Pharmacokinetic and excretion study of eight active constituents in rat by LC-MS/MS after oral administration of the Toddalia asiatica extract.

Toddalia asiatica L., a significant medicinal plant in the family Rutaceae, has been applied to treat rheumatoid diseases for decades. Its pharmacological activities are mainly attributed to the existence of generous coumarins and alkaloids; however, the pharmacokinetics of Toddalia asiatica L. remain unclear. A high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method was established for the simultaneous determination of four coumarins, three alkaloids and one flavonoid (hesperidin, nitidine chloride, chelerythrine, toddalolactone, isopimpinellin, pimpinellin, bergapten and dictamnine) in rat feces as well as four coumarins and one alkaloid (toddalolactone, isopimpinellin, pimpinellin, bergapten and dictamnine) in rat plasma and urine. Chromatographic separation was accomplished on an Agilent ZORBAX SB-C18 column (2.1 × 150 mm, 5 µm) with acetonitrile (containing 0.1% formic acid) and 5 mmol/L aqueous ammonium formate for gradient elution. A correlation coefficient greater than 0.9925 reflected the excellent linearity of the analytical response. The lower limits of quantification were 30.0, 10.0, 10.0, 30.0, 5.0, 10.0, 2.5 and 2.5 ng/mL for hesperidin, nitidine chloride, chelerythrine, toddalolactone, isopimpinellin, pimpinellin, bergapten and dictamnine, respectively. The intra- and inter-day precision were less than 12.7%, and the accuracy was between -11.8% and 12.9%. In summary, this study is the first to study the pharmacokinetics and excretion of T. asiatica extract after oral administration, which may provide a scientific basis for its clinical applications.

The effect of group B streptococcus on maternal and infants' prognosis in Guizhou, China.

Group B Streptococcus (GBS) is a kind of opportunistic pathogenic bacteria and mainly strikes the lower digestive tract and genitourinary tract. It is a major risk factor for neonatal babies, seriously threatening their lives. In the present study, we aimed to detect the GBS colonization in late pregnant women, and to study the effect of GBS on maternal and infants' prognosis. Pregnant women with a gestational age of 35-37 weeks were enrolled in the study. Real-time polymerase chain-reaction (RT-PCR) was used to detect the colonization of GBS in the vaginal and rectal secretions for late pregnant women according to the screening guidelines. Chi-square test was applied to analyze the relationship between GBS colonization and clinical characteristics. A follow-up of 6 weeks was performed on the puerpera and infants after delivery. The positive rate of GBS was 12.6% in late pregnant women. GBS carrier state was positively related to several pregnancy outcomes, including intrauterine infection, premature rupture of membranes, postpartum hemorrhage, fetal distress and puerperal infection, as well as to part neonatal outcomes, containing neonatal infection, neonatal pneumonia and neonatal sepsis (all P < 0.05). GBS infection in late pregnant women results in adverse effects on maternal and neonatal outcomes.

Prophylactic abdominal aortic balloon occlusion in patients with pernicious placenta previa during cesarean section: a systematic review and meta-analysis from randomized controlled trials.

PURPOSE: Pernicious placenta previa induces severe hemorrhage during cesarean section. Abdominal aorta balloon occlusion (AABO) is considered as an effective operation for patients with pernicious placenta previa. The aim of this study was to investigate the clinical application of abdominal aortic balloon occlusion in the placenta previa and cesarean section by systematic review and meta-analysis. METHODS: MEDLINE, EMBASE, Cochrane Library, Web of Science, China National Knowledge Infrastructure (CNKI), WAN-FANG DATA and CQVIP were searched from inception to Jan. 15th, 2019. Operative time, intraoperative blood loss volume, postoperative hospitalization duration, intraoperative blood transfusion volume, hysterectomy rate, lower extremity thrombosis rate, ICU admission rate, adverse reaction rate, neonatal birth weight, Apgar 1-min and 5-min scores were regarded as the endpoints. Randomized controlled trials (RCT) were used for meta-analysis. RESULTS: Fourteen articles were retrieved from total 650 articles, and the results of meta-analysis showed that application of intraoperative AABO had the ability to reduce the operative time (WMD = - 16.581, 95% CI - 26.690 to - 6.472; P = 0.001), the intraoperative blood loss volume (WMD = - 1202.69, 95% CI - 1732.25 to - 673.12; P < 0.001), the intraoperative blood transfusion volume (WMD = - 1202.69, 95% CI - 1732.25 to - 673.12; P < 0.001). The hysterectomy rate (RR = 0.279, 95% CI 0.164-0.474; P < 0.001), postoperative hospitalization duration (WMD = - 1.423, 95% CI - 2.070 to - 0.776; P < 0.001) and the balloon preset time (WMD = - 13.793, 95% CI - 15.341 to - 12.244; P < 0.001; I2 = 0.0%) were also reduced in AABO group. CONCLUSIONS: Application of AABO in patients with pernicious placenta previa is safe and effective, which is worthy of clinical promotion.

