饮料中红花黄色素的固相萃取/超高效液相色谱-质谱测定

利用超高效液相色谱-电喷雾串联四极杆质谱法对从红花中提取纯化的红花黄色素进行结构分析,其包含对羟基红花黄A(34.0%)、红花黄A(27.4%)、脱水红花黄B(23.2%)和红花黄C(8.3%)4种主要成分。建立了测定碳酸类、果汁类饮料中4种色素成分的SPE/UPLC检测方法。利用大孔吸附树脂作为填料,自制SPE固相萃取柱,对饮料中的红花黄色素进行吸附解吸,通过优化固相萃取条件,使得各组分的回收率为84%~116%,相对标准偏差为0.78%~6.6%。各组分在0.40~17 mg/L范围内具有良好的线性关系,线性系数均不小于0.998 8。     

近红外光谱快速测定红花逆流提取过程中羟基红花黄色素A的含量

采用近红外光谱(NIRS)透射法对红花罐组式逆流提取过程中羟基红花黄色素A(Hydroxysafflor yellow A,HSYA)的含量进行快速无损的测定.在红花逆流提取过程中,以高效液相色谱法(HPLC)为对照分析方法,测定提取液中羟基红花黄色素A的含量,运用偏最小二乘(PLS)法建立NIR光谱与羟基红花黄色素A的HPLC分析值之间多元校正模型,并对逆流提取过程的未知样本进行含量预测.校正模型相关系数达到0.982,预测相关系数达到0.965,RMSEC和RMSEP分别为0.053和0.075,RSEC和RSEP分别为3.96%和5.25%.结果表明,NIRS可以作为一种准确、快速、无损的检测方法用于检测中药逆流提取过程有效成分含量变化规律.     

Qualitative evaluation and quantitative determination of 10 major active components in Carthamus tinctorius L. by high-performance liquid chromatography coupled with diode array detector

Flavonoids in the water extract of Carthamus tinctorius L. exhibit potent biological activities such as anti-coagulant, vasodilation, anti-oxidant, neuroprotection and immunosuppressant. A high-performance liquid chromatographic method was established to evaluate the quality of Carthamus tinctorius through a simultaneous quantitation of eight flavonoids, hydroxysafflor yellow A (2), 6-hydroxykaempferol 3,6-di-O-beta-glucoside-7-O-beta-glucuronide (3), 6-hydroxykaempferol 3,6,7-tri-O-beta-glucoside (4), 6-hydroxykaempferol 3-O-beta-rutinoside-6-O-beta-glucoside (6), 6-hydroxykaempferol 3,6-di-O-beta-glucoside (7), 6-hydroxyapigenin 6-O-glucoside-7-O-glucuronide (8), anhydrosafflor yellow B (9), and kaempferol 3-O-beta-rutinoside (10), together with two other compounds named guanosine (1) and syringin (5). Among them, compound 8 was identified as a new compound. The compounds were separated on an Alltech Alltima-C(18) column with gradient elution of acetonitrile and 0.01% trifluoroacetic acid. The detection wavelength was 280 nm. All the compounds showed good linearity (r(2) >or= 0.9989). The recoveries, measured at three concentration levels, varied from 94.9% to 105.2%. This method was also validated with respect to precision, repeatability and accuracy, and was successfully applied to quantify the 10 components in 46 batches of C. tinctorius samples from different areas. Significant variations were found in the contents of these compounds in these samples. Compared with the reported analytical methods of C. tinctorius, this simple and reliable method provided a new basis for overall assessment on quality of C. tinctorius and should be considered as a suitable quality control method.     

动态超声萃取分光光度法在线测定红花中的总红花黄色素

用动态超声萃取分光光度法在线测定了红花中的红花黄色素, 对萃取条件进行了优化. 萃取与测定同时进行, 大大缩短了样品的分析时间, 简化了分析过程. 通过与静态超声萃取、振荡萃取和索式萃取进行比较, 不仅证明该方法耗时最短, 而且样品用量小, 萃取产率较高. 实际样品分析得到了满意的结果.     

