甘草饮片中甘草苷和甘草酸含量分析

目的测定甘草饮片中甘草苷和甘草酸含量,为甘草饮片质量标准制定提供依据。方法以甘草苷和甘草酸为指标,采用HPLC法对十家饮片企业的甘草饮片进行含量测定。结果所测甘草饮片中的甘草苷和甘草酸含量全部达到《中国药典》2005年版一部甘草药材标准要求,且野生甘草饮片明显高于栽培甘草饮片。结论甘草饮片中的甘草苷和甘草酸含量差异较大,建议加强甘草产地适宜性、规范化栽培和饮片质量标准及工艺制备研究。     

乌拉尔甘草HPLC指纹图谱建立及甘草苷和甘草酸的含量测定

摘要：目的:建立乌拉尔甘草HPLC指纹图谱,同时测定不同产地乌拉尔甘草中甘草苷和甘草酸的含量,为经典名方物质基准的研究提供参考。方法:采用色谱柱Agilent 5 TC-C18（250 mm×4.6 mm,5μm）,以流动相为乙腈-0.05%磷酸溶液梯度洗脱,检测波长为237 nm,流速为1 ml·min-1,柱温为35℃。建立10批乌拉尔甘草指纹图谱,并采用HPLC法测定其甘草苷和甘草酸的含量。结果:指纹图谱相似度均大于0.9,标定20个共有峰。含量测定表明甘草苷和甘草酸含量最高分别为宁夏红寺堡和新疆,而甘肃黑虎村的甘草苷和甘草酸含量均较高。结论:通过乌拉尔甘草HPLC指纹图谱及两个有效成分的含量测定,可用于经典名方物质基准的质量控制。     

不同产地甘草有效成分含量分析

目的对不同产地甘草中甘草酸、甘草苷以及甘草总皂苷进行含量测定。方法采用高效液相色谱法测定甘草酸、甘草苷的含量。色谱柱为Thermo sentifi c ODS-2 Hypersil,流动相为乙腈-0.05%磷酸;以甘草酸单铵盐为对照品,香草醛-高氯酸为显色剂,比色法测定甘草总皂苷含量。结果测得不同产地不同种植方式甘草中甘草酸和甘草苷含量符合2010年版药典标准。结论不同产地以及不同种植方式的甘草中,甘草酸、甘草苷及甘草总皂苷的含量差异较大,可为甘草规范化种植提供依据。     

单因素配伍剂量对经方芍药甘草汤药效组分的影响

目的研究中药经方芍药甘草汤中炙甘草或醋白芍单因素配伍剂量变化对其药效组分的影响,为临床合理用药和建立中药药效组分质量评价标准提供科学依据。方法以单因素配伍剂量的醋白芍和炙甘草为载体,以经方芍药甘草汤作为对照,按照汤剂制备供试品溶液,采用HPLC-DAD法,三波长同时测定药效组分氧化芍药苷-芍药内酯苷-芍药苷-苯甲酰芍药苷-甘草酸(λ=230 nm)、甘草苷-甘草素(λ=276 nm)、异甘草苷-异甘草素(λ=360 nm)的含量。结果醋白芍剂量12 g不变时,其药效组分氧化芍药苷-芍药内酯苷-芍药苷-苯甲酰芍药苷的含量随炙甘草的剂量变化如下(mg/m L,n=3):炙甘草12 g,醋白芍组分含量为1.90-0.89-3.45-0.53;炙甘草为1、6、9、15、18 g时,醋白芍组分含量均降低。炙甘草剂量12 g不变时,其药效组分甘草酸-甘草苷-甘草素-异甘草苷-异甘草素的含量随醋白芍的剂量变化如下(mg/m L,n=3):醋白芍12 g,炙甘草的组分含量为9.06-3.19-0.76-0.77-0.15;醋白芍为1、6、9、15、18 g时,炙甘草组分含量均降低。结论 1炙甘草或醋白芍配伍剂量变化时,其药效组分的溶出率与经方比较有显著差异,表明改变中药的配伍剂量其功效会产生变化;2醋白芍-炙甘草剂量比越接近1∶1,其相应的药效组分溶出率越高;3醋白芍的4种药效组分总含量随炙甘草配伍剂量变化规律:炙甘草12 g>9 g>15 g>6 g>18 g>3 g>1 g,药效组分总量为6.77~4.15 mg/m L;4炙甘草的5种药效组分总含量随醋白芍配伍剂量变化规律:醋白芍12 g>9 g>15 g>6 g>18 g>3 g>1 g,药效组分总量为13.93~10.34 mg/m L;5炙甘草剂量对芍药甘草汤药效组分总量的影响大于醋白芍。     

