UHPLC/Q-Orbitrap-MS结合化学计量学方法研究酸枣仁入血成分

建立了一种基于超高效液相色谱-质谱联用技术结合化学计量学快速分析酸枣仁入血成分的方法。以腹腔注射对氯苯丙氨酸(PCPA)建立失眠大鼠模型,给药组连续灌胃给予酸枣仁水提物(30 g/kg)5 d后,分别收集模型组和给药组的血清样品。采用Oasis PRIME HLB 96孔板对血清样品预处理,以超高效液相色谱-静电场轨道离子阱质谱(UHPLC/Q-Orbitrap-MS)进行数据采集,最后采用主成分分析(PCA)、正交偏最小二乘判别分析(OPLS-DA)及变量投影重要性分析(VIP)筛选差异性的原型化合物及代谢产物。共鉴定和推断了20个入血成分,包括11个原型成分及9个代谢产物。研究结果为进一步深入探讨酸枣仁改善失眠的效应物质提供了基础。     

“仙灵骨葆”灌胃给药大鼠后吸收入血成分的HPLC-MS/MS分析

采用高效液相色谱串联质谱法(HPLC-MS/MS),对中药复方制剂"仙灵骨葆"经大鼠灌胃给药后吸收入血的成分进行分析。以甲醇-乙腈(2∶1,体积比)对血清样本进行前处理,采用C18色谱柱,以乙腈-水(各含0.1%甲酸)为流动相进行梯度洗脱,选择正离子采集模式下的一级、二级质谱全扫描,对"仙灵骨葆"给药后血清、空白血清及"仙灵骨葆"制剂进行液相色谱串联质谱对比分析,根据色谱峰保留时间及质谱碎裂规律,结合文献和空白血清加入对照品的相关信息,确认了"仙灵骨葆"13个吸收入血原型成分并推测了2个代谢物,这些入血成份主要来源于淫羊藿、补骨脂、续断和知母四味组方中药,提示这四味中药可能对该复方体内发挥药效的贡献较大。上述研究可为揭示该复方在体内抗骨质疏松药效物质提供一定实验依据。     

新型光敏剂32-(4-甲氧基苯基)-152-天冬氨酸-二氢卟吩e6的药代动力学及组织分布特征

二氢卟吩类衍生物3²-(4-甲氧基苯基)-15²-天冬氨酸-二氢卟吩e6(DYSP-C34)是从程海湖螺旋藻中提取并合成的新型光敏剂。研究DYSP-C34在生物体内的药代动力学及组织分布过程对光动力疗法(PDT)的有效性和安全性至关重要。该文运用高效液相色谱-紫外(HPLC-UV)检测技术,建立了大鼠血浆中DYSP-C34的检测方法。采用沉淀蛋白-液液萃取法处理血浆和组织样品,采用Unitary C₁₈色谱柱(250 mm×4.6 mm, 5 μm)分离,流动相为甲醇-5 mmol/L四丁基磷酸氢铵缓冲盐溶液(70∶30, v/v),流速为1.0 mL/min,进样量为20 μL,检测波长为400 nm,柱温为40 ℃。实验结果表明,大鼠血浆药物质量浓度在1~200 μg/mL范围内线性良好,判定系数(r²)为0.9941。在低、中、高(8、40、120 μg/mL)3个添加水平下的提取回收率分别为74.39%、69.71%和65.89%,日内和日间相对标准偏差(RSD)均在5%以内。运用此方法测定静脉注射DYSP-C34(16 mg/kg)后大鼠血浆中以及荷瘤小鼠组织中的药物浓度,采用DAS 2.0计算出药物半衰期t_1/2z为6.98 h,药-时曲线下面积AUC_(0-∞)为1025.01 h·mg/L,平均驻留时间MRT_(0-∞)为9.19 h。DYSP-C34在荷瘤小鼠体内的分布结果显示,DYSP-C34可以在肿瘤组织中蓄积,并具有一定的滞留作用。综上,该文建立了大鼠血浆中DYSP-C34的HPLC-UV测定方法,并进行了方法学验证,此方法简便、快速,结果准确。阐明了DYSP-C34在静脉给药方式下大鼠体内药代动力学和荷瘤小鼠组织中的分布特征,对临床合理用药和药效学研究具有重要意义。     

