of the toxicity mechanism of MILI and the development of effective strategies to manage MILI-related health problems in the future and accelerate the development of food safety.

[Rapid detection of aflatoxin B_1 in Chinese herbal medicines by indirect and direct competitive enzyme-linked immunosorbent assays: a comparative analysis].

The indirect competitive enzyme-linked immunosorbent assay(ic-ELISA) and direct competitive enzyme-linked immunosorbent assay(dc-ELISA) were performed for the rapid detection of aflatoxin B_1(AFB_1) in Astragali Radix, Zingiberis Rhizoma Recens, Jujubae Fructus, and Nelumbinis Semen with self-made antigens and antibodies. Different extraction methods were investigated to reduce the matrix effects of different medicinal parts in Chinese herbal medicines. The sensitivity of dc-ELISA method was improved by optimizing the molar ratio of AFB_1 to horseradish peroxidase(HRP). In this study, the sensitivity(IC_(50)) of ic-ELISA and dc-ELISA was 0.046 and 0.023 ng·mL~(-1), with the limit of detection(LOD) of 0.007 and 0.004 ng·mL~(-1), respectively. The detection time was 3 h and 50 min for ic-ELISA and dc-ELISA, respectively. The recovery rates were within the range of 62.96%-104.4%, with RSDs of less than 10%. Confirmed by LC-MS/MS, three positive samples of Nelumbinis Semen were detected from 53 samples. Two ELISA methods established in this study were accurate, rapid and sensitive, and can be used for rapid screening of AFB_1 in Chinese herbal medicines such as Astragali Radix, Zingiberis Rhizoma Recens, Jujubae Fructus, and Nelumbinis Semen. In addition, the advantages and limitations of the two methods were compared and discussed, which can provide a reference for the testing institutions to choose the proper method.

Colloidal gold-based lateral flow immunoassay with inline cleanup for rapid on-site screening of carbendazim in functional foods.

In this study, for the first time, we propose a sensitive colloidal gold-based lateral flow immunoassay (LFIA) that can be used to detect carbendazim residues in functional foods. The adoption of inline cleanup LFIA strips effectively improved background interference to reduce misjudgment of results. First, the hapten 2-(methylamino)-1H-benzo[d]imidazole-5-carboxylic acid was used to establish the carbendazim immunoassay method. Subsequently, colloidal gold-mAb preparation and LFIA detection conditions were systematically optimized. For root and fruit samples (ginseng, ginger, jujube, and Chinese wolfberry), the designed strips had a cutoff value of 8 ng/mL. For flower and seed samples (chrysanthemum, coix seed, and malt), the cutoff value was 12 ng/mL. Even in a complex matrix, the established LFIA method demonstrates satisfactory sensitivity and anti-interference ability. This method was successfully applied in detection of carbendazim residues in complex functional foods, and the assay results are consistent with those obtained via liquid chromatography-tandem mass spectrometry. In short, the proposed method is fast and sensitive and has strong anti-interference ability. Furthermore, it provides a new technical method highly relevant to the on-site rapid detection of carbendazim residues in complex sample matrix.

Development of a sensitive indirect competitive enzyme-linked immunosorbent assay for high-throughput detection and risk assessment of aflatoxin B1 in animal-derived medicines.

Animal-derived medicine is an important part of traditional Chinese medicine (TCM). Studies have shown that many animal-derived medicinal products are susceptible to contaminate of aflatoxins, nevertheless, the rapid detection for animal-derived medicine is prone to be ignored. Here we developed a sensitive indirect competitive enzyme-linked immunosorbent assay (icELISA) for rapid screening of aflatoxin B1 (AFB1) in ground beetle, cockroach, silkworm and earthworm. The sensitivity of the icELISA method was significantly enhanced. The IC50 for the four animal-derived medicinal samples ranged from 0.092 to 0.135 ng mL-1; the limit of detection (LOD) was 0.008-0.020 ng mL-1. To obtain high accuracy, the extraction solution and time were evaluated. By using this method, a total of 138 samples were investigated, and the detection rates of AFB1 in ground beetle and earthworm samples were 26.6% and 16.7%, respectively. The result was validated by liquid chromatography combined with tandem mass spectrometry, and an excellent correlation was observed between the two datasets, with a R2 value of 0.999. Our results indicate that the proposed method can be used for the rapid detection of AFB1 in animal-derived medicine. Furthermore, the quantitative risk assessment was conducted for ground beetle and earthworm based on the results, demonstrating that the intake of AFB1 in ground beetle had a slight threat to the risk of cancer.