复方甘草片中甘草酸、异甘草苷、异甘草素的测定

目的：建立复方甘草片中甘草酸、异甘草苷和异苷草素的含量定量分析的高效液相色谱(HPLC)方法。方法：采用色谱柱ODS-C_(18)(4．6mm×250mm,5μm),甘草酸流动相为甲醇：冰醋酸：水(70：1：30)；检测波长254nm。异甘草苷与异甘草素在同一条件下检测,其流动相为甲醇：冰醋酸：水(50：2：50)；检测波长346nm；三者流速均为1ml /min；进样量为10μl；柱温为25℃。结果：复方甘草片中甘草酸的含量为7．51mg/g,异甘草苷和异甘草素的含量分别为3．01mg/g,0．448mg/g,上述3种组分的平均回收率分别为98．32%,99．57%和98．59%。结论：本方法可作为复方甘草片中的甘草酸、异甘草苷和异甘草素的含量质量控制的一种有效方法。     

高效液相色谱?电雾式检测法测定甘草中 4种有效成分的含量

目的建立甘草药材高效液相色谱 ?电雾式检测器( Charged Aerosol Detector,CAD)含量测定方法,测定芹糖甘草苷、甘草苷、芹糖异甘草苷及甘草酸 4种成分的含量。方法采用 Waters XBridge C 18色谱柱( 4.6 mm×250 mm,5 μm)进行分离,以乙腈 ?0.05%甲酸水溶液为流动相,梯度洗脱;流速为 1 mL/min;进样量为 10 μL;柱温: 35 ℃;电雾式检测器检测,雾化器温度为 40 ℃。结果芹糖甘草苷进样量在 0.039 21~0.588 15 μg范围内与峰面积线性关系良好( r＝0.999 9)平均回收率试验为 97.3%,RSD为 1.28%(n＝6);甘草苷进样量在 0.034 29~0.857 20 μg范围内与峰面积线性关系良好( r＝09 0)平均回收率试验为 98.4%,相对标准偏差(RSD)为 0.86%(n＝6);芹糖异甘草苷进样量在 0.032 19~0.643 82 μg范围内与峰面性关系良好(r＝0.999 5)平均回收率试验为 98.9%,相对标准偏差( RSD)为 0.77%(n＝6);甘草酸进样量在 0.169 37~2.540 61 μg范围内与峰面积线性良好( r＝0.999 1)平均回收率试验为 99.9%,RSD为 0.15%(n＝6);甘草药材中芹糖甘草苷含量在 0.26%~ .99,积线,关系,2.27%、甘草苷的含量在 0.33%~5.07芹糖异甘草苷的含量在 0.14%~0.81%、甘草酸的含量在 0.77%~9.76%。结论使用高%、,效液相色谱 ?电雾式检测( HPLC?CAD)方法建立了甘草药材的含量测定方法,并同时测定了芹糖甘草苷、甘草苷、芹糖异甘草苷及甘草酸的含量。该方法简便快捷、重复性好,可用于甘草药材的质量控制。     

HPLC法同时测定炙甘草汤中甘草苷、甘草素、异甘草素的含量

目的:建立同时测定炙甘草汤中甘草苷、甘草素、异甘草素含量的方法。方法:采用高效液相色谱法。色谱柱为Waters Sunfire C18(250mm×4.6mm,5μm),流动相为乙腈-0.5%冰醋酸(50:50,V/V),检测波长为278nm,流速为1.0mL·min-1。结果:甘草苷、甘草素、异甘草素的检测浓度分别在1.2～24.0、64.0～1280.0、50.0～1000.0μg·mL-1范围内与各自峰面积积分值呈良好的线性关系(r分别为0.9998、0.9999、0.9999);三者平均加样回收率分别为100.38%、99.45%、97.46%,RSD分别为0.45%、0.53%、0.96%(n=6)。结论:该方法准确、可靠,可为炙甘草汤的质量控制提供检测依据。     