直链淀粉-三[(S)-1-苯乙基氨基甲酸酯]手性固定相拆分布地奈德对映体及其制剂含量的测定

22R-布地奈德的药物活性比22S-布地奈德的强2~3倍,开发布地奈德对映体拆分和定量分析方法,可为其药物研发及质量控制提供重要依据。目前,主要以反相C₁₈固定相对布地奈德对映体进行拆分,而采用手性固定相对其进行拆分少有报道。通过考察固定相、流动相和柱温对布地奈德对映体拆分的影响,建立了基于直链淀粉-三[(S)-1-苯乙基氨基甲酸酯]手性固定相快速拆分和检测布地奈德对映体的高效液相色谱方法,其色谱条件如下:色谱柱为Chiralpak AS-RH色谱柱(150 mm×4.6 mm, 5.0 μm),流动相为乙腈-水(45∶55, v/v),柱温40 ℃,流速1.0 mL/min,二极管阵列检测器(DAD),检测波长246 nm,进样量10 μL。在该色谱条件下,布地奈德的两个对映体得到较好拆分,22R-布地奈德和22S-布地奈德的保留时间分别6.40 min和7.77 min,分离度为4.64; 22R-布地奈德和22S-布地奈德分别在各自范围内线性关系良好,相关系数(R²)均为0.9999,检出限分别为0.05 μg/mL和0.07 μg/mL,定量限分别为0.16 μg/mL和0.20 μg/mL; 4个添加水平的样品加标回收率为102.63%~104.17%,相对标准偏差(RSD)为0.08%~0.57%(n=6)。将该方法应用于1批次4个吸入用布地奈德混悬液实际样品进行检测,22R-布地奈德和22S-布地奈德的含量分别为283.15~284.63 μg/mL和259.86~261.51 μg/mL。该方法操作简便,分析时间短,重复性好,准确度高,可用于布地奈德对映体的拆分及其制剂的质量控制。     

无花果叶中有效成分鉴别及含量测定

近年来,无花果叶片中所含成分分析主要集中在同类型物质或整类物质,而对多种类型单一有效物质的定性定量研究较少。 本文采用溶剂浸提-超声波辅助法对无花果叶片中的有效成分进行了提取,利用气相色谱-质谱和超高效液相色谱-离子阱质谱对有效成分进行了鉴定。 结果表明,无花果叶中主要成分为绿原酸、芦丁、补骨脂素和佛手苷内酯。 采用单因素实验法,确定了4种成分的最佳提取工艺。 利用高效液相色谱建立了绿原酸、芦丁、补骨脂素和佛手苷内酯的定量分析方法。 绿原酸在1.84~9.2 μg范围内有良好的线性关系(R²=0.9991),回收率为95.2%;芦丁在3.4~17 μg范围内有良好的线性关系(R²=0.9981),回收率为96.8%;佛手内脂在4~20 μg范围内有良好的线性关系(R²=0.9998),回收率为89.5%;补骨脂素在4~20 μg范围内有良好的线性关系(R²=0.9963),回收率为87.1%。     

液相色谱-串联质谱法测定葡萄中链霉素和双氢链霉素

研究系统地优化了样品前处理过程及仪器分析中影响链霉素和双氢链霉素残留分析准确度与响应灵敏度的各主要因素,建立了葡萄中链霉素和双氢链霉素残留的快速精准定量分析方法。葡萄样品经磷酸溶液(pH=2)超声提取、Oasis HLB单固相萃取柱富集净化后,采用Waters HSS T3色谱柱(100 mm×2.1 mm, 1.8 μm)分离,柱温35 ℃,进样量2 μL,以0.1%甲酸水溶液-甲醇溶液(60∶40, v/v)为流动相进行等度洗脱,在正离子、电喷雾电离源多反应监测模式下测定,外标法定量。链霉素和双氢链霉素在2~400 μg/L质量浓度范围内线性关系良好,相关系数(R²)分别为0.9991和0.9997;在5、10、20和40 μg/kg 4个添加水平下的平均回收率为76.8%~91.9%,相对标准偏差为0.4%~10.2%;链霉素和双氢链霉素的检出限(LOD)为1 μg/L,定量限(LOQ)为5 μg/kg。为验证该方法的适用性,将方法适用于无籽红提、新郁葡萄、夏黑葡萄等实际样品中进行添加回收实验,链霉素和双氢链霉素的平均回收率分别为77.2%~83.9%和70.8%~78.9%, RSD为3.0%~15.6%。该法的准确度和精密度均符合葡萄中链霉素和双氢链霉素分析要求,且操作简便、准确,灵敏度高,适用于葡萄中链霉素和双氢链霉素残留量的检测分析。     

