Antitumor Activity of Choerospondias axillaris Fruit Extract by Regulating the Expression of SNCAIP and SNCA on MDA-MB-231 Cells

Objective: Cancer is a huge problem of disease globally. Today, the percentage of people die from cancer is more than a combination of various diseases. In females, most common types of malignancies that occur are breast and cervical. The present focus has been shifted on medicinal plants as a form of therapy and there is a constant need to identify new therapeutic agents. Choerospondias axillaris (C. axillaris), an underutilized fruit, has been used in the remedy of various diseases. In the present communication, we evaluated the molecular mechanism of C. axillaris methanol extract in regulating cell death in human breast cancer cells (MDA-MB-231). Methods: Methanol extract of C. axillaris was prepared and compounds were screened by Gas chromatography-mass spectrometry. The effect of fruit extract was determined on MDA-MB-231 cells by MTT ((3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide) assay and to analyse the molecular mechanism of human breast cancer cells after treating with fruit extract, protein profiling study was performed by two-dimensional gel electrophoresis. Results: A total 9 differentially expressed proteins were identified by matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS/MS) analysis. Among 9 identified proteins, synphilin-1 protein was found to be significantly downregulated, validated by western blot and RT-qPCR analysis. Possible interacting partners of synphilin-1 (SNCAIP) were analyzed for their possible role in cancer by the in-silico method. Conclusion: Our data implicate that the presence of bioactive compound(s) in C. axillaris fruits might play an important role in inhibiting the proliferation of breast carcinoma cells and Synphilin-1 protein may play a role of apoptotic function.     

苓桂术甘汤中多糖结构组成及其抗氧化活性考察

目的:研究苓桂术甘汤中多糖的结构,包括单糖组成以及官能团检测,并对其抗氧化能力进行评估,为体外生物测定法运用于苓桂术甘汤的质量评价提供基础。方法:利用高效凝胶色谱对全方多糖分子量进行研究;进一步采用气相色谱-质谱(GC-MS)柱前衍生化法及红外光谱扫描分析全方多糖结构组成;采用1,1-二苯基-2-三硝基苯肼(DPPH)及羟基自由基测定全方粗多糖及精多糖的抗氧化能力。结果:全方多糖由单一峰组成,且相对分子质量为3 689 Da,主要由阿拉伯糖、甘露糖、葡萄糖、半乳糖、果糖组成,摩尔比为6. 85∶1. 00∶109. 21∶1. 04∶21. 82,其中葡萄糖和果糖为主要组成成分;红外结果显示聚糖结构中存在吡喃糖及糖醛酸,并且糖苷键存在2种立体异构(α-糖苷键及β-糖苷键);抗氧化研究发现全方多糖有一定清除DPPH自由基及羟基自由基的能力,粗多糖的活性优于精多糖。进一步采用LC-Q-TOF-MS对粗多糖中的其他成分进行定性分析,提示这与甘草中五环三萜类成分的吸附有关。结论:苓桂术甘汤中多糖及五环三萜类物质均是苓桂术甘汤抗氧化作用的物质基础,利用体外生物测定法测定抗氧化活性可作为全面控制苓桂术甘汤质量的评价指标。     

