液相色谱-串联质谱法测定柏子仁中黄曲霉毒素残留量

目的建立检测柏子仁中黄曲霉毒素B1,B2,G1,G2残留量的液相色谱-串联三重四极杆质谱法。方法将样品提取液经免疫亲和柱净化,甲醇洗脱,以甲醇/乙腈-0.1 mmol乙酸铵为流动相,经反相色谱柱Agilent Zorbax Extend-C18柱(250 mm×4.6 mm,5μm),用电喷雾正离子二级离子监测扫描模式(MRM)检测。结果黄曲霉毒素G2和B2质量浓度在0.03～3.2 ng/mL范围内、黄曲霉毒素B1和G1在0.1～10 ng/mL范围内与峰面积积分值线性关系良好。检测限(LOD)为0.1 ng/mL,定量限(LOQ)为0.2 ng/mL。3个质量浓度(0.25,1.0,5.0 ng/mL)的加标平均回收率为84.1%～96.8%,RSD为7.7%～13.4%。结论所用方法专属性强、灵敏度高、简便快速、结果准确,可定性定量测定柏子仁中残留的黄曲霉毒素。     

UPLC-MS/MS法与ELISA法测定胖大海中黄曲霉毒素的比较研究

目的 采用超高效液相色谱-串联质谱法(UPLC-MS/MS)和酶联免疫吸附法(ELISA)对胖大海中的黄曲霉毒素进行检测,比较 两种方法的检测结果以及方法学差异性。方法UPLC-MS/MS法测定：样品采用70%甲醇提取经过免疫亲和柱净化,色谱柱为C₁₈(100 mm×2.1 mm,1.7μm),柱温为40℃,流动相为水含0.1%甲酸(A)-甲醇含0.1%甲酸(B),梯度洗脱,流速为0.3 m L·min^-1,多反应检测扫描采集模式,正负离子同时扫描进行数据采集。同时采用ELISA法测定,样品采用甲醇提取,酶联免疫试剂盒反应。结果 UPLC-MS/MS法测定：黄曲霉毒素B₁、B₂、G₁、G₂线性关系良好,检测限分别为0.013 5,0.002 9,0.008 6,0.003 6μg·kg^-1,定量限分别为0.045 0,0.009 8,0.028 6,0.011 9μg·kg^-1,平均回收率分别为98.98%,10...     

UPLC-MS/MS检测浙产延胡索中4种黄曲霉毒素

目的 建立延胡索中黄曲霉毒素（aflatoxin,AF）超高效液相色谱-三重四级杆质谱检测方法。方法 以Agilent Zorbax Eclipse C₁₈（2.1 mm×50 mm,1.8 μm）色谱柱分离,在电喷雾离子源（ESI）正离子模式下电离,多反应监测（MRM）模式下测定。结果 样品用70%甲醇提取,检测20批延胡索中的AFB1、AFB2、AFG1、AFG2。4种AF质谱检测的线性范围宽,相关性好,r ≥ 0.997;回收率78.3%~93.7%;重复性RSD为1.5%~2.7%（n=6）。结论 该方法专属性强,灵敏度高,重复性好,适用于延胡索中AF的检测,鉴于市场上延胡索饮片的AF风险较高,建议中国药典增加延胡索的AF检测。     

尿中黄曲霉毒素代谢产物的高效液相色谱测定法

黄曲霉毒素 (ＡＦ)是一类毒性和致癌性很强的化合物 ,为第一类人类致癌物[1] ,是人类原发性肝癌的主要致病因素之一[2～ 4 ] ,其中毒性和致癌性最强的为ＡＦＢ1,进入机体后除形成ＡＦＭ1、ＡＦＰ1等外 ,也可被活化为ＡＦＢ1 8,9 环氧化物 ,攻击ＤＮＡ形成ＡＦＢ1 Ｎ7 鸟苷 (ＡＦＢ Ｎ7 ＧＵＡ) ,可经尿液排出 ,攻击蛋白质形成ＡＦＢ1 白蛋白加合物而残留于血液中[5] 。机体中ＩＩ相代谢酶如谷胱甘肽转硫酶 (ＧＳＴｓ)可使谷胱甘肽与ＡＦＢ1 8,9 环氧化物结合 ,终以ＡＦＢ 硫醇尿酸 (ＡＦＢ -ＮＡＣ)的形式经尿排出[6] ,许多化学物质可影响机…     