高通量固相萃取-超高效液相色谱-串联质谱法测定人尿中8种环境酚类内分泌干扰物

建立了人尿中8种环境酚类化合物的96孔板固相萃取-超高效液相色谱-串联质谱(96-well SPE LC-MS/MS)检测方法,其中包括7种双酚类化合物和三氯生。尿样解冻到室温,经β-葡萄糖醛酸苷肽酶/芳基磺酸酯酶37 ℃过夜酶解。实验比较了3种96孔板固相萃取柱和不同淋洗条件对人尿样的净化效果和目标化合物的回收率。结果显示,采用Oasis HLB 96孔板(60 mg)对样品进行萃取和用30%(v/v)乙腈水溶液进行淋洗净化的纯化效果最好。纯化后目标物用甲醇溶液洗脱,经氮气吹干,用0.5 mL甲醇-水(1∶1, v/v)溶液定容,目标化合物用UPLC-MS/MS进行检测。比较了2种分析柱(C₁₈和T3分析柱)以及不同的有机流动相对分离样品中目标物的影响。结果显示,以BEH C₁₈(100 mm×2.1 mm, 1.7 μm)作为分析柱,乙腈/水作为流动相,以流速0.3 mL/min梯度洗脱时,目标物的分离效果最好。质谱条件选择串联质谱负离子电喷雾(ESI^-)多反应监测模式(MRM)进行检测。对样品的基质效应进行评估发现,双酚A、双酚F、双酚S、双酚B和双酚AF的绝对基质效应为3.47%~15.32%,不需要补偿措施;四氯双酚A、四溴双酚A和三氯生的绝对基质效应分别是49.58%(中等基质效应)、71.99%和86.93%(强基质效应),均需要补偿效应。因此,该方法采用了一一对应的同位素内标法抵消基质效应。用6份实际尿样基质评估相对基质效应,8种内标的峰面积的相对标准偏差为3.63%~9.06%,说明相对基质效应稳定。在优化条件下,双酚A和双酚AF在0.50~50 μg/L内、四氯双酚A和双酚S在0.05~50 μg/L内、双酚F和四溴双酚A在0.01~50 μg/L内、双酚B在1.00~50 μg/L内、三氯生在5.00~200 μg/L内线性关系良好,相关系数大于0.9995。方法检出限为0.002~1.09 μg/L,定量限为0.007~3.63 μg/L。3个加标水平的加标回收率为81.0%~101.9%,日内精密度为0.4%~19.4%,日间精密度为2.5%~17.8%。应用该方法对2019-2020年采集的北京地区64份尿样进行测定,结果发现8种目标分析物中,除双酚B和双酚AF未检出外,其余均有检出,其中双酚A和双酚S的检出率最高,分别为100%和96.9%。三氯生、四溴双酚A、四氯双酚A和双酚F的检出率分别为57.8%、46.9%、23.4%和21.9%。尿样中8种目标物含量的中位值以降序排列分别为1.44 μg/L(三氯生)、0.69 μg/L(双酚A)、0.086 μg/L(双酚S)、0.0032 μg/L(四溴双酚A)、0.00050 μg/L (四氯双酚A)、0.00 μg/L(双酚F、双酚B和双酚AF)。以上尿样检测结果显示,北京市居民存在普遍的环境酚类化合物暴露,值得关注。该方法操作简单,定量准确,样品需求量小,有机试剂消耗少,适合大批量样本的测定。     

