Simultaneous Determination of Saponins in Radix Glycyrrhizae, Notoginseng and Ginseng by High Performance Liquid Chromatography

To establish a method for determining five saponins(notoginsenoside R1, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1 and ammonium glycyrrhizinate) in Glycyrrhizae, Notoginseng and Ginseng, the high performance liquid chromatography with diode array detector(HPLC-DAD) method was applied to an Inertsil ODS-SP column(4.6 mm×250 mm, 5 μm) with a mobile phase consisting of acetonitrile-0.05% phosphoric acid in a gradient elution manner. The flow rate was 1.0 mL/min. The column temperature was 30 ℃ and the detection wavelengths were 203 nm and 237 nm, respectively. The linear ranges were 0.700,0—7.000,0 μg for R1(r=1.000,0), 0.751,1— 7.511,4 μg for Rg1(r=1.000,0), 0.677,2—6.771,6 μg for Re(r=1.000,0), 0.733,9—7.339,1 μg for Rb1(r= 1.000,0), and 0.540,0—5.399,8 μg for ammonium glycyrrhizinate(r=0.999,9), respectively. In addition, their average recoveries were 100.28%, 105.83%, 104.09%, 99.36% and 98.54%, respectively. The relative standard deviations(RSDs) of precision, reproducibility and recovery were all less than 1.5%. The results indicate that the method is simple, accurate and reproducible so that it can be used for the simultaneous determination of the five saponins in Chinese patent medicines containing the three kinds of herbs.

Determination of AF and AFG in Red Ginseng by High Performance Liquid Chromatography with Evaporative Light Scattering Detector (HPLC-ELSD)

[Objective]The paper was to establish a method for determining AF and AFG in red ginseng.[Method]A new simple,rapid and sensitive method for simultaneous determination of two amadori compounds,arginyl-fructose(AF)and arginyl-fructosyl-glucose(AFG),in extracts of three kinds of ginseng preparations was developed and validated using high performance liquid chromatography with evaporative light scattering detector(HPLC-ELSD).Two target analytes were efficiently separated by Prevail CTM18 column(4.6 mm×250 mm,5μm)at the flow rate of 0.8 mL/min within 15 min of single chromatographic run.[Result]Under optimized conditions,the detection limits were 0.015 and 0.02 mg/mL for AF and AFG,respectively.Calibration curves of peak area for two analytes were linear over three orders of magnitude with the correlation coefficients greater than 0.999.The average recoveries,precision,reproducibility and stability for two analytes(AF and AFG)were 99.5% and 100.9%,0.43% and 0.47%,0.46% and 0.43%,0.41% and 0.49%,respectively.[Conclusion]This method was successfully applied for quantifying AF and AFG in red ginseng and the method was efficient,sensitive and accurate.

Analysis of Low-polar Ginsenosides in Steamed Panax Ginseng at High-temperature by HPLC-ESI-MS/MS

A high performance liquid chromatography coupled with electrospray ionization-tandem mass spectrome try（HPLC-ESI-MS/MS） method was developed for the analysis and identification of ginsenosides in the extracts of raw Panax ginseng（RPG） and steamed Panax ginseng at high temperatures（SPGHT）. A total of 25 ginsenosides were extracted include of which 10 low-polar ginsenosides, such as ginsenosides F4, Rk3, Rh4, 20S-Rg3, 20R-Rg3 and so on, were identified according to their HPLC retention time and MS/MS data. The results indicated that the low polar ginsenosides were seldom found in RPG. For the exploration of the transformation pattern of the ginsenosides in steam processing, the standards of ginsenosides Re, Rg1, Rb1, Rc, Rb2, Rb3 and Rd were selected and hydrolyzed at a temperature of 120 oC. The results show that these polar ginsenosides can be converted to low-polar ginsenosides such as Rg2, Rg6, F4, Rk3 and Rg5 by hydrolyzing the sugar chains.