甘草含有的甘草苷和甘草酸受炮制因素的影响探讨

甘草为益补中药,临床常见甘草饮片入药。不同炮制工艺对甘草药用价值具备一定影响。本文以甘草炮制方法入手,立足于主流的蜜炙工艺,探析炮制手段、炮制要点与不同炮制条件对甘草中的甘草苷与甘草酸的影响,从蜜炙方法、温度控制、时间控制等要素着手,讨论甘草炮制的最佳工艺。     

HPLC法测定糖清胶囊中甘草酸和甘草苷的含量

目的:建立测定糖清胶囊中甘草酸和甘草苷含量的高效液相色谱法。方法:采用Kromasil Cl8色谱柱(250mm×4.6mm,5μm)。甘草酸流动相为甲醇-0.2mo·ll-1醋酸铵溶液-冰醋酸(65:35:1);检测波长250nm;甘草苷乙腈-0.5%冰醋酸(18:82)为流动相;检测波长276nm;两者流速为1.0 ml·min-1,两者柱温为30℃;结果:甘草酸单铵盐在0.52~2.6μg/mg范围内呈现良好的线性关系;甘草苷在0.30~1.5μg/mg范围之内具有良好的线性关系。甘草酸和甘草苷回收率分别为98.1%、97.0%,R SD分别为1.32%、1.67%。结论:该法操作简便、准确,线性相关系数良好,且稳定性和重复性好,适用于糖清胶囊制剂甘草主成分的含量测定。     

替代对照品法测定甘草流浸膏中甘草苷和甘草酸含量

目的 建立以橙皮苷为替代对照品测定甘草流浸膏中甘草苷和甘草酸含量的高效液相色谱法.方法 采用替代对照品内加入法,以橙皮苷为替代对照品,通过测定甘草酸铵与橙皮苷、甘草苷与橙皮苷的相对校正因子(f值),利用f值计算甘草流浸膏中甘草苷和甘草酸含量.色谱条件:Welch Ultimate C18色谱柱(250 mm×4.6 m...     

Variation of glycyrrhizin and liquiritin contents within a population of 5-year-old licorice (Glycyrrhiza uralensis) plants cultivated under the same conditions

Cultivated licorice plants (Glycyrrhiza uralensis FISCH.) contain smaller amounts of the triterpene saponin glycyrrhizin than wild licorice plants. To resolve this problem and to breed strains with high-glycyrrhizin content we determined the glycyrrhizin content of 100 samples of G. uralensis that were propagated from seed and grown under the same conditions in the field for 5 years. There was a 10.2-fold variation in glycyrrhizin content among these plants, ranging from 0.46 to 4.67% (average 2.11±0.90%). There was also a wide variation in liquiritin content, ranging from 0.11 to 2.65% (average 1.00±0.49%). The glycyrrhizin content was positively correlated with that of liquiritin in the taproots (r(2)=0.5525). Our results indicate that there are various genetic strains for glycyrrhizin and liquiritin synthesis within a population of plants propagated from seed. The selected high-glycyrrhizin and liquiritin strains will be useful for licorice production and studies on biosynthetic analysis of glycyrrhizin and liquiritin.     

CYP3A4 inhibitors isolated from Licorice

The extract of licorice (Glycyrrhiza uralensis FISHER, Leguminosae) showed CYP3A4 inhibitory activity with the IC50 value of 0.022 mg/ml. Bioassay-guided purification afforded nine compounds, 3-(p-hydroxyphenyl)propionic acid (1), isoliquiritigenin (2), (3R)-vestitol (3), licopyranocoumarin (4), 4-hydroxyguaiacol apioglucoside (5), liquiritin (6), liquiritigenin 7,4'-diglucoside (7), liquiritin apioside (8), and glucoliquiritin apioside (9). Among these compounds, 3, 7, and 5 showed potent CYP3A4 inhibitory activities with IC50 values of 3.6, 17, and 20 microM, respectively. Glycyrrhizin (10), a main constituent of licorice, however, was inactive for CYP3A4 inhibition.     