Magnetic particles encoding a suspension probe for ultra-sensitive and quantitative determination of atrazine.

As a highly toxic and widely used herbicide, atrazine poses a serious threat to food safety as well as overall environmental and human health. Due to complex matrix interference and the difficulty of signal enrichment, there is an urgent need for a convenient, fast, and ultrasensitive method that detects trace atrazine without concern for matrix effects. Here, we provide the first account of a sensitive and rapid suspension probe based on magnetic microspheres used to detect atrazine in herbs. The self-made magnetic beads featured -COOH groups and were used as the carrier to construct immunofluorescent probes. These probes then conjugated with the atrazine antigen through an activated ester method, ultimately binding to the antibody. Homogeneous detection was ensured using flow cytometry and the microflow optical channel along with allophycocyanin-conjugated goat-anti-mouse secondary antibody (APC-IgG-SecAb) as the fluorescent signal. The magnetic suspension probe allowed for high target enrichment and the inherent two-dimensional selective detection of flow cytometry effectively avoided any matrix interference. This method had good linearity across 1.69-23.19 ng mL-1. The IC50 and LOD values were 4.81 ng mL-1 and 0.95 ng mL-1, respectively; the sensitivity was increased three-fold relative to ELISA. After complete optimization, 2-N-morpholinoeth-anesulfonic acid was used as the coupling solution and maintained good mono-dispersity, stability, and reactivity for the labelled microspheres during the process. The entire experiment was simple, and effectively used reagents; moreover, both the labor required and detection time were greatly reduced. Critically, the strategy presented here greatly reduced interference from complex matrices, and saved preparation for matrix-matched solutions when different herbs were screened. Overall, this strategy was sensitive, rapid, eco-friendly, and labor-saving; collectively, these attributes make it well-suited for on-site screening of atrazine contamination and will allow for increased food safety.

[Preparation of highly sensitive monoclonal antibody against aflatoxin B_1 and its application in rapid detection of contamination in Ziziphi Spinosae Semen].

A highly sensitive monoclonal antibody against aflatoxin B_1(AFB_1) was prepared and an indirect competition enzyme-linked immunosorbent assay(ic-ELISA) was established based on the antibody which was used for high-throughput and rapid screening of AFB_1 contamination in Chinese herbal medicines to ensure the safety of medication. In this study, the structure of AFB_1 was modified by improved oxime method, and the carrier protein was coupled by EDC-NHS method to obtain the complete antigen of AFB_1, which was more convenient and environmental friendly. The Balb/c female mice were immunized using increasing the immunization dose and various ways of injection, and finally the AFB_1 monoclonal antibody was prepared. The AFB_1 monoclonal antibody belongs to IgG_(2 b) immunoglobulin by identifying its immunological characteristics, and its sensitivity(IC_(50)) can reach 0.15 µg·L~(-1), and the affi-nity is 2.81×10~8 L·mol~(-1). The cross-reaction rates of AFB_2, AFG_1, and AFG_2 were 35.07%, 8.75%, and 1.15%, respectively, and there was almost no cross-reactivity with other mycotoxins. Based on the high sensitivity and specificity of the antibody, an ic-ELISA method was established and applied to the determination of AFB_1 contamination in Ziziphi Spinosae Semen. According to the matrix matching standard curve, the linear concentration range for AFB_1 was 0.05-0.58 µg·L~(-1)(R~2=0.992), the recoveries were 88.00%-119.0%, and the detection limit was 1.69 µg·kg~(-1). The AFB_1 in 33 batches of Ziziphi Spinosae Semen samples was determined by ic-ELISA, and the contamination level was 3.62-206.58 µg·kg~(-1). The linear correlation coefficient between the detection results of ic-ELISA and UHPLC-MS/MS was 0.996, and there were no false positive and false negative cases. It indicates that the established ic-ELISA is accurate and reliable, and could provide a simple and effective technique for fast screening of AFB_1 contamination in Ziziphi Spinosae Semen, and also could be considered as the reference for the detection and monitoring of AFB_1 contamination in other Chinese herbal medicines.