超高效液相色谱串联三重四极杆质谱法同时测定舒眠胶囊（片）中4种黄曲霉毒素含量

目的 建立同时测定舒眠胶囊（片）中黄曲霉毒素B1,B2,G1,G2(AFB1,AFB2,AFG1,AFG2)含量的超高效液相色谱串联三重四极杆质谱（UPLC-QQQ/MS）法。方法 以3个市售厂家的舒眠胶囊和舒眠片各6批为样品。色谱柱为Agilent ZORBAX Eclipse Plus C18柱（150 mm×2.1 mm,2.6μm）,流动相为9.0 mmol/L乙酸铵溶液（A）-甲醇-乙腈（1∶1,V/V,B）（梯度洗脱）,流速为0.3 mL/min,柱温为28℃,进样量为2μL;采用电喷雾离子源（ESI+）,多反应监测模式（MRM）。结果 AFB1,AFB2,AFG1,AFG2进样量分别在0.372～37.20 ng(r=0.999 3)、0.132～13.20 ng(r=0.999 1)、0.408～40.80 ng(r=0.999 0)、0.140～14.00 ng(r=0.999 2)范围内与峰面积线性关系良好,检测限分别为1.86,0.66,2.04,0.70 pg/mL,定量限分别为5.44,2.19,6.01,2.24 pg/mL;精密度、重复性、稳定性试验结果的...     

决明子中黄曲霉毒素测定方法的优化

目的:优化决明子中黄曲霉毒素测定方法。方法:采用中普红ODS-H C₁₈色谱柱(250 m×4.6 mm, 5μm),以[甲醇∶乙腈(40∶18)]-水(38∶62)为流动相,流速：1.0 ml·min^-1,用1%吐温80溶液洗脱,激发波长λex=360 nm,发射波长λem=450 nm,光化学衍生法,柱温30℃,进样量20μl。并用HPLC-MS/MS进行验证。结果:有效去除决明子中假阳性的干扰成分,黄曲霉毒素B₁、B₂、G₁、G₂四个成分r值分别为0.999 7,0.999 9,0.999 9,0.999 9,线性范围分别为：0.006 48～0.054 0 ng、0.001 98～0.016 5 ng、0.006 06～0.050 5 ng、0.001 80～0.015 0 ng,平均回收率分别为92.31%,91.45%,93.27%,93.34%(RSD分别为0.92%,1.15%,1.08%,0.78%,n=9)。液质与液相测定结果一致。结论...     

液-质串联法测定软胶囊中药用油辅料内黄曲霉毒素B1的含量

摘 要 目的： 尝试利用液相色谱 串联质谱法对软胶囊中药用油辅料内黄曲霉毒素B1含量进行测定。方法: 以甲醇-0.1%甲酸水溶液为溶剂提取软胶囊中花生油所含黄曲霉毒素B 1,离心后取上清液过中性氧化铝固相萃取小柱净化,浓缩后进样测定,以甲醇-0.1%甲酸流动相梯度洗脱,流速为0.3 ml·min^-1,柱温：30℃,进样量：25 μl。采用电喷雾离子化四极杆串联质谱,多反应监测方式测定样品的浓度。结果: 黄曲霉毒素B1在0.098～1.960 μg·L^-1范围内线性关系良好（r=0.999 5）,检出限为0.05 μg·L^-1,平均加样回收率为97.73%,RSD=4.625％(n= 6)。结论:此法灵敏准确,专属性强,干扰少,重现性佳,对含油药物制剂中黄曲霉毒素B1含量检测有参考意义。     