Determination of Seven Major Ginsenosides in Different Parts of Panax quinquefolius L.(American Ginseng) with Different Ages

Ginsenosides Rgl, Re, Rb1, Rc, Rb2, Rb3, and Rd in different parts of the American ginseng plant were investigated. The extraction process was a pressurized microwave-assisted extraction（PMAE）. The seven ginsenosides were separated and determined by high-performance liquid chromatography（HPLC） with a ultraviolet（UV） detector, at 203 nm. The experiment results showed significant variations in the individual ginsenoside contents of the American ginseng in different parts and ages of the plant. The results demonstrated that the leaves, root hairs, and rhizomes of Panax quinquefolius L. contained higher ginsenoside contents, followed by the main roots and stems. The leaves contained dramatically higher levels of ginsenoside Rg1 Rb3, and Rd than the other four parts. Higher contents of Rb1 and Re were present in the main roots, root hairs, and rhizomes. The amount of ginsenoside content in the stems was the lowest. The total content of the seven ginsenosides in main roots, root hairs and rhizomes increased with the age of the plant. In contrast, the ginsenoside contents in the leaves and stems decreased with a year of growth.

Rapid Determination of Amino Acids in Ginseng by High Performance Liquid Chromatography and Chemometric Resolution

Ginseng is one of the most important traditional Chinese medicines and functional foods.A method for the fast determination of amino acids in ginseng samples using high performance liquid chromatography（HPLC） was developed,in which strong isocratic elution was employed for simplifying the separation and speeding up the analysis.All amino acids were eluted within 3 min with the chromatogram composed of overlapped peaks from the interferences.Then,non-negative immune algorithm（NNIA） was adopted to resolve the chromatographic signals of the components from the chromatogram measured.The results show that the signals of the amino acids can be correctly extracted by NNIA and the signal extracted can be used for the quantitative analysis.The method was validated via determining six amino acids of four different samples of ginseng.The recoveries of the spiked samples are in a range of 96.6％-106.3％.

高效液相色谱-串联四极杆质谱法测定人参组培不定根中11种皂苷成分

建立高效液相色谱-串联四极杆质谱(HPLC-MS/MS)法定量分析人参组培不定根中11种皂苷成分。样品经粉碎研磨,以70%甲醇水溶液为溶剂进行超声提取,离心过滤后测定。采用C_(18)色谱柱(100 mm×2.1 mm,1.8μm),用水和乙腈进行梯度洗脱,流量为0.3 mL/min,柱温为35℃。质谱采用电喷雾电离源,负离子扫描,多反应监测模式进行检测。11种皂苷的质量浓度在0.1~10μg/mL(Rb1为0.2~10μg/mL)范围内和响应强度线性相关,相关系数均大于0.995,各皂苷的定量限为0.001~0.010 g/kg,加标回收率为90.43%~97.82%,相对标准偏差为1.93%~6.33%(n=6)。该方法操作简单,分析时间短,灵敏度高,准确可靠,适用于人参组培不定根中多种皂苷类成分的同时测定。

Qualitative Analysis of Xinyue Capsules(心悦胶囊)by High-Performance Liquid Chromatography:Preliminary Evaluation of Drug Quality in A Sino-Austrian Joint Study

ABSTRACT Objective： To develop a reliable method to assess the stability of Xinyue Capsules （心悦胶囊） containing Panax quinquefolius saponins according to European quality standards. Methods： An efficient high-performance liquid chromatography ultraviolet （HPLC-UV） method was established to analyse six main ginsenosides （Rb1, Rb2, Rc, Rd, Re and Rg1） in six different batches （120 capsules/batch） from the same lot of Xinyue Capsules and in one batch measured six times within one day. The six ginsenosides were separated on a Hypersil BDS-C18 column （3 μm, 100 mm × 3 mm） at a flow rate of 0.5 mL/min. Gradient elution was performed using a mobile phase gradient of acetonitrile-water modified with 0.01% formic acid. The HPLC chromatograms were analyzed with ＂LC data comparison＂ using Lab Solutions software. Results： The HPLC peaks were identified by comparing their retention times （Rg1： 23.44 min, Re： 23.77 rain, Rb1： 35.24 min, Rc： 36.18 min, Rb2：38.55 min and Rd： 40.88 min） with those of the standards under the same chromatographic conditions, which showed similar results among the samples of six different batches and among the samples from one batch detected six times within one day. Conclusions： Xinyue Capsules have good drug intra-day consistency at room temperature and exhibit a consistent quality between different batches. This study established a reliable method to assess the stability of Xinyue Capsules, which is suitable for further qualitative analysis and may assist in promoting the safe and effective use of Chinese herbal medicine.