Comprehensive assessment for the residual characteristics and degradation kinetics of pesticides in Panax notoginseng and planting soil.

Incorrect and excess usage of pesticides during crop cultivation poses a serious threat to human health and ecosystems. In this study, we tested for the presence of 201 pesticide residues in 90 batches of Panax notoginseng (P. notoginseng) and 10 batches of planting soil. Pesticide residue characteristics and the relationship between pesticides present in P. notoginseng and the soil were discussed. Twenty-nine pesticides were detected in P. notoginseng samples and 15 pesticides were found in the soil samples. In P. notoginseng samples, the 68.9% of the identified pesticides were fungicides, and six fungicides (procymidone, iprodione, pyrimethanil, propiconazole, dimethomorph and tebuconazole) were found in >90% of the samples. Nine insecticides were found, with one insecticide, chlorpyrifos, detected in 93.3% of the P. notoginseng samples. The residual concentrations of 17 pesticides were found at levels exceeded the "non-Chinese" maximum residue levels (MRLs) for Ginseng and 17 pesticides were found at levels exceeding the MRLs set by China for "pollution-free" P. notoginseng. We observed no significant differences in pesticide residues were found on P. notoginseng from different cultivation areas. We also analyzed the degradation kinetics of pesticides in the soil, as well as their bioconcentration factors (BCFs), and found that the fungicides iprodione and myclobutanil displayed strong uptake from the soil to the root of P. notoginseng. Together, our data suggest that fungicides should be considered as key monitoring substances in P. notoginseng and planting soil.

A comprehensive analysis of 201 pesticides for different herbal species-ready application using gas chromatography-tandem mass spectrometry coupled with QuEChERs.

Developing an analysis of multi-pesticide residues for different herbal species-ready applications is a challenge. In the present work, a comprehensive analysis was proposed for rapid detection of 201 pesticides in various medicinal herbs. Samples were extracted and cleaned up with a high throughput pretreatment approach (modified QuEChERS), and then detected by gas chromatograph coupled to an electron impact ionization triple quadrupole mass spectrometer (GC-EI-MS/MS). The clean-up procedure has been optimized using four types of representative medicinal herbs with different primary or secondary metabolites. Moreover, a mixture of analyte protectants (APs) was to improve the peak shape and intensity of some compounds. The performance of the method was validated according to the European Union SANTE/11813/2017 regulatory guidelines. The limit of quantification (LOQ) was determined to be ≤10 ng mL-1, and the recovery was between 70.0%-120.0%, with ≤20% RSD for the majority of pesticides. Sixty samples belonging to different species of medicinal herbs (such radix, flos, cortex, fructus, and seeds) were analyzed to evaluate the applicability of the optimized method. High frequency of chlorpyrifos was found in Citri reticulatae pericarpium, Crataegi fructus and Cuscutae semen samples.

Screening techniques for the identification of bioactive compounds in natural products.

Natural products (NPs) have a long history of clinical use and are rich source of bioactive compounds. The development of tools and techniques for identifying and analyzing NP bioactive compounds to ensure their quality and discover new drugs is thus very important and still in demand. Screening techniques have proven highly useful for screening and analyzing active components in complex mixtures, which rely on cell culture, dialysis, ultrafiltration, chromatographic methods and target molecule immobilization, using biological targets to identify the active compounds. The recent progress in biological screening techniques in the field of natural products is reviewed here. This includes a review on the strategy and application of the screening methods, their detailed description and discussion of their existing limitations of the different models along with prospective in future development of screening techniques.

[Simultaneous determination of aflatoxin B_1,B_2,G_1,G_2,M_1,M_2 in Eupolyphaga Steleophaga by immunoaffinity column clean-up and HPLC-FLD with post-column photochemical derivatization].