Separation and determination of four active anthraquinones in Chinese herbal preparations by flow injection-capillary electrophoresis

A simple, rapid, and accurate method for the separation and determination of physcion, chrysophanol, aloe-emodin, and emodin in Rhubarb, Juemingzi, and Chinese herbal preparations was developed by combination of flow injection-capillary zone electrophoresis for the first time. The analysis was carried out using an unmodified fused-silica capillary (75 mm x 50 microm ID x 375 microm OD, effective separation length of 48 mm) and direct ultraviolet detection at 254 nm. By a series of optimization, the sample solvent consisted of NaOH (100 mmol/L) and ACN (1:1 v/v), and a running buffer composed of 15 mmol/L sodium borate - 12.5 mmol/L sodium dihydrogen phosphate - 42% v/v ACN (pH 10.1) was applied for the separation of the four anthraquinones. The separation was rapid and highly reproducible, with complete resolution of all four compounds within 6 min. The sample throughput rate could reach up to 12 per h. The repeatability (defined as relative standard deviation) was 4.45, 4.44, 4.34, 0.61% with peak height evaluation and 1.62, 0.89, 2.49, 2.19% with peak area evaluation for physcion, chrysophanol, aloe-emodin, and emodin, respectively.     

Characterization of C-glycosyl quinochalcones in Carthamus tinctorius L. by ultraperformance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry

C-Glycosyl quinochalcones are unique components in Carthamus tinctorius L. The reported C-glycosyl quinochalcones have the same quinochalcone skeleton with a hydroxyl group at the 5'-position and a glucose linked to this position with a carbon-carbon bond. In this study, the standard hydroxysafflor yellow A and water-extracted fraction of Carthamus tinctorius L. were analyzed by ultraperformance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry (UPLC/Q-TOFMS) in both positive and negative ion modes. The fragmentation pathways of C-glycosyl quinochalcones were interpreted and validated by accurate mass measurement. Their fragmentation showed a special cleavage at the C-C bond except for the typical internal cleavage at the sugar moiety of other C-glycosyl flavonoids. In positive ion mode, cleavage of the 5'-glucose produced an [M+H-162](+) ion by a neutral loss, while cleavage of the 5'-glucose in negative ion mode led to an [M-H-163](-.) ion by radical cleavage. The cleavage from the carbonyl group produced fragment ions containing an A or a B ring. The fragment ions containing an A ring were common product ions of seven compounds in both ion modes, and fragment ions containing the B ring were used to judge the different substituent groups at the 3'-position. The fragmentation patterns of seven structurally related C-glycosyl quinochalcones were analyzed systematically and the formation of the fragment ions in two modes is explained in detail in this report. UPLC/Q-TOFMS is an effective tool for characterizing a complex sample, which gives higher resolution separation and generates accurate mass measurement of the product ions.     

Separation and determination of isoflavones in red clover by micellar electrokinetic capillary chromatography

A micellar electrokinetic capillary chromatography (MECC) method has been developed for the determination of the four isoflavones, i.e. biochanin A, formononetin, genstein and daidzein in red clover (Trifolium Pratense L.). The effect of running buffer pH and concentration were investigated. An electrolyte composed of 30 mm borate, 20 mm sodium dodecyl sulfate (SDS) and 4 mg/mL HP-beta-CD containing 5% (v/v) ethanol at pH 10.1 provides a satisfactory separation for all the analytes. The applied voltage was 25 kV, and the capillary temperature was kept constant at 25 degrees C with a UV detection at 254 nm. The relative standard deviations (RSD) of the migration time and peak area were less than 1.73 and 3.94% (intra-day), and 2.29 and 4.38% (inter-day), respectively, under the optimized separation conditions. Regression equations revealed a good linear relationship between the peak area of each compound and its concentration. The contents of the four compounds in red clover were successfully determined with satisfactory repeatability and recovery.     

毛细管电泳法同时测定复方化学消毒剂中醋酸洗必泰和苯扎氯铵

建立了复方化学消毒剂中常用有效成分醋酸洗必泰和苯扎氯铵(C12-BAC、C14-BAC及C16-BAC)同时分离测定的毛细管电泳(CE)方法。以37 cm×50 μm未涂层熔融石英毛细管为分离柱,以150 mmol/L磷酸二氢钠-62.5 mmol/L磷酸(pH 2.5)缓冲液(含体积分数为40%的乙腈)为分离缓冲溶液,50 mmol/L醋酸-乙腈(体积比为1:1)为样品介质,检测波长为214 nm。方法的日内及日间精密度分别小于3.0%及3.7%。醋酸洗必泰、C12-BAC、C14-BAC及C16-BAC的检出限(信噪比为3)分别为0.3、0.5、0.5、0.5 mg/L,定量限(信噪比为10)分别为1.0、1.5、1.5和1.5 mg/L,在1.0～400、1.5～200、1.5～200和1.5～200 mg/L范围内,4种有效成分的校正峰面积与相应质量浓度均具有良好的线性关系,相关系数分别为0.9995、0.9998、0.9997和0.9998。加标回收率为93.83%～104.97%。将该法用于实际样品分析,并与液相色谱的分析结果进行比对,获得满意结果。     

