Doping Control Analysis of 16 Non-Steroidal Anti-Inflammatory Drugs in Equine Plasma Using Liquid Chromatography-Tandem Mass Spectrometry

Non-steroidal anti-inflammatory drugs (NSAIDs) are classified as Class 4 agents by the Association of Racing Commissioners International and are banned in racehorses during competition in Pennsylvania (PA). To control the abuse of these agents in racehorses competing in PA, a forensic method for screening and confirmation of the presence of these agents is needed. Equine plasma (0.5 mL) was acidified with 75 μL 1M H3PO4 to increase recovery of the analytes by liquid-liquid extraction using methyl tert-butyl ether (MTBE). Extracted analytes were separated by reversed-phase liquid chromatography using a C8 column under gradient condition. All 16 analytes were detected, quantified and confirmed using a triple quadrupole tandem mass spectrometry with selected reaction monitoring (SRM) in both negative and positive electrospray ionization modes. The limit of detection, quantification and confirmation of the analytes were 1.0 - 5.0 ng/mL, 1.0 - 5.0 ng/mL and 1.0 - 20 ng/mL, respectively. The linear dynamic range of quantification was 5.0 - 200 ng/mL. The method is routinely used in anti-doping analysis to control the abuse of NSAIDs in racehorses competing in PA.

The analgesic, anti-inflammatory, and anti-infection effects of Chai Ge fever relief oral liquid

Backgroud:To explore the analgesic,anti-inflammatory,and anti-infective effects of Chai Ge fever relief oral liquid and provide evidence for clinical application of Chai Ge fever relief oral liquid.Methods:In this study,four groups of experiments were designed:analgesic,anti-inflammatory,antiviral,and antibacterial.In the mouse pain model,the analgesic effect of Chai Ge fever relief oral liquid was studied by the writhing method and pain threshold;the anti-inflammatory effect by measuring the level of capillary permeability in the abdominal cavity of mice in different dose groups and the weight of cotton ball granuloma formation in rats;the antiviral effect by measuring the lung index of a pneumonia model of mouse infected with influenza virus;and the antibacterial effect by comparing the difference in the death protection ratio between each dose group and the model group result.Results:In the analgesic experiment,the high and medium dose of Chai Ge fever relief oral liquid could significantly reduce the number of writhing in mice caused by acetic acid,and the pain threshold of mice in the high-and medium-dose groups was significantly increased for 1–3 hours.In the anti-inflammatory experiment,the medium-dose group could significantly inhibit the increase of capillary permeability in the abdominal cavity of mice caused by acetic acid,and the low-dose group could significantly reduce the weight of rat cotton ball granuloma.In the antiviral experiment,the high-and medium-doses of Chai Ge fever relief oral liquid could significantly reduce the lung index of normal mouse pneumonia model of influenza virus infection and achieve a higher inhibition rate.In the anti-infective experiment,the death protection rate of the high-dose group was significantly different from that of the model control group.All three dose groups could significantly prolong the survival days of infected mice.Conclusion:These experimental results prove that in addition to its antipyretic effect,Chai Ge fever relief oral liquid also has analgesic,anti-inflammatory,antiviral,and antibacterial effects.

Solid and liquid state characterization of tetrahydrocurcumin using XRPD, FT-IR, DSC, TGA, LC-MS, GC-MS, and NMR and its biological activities

Tetrahydrocurcumin(THC)is one of the major metabolites of curcumin(CUR),an ancient bioactive natural polyphenolic compound.This research article describes both the solid and liquid state characterization of THC using advanced spectroscopic and thermo-analytical techniques.Anti-inflammatory,anti-oxidant,and neuroprotective activities of THC were investigated using in vitro cell lines.Liquid chromatography-mass spectrometry analysis revealed that our sample comprised 95.15%THC,0.51%tetrahydrodemethoxycurcumin(THDC),3.40%hexahydrocurcumin,and 0.94%octahydrocurcumin.Gas chromatography-mass spectrometry analysis indicated the presence of 96.68%THC and 3.32%THDC.THC in solution existed as keto-enol tautomers in three different forms at different retention time,but the enol form was found to be dominant,which was also supported by nuclear magnetic resonance analysis.THC was thermally stable up to 335.55
C.THC exhibited more suppression of cytokines(TNF-a,IL-1b,and MIP-1a)than CUR in a concentration-dependent manner in mouse splenocytes,while NK-cell and phagocytosis activity was increased in macrophages.THC showed a significant reduction of free radicals(LPO)along with improved antioxidant enzymes(SOD and catalase)and increased free radical scavenging activity against ABTSt radicals in HepG2 cells.THC displayed higher protection capability than CUR from oxidative stress and neuronal damage by improving cell viability against H2O2 induced HepG2 cells and MPPt induced SH-SY5Y cells,respectively,in a concentration-dependent manner.Thus,a variation of the biological activities of THC might rely on its keto-enol form and the presence of other THC analogs as impurities.The present study could be advantageous for further research on THC for better understanding its physicochemical properties and biological variation.