高效液相色谱法测定人参保健酒中11种人参皂苷含量

该研究建立了高效液相色谱法(HPLC)同时测定人参保健酒中11种人参皂苷(人参皂苷Rg1、Re、Rf、Ra2、Rb1、Rc、Ra1、Ro、Rb2、Rb3、Rd)的方法。采用Agilent SB C_(18)色谱柱(4.6 mm×250 mm,5μm),以乙腈-0.1%磷酸为流动相梯度洗脱,流速1.0 mL/min,柱温28℃,人参皂苷的检测波长203 nm,进样量10μL的条件进行测定。结果表明,11种人参皂苷在各自浓度范围内线性关系良好(R^(2)≥0.9999),检出限在1.52~3.03 mg/L,定量限在4.59~9.18 mg/L,该方法精密度试验、稳定性试验、重复性试验结果相对标准偏差(RSD)均小于5%,平均加标回收率95.39%~101.96%,RSD为0.07%~2.46%。该方法操作简单,准确度高,重复性好,可用于人参保健酒中11种人参皂苷含量测定。

超高效液相色谱-四级杆飞行时间质谱测定三七中九种真菌毒素

建立一种三七中9种真菌毒素的超高效液相色谱-四极杆飞行时间质谱筛查与确证方法,快速筛查三七中9种真菌毒素(黄曲霉毒素B_(1)、黄曲霉毒素B_(2)、黄曲霉毒素G_(1)、黄曲霉毒素G_(2)、伏马菌素B_(1)、伏马菌素B_(2)、脱氧雪腐镰刀菌烯醇、玉米赤霉烯酮、赭曲霉毒素A)的残留量。色谱柱为Phenomenex Kinetex C18(100 mm×2.1 mm,2.6μm),以0.1%甲酸水溶液和0.1%甲酸甲醇水溶液为流动相进行梯度洗脱,正离子扫描,基质匹配外标法定量。结果表明,9种真菌毒素分别在0.1~100.0、5.0~1 000.0、0.5~500.0μg/L浓度范围内具有良好的线性关系,相关系数均大于0.990 00,方法定量限为0.3~15.0μg/kg,在高、中、低3个浓度加标水平下,回收率为71.22%~98.57%,相对标准偏差为2.31%~6.72%。该方法在没有对照品的情况下,可与质谱数据库中一级、二级质谱的精确质量数和碎片离子信息等进行匹配,降低了试验成本,具有简便、快速、高效、准确等优点,适用于三七中真菌毒素残留的快速筛查和测定。

黑参炮制工艺优化及成分变化分析

以人参为原料,采用高温高压法炮制黑参,加工过程中使用蒸制与焖制结合的方法,缩短黑参的炮制时间,利用单因素试验和正交法优化黑参的炮制工艺,以感官评价为参考值,考查蒸制时间、烘干时间、烘干温度3个因素对黑参品质的影响。结果表明,在蒸制时间120 min,烘干时间8 h,烘干温度60℃时,炮制出的黑参的品质最佳;其含水量平均为10.132 513%,人参皂苷含量分别为0.033 g/100 g (Re),0.141 g/100 g (20(S)-Rg3);0.256 g/100 g(20(R)-Rg3)。与原参相比,黑参的稀有人参皂苷Rg3含量增加,人参皂苷Re含量减少。