Simultaneous qualitative and quantitative evaluation of Toddalia asiatica root by using HPLC-DAD and UPLC-QTOF-MS/MS.

INTRODUCTION: Coumarin and alkaloids are the major bioactive constituents of Toddalia asiatica, playing an important role in various biological activities such as anti-inflammatory, analgesic, anti-bacterial and anti-tumour. OBJECTIVE: To establish a method that will simultaneously determine the coumarins and alkaloids compounds in T. asiatica and identify their characteristic fragmentation patterns, while combining fingerprints and chemical identification with chemometrics for discrimination and quality assessment of T. asiatica samples. METHODOLOGY: Qualitative characterisation of coumarins and alkaloids compounds in the methanol extracts of T. asiatica was determined by ultra-high-performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry (UPLC-QTOF-MS/MS). Quantitative analysis relies on high-performance liquid chromatography with a diode array detector (HPLC-DAD). RESULTS: A total of 59 components were characterised by UPLC-QTOF-MS/MS, including 29 coumarin, 25 alkaloids, one phenolic acid and four flavonoids. While the 19 characteristic components out of 23 common peaks in the chromatographic fingerprints of T. asiatica were confirmed. Quantitative analysis of seven major compounds from 18 samples were simultaneously detected by HPLC-DAD at wavelengths of 280 nm. The samples were classified into three groups by hierarchical clustering analysis (HCA) combined with principal component analysis (PCA), and orthogonal partial least squares discriminant analysis (OPLS-DA) which screened out the main chemical markers responsible for the samples differences. CONCLUSION: Fingerprints combined with chemometrics and chemical identification are a simple, rapid and effective method for the quality control of T. asiatica.

Multi-Component Analysis of Ilex Kudingcha C. J. Tseng by a Single Marker Quantification Method and Chemometric Discrimination of HPLC Fingerprints.

The quantitative analysis of multiple components with a single marker (QAMS) method was firstly established for simultaneous determination of 18 active components in Ilex kudingcha C. J. Tseng by HPLC. Using rutin, isochlorogenic acid A and kudinoside A as internal refererence substances (IRS), compatibility results showed that the relative correction factors (RCFs) of all compounds showed good reproducibility under different chromatographic conditions. On the basis of previous studies, the accuracy of the QAMS method was systematically evaluated by investigating the influences of curve intercept, analytes and IRS concentration. The results showed that the concentration (especially at low level) of analytes and curve intercept were the major influencing parameters for the LRG-QAMS method (LRG = linear regression), whereas the influence of IRS concentration seemed more apparent in terms of the AVG-QAMS method (AVG = average). The two approaches were complementary with each other. In addition, hierarchical clustering analysis (HCA), principal components analysis (PCA) and similarity analysis (SA) were performed to differentiate and classify the samples based on the contents of 18 marker compounds. The results of the different chemometric analyses were completely consistent with each other, and could be supported by the quantification results.

Simultaneous qualitative and quantitative evaluation of Ilex kudingcha C. J. tseng by using UPLC and UHPLC-qTOF-MS/MS.

In this study, a systematic method was established for the holistic quality control of Ilex kudingcha C. J. Tseng, a popular functional drink for adjuvant treatment of diabetes, hypertension, obesity and hyperlipidemia. Both qualitative and quantitative analyses were conducted. For qualitative analysis, an ultra high performance liquid chromatography (UHPLC) coupled with an electrospray ionization quadrupole time-of-flight mass spectrometry (ESI-qTOF-MS) method was established for rapid separation and structural identification of the constituents in Ilex kudingcha. Samples were separated on an ACQUITY UPLC HSS T3C18 column (2.1 mm × 100 mm, 1.8 µm) by gradient elution using 0.1% (v/v) formic acid (solvent A) and acetonitrile (solvent B) as mobile phases at a flow rate of 0.25 mL min-1. The chromatographic profiling of Ilex kudingcha by UHPLC-qTOF-MS/MS resulted in the characterization of 53 compounds, comprising 18 compounds that were unambiguously identified by comparison with reference standards. For quantitative analysis, 18 major compounds from 15 batches of Ilex kudingcha samples were simultaneously detected by UPLC-DAD at wavelengths of 210 nm, 260 nm, and 326 nm. The method was validated with respect to precision, linearity, repeatability, stability, accuracy, and so on. The contents of the 18 target compounds were applied for hierarchical clustering analysis (HCA) and principal component analysis (PCA) to differentiate between the samples. The results of HCA and PCA were consistent with each other. Sample No. 1 differed significantly based on HCA and PCA, and the differentiating components were confirmed to originate from different batches of samples. Phenolic acids and triterpenes were found to be the main ingredients in Ilex kudingcha. This strategy was effective and straightforward, and provided a potential approach for holistic quality control of Ilex kudingcha.

