门氏柴胡理中颗粒的薄层鉴别及含量测定

目的:建立门氏柴胡理中颗粒定性定量质量标准控制方法。方法:采用薄层色谱法对门氏柴胡理中颗粒制剂中柴胡、黄芩进行薄层鉴别,采用高效液相色谱法(HPLC)对柴胡理中颗粒中的黄芩苷进行含量测定,色谱柱为Diamonsil-C_(18)色谱柱(200 mm×4.6 mm,5.0μm),流动相为甲醇-0.1%醋酸水溶液进行梯度洗脱,流速为1.0 mL/min,检测波长为280 nm,柱温为25℃,进样量为10μL.结果:柴胡、黄芩薄层色谱斑点分离清晰,阴性均无干扰;回归方程为y=2 907 186.561 0x+21 224.436 6(r=0.999 8),黄芩苷在0.12～1.20μg范围内具有良好的线性关系。平均加样回收率为99.45%,RSD为2.48%。结论:建立的方法方便、可靠,可作为门氏柴胡理中颗粒的薄层鉴别和含量测定质量控制方法。     

UPLC法测定黄芩药材中黄芩苷的含量

目的:用超高效液相色谱法测定黄芩中黄芩苷的含量。方法:参考已有文献方法对原高效液相色谱方法进行转换并优化为超高效液相色谱方法。色谱柱为Shim-pack XR-ODS C18柱(4.6mm×150mm,2.2μm);流动相:甲醇-水-冰醋酸(45∶55∶1);流速:0.6mL.min-1;检测波长:274nm;柱温:45℃。结果:黄芩苷的线性范围为29.86～238.85ng(r=0.9999);平均回收率(n=6)为101.2%,RSD为1.8%;1个分析流程大约需要3min。结论:与高效液相色谱法相比,超高效液相色谱法测定黄芩中黄芩苷含量,加快了分析速度,提高了分析效率,减少了有机溶剂的使用,并且得到了更高的分析灵敏度。     

高效液相色谱法测定黄芩颗粒饮片中黄芩苷含量

目的:应用高效液相色谱法测定黄芩颗粒饮片中黄芩苷含量。方法:色谱柱为Alltech色谱柱(4 6mm×150mm,5μm),流动相为甲醇-水-磷酸( 50∶50∶0 2 ),检测波长为274nm,流速为1 0ml/min,柱温30℃。结果:黄芩苷在0 .094 ~0. 564μg范围内进样量与峰面积呈良好线性关系(r=0. 9996),平均加样回收率为97. 63%,RSD=0 .45% (n=5)。结论:高效液相色谱法测定结果准确、灵敏、重现性好。     

HPLC测定小儿清肺化痰颗粒中黄芩苷的含量

目的:建立高效液相色谱法测定小儿清肺化痰颗粒中黄芩苷的含量方法。方法:Agilent XDB C18色谱柱(4.6 mm×250 mm,4.6μm);流动相:甲醇-0.4%磷酸溶液(47∶53);流速1.0 mL.min-1;检测波长280 nm柱温:40℃。结果:回归方程为A=34 884.9C-19 478.8,R2=1.000 0。黄芩苷在1.482～148.2 mg.L-1线性关系良好,平均回收率为97.8%,RSD 1.9%(n=6)。结论:该方法简便、准确、重复性好,可作为小儿清肺化痰颗粒黄芩苷含量的测定。     

HPLC测定芩栀胶囊中黄芩苷和栀子苷的含量

目的:建立芩栀胶囊中黄芩苷和栀子苷的含量测定方法。方法:采用高效液相色谱法进行测定。黄芩苷的测定条件为:色谱柱:KR100-5 C18柱(5μm,4.6 mm×250 mm);流动相:甲醇-水-磷酸(47∶53∶0.2),流速:1 mL/m in;检测波长280 nm。栀子苷的测定条件为:色谱柱:KR100-5C18柱(5μm,4.6 mm×250 mm);流动相:乙腈-水(14∶86):流速:1 mL/m in;检测波长:238 nm。结果:黄芩苷在0.300 4~1.502 0μg范围内线性关系良好,r=0.999 920,平均加样回收率为99.07%,RSD=0.93%。栀子苷在0.150 2~0.751 0μg范围内线性关系良好,r=0.999 986,平均加样回收率为99.06%,RSD=1.09%。结论:该方法简便、快速、准确,可用于芩栀胶囊的质量控制。     