冷诱导液液萃取-分散固相萃取净化-超高效液相色谱-串联质谱法同时测定鸡蛋中白僵菌素和4种恩镰孢菌素残留

新型生物毒素白僵菌素(BEA)和恩镰孢菌素(ENNs)是由镰刀菌种产生的有毒代谢产物,主要污染谷物及其制品,会威胁人类健康,因此受到人们越来越多的关注。该工作建立了冷诱导液液萃取-分散固相萃取净化-超高效液相色谱-串联质谱法(CI-LLE-DSPE-UPLC-MS/MS)同时测定鸡蛋中白僵菌素和4种恩镰孢菌素残留的分析方法。以乙腈-水-乙酸(79∶20∶1, v/v/v)为提取溶剂,采用冷诱导液液萃取与分散固相萃取净化相结合的方法进行样品处理,同时,对影响待测物提取与净化效率的提取溶剂、冷冻萃取温度与时间、净化剂用量等因素和色谱条件进行了优化。样品经20 mL提取液涡旋提取20 min,放入-40 ℃冰箱静置30 min后,取2 mL上层溶液经70 mg C18粉末净化,离心,上清液于40 ℃浓缩至近干,残留物用1 mL 80%(v/v)乙腈水溶液溶解,进样分析。以乙腈与5 mmol/L甲酸铵溶液作为流动相进行梯度洗脱,经ACQUITY UPLC BEH C18色谱柱(100 mm×2.1 mm, 1.7 μm)分离,采用ESI⁺电离,在多反应监测模式下采集,白僵菌素采用稳定同位素内标法定量,4种恩镰孢菌素采用基质匹配曲线外标法定量。结果表明,5种待测物在0.1~50.0 μg/L范围内线性关系良好,相关系数(r²)为0.9983~0.9997,该方法的检出限(LOD)为0.05~0.15 μg/kg,定量限(LOQ)为0.20~0.50 μg/kg。以阴性鸡蛋样品为基质,在低、中、高3个浓度水平(0.5、5.0、25.0 μg/kg)下进行加标试验考察方法的准确度与精密度,各待测物的平均回收率为81.1%~106%,相对标准偏差(RSD)为0.27%~9.79%。采用所建立的方法对农村散养鸡蛋与市售鸡蛋进行检测,结果表明,BEA在散养鸡蛋的检出率为30.4%, 4种ENNs均未被检出。该方法灵敏度高,稳定性好,回收率高,定量准确,简单易操作,适用于禽蛋食品中白僵菌素与恩镰孢菌素的同时快速测定。     

高效液相色谱-串联质谱法检测婴幼儿血清中4种双酚类环境激素残留

目前,双酚类化合物是重要的工业原料,常用来制造塑料(奶)瓶、幼儿用吸口杯、食品和饮料(奶粉)罐内侧涂层,其具有类似雌激素的作用,摄入低剂量的双酚类物质便会引起机体尤其婴幼儿体内激素水平的调节。建立了一种同时测定婴幼儿血清中双酚A(BPA)、双酚B(BPB)、双酚F(BPF)、双酚S (BPS)4种双酚类环境激素的高效液相色谱-串联质谱法(HPLC-MS/MS)。以甲基叔丁基醚(MTBE)为提取溶剂,采用液液萃取的方法进行样品处理,同时对影响4种双酚类环境激素提取效率的提取溶剂、提取时间、提取溶剂体积等影响因素进行了优化。100 μL血清样本在40 ℃下经400 μL MTBE提取15 min后,采用Waters ACQUITY UPLC BEH C18柱(50 mm×2.1 mm, 1.7 μm)进行分离,以超纯水和含0.5 mmol/L乙酸铵的甲醇溶液为流动相进行梯度洗脱,流速为0.2 mL/min,采用电喷雾电离、负离子模式扫描,在多反应监测(MRM)模式下测定。BPA、BPB、BPS在0.25~100 μg/L范围内线性关系良好,BPF在1~00 μg/L范围内线性关系良好,相关系数为0.9929~0.9959;方法的检出限分别为0.05、0.05、0.05、0.5 μg/L;在3个添加水平(5、20、100 μg/L)下,4种目标化合物的回收率为84.56%~104.43%,相对标准偏差小于10%。应用该方法对150例婴幼儿血清样品进行检测,结果表明:BPA、BPF、BPS在男童和女童血清中的检出率分别为90.67%和89.33%、6.67%和1.33%、5.33%和16.00%, BPB未被检出。该方法操作简单,回收率好,精密度高,适用于婴幼儿血清中4种双酚类环境激素的同时测定。     