HPLC法测定虫草发酵粉中黄曲霉毒素残留量

目的:建立虫草发酵粉中黄曲霉毒素B1、B2、G1、G2的残留量测定方法。方法:样品经有机溶剂提取、免疫亲和柱净化后,利用高效液相色谱-光化学衍生-荧光检测器进行分析测定。结果:黄曲霉毒素G2、B2在1.5～150 pg范围内线性关系良好,黄曲霉毒素G1、B1在5～500 pg范围内线性关系良好,r>0.9998。回收率在80%～93%之间。结论:该方法快速简便,准确,可作为虫草发酵粉黄曲霉毒素的残留量测定方法,亦可作为原料药的黄曲霉毒素筛查方法,以达到控制产品质量的目的。     

基于HPLC研究柏子仁中黄曲霉毒素的污染状况与风险分析

目的 基于HPLC研究柏子仁中4种黄曲霉毒素的污染情况,并进行风险分析,评估柏子仁的用药安全。方法 采用Agilent Eclipse Plus C₁₈(4.6×250 mm 5μm)为色谱柱,柱后光化学衍生法检测,以甲醇∶乙腈∶水(35∶10∶55)为流动相,流速1.0 mL·min^-1;柱温：40℃;荧光检测器检测。结果 黄曲霉毒素B₁、B₂、G₁、G₂分别在0.35～20.8μg·L^-1、0.13～7.6μg·L^-1、0.36～21.6μg·L^-1、0.13～7.6μg·L^-1范围内呈良好的线性关系,黄曲霉毒素B₁、B₂、G₁、G₂平均回收率分别为90.6%、83.46%、87.84%、86.58%,相对偏差分别为2.9%、3.6%、3.6%、4.7%。23批柏子仁中有14批检...     

HPLC法测定中药材覆盆子中黄曲霉毒素的含量及安全性评价

目的 测定覆盆子中黄曲霉毒素含量并对其进行安全性评价。方法 覆盆子经取样、粉碎、过筛、70%甲醇匀浆提取、稀释、离心、免疫亲和柱富集、除杂、甲醇洗脱制样,使用C18色谱柱分离,光化学衍生器柱后衍生,并由荧光检测器测定结果。结果 方法线性良好（r>0.999）,平均回收率为93.8%～97.7%,RSD≤1.6%。结论 中药材覆盆子被黄曲霉毒素污染的风险低,安全性较好。     

黑豆、半成品和成品淡豆豉中6种异黄酮含量变化比较

目的 比较发酵前的黑豆及半成品和成品淡豆豉中大豆苷、黄豆黄苷、染料木苷、大豆苷元、黄豆黄素和染料木素6种异黄酮的含量变化。方法 采用高效液相色谱法测定异黄酮的含量,色谱条件为DiamonsilC₁₈色谱柱(4.6×250 mm,5μm),柱温30℃,检测波长为260 nm,流动相为0.2%醋酸水(A)-甲醇(B),梯度洗脱,流速为1.0 ml/min。结果 该方法测定6种异黄酮成分在测定范围内线性良好(r≥0.999 3),且回收率符合要求,黑豆经发酵后其中的6种异黄酮含量明显提升,且成品淡豆豉比半成品淡豆豉中含量更多。结论 本研究建立的高效液相色谱法能够准确测定淡豆豉中6种异黄酮的含量。发酵可提升黑豆中的异黄酮的含量,且在发酵过程中结合型异黄酮向游离型异黄酮苷元转化。     

Development of a Novel UPLC-MS/MS Method for the Simultaneous Determination of 16 Mycotoxins in Different Tea Categories