顶空气相色谱-质谱法测定布南色林原料药中遗传毒性杂质

目的：建立顶空气相色谱-质谱法测定布南色林原料药中甲磺酸甲酯、甲磺酸乙酯和甲磺酸异丙酯3种遗传毒性杂质含量的方法。方法：采用顶空气相色谱-质谱法，以甲磺酸丁酯为内标，按内标标准曲线法进行甲磺酸甲酯、甲磺酸乙酯和甲磺酸异丙酯的含量测定。色谱条件：DB-WAX毛细管色谱柱（30 m×0.25 mm×0.25 μm）；程序升温，初始柱温为40℃，维持3 min，升温速率为30℃·min^-1，终止温度150℃，保持2 min；进样口温度为110℃；载气（He）流速为0.6 mL·min^-1；进样量为1 mL；进样方式为分流进样，分流比为20：1。质谱条件：电子轰击离子源（EI），扫描方式为选择性离子检测；离子源温度为200℃；接口温度为150℃；电子能量为70 eV；溶剂延迟1 min。结果：3种杂质成分之间的分离度均大于2.0；甲磺酸甲酯、甲磺酸乙酯、甲磺酸异丙酯检测质量浓度线性范围均为0.025~3.0 μg·mL^-1（r ≥ 0.998 5）；精密度、稳定性、重复性试验的RSD<5%；加样回收率分别为93.40%~101.40%（RSD为3.2%，n=9）、92.80%~99.70%（RSD为2.5%，n=9）和96.30%~100.75%（RSD为1.6%，n=9）。结论：该方法简便、准确、灵敏、迅速，可用于布南色林原料药中3种遗传毒性杂质的测定。     

Determination of polycyclic aromatic hydrocarbons (PAHs) in cosmetic products by gas chromatography-tandem mass spectrometry

A gas chromatography coupled with tandem mass spectrometry (GC-MS/MS) method is developed to determine 18 representative polycyclic aromatic hydrocarbons (PAHs) in cosmetics, including Benzo[a]pyrene (BaP) and others. The method offers high sensitivity and selectivity under selected reaction monitoring (SRM) mode to satisfy the requirements of both quantitation and qualitation. The extraction solvent system used in this study is acetone/hexane 1:1 (v/v) and other purification procedure is unnecessary. The linearities of 18 PAHs are validated in different concentration in the range of 0.25–20 ng/mL individually with coefficient correlation (r) higher than 0.996. The recoveries for spiking 3 different concentrations are from 87.40% to 120.44% for 18 PAHs and the coefficient of variation (CV) are below 12.32%. Limit of quantification (LOQ) of 18 PAHs is in the range of 0.05–0.2 mg/kg. A matrix enhancement effect is observed and can be compensated with deuterated internal standard. The method has been successfully applied to 73 samples, over 40 of them are lipsticks. The results show none of the samples detect Benzo[a]pyrene (BaP) and Dibenzo[a,h]anthracene (DBA), both are classified as the most carcinogenic. 8 PAHs are detected and the average value between 0.08 and 0.27 mg/kg. This study offers a sensitive and simple method to analyze 18 representative PAHs successfully and can be applied to cosmetic products and raw materials.     

山稔不同部位挥发油成分GC-MS分析

目的:分析山稔根、茎、叶中挥发油的化学成分。方法:采用水蒸气蒸馏法分别提取山稔不同部位的挥发油,通过气相色谱-质谱联用法(GC-MS)对根、茎、叶3个部位挥发油的化学成分进行对比研究,并用面积归一化法获得各化合物的相对含量。结果:从山稔根、茎、叶中分别鉴定化学成分22、32和36个,分别占挥发油总量的52.4%、76.2%和86.2%。结论:山稔根、茎、叶中挥发油的化学成分在种类及主要成分的含量上都存在较大差异,为山稔的进一步开发利用提供了实验依据。     

树发挥发油成分和脂溶性成分的GC-MS分析

目的:研究树发的挥发油和脂溶性成分。方法:利用气相色谱-质谱联用(GC-MS)技术对树发挥发油和脂溶性成分进行分析。结果:共鉴定出34个挥发油成分,占总量的17.93%,其成分类型主要包括萜类(6.71%),酮类(4.19%)及醇类(3.27%),主要成分为4-戊烯-2-酮(4.19%),cis-7-十四烯醇(3.03%),3,7-二甲基辛烯(1.30%),新己烷(1.28%)和大牦牛儿烯D(1.01%);共鉴定出33个脂溶性成分,占总量的89.63%,其成分类型主要包括脂肪酸类(81.65%),烷烃类(4.49%)和醇类(1.67%),主要成分为反亚油酸甲酯(34.98%),油酸甲酯(10.94%),14-甲基-十五烷酸甲酯(7.40%)和硬脂酸甲酯(4.13%)。结论:首次从树发中提取并鉴定了挥发油和脂溶性成分,为树发的开发利用提供了科学依据。     