参麦颗粒中红参掺伪研究

目的建立参麦颗粒中红参掺伪的检查方法。方法采用固相萃取-超高效液相色谱串联质谱(SPE-UPLC-MS/MS)法,色谱柱为Shimadzu Shim-pack GIST C18柱(100 mm×2.1 mm,2μm),流动相为乙腈-水(梯度洗脱),流速为0.35 mL/min,柱温为40℃,进样量为5μL。电喷雾离子源、负离子模式(ESI-),多反应监测(MRM)模式。结果拟人参皂苷F11质量浓度在9.030~344.0 ng/mL范围内与峰面积线性关系良好(r=0.9992,n=5),检测限和定量限分别为1.1288,22.8 ng/mL;精密度、稳定性、重复性试验结果的RSD均小于4.0%;平均加样回收率为108.17%,RSD为4.24%(n=6)。以0.051μg/g为掺伪判定限度,3家企业的85批样品中有9批超出限度(检出率为10.59%)。结论所建立的方法简便快速、准确度高、专属性强,可作为参麦颗粒中红参掺伪的检查方法。

基于UPLC-MS/MS和感官评价对人参提取物中主要苦味物质分析研究

采用高效液相色谱法、高效制备液相色谱法、超高效液相色谱-串联质谱法(UPLC-MS/MS)以及感官评价对人参提取物中的主要苦味物质进行分离、纯化和鉴定。通过滋味稀释分析法对人参提取物中的主要苦味物质进行贡献度排序。结果表明,人参皂苷Rg1、人参皂苷Rf、人参皂苷Rb1、20(S)-人参皂苷Rg2和人参皂苷Rb3是人参提取物样品中的苦味物质,其中人参皂苷Rb1和20(S)-人参皂苷Rg2滋味稀释因子最高,是其主要的苦味物质,且与20(S)-人参皂苷Rg2相比,人参皂苷Rb1具有更小的苦味识别阈值,是人参提取物中最关键的苦味物质。

气相色谱——三重四极杆串联质谱法及高效液相色谱——串联质谱检法测定人参中禁用农药残留物

目的:建立使用气相色谱-三重四极杆串联质谱法及高效液相色谱-串联质谱检法测定人参中禁用农药残留物的测定方法。方法:参照2020年版《中国药典》通则2341第五法4.1方式操作步骤,分别采用岛津气相色谱-串联质谱GC-MSTQ8040、高效液相色谱-串联质谱Waters XEVO TQMS进行检测分析。结果:对空白样品按低中高浓度分别加入混合对照品溶液500μL、1000μL、1500μL(添加浓度以灭线磷计分别为:0.04 mg/kg;0.08 mg/kg;0.12 mg/kg)后,按照文中前处理方法处理后,在岛津气相色谱-串联质谱GC-MSTQ8040上机,每个浓度平行3份样品考察回收率和RSD,结果显示,低中高浓度加标回收率分别为:86.83%~108.43%;91.39%~112.99%;88.47%~115.09%。,RSD分别为:0.54%~9.74%;2.24%~4.20%;1.07%~2.96%。回收率高,重现性好。对空白样品按低中高浓度分别加入混合对照品溶液500μL、1000μL、1500μL(添加浓度以灭线磷计分别为:0.1 mg/kg;0.2 mg/kg;0.3 mg/kg)后,按照文中前处理方法处理后,在高效液相色谱-串联质谱WatersXEVOTQMS上机,每个浓度平行3份样品考察回收率和RSD,结果显示,低中高浓度加标回收率分别为:74.33%~103.29%;72.55%~105.16%;81.54%~103.41%。RSD分别为:0.29%~6.01%;1.07%~8.32%;0.41%~7.74%。回收率高,重现性好。结论:该方法便捷,回收率和稳定性良好,适用于人参中禁用农药残留物的测定。