The contamination of potential mycotoxins in tea production and consumption has always been a concern. However, the risk monitoring on multiple mycotoxins remains a challenge by existing methods due to the high cost and complex operation in tea matrices. This research has developed a simple ultra-performance liquid chromatography-tandem mass spectrometry strategy based on our homemade purification column, which can be applied in the detections of mycotoxins in complex tea matrices with high-effectively purifying and removing pigment capacity for 16 mycotoxins. The limits of detection and the limits of quantification were in the ranges of 0.015~15.00 and 0.03~30.00 µg·kg⁻¹ for 16 mycotoxins, respectively. Recoveries from mycotoxin-fortified tea samples (0.13~1200 µg·kg⁻¹) in different tea matrices ranged from 61.27 to 118.46%, with their relative standard deviations below 20%. Moreover, this method has been successfully applied to the analysis and investigation of the levels of 16 mycotoxins in major categories of tea and the monitoring of multiple mycotoxins in processed samples of ripened Pu-erh. In conclusion, the proposed strategy is simple, effective, time-saving, and low-cost for the determination of a large number of tea samples.     

肺癌患者血浆中安罗替尼浓度与不良反应的相关性研究

目的:建立超高效液相色谱-串联质谱法(UPLC-MS/MS)测定安罗替尼血药浓度的方法,探讨安罗替尼血药浓度与不良反应的相关性。方法:血浆样品采用乙腈沉淀蛋白进行前处理,以[D5]-氘代安罗替尼为内标;色谱柱:ACQUITY UPLC BEH C₁₈(2.1 mm×50 mm,1.7 μm);流动相:0.01%氨水-甲醇;梯度洗脱,流速0.5 mL·min^-1;柱温40 ℃;进样量0.5 μL;电喷雾离子源,正离子方式,多反应监测模式。对UPLC-MS/MS进行方法学考察,将其用于测定肺癌患者血浆中安罗替尼浓度,收集相关病例的不良反应信息,分析两者之间的相关性。结果:血浆中安罗替尼在5~250 ng·mL^-1内线性关系良好,定量下限为5 ng·mL^-1,标准曲线为y=0.007 2x+0.004 9(r=0.999);批内和批间精密度、提取回收率及稳定性考察均符合要求。共测定了51例患者血浆中安罗替尼的稳态谷浓度,其中8 mg组27例患者的平均质量浓度为(29.08±8.35)ng·mL^-1;12 mg组24例患者的平均质量浓度为(40.79±23.91)ng·mL^-1。发生高血压、蛋白尿、手足综合征或促甲状腺激素升高的患者平均血药浓度均高于未发生相关不良反应患者的平均血药浓度(P＜0.05),相关风险的阈值浓度分别为31.22,23.13,43.15 ng·mL^-1和45.59 ng·mL^-1。结论:该方法准确简便、快速灵敏、重现性好,可用于测定肺癌患者体内安罗替尼的血药浓度。较高的安罗替尼稳态谷浓度会增加患者高血压、蛋白尿、手足综合征和促甲状腺激素升高的发生风险。     

超高效液相串联质谱法同时测定保健食品中10种水溶性维生素

目的：建立超高效液相色谱-串联质谱检测方法同时测定保健食品中10种水溶性维生素。方法：样品经过前处理后,采用Waters ACQUITY UPLC HSS T3（2.1×100 mm,1.8μm）色谱柱分离;以0.1%甲酸的甲醇溶液和0.1%甲酸的水溶液为流动相进行梯度洗脱,通过电喷雾离子源多反应监测（MRM）模式进行检测。结果：10种水溶性维生素在6 min内完成分离,线性关系良好（r≥0.997）,方法检出限在5~250μg·kg^-1之间。3个添加水平的回收率和相对标准偏差RSD（n=6）分别为85.9%~109.5%和1.09%~0.79%。结论：该方法简便、快速、准确、灵敏,适用于保健食品中水溶性维生素的测定。     

Qualitative and quantitative characterization of secondary metabolites and carbohydrates in Bai-Hu-Tang using ultraperformance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry and ultraperformance liquid chromatography coupled with photodiode array detector