混合型离子交换液相色谱-串联质谱法检测鸡蛋中10种氨基糖苷类药物残留

该研究系统地优化了样品前处理过程及仪器分析中影响氨基糖苷残留分析准确度与灵敏度的各主要因素,建立了鸡蛋中10种氨基糖苷类药物(链霉素、双氢链霉素、潮霉素B、卡那霉素、阿米卡星、妥布霉素、安普霉素、大观霉素、新霉素、庆大霉素)残留量的混合型离子交换液相色谱-串联质谱分析方法。样品经10 mmol/L乙酸铵缓冲溶液(含0.4 mmol/L EDTA和50 g/L三氯乙酸)超声提取,调节pH至6~7后,经PRiME HLB固相萃取柱富集净化,采用SIELC Obelisc R色谱柱分离,以乙腈和1.0%(v/v)甲酸水溶液(含1 mmol/L甲酸铵)为流动相进行梯度洗脱,在正离子、多反应监测模式下经串联质谱仪测定,外标法定量。该方法在5~200 μg/L质量浓度范围内线性关系良好,相关系数(r²)均大于0.99;方法的检出限(LOD, S/N≥3)为2~5 μg/kg,定量限(LOQ, S/N≥10)为5~10 μg/kg。在空白鸡蛋中进行LOQ、20 μg/kg、100 μg/kg 3个水平的加标回收实验,方法的平均回收率(n=6)为68.1%~111.3%,相对标准偏差为1.2%~12.3%。利用该方法对市售的20批次鸡蛋样品进行测定,均未检出目标物。本方法简单、灵敏、准确,可实现鸡蛋中10种氨基糖苷类药物残留的批量检测。     

基于非靶向代谢组学的茄梨和红茄梨成熟期果皮代谢产物的差异分析

为了探究西洋梨品种茄梨及其红色芽变红茄梨成熟期果皮代谢产物差异,采用超高效液相色谱-质谱联用技术,对茄梨和红茄梨成熟期果皮进行非靶向代谢组学研究。通过主成分分析和正交偏最小二乘判别分析,构建了多变量统计分析模型,结合模型和变量重要性投影与最大差异倍数值,基于精确质量数、二级碎片以及同位素分布,使用PMDB(Plant Metabolome Database)数据库进行定性,筛选并鉴定出茄梨和红茄梨果皮中显著性变化(P<0.05, VIP(variable importance in project)≥1)的差异代谢物有83种,主要包括酚酸类、黄酮类和氨基酸类物质,涉及类黄酮代谢、氨基酸代谢、苯丙烷类代谢等代谢途径,其中53种物质含量上调,30种物质表达下调。通过KEGG(Kyoto Encyclopedia of Genes and Genomes)数据库进一步对差异代谢物质进行通路富集分析,差异代谢物主要分布在20条代谢途径中,P<0.05的代谢途径有6条,分别是类黄酮生物合成、黄酮和黄酮醇生物合成、苯丙烷生物合成、丁酸酯代谢、苯丙氨酸代谢、酪氨酸代谢。这些差异代谢物的变化可能是导致茄梨和红茄梨果皮色泽不同的原因。该研究从植物代谢组学角度初步揭示了茄梨和红茄梨成熟期果皮的代谢产物差异性。     

Effect of wine processing and acute blood stasis on the serum pharmacochemistry of rhubarb: A possible explanation for processing mechanism

As a specific item mentioned in traditional Chinese medicine theory, processing can fulfill different requirements of therapies. Crude and wine‐processed rhubarbs are used as drastic and mild laxatives, respectively. In this study, a practical method based on ultra‐fast liquid chromatography coupled with diode‐array detection and ion trap time‐of‐flight mass spectrometry was developed to screen and analyze multiple absorbed bioactive components and metabolites in the serum of both normal and acute blood stasis rats after oral administration of crude or wine‐processed rhubarbs. A total of 16 compounds, mainly including phase II metabolites, were tentatively identified. Possible explanations for the processing‐induced changes in pharmacological effects of traditional Chinese medicines were first explored at serum pharmacochemistry level.     