HPLC-MS法鉴定甘草的指纹图谱

目的：对甘草液相色谱指纹图谱主要色谱峰进行定性,了解甘草甲醇-水提取物的化学成份。方法：测定甘草甲醇提取物的HPLC—DAD及HPLC—MS指纹图谱。结果：结合文献对指纹图谱主要色谱峰进行鉴定,推断出17个色谱峰中19个可能的成分。结论：本研究给甘草质量评价等各类研究提供较全面的化学依据。     

Constituent properties of licorices derived from Glycyrrhiza uralensis, G. glabra, or G. inflata identified by genetic information

Constituent properties of licorices derived from Glycyrrhiza uralensis, G. glabra, and G. inflata are revealed by comparing 117 of licorice identified using four genetic markers; internal tracscribed spacer (ITS) on nuclear ribosomal DNA, rbcL gene, matK gene, and trnH-trnK1 intergenic region on chloroplast DNA. Regarding six main constituents of licorice; glycyrrhizin, liquiritin, liquiritin apioside, isoliquiritin, isoliquiritin apioside, and liquiritigenin, the constituent property of G. glabra resembles to that of G. inflata. On the other hand, the constituent property of G. uralensis is not similar to that of G. glabra or G. inflata and is characterized by a wide content variation of the six constituents compared to those of G. glabra and/or G. inflata. The mean contents of liquiritin, isoliquiritin, or liquilitigenin in G. uralensis are significantly higher than those of G. glabra or G. inflata. Therefore, the licorice species should be selected depending on these constituent properties for the traditional Chinese medicines or the Japanese Kampo medicines. Additionally, glycycoumarin, glabridin, and licochalcone A were reconfirmed as the species-specific typical constituents of G. uralensis, G. glabra, and G. inflata respectively. Therefore, it is resulted that the determination of the three species-specific constituents may be useful for the species identification of licorice. However, since 6% of licorice examined and hybrids were exceptions to the rule, their genetic information is necessary for the accurate species identification of licorice.     

Risk and safety assessment on the consumption of Licorice root (Glycyrrhiza sp.), its extract and powder as a food ingredient, with emphasis on the pharmacology and toxicology of glycyrrhizin

Licorice (or 'liquorice') is a plant of ancient origin and steeped in history. Licorice extracts and its principle component, glycyrrhizin, have extensive use in foods, tobacco and in both traditional and herbal medicine. As a result, there is a high level of use of licorice and glycyrrhizin in the US with an estimated consumption of 0.027-3.6 mg glycyrrhizin/kg/day. Both products have been approved for use in foods by most national and supranational regulatory agencies. Biochemical studies indicate that glycyrrhizinates inhibit 11beta-hydroxysteroid dehydrogenase, the enzyme responsible for inactivating cortisol. As a result, the continuous, high level exposure to glycyrrhizin compounds can produce hypermineralocorticoid-like effects in both animals and humans. These effects are reversible upon withdrawal of licorice or glycyrrhizin. Other in vivo and clinical studies have reported beneficial effects of both licorice and glycyrrhizin consumption including anti-ulcer, anti-viral, and hepatoprotective responses. Various genotoxic studies have indicated that glycyrrhizin is neither teratogenic nor mutagenic, and may possess anti-genotoxic properties under certain conditions. The pharmacokinetics of glycyrrhizin have been described and show that its bioavailability is reduced when consumed as licorice; this has hampered attempts to establish clear dose-effect levels in animals and humans. Based on the in vivo and clinical evidence, we propose an acceptable daily intake of 0.015-0.229 mg glycyrrhizin/kg body weight/day.     