HPLC法测定小儿咳喘颗粒中黄芩苷

目的建立HPLC法测定小儿咳喘颗粒中黄芩苷的方法,控制产品的质量。方法采用高效液相色谱法对小儿咳喘颗粒中的黄芩苷进行测定。色谱柱:Hypersil C18色谱柱(250 mm×4.6 mm,5μm),流动相为甲醇-水-磷酸(47∶53∶0.2),体积流量1 mL/min,检测波长280 nm;柱温室温。结果黄芩苷的线性范围分别为0.049 5～1.485 0μg,平均回收率为99.99%,RSD为0.52%。结论方法可行、重现性好,能有效地控制小儿咳喘颗粒的质量。     

福乐欣口服液中黄芩苷含量的测定研究

目的建立福乐欣口服液中黄芩苷含量测定方法。方法采用反相高效液相色谱法Symmetry C18柱(150 mm×4.6 mm5μm)分析柱;流动相:甲醇-0.006%磷酸(50∶50);流速1.0 ml.min-1;色谱柱温度25℃;检测波长278 nm;进样体积10μl;外标法计算含量。结果黄芩苷进样量在0.412-4.12μg范围内与色谱峰面积有良好的线性关系,r=0.999 5,回收率为99.48%,RSD=1.70%(n=5)。结论该方法简便、灵敏、可靠,可用于控制福乐欣口服液中黄芩苷含量。     

高效液相色谱法测定复方黄芩消炎片中黄芩苷的含量

目的:建立用高效液相色谱法测定复方黄芩消炎片中黄芩苷含量的方法。方法:采用Lichrospher色谱柱(C185μm,4.60 mm×250 mm),以甲醇-水-磷酸(47∶53∶0.20)为流动相,流速为1.00 mL.min-1,检测波长为280 nm。结果:黄芩苷的线性范围为(0.40-2.41)μg(r=0.999 9),平均回收率(n=5)为97.19%(RSD=2.00%)。结论:方法准确、可靠、重复性好。     

三黄栓质量标准研究

目的建立三黄栓的质量控制方法。方法采用薄层色谱法鉴别三黄栓中的黄连、黄芩、黄柏。采用高效液相色谱法测定制剂中黄芩苷、盐酸小檗碱的含量,色谱柱为Aglient ZORBAXSB-C18(4.6 mm×250 mm,0.45μm),甲醇-水-磷酸(47∶53∶0.2)为流动相,流速为1.0 mL/min,检测波长黄芩苷为280 nm、盐酸小檗碱为365 nm,柱温30℃。结果定性鉴别色谱特征明显。黄芩苷在0.248~2.48μg线性关系良好(r=0.999 9),平均回收率为99.77%,RSD=1.05%;盐酸小檗碱在0.336~1.68μg线性关系良好(r=0.999 9),平均回收率为97.74%,RSD=1.48%。结论该定性定量检测方法简便可行,可有效控制该制剂质量。     

清开灵片中黄芩苷体内药代动力学研究

目的 建立测定人血浆中黄芩苷浓度的高效液相色谱法.方法 采用HPLC法测定血浆中黄芩苷浓度.色谱柱:Agilent HC-C 18柱(250 mm×4.6 mm,5μm);流动相:甲醇-水-磷酸(47∶53∶0.2);检测波长280 nm;汉黄芩素作为内标,用HPLC法测定人血浆中黄芩苷浓度.结果 黄芩苷线性范围为0....     

HPLC-ELSD法测定麦冬药材及其提取物中果糖的含量

目的测定不同产地的川麦冬、浙麦冬药材及麦冬提取物中果糖的含量,为麦冬药材及其提取物的质量控制提供科学依据。方法采用HPLC-ELSD法测定麦冬药材及提取物中果糖的含量。结果川麦冬和浙麦冬药材中果糖的含量分别为11.32%～19.96%、3.85%～10.19%,川麦冬和浙麦冬提取物中果糖的含量分别为23.48%～25.82%、21.39%～23.29%。结论川麦冬药材中果糖的含量明显高于浙麦冬药材,川麦冬提取物中果糖含量与浙麦冬提取物基本一致。     

不同性别大鼠中药含药血清对成骨细胞增殖的影响

[目的]观察不同性别大鼠的中药续断含药血清对成骨细胞增殖的影响.[方法]1)制备大鼠含药血清:取雌性与雄性Wistar大鼠各12只,按性别随机分为4组,分别为:雄性给药组,雄性对照组,雌性给药组,雌性对照组,每组各6只,给药组按体质量(4.5g/kg)分别给予续断水提液灌胃、对照组按体质量分别给予等量生理盐水.灌胃1周后取血,离心提取血清.2)分离、培养大鼠的原代成骨细胞.3)将各组大鼠血清稀释为2.5%,5%和10%3个浓度分别培养大鼠成骨样细胞,采用噻唑蓝法(MTT)检测细胞增殖能力.[结果]血清浓度为2.5%时,雌性给药组(0.569 0±0.029 5)与雄性给药组(0.361 3±0.098 3)比较,有显著性差异(P<0.05),雌性给药组与雌性对照组(0.378 7±0.047 6)比较,也有显著性差异(P<0.01);当血清浓度为5%时,雌性给药组(0.879 3±0.100 7)与雄性给药组(O.356 0±0.048 2)比较,有显著性差异(P<0.01),雌性给药组与雌性对照组(0.386 3±0.018 0)比较,也存在显著性差异(P<0.01);当血清浓度为10%时,雌性给药组(0.856 0±0.017 5)与雌性对照组(0.381 7±0.198 6)比较,存在显著性差异(P<0.05).在各浓度的血清中,雄性给药组与雄性对照组之间均无显著性差异.[结论]大鼠的续断含药血清具有刺激成骨细胞增殖的作用.这种作用在雌性大鼠的血清中表达强于雄性大鼠.     