The contamination of aflatoxin B_1,B_2,G_1,G_2,M_1 and M_2 in Eupolyphaga Steleophaga was determined by immunoaffinity column clean-up and HPLC-FLD with post-column photochemical derivatization. Chromatographic separations were carried out using a Cloversil C_(18) column( 4. 6 mm×250 mm,5 µm) that were eluted in isocratic with methanol-acetonitrile-water( 20 ∶ 20 ∶ 60) as the mobile phase. The excitation wavelength and the emission wavelength of fluorescence detector were maintained at 360 nm and 450 nm,respectively. The flow rate was 0. 8 m L·min~(-1),and the column temperature was 30 ℃ ． The sample was prepared using the immunoaffinity column,then the recovery was measured with 75. 47%-101. 8% with RSD values lower than 6. 7%. A total of 20 batches of Eupolyphaga Steleophaga samples were assayed. According to the Chinese Pharmacopoeia( 2015 edition,part 1),the aflatoxin B_1 limit should be less than 5 µg·kg~(-1),and the sum of aflatoxins( AFB_1,AFB2,AFG_1,AFG_2) should be less than 10 µg·kg-1. Therefore,the positive rate of the 20 samples was 50. 0%,and 7 batches of samples exceeded the standard,and the over-standard rate was as high as 70. 0%. Among them,aflatoxins B_1,B_2,G_1,G_2,M_1,and M_2 were detected in three batches( SD-1,AH-1,AH-3),and aflatoxins B_1,B2,G1,G2,and M1 were detected in one batch( AH-7). The results showed that the newly developed method in this work is suitable for the simultaneous determination of six aflatoxins in Eupolyphaga Steleophaga,and also suggested that it should be of high values to take the contamination with aflatoxins into concerns.

[Survey on pesticide use in Crataegi Fructus and analysis of pesticide residues based on LC-ESI-MS/MS].

In order to study the pesticide residues of the medicinal Crataegi Fructus,this study aims to establish an analysis method for pesticide residues( mainly containing insecticides and fungicides) suitable for the actual situation of medicinal Crataegi Fructus based on the survey of the pesticides of the Crataegi Fructus base,combined with the blind screening results of the LC-ESI-MS/MS pesticide screening platform established by the research team in the early stage. Then,the pesticide residues in medicinal Crataegi Fructus from Shandong,Hebei,Henan,Shanxi,and Liaoning( main cultivation areas) were analyzed. The samples were pretreated by the modified Qu ECh ERS method,i.e.,extracted with acetonitrile-water( 9 ∶1),purified by PSA,C_(18),GCB,silica gel. The detection of pesticides was performed by LC-MS/MS. The ion source was ESI with positive scanning mode,and the linearity of 11 kinds of pesticides in the range of 5-300 µg·kg~(-1) was acceptable( R~2>0. 996 9). All the recoveries of pesticides were within 70. 02%~(-1)12. 0% in the low,medium and high levels,with RSD≤17%. The results showed that the detection rate of carbendazim,chlorpyrifos and difenoconazole is 79%,82%,56%,respectively. Besides,the prohibition pesticide carbofuran were detected in some of the batches,indicating the security risk. This study provides methodological references and basic data for risk assessment of Crataegi Fructus and government regulation.

[QuEchERS pretreatment coupled to gas chromatography and tandem mass spectrometry to fast determination of 34 pesticide residues in Glycyrrhizae Radix et Rhizoma].