Screening and analysis of the multiple absorbed bioactive components and metabolites in rat plasma after oral administration of Jitai tablets by high‐performance liquid chromatography/diode‐array detection coupled with electrospray ionization tandem mass spectrometry

Based on the serum pharmacochemistry technique and high‐performance liquid chromatography/diode‐array detection (HPLC/DAD) coupled with electrospray tandem mass spectrometry (HPLC/ESI‐MS/MS), a method for screening and analysis of the multiple absorbed bioactive components and metabolites of Jitai tablets (JTT) in orally dosed rat plasma was developed. Plasma was treated by methanol precipitation prior to liquid chromatography, and the separation was carried out on a Symmetry C₁₈ column, with a linear gradient (0.1% formic acid/water/acetonitrile). Mass spectra were acquired in negative and positive ion modes, respectively. As a result, 26 bioactive components originated from JTT and 5 metabolites were tentatively identified in orally dosed rat plasma by comparing their retention times and MS spectra with those of authentic standards and literature data. It is concluded that an effective and reliable analytical method was set up for screening the bioactive components of Chinese herbal medicine, which provided a meaningful basis for further pharmacology and active mechanism research of JTT. Copyright © 2010 John Wiley & Sons, Ltd.     

Screening out Potential Cardio-Toxic Components of Chinese Herb Radix Aconiti Lateralis in Rat Dosed Plasma by High Performance Liquid Chromatography/Electrospray Ionization Quadrupole Time-Of-Flight Mass Spectrometry

This study established a new, rapid, and accurate method for bioactive compound screening by the combination of serum pharmacochemistry and serum pharmacology based on a spectrum-effect relationship model. HPLC/ESI-Q-TOF-MS was used for the chemical profiling of samples of dosed plasma, control plasma, and Radix Aconiti Lateralis decoction. A comparison was made of the resulting chemical profiles, which totaled 20 compounds including 16 prototype compounds and 4 metabolites derived from SF decoction that were detected as potential bioactive components. Using MS/MS analysis and accurate molecular weight assessments of TOF, 17 components in 20 potential bioactive compounds were structurally identified. Moreover, the pretreated plasma samples were injected into venous sinus of isolated toad hearts to investigate the cardio-toxicity according to heart ratio (HR) situation and electrocardiograph (ECG) parameters, including PR interval, QRS interval, and QT interval. Finally, 4 compounds in these 20 potential bioactive components that had closest correlation with 4 cardio-toxic experimental indexes respectively were investigated by spectrum-effect relationship model for the first time. The 4 compounds were considered as the real cardio-toxic components. This study proposed a series of potential bioactive components in Radix Aconiti Lateralis and provided a general research pattern to identify the potential bioactivity compounds in Chinese herbs or natural pharmacy.     