HPLC determination of safflor yellow A and three active isoflavones from TCM Naodesheng in rat plasma and tissues and its application to pharmacokinetic studies

A high-performance liquid chromatographic method was developed for the simultaneous determination and pharmacokinetic studies of safflor yellow A, puerarin, 3'-methoxyl-puerarin, and puerarinapioside in the plasma and tissues of rats that had been administered with the traditional Chinese medicine (TCM) preparation Naodesheng via the caudal vein. Samples taken from rats were subjected to protein precipitation with acetone. Separation of these four compounds was accomplished on a Kromisil C18 stationary phase using a mobile phase of acetonitrile-0.1% phosphoric acid-tetrahydrofuran (8:92:2, v/v/v) at a flow-rate of 1.0 mL/min. The detection wavelength was set at 250 nm. The calibration curves of the four components were linear in the given concentration ranges. The intra- and inter-day precisions in plasma and tissues were less than 15% and the extraction recoveries were higher than 60%. The lower limits of quantitation of four components were low enough to determine the four components. These four components all exhibited kinetics that fitted a two-compartment model in rats. The elimination half-life was 1.19 h for safflor yellow A, 2.69 h for puerarin, 2.94 h for 3'-methoxyl-puerarin, and 0.87 h for puerarinapioside, respectively. Following administration of a single injection of Naodesheng, the concentration (C) of the four components in the tissues showed C(kidney) > C(lung), C(liver) > C(spleen), C(stomach), C(heart), approximately. The method is a reliable tool for performing studies of safflor yellow A and three puerarin isoflavones in different biological material.     

毛细管区带电泳法测定板蓝根注射液中四种核苷的含量

采用毛细管区带电泳法测定了板蓝根注射液中胞苷、腺苷、鸟苷和尿苷的含量。电泳条件:采用未涂层石英毛细管(32.5 cm×50 μm i.d.,有效长度23.5 cm),以60 mmol/L硼砂溶液-10%(体积分数)异丙醇-20%（体积分数）乙腈为运行缓冲液,在25 ℃下以20 kV恒压电泳分离,压力进样 (1 kPa×10 s),检测波长254 nm。对电泳条件各因素进行了讨论,如缓冲液的种类、浓度和pH值,有机改性剂的种类和浓度,分离电压和毛细管温度等。样品经0.45 μm微孔滤膜过滤后直接进样;采用外     

胶束电动毛细管色谱法同时测定复方化学消毒剂及日化产品中邻苯二甲醛、对氯间二甲基苯酚及三氯生

建立了邻苯二甲醛(OPA)、对氯间二甲基苯酚(PCMX)和三氯生3种杀菌剂同时分离测定的胶束电动毛细管色谱(MEKC)新方法。详细研究了影响上述3种杀菌剂同时分离与准确定量的因素: 如分离缓冲溶液的浓度及pH,十二烷基硫酸钠(SDS)浓度、样品缓冲溶液等。以40.2 cm (有效长度: 30 cm)×50 μm未涂层熔融石英毛细管为分离柱,20 mmol/L硼砂-80 mmol/L SDS(无需调pH)为分离缓冲溶液,2 mmol/L硼砂-8 mmol/L SDS (含体积分数为10%甲醇)为样品缓冲溶液,检测波长为214 nm。3种杀菌剂的校正峰面积的相对标准偏差(RSD)在1.1%～ 3.8%范围内,迁移时间的RSD均小于0.9%, OPA、PCMX和三氯生的检出限(LOD,信噪比为3)分别为4.0、0.4、0.4 mg/L,定量限(LOQ,信噪比为10)分别为12、1.2、1.2 mg/L,校正峰面积与相应的质量浓度分别在12～2 000 mg/L、1.2～200 mg/L和1.2～200 mg/L范围内具有良好的线性关系,相关系数分别为0.9994、0.9993和0.9995。该法前处理简单,可快速、准确地同时测定3种组分,非常适合常规实验室分析。     

An HPLC/MS method for identifying major constituents in the hypocholesterolemic extracts of Chinese medicine formula 'Xue-Fu-Zhu-Yu decoction'