This study proposed a quantitative method for 34 pesticides including organochlorine,organophosphorus and pyrethroids in Glycyrrhizae Radix et Rhizoma herbs and medicinal slices,and analyzed the pesticide residues of collected Glycyrrhizae Radix et Rhizoma samples from different regions. With acetonitrile extraction and optimized Qu Ech ERS purification,the 32 batches of Glycyrrhizae Radix et Rhizoma herbs and medicinal slices were analyzed by matrix matching standard curve quantitative analysis under GC-MS/MS multi-response monitoring( MRM) mode. This study investigated the pretreatment of Glycyrrhizae Radix et Rhizoma samples based on the Qu Ech ERS method of Chinese Pharmacopoeia( 2015 edition,4),and the result showed that the recoveries of some pesticide was low and pigment has a strong interference in analysis,which result in worse purification effect. Therefore,this paper further optimized the Qu Ech ERS method and corrected the matrix matching standard curve method,and compensated the qualitative and quantitative effects of matrix effects on the detected target compounds in Glycyrrhizae Radix et Rhizoma. The results showed that 34 kinds of pesticide had good linear( R~2 of 0. 996 4 or higher) within a covering 0. 01-0. 2 mg·kg~(-1) concentration range. The limits of quantitation are less than 0. 01 mg·kg~(-1). This method was further applied to the simultaneous determination of 34 pesticide residues of typical organochlorine,organophosphorus and pyrethroids in 32 batches of Glycyrrhizae Radix et Rhizoma herbs and medicinal slices. Six batches containing beta-endosulfan,thiosulphate,o,p'-DDD and thrta-cypermethrin were detected,but none of them exceeded the limit of pesticide residues stipulated in the Chinese Pharmacopoeia and the EU Pharmacopoeia. This study indicates that the established method is rapid,convenient,accurate,and sensitive,which provides a rapid and efficient method for the simultaneous determination of typical organochlorine,organophosphorus and pyrethroids in Glycyrrhizae Radix et Rhizoma.

Occurrence of multi-class mycotoxins in Menthae haplocalycis analyzed by ultra-fast liquid chromatography coupled with tandem mass spectrometry.

To determine the presence of multi-class mycotoxins in Menthae haplocalycis, a sample processing procedure based on a modified quick, easy, effective, rugged, and safe method, and a rapid and accurate testing method using ultra-fast performance liquid chromatography coupled with tandem mass spectrometry, was developed and validated. We systematically evaluated the methodology for multi-mycotoxin analysis in the Menthae haplocalycis samples, and chose matrix-matched calibration curves as a reference to calculate the recoveries. Overall, the average recoveries varied between 67.1 and 103%, with relative standard deviations ranging from 0.34 to 10.3%. The optimized and validated method was applied to detect the presence of the target mycotoxins in 40 batches of Menthae haplocalycis samples. Results showed that the levels of mycotoxins varied among the samples. The most prevalent mycotoxin was tentoxin, followed by alternariol, alternariol monomethyl ether, zearalenone, fumonisin B2 , fumonisin B1 , ochratoxin A, aflatoxin B1 , aflatoxin B2 , aflatoxin G1 , and T-2 toxin. The analytical method developed herein could be applied for the routine monitoring of multi-mycotoxins in Menthae haplocalycis.

6'-Hydroxy Justicidin B Triggers a Critical Imbalance in Ca2+ Homeostasis and Mitochondrion-Dependent Cell Death in Human Leukemia K562 Cells.

Justicia procumbens (J. procumbens) is a traditional Chinese herbal medicine which was used for the treatment of fever, pain, and cancer. A compound 6'-hydroxy justicidin B (HJB) isolated from J. procumbens exhibits promising biological properties. However, the mechanism of action and the in vivo behavior of HJB remain to be elucidated. In this study, we investigated the mechanism of action of HJB on human leukemia K562 cells and its pharmacokinetic properties in rats. The results demonstrated that HJB significantly inhibited the proliferation of K562 cells and promoted apoptosis. Besides, HJB resulted in decreased mitochondrial membrane potential deltaPSIm, increased the level of the calcium homeostasis regulator protein TRPC6 and cytosolic calcium. The activity of caspase-8, caspase-9 and the expression of p53 were significantly increased after treatment with HJB. Additionally, HJB has rapid absorption rate and relative long elimination t1/2, indicating a longer residence time in vivo. The results indicate that HJB inhibited the proliferation of K562 cells and induced apoptosis by affecting the function of mitochondria and calcium homeostasis to activate the p53 signaling pathway. The pharmacokinetic study of HJB suggested it is absorbed well and has moderate metabolism in vivo. These results present HJB as a potential novel alternative to standard human leukemia therapies.