Pharmacodynamic study in multi-animal models on the efficacy and optimal dosage of antiviral oral liquid for children

Antiviral Oral Liquid(AOL)is an adult medicine in the Chinese Pharmacopoeia used to treat upper respiratory infections such as influenza.It has shown promising clinical efficacy in relieving flu-like symptoms such as fever,inflammation,and pharyngalgia both in adults and children.However,the instruction manual does not specify the exact usage and dosage of AOL for children.In this article,we set 6 dosage ranges:0.2,0.5,0.7,0.9,1.1,1.4 mL/kg/d,according to its dosage for adults and the conversion method between adult and children dosage.And six animal models were used to evaluate the effectiveness of AOL in different doses.The results indicated that AOL could reduce the lung index,virus load,and expression of proinflammatory cytokines in the lung.AOL could improve pathological changes and prolong the survival time of mice infected by the Influenza A virus(H1N1)A/PR/8/34 strains at 0.5–0.9 mL/kg/d concentrations in different degrees.The four dose groups of 0.7–1.4 mL/kg/d could significantly inhibit the ear shell swelling caused by xylene and reduce the rabbit body temperature induced by lipopolysaccharide(P<0.01,P<0.05).All the five dosage groups of 0.2–1.1 mL/kg/d could inhibit the increase of peritoneal capillary permeability induced by glacial acetic acid(P<0.01).AOL at 0.7 and 0.9 mL/kg/d reduced the painful writhing times in young mice induced by glacial acetic(P<0.01).These results indicated that the optimal dose of AOL in antiviral,antipyretic,anti-inflammatory,and analgesic effects is 0.7 mL/kg/d.

丹芍消炎合剂联合静脉抗生素预防盆腔炎性疾病后遗症

目的:探讨丹芍消炎合剂联合静脉抗生素预防盆腔炎性疾病后遗症的临床疗效。方法:83例亚急性盆腔炎患者,随机分为治疗组42例(丹芍消炎合剂联合静脉抗生素治疗)和对照组41例(单纯静脉抗生素治疗),经治疗后,对比两组在2个月后盆腔积液和盆腔包块的改善情况、两组中初发患者的超声输卵管造影结果,以及两组在半年后的复发率和生活质量评分情况。结果:在2个月后,治疗组在减少盆腔积液方面疗效达90.6%,显著高于对照组(疗效69.0%),且两组比较差异有统计学意义(P<0.01);治疗组在缩小盆腔包块方面疗效达61.8%,显著高于对照组(疗效32.9%),且两组比较差异有统计学意义(P<0.01);治疗组的输卵管通畅程度(有效通畅程度71.9%)高于对照组(有效通畅程度37.5%),且差异有统计学意义(P<0.05);在半年后,治疗组复发率为9.5%(2/42)低于对照组复发率19.5%(9/41),且差异有统计学意义(P<0.05);治疗组患者半年后的生活质量评分高于对照组,且差异有统计学意义(P<0.01)。结论:丹芍消炎合剂联合静脉抗生素治疗能显著减少盆腔积液、缩小盆腔包块,有效减少输卵管黏连的发生,并且可以降低盆腔炎的复发率、改善患者的生活质量,能有明确预防盆腔炎性疾病后遗症的作用,值得临床推广应用。

院内制剂丹芍清胃颗粒中芍药苷含量的HPLC测定

目的研究并建立丹芍清胃颗粒含量测定方法,为质量控制提供实验依据。方法以十八烷基硅烷键合硅胶为填充剂的色谱柱;以乙腈-0.1%磷酸水(14:86)为流动相;检测波长230 nm。结果该条件下芍药苷峰面积和质量浓度之间线性关系良好,r为0.9999,精密度、稳定性均符合要求。平均加样回收率为98.17%,RSD为1.75%。结论本研究建立的HPLC芍药苷含量测定方法,可以作为丹芍清胃颗粒质量控制的依据。

In vitro anti-inflammatory effects of different solution fractions of ethanol extract from Melilotus suaveolens Ledeb