芍药及不同炮制品中芍药苷含量的比较研究

目的：比较赤芍（Radix Paeoniae Rubra）与白芍（Radix Paeoniae Alba）及不同加工炮制方法对赤芍、白芍饮片中芍药苷含量的影响。方法：采用HPLC法,色谱柱：SHIMADCIS ODSC18。流动相：乙腈-0．1％磷酸水溶液（15：85）。检测波长：230nm,流速：1．0ml／min。结果：芍药苷的含量：赤芍〉炒赤芍〉醋炙白芍〉酒炙白芍〉炒白芍〉白芍〉去皮白芍〉土炒白芍,白芍与赤芍芍药苷含量存在显著差异,白芍经酒炙、醋炙后含量显著高于生品,其中以醋炙品最高;炒黄品芍药苷略有升高,土炒品略有降低,去皮白芍低于不去皮白芍,但四者均无显著性差异。结论：赤芍与不去皮白芍中芍药苷含量存在显著差异,故以不去皮白芍作为赤芍代替品使用缺乏合理性。而白芍炮制方法中的醋炙和酒炙对芍药苷溶出有明显影响,清炒和土炒的影响并不显著。     

Enzyme-Linked Immunosorbent Assay and Immunochromatographic Strip for Rapid Detection of Atrazine in Three Medicinal Herbal Roots

Objectives: An enzyme?linked immunosorbent assay (ELISA) and colloidal gold?based immunochromatographic (ICG) strip assay will be developed for the rapid and high-throughput detection of atrazine (ATZ) in medicinal herbs. Methods: A monoclonal antibody against ATZ was obtained after the immunization of mice, cell fusion, and hybridoma screening, and the antibody was used to develop direct competitive ELISA (dcELISA) and the ICG strip assay. Results: Both dcELISA and ICG strip methods were established, optimized, and validated for the detection of ATZ in Salviae miltiorrhizae radix et rhizome, Astragali radix, and Isatidis radix. dcELISA had a half-maximum inhibition concentration of 10.56 ng/mL (Salviae miltiorrhizae radix et rhizome), 7.6 ng/mL (Astragali radix), and 8.15 ng/mL (Isatidis radix). The limit of detection (LOD) of the ICG strip assay was 12.5 ng/mL (Salviae miltiorrhizae radix et rhizome), 12.5 ng/mL (Astragali radix), and 6.25 ng/mL (Isatidis radix) in different herb matrices. Due to the recognition characteristics of the monoclonal antibody for the pesticides ATZ, propazine, sebuthylazine, and prometryn, the detection results of realsamples by the two immunoassays were confirmed by liquid chromatography–tandem mass spectrometry, which proved the accuracy and reliability of the established methods. Conclusion: The proposed dcELISA and ICG strip methods were suitable for the rapid, convenient, and high-throughput detection of ATZ in these medicinal herbs.     

肠安口服液的质量标准研究

目的：制定肠安口服液的质量标准。方法：采用薄层色谱法定性鉴别黄芩、芍药和甘草，用高效液相色谱法测定制剂中黄芩苷的含量。以C18化学键合硅胶为固定相，以甲醇-水-磷酸（50：50：0．3）为流动相，检测波长为280nm。结果：定性鉴别色谱特征明显，重现性好。黄芩苷在9．6～57．60μg／mL范围内呈良好的线性关系，r=0．9998，平均回收率为98．70％，RSD=3．10％。结论：上述方法可靠、实用，可有效地控制该制荆的质量。     

肠安口服液的质量标准研究

目的:制定肠安口服液的质量标准。方法:采用薄层色谱法定性鉴别黄芩、芍药和甘草,用高效液相色谱法测定制剂中黄芩苷的含量。以C18化学键合硅胶为固定相,以甲醇-水-磷酸(50∶50∶0.3)为流动相,检测波长为280nm。结果:定性鉴别色谱特征明显,重现性好。黄芩苷在9.6~57.6μg/mL范围内呈良好的线性关系,r=0.9998,平均回收率为98.70%,RSD=3.10%。结论:上述方法可靠、实用,可有效地控制该制剂的质量。     