In this paper, HPLC-ESI-MS technique was used to analyze the hypocholesterolemic extracts of 'Xue-Fu-Zhu-Yu decoction', a traditional Chinese medicine consisting of six crude drugs (i.e. Paeonia lactiflora, Ligusticum chuanxiong, Citrus aurantium, Carthamus tinctorius, Prunus persica and Bupleurum falcatum). A total of 17 compounds were identified and their plant derivations were also distinguished. Nine compounds among them were unambiguously determined as oxypaeoniflorin, amygdalin, albiflorin, paeoniflorin, ferulic acid, naringin, hesperidin, senkyunolide I and neohesperidin by comparing the retention times (tR), UV spectra and m/z values with those of the reference compounds. The other eight compounds were tentatively identified as prunasin, 6-hydroxy-kaempferol 3,6-diglucoside, 6-hydroxykaempferol 3-rutinoside-6-glucoside, galloylpaeoniflorin, 6-hydroxy-kaempferol 3-glucoside, anhydrosafflor yellow B, narirutin and kaempferol 3-rutinoside by MS2 spectra and the comparison of their UV spectra and MS spectra with literature data.     

毛细管区带电泳法同时测定饮料中16种食品添加剂

建立了毛细管区带电泳法测定饮料中酸性红92、专利蓝V、荧光素二钠、酸性红1、靛蓝胭脂红、亮黑、丽春红6R、日落黄、苋菜红、柠檬黄等10种人工合成色素和苯甲酸、山梨酸、对羟基苯甲酸甲酯、对羟基苯甲酸乙酯、对羟基苯甲酸丙酯、对羟基苯甲酸丁酯等6种防腐剂的分析方法。考察了缓冲溶液种类、浓度、pH值及运行电压、温度等对分离的影响,确定最佳电泳条件为: 未涂层弹性石英毛细管柱(46 cm×50 μm),缓冲溶液为70 mmol/L硼酸(pH=9.5)(含体积分数为4%的乙腈),检测波长220 nm,电泳电压30 kV,进样时间5 s,电泳温度25 ℃。该法用于测定市售饮料样品得到满意结果: 在1～250 mg/L范围内线性关系良好,相关系数不小于0.9938,回收率在95.8%与108.7%之间。该法简便、准确,能够满足食品添加剂的常规检测要求。     

高效液相色谱-串联质谱法同时测定食品中五种黄色化工染料

采用高效液相色谱-串联质谱法(HPLC-MS/MS)建立了食品中非法添加的碱性橙、碱性嫩黄、酸性橙I、酸性橙II和酸性黄36这5种黄色工业染料的定量定性分析方法。使用Agilent ODS C18分离柱(50 mm×2.0 mm, 1.8 μm),以5 mmol/L乙酸铵水溶液(0.1%甲酸)-乙腈(3:2, v/v)为流动相,流速为0.3 mL/min。采用电喷雾离子化源,以多反应监测(MRM)方式分别在正、负离子模式下进行检测。在最佳检测条件下,得到了较宽的线性范围和较低的定量检出限。碱性橙和碱性嫩黄的线性范围均为5.0～80.0 mg/L;酸性橙I、酸性橙II及酸性黄36的线性范围均为10.0～160.0 μg/L。食品中碱性橙、碱性嫩黄、酸性橙I、酸性橙II及酸性黄36的定量限分别为20、20、40、40、40 ng/g。该方法重现性较好,保留时间和峰面积的相对标准偏差分别不大于0.50%和2.14%。本研究还测定了鸡肉、豆制品和黄鱼中添加的5种化工染料,回收率在79.8%~95.2%之间,结果令人满意。     

反向电渗流非水毛细管电泳法快速测定微量赤霉素

赤霉素是一类重要的植物激素,是结构相似的弱酸性二萜类化合物。针对毛细管电泳法分离、测定赤霉素速度慢、效率不佳等问题,该文发展了一种可快速测定微量活性赤霉素的非水毛细管电泳-紫外检测法。以聚环氧乙烷动态涂布毛细管,利用正离子使电渗流反向,将含10 mmol/L醋酸铵的甲醇-水(95∶5,v/v)作为缓冲体系,在0.08%(v/v)醋酸含量下,分离8种内源性赤霉素。结果表明,赤霉素出峰时间的相对标准偏差(RSD)≤2.1%(日内)或≤4.3%(日间),峰面积的RSD≤4.5%(日内)或≤6.9%(日间),检出限(S/N=3)为1.04~2.20 mg/L,相关系数为0.998 2~0.999 3,回收率为87.2%~93.5%。该方法简单、快速、稳定,以含醋酸铵的甲醇水溶液为缓冲体系,预先考虑了质谱测定的需要,可用于实际样品如麦芽中赤霉素的分析,具有一定的应用价值。     