Background Melilotus suaveolens Ledeb （M. suaveolens Ledeb） has long been used as a folk medicine in inflammation-related therapy. This study was undertaken to determine the anti-inflammatory effect of the plant. Methods Petroleum ether fraction, ethyl acetate fraction, n-butanol fraction, aqueous fraction were obtained from ethanol extract of M. suaveolens Ledeb and evaluated by high performance liquid chromatography （HPLC）. While dexamethasone （DM） was used as a positive control, the effects of different solution fractions of ethanol extract on tumor necrosis factor α （TNF-α） mRNA, cyclooxygenase 2 （COX-2） mRNA, COX-2 and nuclear factor KB （NF-KB） of LPS-stimulated RAW 264.7 cells were studied by real-time PCR, Westem blot analysis and immunocytochemical assay, respectively. Results Coumarin was one of the main ingredients in different solution fractions of ethanol extract except the aqueous fraction with no inflammatory effect. The petroleum ether fraction, ethyl acetate fraction and n-butanol fraction of ethanol extract could inhibit the production of TNF-α mRNA, COX-2 mRNA and NF-KB to some extent. Conclusions Different solution fractions of ethanol extract from M. suaveolens Ledeb had similar anti-inflammatory effect as did dexamethasone except the aqueous fraction. Coumarin was likely to be essential to the anti-inflammatory effect, and other ingredients might attribute to their different anti-inflammatory effects from the HPLC fingerprint.

Simultaneous and interference-free determination of eleven nonsteroidal anti-inflammatory drugs illegally added into Chinese patent drugs using chemometrics-assisted HPLC-DAD strategy

In this work, a smart strategy that combines three-way high performance liquid chromatography-diode array detection(HPLCDAD) data with second-order calibration method based on alternating trilinear decomposition(ATLD) algorithm was proposed for simultaneous determination of eleven non-steroidal anti-inflammatory drugs(NSAIDs) illegally added into Chinese patent drugs and health products. All target analytes were rapidly eluted out within 14.5 min under a simple gradient elution. With the aid of the prominent "second-order advantage" of the ATLD algorithm, three HPLC problems, i.e. peak overlaps, unknown interferences and baseline drift, could be mathematically calibrated, and pure signals of target analytes could be extracted out from heavy-interference but information-rich HPLC-DAD data. The average spiked recoveries for all target analytes were in the range of 95.9%–106.4% with standard deviations lower than 7.5%. Validation parameters including sensitivity(SEN), selectivity(SEL), limit of detection(LOD), limit of quantitation(LOQ) and precisions of intra-day and inter-day were calculated to validate the accuracy of the proposed method, quantitative results were further confirmed by the classic HPLC method, which proved that chemometrics-assisted HPLC-DAD analytical strategy was highly efficient, accurate and green for drug-abuse monitoring of NSAIDs in Chinese patent drugs and health products.

Ionic liquid mediated morphologically improved lanthanum oxide nanoparticles by Andrographis paniculata leaves extract and its biomedical applications

Greener synthesis of ionic liquid(IL) assisted lanthanum oxide(La2O3) nanoparticles(NPs) was followed using Andrographis paniculata leaves extract by hydrothermal method at low temperature,The IL assisted La2O3(La2O3-IL) NPs were characterized by various properties such as structural,spectral,optical,morphological,and biological studies.Powder X-ray diffraction studies confirm that La2O3-IL NPs crystallize in the form of body-centered cubic structure and average crystalline size is shown-43 nm.FTIR and Laser Raman spectroscopy confirm the La-O,O-H and various functional groups.The La-O stretching vibration band attributes at 639 cm-1.The optical property of La2O3-IL NPs was characterized by UV-Vis and photo luminesce nce spectroscopy.The sharp and intense absorbance peak observed in the UV region at 288 nm can be assigned.In the PL spectrum,we observe two different emission bands like blue and green emission.The morphological and particle size determination were investigated by SEM with EDX spectra and TEM analyses.The agglomerated particles of hexagonal,spherical,and parallelogram shape are observed in the TEM images.The chemical elements,binding energies as well as La metal states on the surface of these synthesized NPs were determined using XPS.La2O3 NPs for good biological activities regarding antibacterial and anti-inflammatory potentials could be utilized in different biological applications to the food and biomedical industries.La2O3-IL has a high percentage of inhibitions than La2O3 and standard diclofenac.

丹芍消炎合剂联合抗生素治疗亚急性盆腔炎的疗效观察

目的探讨丹芍消炎合剂联合静脉抗生素治疗湿热型亚急性盆腔炎的临床效果。方法 83 例确诊为亚急性盆腔炎患者随机分为2 组:对照组41 例,每12 h 静脉滴注头孢西丁2 g(头孢类过敏者改用克林霉素1 g),使用至临床症状、体征改善后的24 h,同时po 强力霉素100 mg,2 次/d,共2 周;治疗组42 例,在对照组基础上po 芍消炎合剂35 mL/次,3次/d,连续2 周,比较两组患者的临床疗效与不良反应。结果治疗组的总有效率为 97.6%、愈显率76.1%,静脉抗生素使用(3.52±0.94)d;对照组总有效率90.24%、愈显率29.2%,静脉抗生素使用(4.95±1.18)d,两组比较差异有统计学意义(P<0.01);且两组均未发生不良反应。结论丹芍消炎合剂联合静脉抗生素治疗亚急性盆腔炎疗效确切,能明显改善患者的临床症状、缩短静脉抗生素的使用时间,值得临床推广应用。