[HPLC-UV fingerprints and chemical pattern recognition of Ilicis Pubescentis Radix].

The present study is to establish the fingerprints for the quality evaluation of Ilicis Pubescentis Radix by HPLC-UV. The chromatographic conditions were defined as Phenomenex Luna C18(4.6 mm × 250 mm, 5 µm). Mobile phase was acetonitrile-0.05% phosphoric acid in gradient elution, and the flow rate was 0.8 mL·min⁻¹.Column temperature was 30 °C and the injection volume was 10 µL．The detection wavelength was 210 nm. According to the similarity evaluation, the chemometric method was used to assess the quality of Ilicis Pubescentis Radix. The fingerprints of 16 batches of Ilicis Pubescentis Radix were established. There were 29 common peaks in the fingerprints and 12 common peaks were identified by reference substances. Fingerprints similarity of samples were greater than 0.92. The samples were classified into three groups by hierarchical cluster analysis combined with principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA), and seven components were the main markers that cause differences in the different batches of samples. By comparing the on-line UV spectra of chromatographic peaks, the chromatographic fingerprint was divided into three regions: region A showed seventeen main peaks (mainly lignans and phenolic acids); region B showed eight main peaks, which were proved as saponins; region C showed four main peaks, which were proved as other components. The established HPLC-UV fingerprint is highly specific, and can be used to evaluate the quality consistency of different batches of Ilicis Pubescentis Radix.

Study of Absorption Characteristics of the Total Saponins from Radix Ilicis Pubescentis in an In Situ Single-Pass Intestinal Perfusion (SPIP) Rat Model by Using Ultra Performance Liquid Chromatography (UPLC).

In contrast to the extensively reported therapeutic activities, far less attention has been paid to the intestinal absorption of the total saponins from Radix Ilicis Pubescentis (in Chinese Mao-Dong-Qing, MDQ). This study aimed to investigate the intestinal absorption characteristics of ilexgenin A (C1), ilexsaponin A1 (C2), ilexsaponin B1 (C3), ilexsaponin B2 (C4), ilexsaponin B3 (DC1), and ilexoside O (DC2) when administrated with the total saponins from MDQ (MDQ-TS). An UPLC method for simultaneous determination of C1, C2, C3, C4, DC1, and DC2 in intestinal outflow perfusate was developed and validated. The absorption characteristics of MDQ-TS were investigated by evaluating the effects of intestinal segments, drug concentration, P-glycoprotein (P-gp) inhibitor (verapomil), endocytosis inhibitor (amantadine) and ethylene diamine tetraacetic acid (EDTA, tight junction modulator) on the intestinal transportation of MDQ-TS by using a single-pass intestinal perfusion (SPIP) rat model, and the influence of co-existing components on the intestinal transport of the six saponins was discussed. The results showed that effective apparent permeability (Papp) of C1, C2, C3, C4, and DC2 administrated in MDQ-TS form had no segment-dependent changes at low and middle dosage levels. C1, C2, C3, D4, DC1, and DC2 administrated in MDQ-TS form all exhibited excellent transmembrane permeability with Papp > 0.12 × 10-2 cm·min-1. Meanwhile, Papp and effective absorption rate constant (Ka) values for the most saponins showed concentration dependence and saturation characteristics. After combining with P-gp inhibitor of verapamil, Papp of C2, C3, and DC1 in MDQ-TS group was significantly increased up to about 2.3-fold, 1.4-fold, and 3.4-fold, respectively in comparison to that of non-verapamil added group. Verapamil was found to improve the absorption of C2, C3, and DC1, indicating the involvement of an active transport mechanism in the absorption process. Compared with the non-amantadine added group, the absorption of C1, C2, C4, DC1, and DC2 were decreased by 40%, 71%, 31%, 53%, and 100%, respectively. Papp for the six target compounds increased up to about 1.2-2.1-fold in comparison with the non-EDTA added, respectively. The gastrointestinal transport of MDQ-TS could be greatly promoted by EDTA, and inhibited by amantadine, implying that the intestinal absorption of MDQ-TS was by passive diffusion and endocytosis process. Compared with monomer administration group, the intestinal absorption of C3, C4, DC1, and DC2 was significantly improved by co-existing components in MDQ-TS, and the non-absorbable saponins of C4, DC1, and DC2 unexpectedly showed sufficient intestinal permeability with Papp > 0.12 × 10-2 cm·min-1. This suggested that compounds orally administrated in TCM extract forms displayed unique intestinal absorption characteristics different from those of monomers, and the enhancing intestinal absorption of MDQ-TS reflected a holistic and specific view of traditional Chinese medicines (TCMs).