Comparative study of bioactive constituents in crude and processed Glycyrrhizae radix and their respective metabolic profiles in gastrointestinal tract in vitro by HPLC-DAD and HPLC-ESI/MS analyses

Two HPLC methods with diode array detection (HPLC-DAD) and electrospray ionization-mass spectrometry (HPLC-ESI/MS), respectively, were developed to investigate the differences of chemical constituents and their metabolism in gastrointestinal tract in vitro between two decoctions of crude and processed Glycyrrhizae radix. Total of eleven constituents (liquiritin apioside, liquiritin, licuraside, isoliquiritin, ononin, glycyrrhizin, liquiritigenin-7,4??-diglucoside, licorice saponin A3, 22??-acetoxylglycyrrhizic acid, licorice saponin G2, and yunganoside E2) were identified in the two decoctions, whereas lower contents of these constituents were usually found in the decoction of processed G. radix. Furthermore, these constituents were metabolized into their respective aglycons in human intestinal bacteria juice, and the metabolism ratios were all higher in processed G. radix decoction. No change was found in artificial gastric or intestinal juice. This study revealed that the processing can alter the contents of main constituents in crude G. radix and their metabolism in gastrointestinal tract, in which intestinal bacteria play an important role in the metabolism of licorice constituents.     

通过包合工艺改善制剂中甘草的特有甜味

目的:应用正交实验设计方法研究中药制剂中β-环糊精包合甘草的最佳工艺.方法:采用高效液相色谱法,以甘草酸含量测定[1]为指标,考察β-环糊精包合甘草工艺的最佳条件.结果:每片药物中包合甘草的β-环糊精最佳用量为0.008g、包合温度85℃、包合时间4小时,并能掩盖甘草的不良口味,不影响甘草酸的含量测定,该工艺可靠、简便...     

Anticariogenic activity of licorice and glycyrrhizine I: Inhibition of in vitro plaque formation by Streptococcus mutans

The effect of licorice and its active sweet component glycyrrhizin was tested on the growth and adherence to glass of the cariogenic Streptococcus mutans. Neither licorice nor glycyrrhizin promoted growth or induced plaque formation. In the presence of sucrose, glycyrrhizin did not affect bacterial growth, but the adherence (plaque formation) was markedly inhibited. At 0.5-1% glycyrrhizin, inhibition was almost complete. These results support our previous suggestions that glycyrrhizin might serve as an efficient vehicle for topical oral medications.     

甘草主成分及其提取物肠吸收差异的多元化分析

考察甘草苷、甘草酸及甘草提取物中相应成分的大鼠在体肠吸收差异。运用大鼠在体肠灌流模型,采用UPLC和HPLC法同时测定肠灌流液、胆汁和颈静脉血样中甘草苷和甘草酸浓度,并结合肠道酶共孵育实验及对两种成分表观油水分配系数的测定,多元化比较分析甘草中甘草苷、甘草酸及提取物中相应成分在大鼠体内的吸收代谢特性。结果发现,甘草苷、甘草酸及甘草提取物中相应成分在大鼠肠道的有效渗透系数Peff值均小于0.3,表明两种成分肠道吸收均较差。各成分在十二指肠、空肠、回肠和结肠中吸收均无显著性差异;各成分单体与甘草提取物中相应成分比较发现,甘草提取物中甘草酸的各肠段Peff值略有增加,但无显著性差异;甘草提取物中甘草苷在回肠段的Peff值有显著性提高(P<0.05),其余肠段均无显著性差异,提示甘草提取物中其他成分促进甘草苷的吸收,但对甘草酸无明显影响。同时收集的提取物及单体的胆汁、血浆及肠道酶共孵育样品中均未检测到甘草苷、甘草酸及代谢产物,表明提取物中其他成分对于两种成分的影响在短时间(1～3 h)内可能未能提高其吸收入血及胆汁中的含量。     

甘草中异戊烯基类黄酮的富集及高效液相色谱-质谱联用分析

目的:对甘草中异戊烯基类黄酮部位的富集工艺研究及色谱峰定性分析,了解甘草中异戊烯基类黄酮的化学成分。方法:以浸膏得率及总黄酮含量为衡量指标,对甘草中的异戊烯基类黄酮进行富集;通过高效液相色谱-质谱联用法对甘草中异戊烯基类黄酮进行分析。结果:结合文献对图谱的色谱峰进行鉴定,推断出具异戊烯基基团的黄酮有23种,确定化学结构的有12种。为下一步甘草异戊烯基类黄酮部位研究打下了基础。