Bai-Hu-Tang (BHT), a classic traditional Chinese medicine (TCM) formula used for clearing heat and promoting body fluid, consists of four traditional Chinese medicines, i.e., Gypsum Fibrosum (Shigao), Anemarrhenae Rhizoma (Zhimu), Glycyrrhizae Radix et Rhizoma Praeparata cum Melle (Zhigancao), and nonglutinous rice (Jingmi). The chemical composition of BHT still remains largely elusive thus far. To qualitatively and quantitatively characterize secondary metabolites and carbohydrates in BHT, here a combination of analytical approaches using ultraperformance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry and ultraperformance liquid chromatography coupled with photodiode array detector was developed and validated. A total of 42 secondary metabolites in BHT were tentatively or definitely identified, of which 10 major chemicals were quantified by the extracting ion mode of quadrupole time-of-flight mass spectrometry. Meanwhile, polysaccharides, oligosaccharides, and monosaccharides in BHT were also characterized via sample pretreatment followed by sugar composition analysis. The quantitative results indicated that the determined chemicals accounted for 35.76% of the total extract of BHT, which demonstrated that the study could be instrumental in chemical dissection and quality control of BHT. The research deliverables not only laid the root for further chemical and biological evaluation of BHT, but also provided a comprehensive analytical strategy for chemical characterization of secondary metabolites and carbohydrates in traditional Chinese medicine formulas.     

UHPLC—MS／MS法在医院制剂多成分含量测定中的应用

射干合剂和甘地胶囊是新华医院的两个院内中药复方制剂,本文建立了UHPLC—MS／MS方法同时测定这两个制剂中的麻黄碱、咖啡酸、阿魏酸、芦丁、野黄芩苷、射千苷、黄芩苷、黄芩素、黄芪甲苷、次野鸢尾黄紊、汉黄芩素等11种中药成分的含量。色谱柱采用ZORBAXSB-C18（2．1mm×50mm,1．8μm）,流动相为0．1％甲酸水溶液-乙腈,梯度沈脱,流速为0．3mL／min,柱温35℃;质谱采用电喷雾离子源（ESI）,多反应离子监测（MRM）,并结合正负离子扫描切换,其中咖啡酸、阿魏酸、野黄芩苷、射干苷采用负离子模式检测,麻黄碱、芦丁、黄芩苷、黄芩素、黄芪甲苷、次野鸢尾黄素、汉黄芩素采用正离子模式检测。结果显示麻黄碱、咖啡酸、阿魏酸、芦丁、野黄芩苷、射干苷、黄芩苷、黄芩素、黄芪甲苷、次野鸢尾黄素、汉黄芩素的定量限分别为4．90×10^-3ng／mL、7．80ng／mL、6．8ng／mL、5.3×10^-2ng／mL、4．20×10^-3ng／mL、4．6×10^-2ng／mL、1．44×10^-4ng／mL、4．85ng／mL、0．23ng／mL、3．18×10^-4ng／mL、2．95×10^-4ng／mL,检测限分别为2．90×10^-4ng／mL、0．77ng／mL、2．0ng／mL、0．016ng／mL、1．3×10^-3ng／mL、3．33×10^-4ng／mL、4．32×10^-5ng／mL、1．46ng／mL、0．07ng／mL、9．5×10^-5ng／mL、8．84×10^-5ng／mL。在相应的线性范围内R^2〉0．99;日内和日问精密度（RSD）均小于5％,平均同收率均在80％-120％。本方法在20min内实现这11种目标化合物的分离和测定,简单、快速、灵敏、准确,可用于射干合剂和甘地胶囊的指标成分含量测定,为这两个制剂的质量控制提供依据。     

超高效液相色谱串联三重四极杆质谱法检查脑立清制剂中水麦冬酸成分

目的 建立脑立清制剂中水麦冬酸成分的检测方法。方法 样品经萃取富集后,利用超高效液相色谱-三重四极杆质谱(UPLC-MS/MS),以水麦冬酸为指标成分,对脑立清制剂中常见掺伪虎掌南星情况进行监控。采用Waters ACQUITY UPLC HSS T3 C₁₈(2.1 mm×100 mm, 1.8μm)色谱柱,以乙腈-0.1%甲酸溶液为流动相,梯度洗脱,体积流量0.2 mL·min^-1;柱温30℃,进样量2μL;质谱条件采用ESI^-多反应监测(MRM)模式扫描,选择m/z 187.0→143.3和m/z 187.0→98.9作为检测离子对。结果 以所建立的方法对88批次脑立清制剂进行检测,结果5个厂家的15批样品检查出水麦冬酸成分。结论 所建立的方法准确可靠,专属性强,可用于脑立清制剂中的水麦冬酸成分的检查;同时也为其他含半夏制剂中掺伪虎掌南星的监控提供参考。     