野葛与粉葛总黄酮提取方法的比较

目的：野葛与粉葛总黄酮提取方法的比较。方法：分别采用传统回流提取法、超声提取法、微波辅助提取法对中药野葛和粉葛中总黄酮进行提取;以葛根素为对照品,采用紫外分光光度法测定提取物中总黄酮的含量;以总黄酮的含量为考察指标,优选出适宜的提取方法。结果：3种提取方法中,野葛超声提取法总黄酮含量最高,粉葛微波辅助提取法总黄酮含量最高,但回流提取法和超声提取法提取野葛与粉葛总黄酮时没有显著性差异。结论：提取野葛与粉葛黄酮时应采用不同的提取方法。     

陈修园女科用药“四害”

陈修园为清代著名医家,对于女科疾病研究有着深厚的造诣,著有《女科要旨》一书,书中认为唐代以后,许多用药之法,早已失其宗旨,与古法相异,若不识其药性,颠倒而用,则良药亦会变成毒药。丹参、益母草、何首乌、郁金四味为女科常用药,陈修园认为丹参若是用于产后调理,则容易攻乏无过,伤及脏器;益母草药力则不足,若是症重药轻,最终将不可治也;何首乌只适合截疟或坚肠;郁金并无解郁的作用,因此陈修园将此四味药列为女科四大害人之品。     

野葛与粉葛总黄酮提取方法的比较△

目的:野葛与粉葛总黄酮提取方法的比较。方法:分别采用传统回流提取法、超声提取法、微波辅助提取法对中药野葛和粉葛中总黄酮进行提取;以葛根素为对照品,采用紫外分光光度法测定提取物中总黄酮的含量;以总黄酮的含量为考察指标,优选出适宜的提取方法。结果:3种提取方法中,野葛超声提取法总黄酮含量最高,粉葛微波辅助提取法总黄酮含量最高,但回流提取法和超声提取法提取野葛与粉葛总黄酮时没有显著性差异。结论:提取野葛与粉葛黄酮时应采用不同的提取方法。     

Antispasmodic effect of shakuyakukanzoto extract on experimental muscle cramps in vivo: Role of the active constituents of Glycyrrhizae radix

Ethnopharmacological relevance

Shakuyakukanzoto (SKT) composed of Glycyrrhizae radix (G. radix) and Paeoniae radix (P. radix) has been traditionally used in Japan, Korea and China as an antispasmodic drug for the treatment of skeletal muscle cramps and intestinal cramps.

Aim of this study

To evaluate the antispasmodic activity of SKT and its two components, as well as to identify the key constituents of the components which mediate this effect in skeletal muscles in vivo.

Materials and methods

An experimental cramp model was constructed to evaluate the effects of peripherally-acting muscle relaxants on electrically-induced cramps under physiological conditions. This was accomplished by surgically isolating the motor supply to the gastrocnemius muscle in an anesthetized rat and delivering electrical stimuli to an isolated tibial nerve to induce tetanic contractions. We first tested dantrolene, a well-known peripherally-acting relaxant, to determine the sensitivity and reliability of our experimental model. We then evaluated the effects of SKT, P. radix, G. radix, and the eight active constituents of G. radix against tetanic contractions.

Results

We found that dantrolene (10 and 30 mg/kg, i.d.) rapidly and significantly inhibited tetanic contractions (P<0.01) irrespective of dose. SKT (0.5, 1.0, and 2.0 g/kg, i.d.) and G. radix (0.5 and 1.0 g/kg, i.d.) also significantly inhibited tetanic contractions (P<0.01) but in a dose-dependent manner owing to the actions of six of the eight active constituents in G. radix (liquiritin apioside, liquiritigenin, isoliquiritin apioside, isoliquiritigenin, glycycoumarin, and glycyrrhetinic acid, 20 μmol/kg, i.v.). These constituents, which include flavonoids, a triterpenoid, and a courmarin derivative, demonstrated temporal variations in their inhibitory activity. In contrast, P. radix (0.5 and 1.0 g/kg, i.d.) did not show a statistically significant antispasmodic effect in our study; however, we previously found that it had a significant antinociceptive effect.

Conclusions

Our findings show that SKT inhibits tetanic contractions in vivo and that G. radix is the main antispasmodic component due to the actions of its active constituents, thus supporting the traditional use of SKT. We further propose that SKT containing the antispasmodic G. radix and antinociceptive P. radix is a pharmaceutically elegant option for muscle cramps as treatment requires a two-pronged approach, i.e., inhibition of hyperexcitable skeletal tissues and modulation of the pain accompanying cramps.