GC-MS分析王不留行、决明子、酸枣仁炒制前后脂肪油成分的变化

目的:分析和鉴定王不留行、决明子、酸枣仁3种药材炒制前后脂肪油类成分的变化,为探索种子类药材"逢子必炒"的炮制机制奠定基础。方法:以石油醚为溶剂,采取索氏提取器提取脂肪油,甲酯化后用气相色谱-质谱联用技术(GCMS)分离、鉴定其组成和相对质量分数。HP-5毛细管柱(0.25 mm×30 m,0.25μm),载气99.999%高纯氮,流速1 mL·min~(-1),采用程序升温方式(初始温度50℃,以5℃·min~(-1)升至170℃,停留4 min,5℃·min~(-1)升至190℃,0.5℃·min~(-1)升至200℃,10℃·min~(-1)升至240℃,保持4 min),进样量1μL,电子轰击离子源(EI),离子源温度220℃,电子能量70 eV。结果:王不留行、决明子、酸枣仁3种药材生品脂肪油提取率分别为3.34%,6.16%,20.66%,炒制品脂肪油提取率分别为4.67%,7.40%,26.35%。王不留行生品鉴定出7个化合物,炒制品鉴定出10个化合物;决明子生品鉴定出8个化合物,炒制品鉴定出9个化合物;酸枣仁生品鉴定出10个化合物,炒制品鉴定出14个化合物。结论:炒制能提高脂肪油的提取效率,炒制后3种药材脂肪油的组成出现了一些共性变化。     

GC-MS/MS测定替米沙坦片中10种亚硝胺类基因毒性杂质

目的 建立GC-MS/MS同时测定替米沙坦片中N-亚硝基二甲胺(NDMA)、N-亚硝基甲乙胺(NMEA)、N-亚硝基二乙胺(NDEA)、N-亚硝基-N-乙基异丙胺(NEIPA)、N-亚硝基二异丙胺(NDIPA)、N-亚硝基二正丙胺(NDPA)、N-亚硝基二正丁胺(NDBA)、N-亚硝基哌啶(NPIP)、N-亚硝基吡咯烷(NPYR)和N-亚硝基吗啉(NMOR)10种亚硝胺类基因毒性杂质的含量。方法样品经甲醇提取,经Agilent VF-WAXms(30 m×0.25 mm,1μm)毛细管气相色谱柱分离,采用多反应离子监测(MRM)模式进行定量分析。结果 NDMA、NMEA、NDEA、NEIPA、NDIPA、NDPA、NDBA和NPIP在0.2～50 ng·mL^–1线性关系良好,相关系数均为1.000 0;检测限均为0.05 ng·mL^–1,定量限均为0.2 ng·mL^–1,平均回收率分别为103%(RSD=9.2%,n=9),108%(RSD=6.1%,n=9),107%(RSD=5.3%,n=9),106%(RSD=3....     

安徽省不同药用菊花中挥发油成分的GC-MS分析

目的分析研究安徽省不同药用菊花中的挥发油成分,对挥发油的含量、成分等进行比较研究,为安徽省不同药用菊花的质量评价提供理论依据。方法在适宜的采收期采集新鲜菊花,通过传统的干燥方法干燥得到实验样品,采用水蒸气-蒸馏法提取挥发油,用气相-质谱(GC-MS)联用的方法分析并鉴定挥发油中的化学成分。结果从贡菊、七月菊、滁菊、小亳菊和大马牙挥发油中分别检出210、100、202、205和218个色谱峰,分别鉴定了31、17、26、42和53个化合物,分别占挥发油总量的72.53%、85.62%、79.97%、78.73%和82.49%。结论安徽省不同药用菊花的挥发油成分之间差异较大,本研究结果为安徽省不同药用菊花的质量评价提供了物质基础。