保心宁片(胶囊)中枳壳伪品香圆检查方法的研究

目的：建立保心宁片(胶囊)中枳壳伪品香圆的检查方法,为打击中药掺伪提供技术支撑。方法：采用超高效液相色谱-四极杆飞行时间质谱(UPLC-Q-TOF-MS),Agilent SB C₁₈(100 mm×2.1mm,1.8μm)色谱柱,以乙腈和1%甲酸梯度洗脱,流速0.4 m L·min^-1,柱温30℃;电喷雾离子化(ESI)正离子模式进行检测。采用多元分析统计软件筛选查找枳壳常见伪品香圆的特征性成分水合氧化前胡素,并应用超高效液相色谱-串联三重四极杆质谱(UPLC-QQQ-MS)建立保心宁片(胶囊)中香圆的检查方法。结果：水合氧化前胡素在0.17～1.02 ng范围内线性良好,回归方程：Y=24451X+529.43,r=0.999,精密度RSD为1.8%,稳定性RSD为2.1%,回收率范围为81.1%～86.9%,检测限为0.028 mg·kg^-1,定量限为0.092 mg·kg^-1。6批样品中有5批检出水合氧化前胡素,1批未检出。保心宁片(胶囊)中枳壳的处方量为每片(粒)0.5010 g,按照掺伪...     

Recent applications of liquid chromatography-mass spectrometry in natural products bioanalysis

Natural flavonoids, alkaloids, saponins and sesquiterpenoids have been extensively investigated because of their biological and physiological significances, as well as their promising clinical uses. It is necessary to monitor them or their metabolites in biological fluids for both pre-clinical studies and routine clinical uses. The successful hyphenation of LC and MS, which was thought as "the bird wants to marry with fish", has been conducted widely in biological samples analysis. This present paper reviewed the feasibility of LC-MS techniques in the identification and quantification of natural products (flavonoids, alkaloids, saponins and sesquiterpenoids) in biological fluids, dealing with sample preparation, LC techniques, suitability of different MS techniques. Perspective of LC-MS was also discussed to show the potential of this technology. The citations cover the period 2002-2006. We conclude that LC-MS is an extremely powerful tool for the analysis of natural products in biological samples.     

Shenmai-Yin decreased the clearance of nifedipine in rats: The involvement of time-dependent inhibition of nifedipine oxidation

The traditional Chinese herbal formula Shenmai-Yin (SY) and nifedipine have both been used to treat patients with cardiovascular disorders. Nifedipine is primarily oxidized by cytochrome P450 (CYP) 3A. The oxidation and pharmacokinetics of nifedipine were studied in rats in vitro and in vivo to illustrate the interaction of SY with nifedipine. Schisandrol A, schisandrin A and schisandrin B were identified as the main lignans in SY. In the study in vitro, the ethanolic extract of SY was used due to the solubility and the extract inhibited nifedipine oxidation (NFO) activity in a time-dependent manner. Among lignans, schisandrin B caused the most potent inhibition. According to the time-dependent inhibition behavior, rats were treated with SY 1 h before nifedipine administration. After oral treatment with 1.9 g/kg SY, nifedipine clearance decreased by 34% and half-life increased by 142%. SY treatment decreased hepatic NFO activity by 49%. Compared to the change caused by ketoconazole, the SY-mediated reduction of nifedipine clearance was moderate. These findings demonstrate that SY causes a time-dependent inhibition of NFO and schisandrin B contributes to the inhibition. The decreased nifedipine clearance by SY in rats warrants further human study to examine the clinical impact of this decrease.