32种氧化型染料的高效液相色谱定量及高效液相色谱-串联质谱确证方法北大核心CSCD

染发类产品中氧化型染料种类多,实际样品测定时干扰多,建立染发类产品中多种常用染料的测定方法,为该类产品的有效监管提供技术手段十分必要。该研究根据染料使用频率分组,采用能够屏蔽硅羟基和金属离子影响的C_(18)柱,优化了《化妆品安全技术规范》(2015年版)中32种染料的高效液相色谱法(HPLC)并建立了高效液相色谱-串联质谱(HPLC-MS/MS)确证方法。样品以10 g/L亚硫酸氢钠水溶液为抗氧化剂,用无水乙醇-水(1∶1,v/v)混合溶液冰浴超声提取10 min。HPLC方法采用甲醇、乙腈和磷酸盐缓冲液为流动相分两个液相色谱条件进行梯度洗脱分离,于280 nm波长下检测,其中一个HPLC条件中的相互干扰组分均在另一个HPLC条件下完全分离,避免了实际样品检测时组分间的干扰,并排除了32种以外的其他15种常用染料的干扰。HPLC-MS/MS方法分别采用5 mmol/L乙酸铵水溶液-乙腈和5 mmol/L乙酸水溶液-乙腈为正离子和负离子模式下的流动相,电喷雾离子模式下用多反应监测(MRM)模式进行定性和定量分析。HPLC和HPLC-MS/MS两个方法中,日内精密度和48 h内稳定性的相对标准偏差(RSD)<10%,回收率为82.6%~114.9%(RSD<10%)。HPLC方法中32种染料在大约10~500 mg/L范围内线性关系良好(r^(2)>0.99),检出限为9.7~40.1μg/g;HPLC-MS/MS方法中氢醌线性范围为2.0~79.7 mg/L,检出限为8.0μg/g,其他组分线性范围约为0.1~4 mg/L,检出限为0.01~0.4μg/g。采用HPLC、HPLC-MS/MS两个方法和《化妆品安全技术规范》方法同时测定实际样品,共检出16种染料,检出含量范围为58~25160μg/g。3个方法检测结果的RSD为1.9%~10.1%。该研究增加了HPLC-MS/MS确证方法,适应化妆品法定检验中的未知物确认程序;方法简便快速,结果准确,专属性强,具有较好的通用性和可操作性。     

Plasma pharmacochemistry combined with pharmacokinetics and pattern recognition analysis to screen potentially bioactive components from Daming capsule using ultra high performance liquid chromatography with electrospray ionization quadrupole time‐of‐flight mass spectrometry

Daming capsule is a traditional Chinese medicine for hyperlipidemia treatment. However, the vague understanding of the bioactive components of Daming capsule hampers its modernization and internationalization. This work first developed a high‐throughput, high‐resolution, and high‐sensitivity ultra high performance liquid chromatography with electrospray ionization quadrupole time‐of‐flight tandem mass spectrometry method for identifying the absorbed compounds and monitoring the pharmacokinetics of Daming capsule. A high‐throughput strategy integrating plasma pharmacochemistry, pharmacokinetics, and pattern recognition analysis was also established for screening the bioactive components of Daming capsule in vivo. The established strategy based on ultra high performance liquid chromatography with electrospray ionization quadrupole time‐of‐flight tandem mass spectrometry was successfully applied to screen the bioactive components of Daming capsule. Up to 53 absorbed compounds were identified. Six anthraquinones with fast and high absorption, namely, emodin‐O‐glucoside, aurantio‐obtusin, aloe‐emodin, rhein, emodin, and chrysophanol, were screened as potentially bioactive components of Daming capsule. The plasma pharmacochemistry and pharmacokinetics of Daming capsule were reported for the first time. Notably, the high‐throughput and reliable strategy facilitated the screening and identification of bioactive components of traditional Chinese medicine, thereby providing novel insights into the research and development of new drugs.     

Analysis of plasma migration components in Patrinia villosa (Thunb.) Juss. effective parts by UPLC–Q-TOF–MS

Patrinia villosa (Thunb.) Juss. (PVJ) is described as pungent, bitter and slightly cold in Chinese medicine, and is associated with the large intestine, stomach and liver meridians. The preliminary experiments of our research team proved that PVJ total flavonoids have excellent inhibitory effects on liver cancer cells. The present experiment uses the UPLC–Q-TOF–MS technology and serum pharmacochemistry methods to analyze the chemical components in vitro and in vivo of PVJ antiliver tumors. A total of 14 chemical components were identified in the total flavonoids extract of PVJ, and it is mainly composed of flavonoids, flavonones, flavonols and phenolic acids. At the same time, seven prototypical components and seven metabolic components were detected in the drug-containing plasma. Hydrocaffeate and scutellarein are the phase I metabolites of caffeic acid and scutellarin, respectively. Sulfated apigenin, sulfated luteolin, sulfated kaempferol and methylated kaempferol are the II phase metabolites of apigenin, luteolin, kaempferol, respectively. The experiment provides a reference for the research and development of antitumor drug candidates, and provides a basis for revealing the bioactive components of PVJ and the antitumor mechanism.     