Crystal structure of diethyl [(4-chloro-anilino)(4-hy-droxy-phen-yl)meth-yl]phospho-nate N,N-di-methyl-formamide monosolvate.

In the title compound, C17H21ClNO4P·C3H7NO, the dihedral angle formed by the aromatic rings is 83.98 (7)°. In the crystal, O-H⋯O, N-H⋯O and C-H⋯O hydrogen bonds link the mol-ecules into double layers parallel to (011).

Development of a high analytical performance-xanthine biosensor based on layered double hydroxides modified-electrode and investigation of the inhibitory effect by allopurinol.

The determination of xanthine has considerable importance in clinical and food quality control. Therefore, in this present work, we developed a novel xanthine biosensor based on immobilization of xanthine oxidase (XnOx) by attractive materials layered double hydroxides (LDHs). Amperometric detection of xanthine was evaluated by holding the modified electrode at 0.55V (versus saturated calomel electrode (SCE)). Due to the special properties of LDHs, such as chemical inertia, mechanical and thermal stability, anionic exchange ability, high porosity and swelling properties, XnOx/LDHs-modified electrode exhibited a developed analytical performance. The biosensor provided a linear response to xanthine over a concentration range of 1 x 10(-6)M to 2 x 10(-4)M with a sensitivity of 220 mAM(-1)cm(-2) and a detection limit of 1x10(-7)M based on S/N=3. In addition, the immobilized XnOx layers have been characterized using atomic force microscopy under both air atmosphere and liquid environment, which exhibited the interesting swelling phenomenon of LDHs. The investigation of inhibition of XnOx by allopurinol was carried out using this XnOx/LDHs-modified electrode. The experimental results indicated that inhibitory effect could be achieved by allopurinol with a quasi-reversible competitive type.

Calcium carbonate nanoparticles: a host matrix for the construction of highly sensitive amperometric phenol biosensor.

We reported on the utilization of a novel attractive nanoscaled calcium carbonate (nano-CaCO(3))-polyphenol oxidase (PPO) biocomposite to create a highly responsive phenol biosensor. The phenol sensor could be easily achieved by casting the biocomposite on the surface of glassy carbon electrode (GCE) via the cross-linking step by glutaraldehyde. The special three-dimensional structure, porous morphology, hydrophilic and biocompatible properties of the nano-CaCO(3) matrix resulted in high enzyme loading, and the enzyme entrapped in this matrix retained its activity to a large extent. The proposed PPO/nano-CaCO(3) exhibited dramatically developed analytical performance such as such as a broad determination range (6 x 10(-9) -2 x 10(-5)M), a short response time (less than 12 s), high sensitivity (474 mA M(-1)), subnanomolar detection limit (0.44 nM at a signal to noise ratio of 3) and good long-term stability (70% remained after 56 days). In addition, effects of pH value, applied potential, temperature and electrode construction were investigated and discussed.

Development of amperometric biosensor for glucose based on a novel attractive enzyme immobilization matrix: calcium carbonate nanoparticles.

Calcium carbonate nanoparticles (nano-CaCO3) may be a promising material for enzyme immobilization owing to their high biocompatibility, large specific surface area and their aggregation properties. This attractive material was exploited for the mild immobilization of glucose oxidase (GOD) in order to develop glucose amperometric biosensor. The GOD/nano-CaCO3-based sensor exhibited a marked improvement in thermal stability compared to other glucose biosensors based on inorganic host matrixes. Amperometric detection of glucose was evaluated by holding the modified electrode at 0.60 V (versus SCE) in order to oxidize the hydrogen peroxide generated by the enzymatic reaction. The biosensor exhibited a rapid response (6s), a low detection limit (0.1 microM), a wide linear range of 0.001-12 mM, a high sensitivity (58.1 mAcm-2M-1), as well as a good operational and storage stability. In addition, optimization of the biosensor construction, the effects of the applied potential as well as common interfering compounds on the amperometric response of the sensor were investigated and discussed herein.