高效液相色谱-二酰胺基色谱柱HILIC模式下对人参口服液中三种人参皂苷的测定

建立高效液相色谱-酰胺基色谱柱HILIC模式,对人参口服液进行含量测定。采用Venusil HILIC为色谱柱(4.5 mm×250 mm),以水和乙腈为流动相进行等度洗脱,建立数学模型,分析整个测定流程。结果表明,人参皂苷Rg_(1)、人参皂苷Re、人参皂苷Rb_(1)在10.0~500μg/mL范围内线性关系良好,相关系数(r^(2))均大于0.9998,系统精密度实验RSD分别为1.01%,1.21%,1.05%,三种人参皂苷的回收率均较好。研究为人参口服液中人参皂苷Rg_(1)、人参皂苷Re、人参皂苷Rb_(1)的检测提供参考和借鉴。

分散固相萃取结合UPLC-MS/MS测定不同人参加工品中46种皂苷类化合物

运用基质分散固相萃取净化,建立一种同时测定人参不同加工品中46种皂苷类化合物的超高效液相色谱-质谱分析方法。样品经体积分数80%甲醇溶液超声提取,通过N-丙基乙二胺(primary secondary amine,PSA)分散固相萃取净化后,采用KINETEX XB-C18色谱柱(2.1 mm×100 mm,2.6μm)分离,以0.1%甲酸溶液(A)和乙腈(B)为流动相进行梯度洗脱,采用负离子多反应监测模式分析和外标法定量。结果表明,46种皂苷类化合物在35 min内完成检测,在0.02~50μg/mL质量浓度范围内有较好的线性关系,所对应的相关系数(R2)均大于0.99,方法检出限为0.4~3.2 mg/kg,定量限为1.4~10.5 mg/kg,在低、中、高3个添加水平下的平均回收率为80.5%~111.6%,相对标准偏差(relative standard deviations,RSD)为1.3%~6.8%。方法日内和日间精密度的RSD分别为0.5%~4.5%(n=6)和1.1%~6.6%(n=6),重复性的RSD在1.5%~7.3%(n=6)之间,24 h内样品稳定性的RSD在0.9%~5.8%(n=6)之间。方法优化了分散固相萃取剂的种类和用量,并考察分散固相萃取净化前后对基质效应的影响,结果表明,样品前处理通过PSA分散固相萃取净化,有效降低了基质效应,且操作简便、高效、节约成本。将该方法用于3种不同人参加工产品生晒参、红参和黑参样品中46种皂苷类化合物的分析测定,结果发现红参和黑参经过蒸制加工处理后,部分原有的皂苷发生了转化,产生了稀有皂苷,且黑参中产生的稀有皂苷含量远高于红参。该方法准确可靠、快速、易操作,杂质干扰小,适合批量人参加工品中46种皂苷类化合物的同时定量分析。

超高效液相色谱-串联质谱法快速检测人参制品中39种农药残留

本研究建立并验证了一种快速检测4种人参制品(人参蜜片、人参水煎液、人参功能饮料、人参酒)中39种农药残留的方法。样品采用QuEChERS法提取,磁性氧化锆(Fe_(3)O_(4)-ZrO_(2))净化,超高效液相色谱-串联质谱法(UHPLC-MS/MS)检测,外标法定量。结果表明:39种农药在0.005~0.1 mg/L范围内线性良好(r>0.99);在4种人参制品基质中的平均回收率为72%~117%,相对标准偏差(RSD)小于18%;定量限(LOQ)为0.01 mg/kg。通过对4种人参制品实际样品的筛查,未检出方法中所包含的农药。