Identification of the absorbed components and their metabolites of Tianma‐Gouteng granule in rat plasma and bile using ultra‐high‐performance liquid chromatography combined with quadrupole time‐of‐flight mass spectrometry

Tianma‐Gouteng granule (TGG), a Chinese herbal formula preparation, is clinically used for the treatment of cardio‐cerebrovascular diseases such as hypertension, cerebral ischaemia, acute ischaemic stroke and Parkinson's disease. Although few reports have been published concerning the absorbed prototype components of TGG, the possible metabolic pathways of TGG in vivo remain largely unclear. In this study, a method using UPLC–Q/TOF MS was established for the detection and identification of the absorbed prototype components and related metabolites in rat plasma and bile after oral administration of TGG at high and normal clinical dosages. A total of 68 components were identified or tentatively identified in plasma and bile samples, including absorbed prototypes and their metabolites. The major absorbed components were gastrodin, isorhynchophylline, rhynchophylline, isocorynoxeine, corynoxeine, geissoschizine methyl ether baicalin, baicalein, wogonoside, wogonin, geniposidic acid, leonurine, 2,3,5,4′‐tetrahydroxystilbene‐2‐O‐β‐d ‐glucoside and emodin. The main metabolic pathways of these components involved phase I (isomerization, hydrolysis and reduction) and phase II (glucuronidation and sulfation) reaction, and the phase II biotransformation pathway was predominant. The present study provides rich information on the in vivo absorption and metabolism of TGG, and the results will be helpful for further studies on the pharmacokinetics and pharmacodynamics of TGG.     

Comprehensive investigation of mechanism and effective ingredients of Fangji Huangqi Tang by serum pharmacochemistry and network pharmacology

Fangji Huangqi Tang (FHT), has been reported to show effects on nephrotic syndrome, but its mechanism of action and bioactive components have not yet been determined. In this study, a method using UPLC–HRMS/MS was established for the detection and identification of the chemical constituents and metabolites absorbed into the blood. Absorbed components in serum were then used for the network pharmacology analysis to deduce the mechanism and effective components. A total of 86 compounds were identified or tentatively characterized. Based on the same instrumental conditions, 85 compounds were found in rat serum after oral administration of FHT, including 22 prototypes and 63 metabolites. Network pharmacology analysis showed that absorbed components, such as (3R)-2′,3′,4′,7-tetrahydroxyisoflavan, astrapterocarpan, cycloastragenol, 7,2′-dihydroxy-3′,4′-dimethoxyisoflavan, astragaloside IV, astrapterocarpan glucoside and glycyrrhetinic acid, could be responsible for the pharmacological activity of nephrotic syndrome by regulating the VEGF signaling pathway, focal adhesion and MAPK signaling pathway. Furthermore, the pathway-target network showed that the MAPK1, AKT2 and CDC42 were involved in the signal pathways above. This study provides a scientific basis for the mechanism and effective ingredients of FHT.     

Strategies for determining the bioactive ingredients of honey‐processed Astragalus by serum pharmacochemistry integrated with multivariate statistical analysis

Honey‐processed Astragalus is a widely used traditional Chinese medicine that has a better effect on reinforcing “Qi” (vital energy) than the raw one. A comparative study of metabolites analysis between them in rat serum for finding the bioactive ingredients was carried out using serum pharmacochemistry and multivariate statistical analysis. The blood collection methods and time were optimized first. Then the prototypes and metabolites in serum samples after oral administration were investigated by ultra‐high‐performance liquid chromatography coupled with electrospray ionization quadrupole time‐of‐flight mass spectrometry integrated with principal component analysis and orthogonal partial least squares discriminant analysis. The contents of metabolites were also analyzed to evaluate the metabolic profile differences. As a result, nine prototypes and 36 metabolites were identified. Only two prototypes and 15 metabolites were different between raw and honey‐processed Astragalus. Their biotransformation reactions contained the process of oxidation, demethylation, and hydrolysis in phase I and glucuronide conjugation or sulfate conjugation in phase II. Most of the detected metabolites were transformed from isoflavones and isoflavanes. Our results expand the knowledge about the influence of honey‐processing on Astragalus and suggest the different curative effects between raw and honey‐processed Astragalus might due to their therapeutic material discrepancy.