毛细管电泳-电致化学发光检测法分离测定中药马尿泡中的托烷类生物碱成分

以铕离子掺杂类普鲁士蓝(Eu-PB)化学修饰铂电极为工作电极,采用毛细管电泳-电致化学发光检测法对4种托烷类生物碱成分（如山莨菪碱、东莨菪碱、阿托品和樟柳碱）进行了分离检测。考察了氧化电位值、运行缓冲液酸度、盐浓度和甲醇含量等实验条件对电泳分离效果及检测灵敏度的影响。在优化的实验条件下,以20 mmol/L的磷酸盐(pH 8.0)-7%(体积分数）甲醇为运行液,各组分在6 min内可达到基线分离,其峰面积的相对标准偏差小于5.0%,迁移时间的相对标准偏差小于1.1% (n=12)。并将该法成功地应用于测定中药马尿泡根茎中的山莨菪碱和东莨菪碱的含量,其含量平均值分别为27.8 g/kg和4.43 g/kg。样品的加标回收率为97.8%～102%。     

Chemical constitution of safflor yellow B,a quinochalcone c-glycoside from the flower petals of carthamus tinctorius L.

The chemical constitution of safflor yellow B, a quinochalcone glycoside from the flower petals of $\text{Carthamus tinctorius L}$, has been characterized as 1, mainly on the basis of spectral means.     

Identification and determination of effective components in Euphrasia regelii by capillary zone electrophoresis

A capillary zone electrophoresis (CZE) method has been developed for simultaneous determination of eukovoside, cinnamic acid and ferulic acid in Euphrasia regelii for the first time. The electrophoresis buffer was 20 mmol/L sodium borate containing 10% (v/v) methanol (pH 8.50). The effects of concentration of borate and electrolyte pH on electrophoretic behavior and separation were studied. Regression equations revealed linear relationships (correlation coefficients 0.9995-0.9998) between the peak area of each analyte and the concentration. The levels of analytes in the different parts of Euphrasia regelii were easily determined with recoveries ranging from 95.5 to 104.2%.     

Method development and validation for the simultaneous determination of four coumarins in Saussurea superba by capillary zone electrophoresis

A capillary zone electrophoretic method has been developed for the determination of four coumarins--skimmin, scopolin, scopoletin, and umbelliferone-in Saussurea superba with UV detection at 254 nm. The capillary temperature was kept constant at 25 degrees C. Effects of buffer pH, electrolyte concentration, organic modifier, and applied voltage on migration behavior were studied systematically. The optimum conditions for separation were achieved by using 30 mM borate buffer at pH 9.02 containing 15% (v/v) methanol as the electrolyte and 25 kV as the applied voltage. For all analytes a good linear regression relationship (r > 0.999) was obtained between peak area and concentration over a relatively wide range. The method was validated for repeatability, precision, and accuracy. The validated method was successfully applied to the simultaneous determination of the four analytes in S. superba.     

Determination of bavachin and isobavachalcone in Fructus Psoraleae by high-performance liquid chromatography with electrochemical detection

A simple, sensitive and selective method of high-performance liquid chromatography with electrochemical detection (HPLC-ECD) has been developed for simultaneous determination of bavachin and isobavachalcone in Fructus Psoraleae. At optimized conditions, bavachin and isobavachalcone could be well separated within 15 min at a detection potential of +0.80 V with 0.03 mol/L acetate buffer solution (pH 5.17)/acetonitrile (2:3, v/v) as the mobile phase. The relationships between peak areas and concentrations were linear from 8.26 × 10(-7) to 1.21 × 10(-4) mol/L for bavachin, and from 1.01 × 10(-8) to 1.61 × 10(-4) mol/L for isobavachalcone, respectively. The method offered excellent linearity with regression coefficient R(2) >0.995. The method presented detection limits (S/N = 3) of 8.81 × 10(-9) mol/L for bavachin and 1.17 × 10(-10) mol/L for isobavachalcone. It indicates that the sensitivity of electrochemical detection is ten times higher than that of diode array detection (DAD). The mean recoveries around 98% with a relative standard deviation less than 3.1% for the two analytes have been obtained. The proposed separation and detection procedures were successfully applied to the simultaneous determination of bavachin and isobavachalcone in traditional Chinese medicine.