超高效液相色谱法测定三七总皂苷喷雾给药大鼠皮肤中皂苷成分的含量

目的建立同时测定大鼠皮肤中4种皂苷成分(三七皂苷R_(1)、人参皂苷Rg_(1)、人参皂苷Re、人参皂苷Rb_(1))含量的超高效液相色谱法,并对2种皮肤外用制剂(三七总皂苷传递体喷雾剂和三七总皂苷脂质体喷雾剂)中4种皂苷成分的皮肤滞留特性进行初步研究。方法采用Waters BEH C_(18)(2.1 mm×100 mm,1.7μm)色谱柱,以乙腈(A)-水(B)为流动相梯度洗脱(0~8 min,19%→25%A;8~9 min,25%→42%A;9~12 min,42%→55%A),体积流速为0.5 mL·min^(-1),检测波长为203 nm,进样量为1μL,柱温为30℃。结果三七皂苷R_(1)、人参皂苷Rg_(1)、人参皂苷Re、人参皂苷Rb_(1)分别在0.208~20.800,0.852~85.200,0.116~11.600,0.904~90.400μg范围内呈良好的线性关系(r>0.9990)。该方法日内、日间精密度RSD<15%,样品提取回收率为90.31%~102.70%,室温、冻融及长期冷冻稳定性RSD<15%。大鼠皮肤给药12 h后,三七皂苷R_(1)、人参皂苷Rg_(1)、人参皂苷Re在全皮及角质层下的滞留量为三七总皂苷脂质体喷雾剂>三七总皂苷传递体喷雾剂,而人参皂苷Rb 1则恰好相反。结论采用超高效液相色谱法同时检测大鼠皮肤中4种皂苷成分的含量能大大缩短检测时间,该方法操作简单、准确、灵敏,可为三七总皂苷外用制剂的皮肤渗透研究提供科学依据。

昭通产三七不同部位总皂苷含量比较研究

目的:通过建立高效液相色谱(HPLC)法,测定昭通产三七剪口、根茎、须根以及三七茎叶中的5种皂苷:三七皂苷R_(1)、人参皂苷Rg_(1)、人参皂苷Re、人参皂苷Rb_(1)和人参皂苷Rd的含量,为昭通产三七药材质量控制奠定基础。方法:采用HPLC法,以乙腈-水为流动相,进行梯度洗脱。结果:在所建立的色谱条件下,三七皂苷R_(1)、人参皂苷Rg_(1)、人参皂苷Re、人参皂苷Rb_(1)和人参皂苷Rd有较好的分离度,线性关系、稳定性、精密度和加样回收试验均符合中药质量分析的要求;昭通三七剪口中以上5种皂苷含量最高,其总含量为6.709%,其中人参皂苷Rg_(1)、人参皂苷Rb_(1)以及三七皂苷R_(1)3种皂苷总含量为5.720%,茎叶中仅含有人参皂苷Rb_(1)、人参皂苷Rd两种皂苷,其总含量为0.2539%。结论:昭通产三七剪口的总皂苷含量符合现行版《中国药典》要求、根茎总皂苷含量略低于《中国药典》标准,昭通引种三七值得进一步探讨。

人参种植土壤中10种农药残留的高分辨质谱筛查方法

建立了一种能同时检测参地土壤中10种农药残留的超高液相色谱-高分辨质谱联用方法。土样加入乙腈提取,N-丙基乙二胺(PSA)萃取净化,C18柱分离富集得到待测液,经Accucore aQ色谱柱洗脱后,通过超高液相色谱-高分辨质谱进行快速筛查分析。检测结果表明,在0.01~1mg/L浓度范围内,10种农药均具有良好的线性相关性(r>0.9986),定量限为10μg/kg,RSD在2.3%~8.9%之间,平均回收率为71.0%~109.6%,符合要求。本方法灵敏度和准确度较好,可用于参地土壤环境中这10种农药残留量快速筛查。同时可以降低前处理时间,提高工作效率。

天麻洋参含片中天麻素和人参总皂苷的含量测定

采用高效液相色谱法(HPLC)测定天麻洋参含片中天麻素的含量。使用ULtimate AQ-C_(18)色谱柱,流动相为甲醇-磷酸盐缓冲液一水(3.0:4.5:92.5),流速为1.0 mL/min,检测器为DAD,检测波长为220nm。天麻素线性范围为Y=655428X-651265,相关系数为R^(2)=0.9997,平均加样回收率为97.6%,相对偏差为0.46%。采用紫外分光光度法(UV)测定天麻洋参含片中天麻素的含量。人参总皂苷的线性范围为y=0.4821x-0.0592,相关系数为R^(2)=0.9995,平均加样回收率为96.8%,相对偏差RSD为0.9%。