正交试验法优选金茵利胆口服液水提工艺

目的:优选金茵利胆口服液的水提工艺。方法:采用正交试验法,以加水量、提取时间、提取次数、浸泡时间为考察因素,出膏率、绿原酸、柚皮苷和新橙皮苷为考察指标,采用高效液相色谱法对绿原酸、柚皮苷和新橙皮苷进行含量测定,以筛选出最佳水提工艺。结果:金茵利胆口服液的最佳水提工艺为:处方量药材加8倍量水,浸泡1 h,煎煮2次,每次1.5 h。结论:该水提工艺稳定、合理,为该制剂的生产工艺和质量控制研究提供研究基础。     

Effect of hesperidin and neohesperidin from bittersweet orange (Citrus aurantium var. bigaradia) peel on indomethacin-induced peptic ulcers in rats

Hesperidin and neohesperidin are the major flavanones isolated from bittersweet orange. It was recently reported that they have potent anti-inflammatory effects in various inflammatory models. In the present study, the effects of hesperidin and neohesperidin on indomethacin-induced ulcers in rats and the underlying mechanisms were investigated. Gastric ulcers were induced in rats with a single dose of indomethacin. The effects of pretreatment with hesperidin and neohesperidin were assessed in comparison with omeprazole as reference standard. Ulcer index, gene expression of gastric cyclooxygenase-2 (COX-2), tumor necrosis factor alpha (TNF-α), lipid peroxidation product, malondialdhyde (MDA), and reduced glutathione (GSH) content in stomach were measured. Furthermore, gross and histopathological examination was performed. Our results indicated that both hesperidin and neohesperidin significantly aggravated gastric damage caused by indomethacin administration as evidenced by increased ulcer index and histopathological changes of stomach.     

UPLC同时测定清热止咳颗粒中7种黄酮类成分含量

目的 建立同时测定清热止咳颗粒中芸香柚皮苷、柚皮苷、橙皮苷、新橙皮苷、黄芩苷、木蝴蝶素A-7-葡萄糖醛酸苷、汉黄芩苷含量的方法。方法 采用超高效液相色谱法。采用ACQUITY UPLC BEH C₁₈色谱柱（2.1 mm×100 mm,1.7 μm）,流动相乙腈-0.1%甲酸水溶液(梯度洗脱) ,流速0.3 mL·min^-1,检测波长280 nm,柱温30 ℃。结果 芸香柚皮苷、柚皮苷、橙皮苷、新橙皮苷、黄芩苷、木蝴蝶素A-7-葡萄糖醛酸苷、汉黄芩苷分别在0.324～6.480（r=0.999 9）,1.337～26.75（r=0.999 7）,0.687～13.80（r=0.999 6）,1.347～26.95（r=0.999 8）,2.770～55.40（r=0.999 2）,0.330～6.600（r=0.999 8）,0.242～4.840（r=0.999 6）μg·mL^-1范围内与峰面积呈良好的线性关系;并且芸香柚皮苷、柚皮苷、橙皮苷、新橙皮苷、黄芩苷、木蝴蝶素A-7-葡萄糖醛酸苷、汉黄芩苷平均加样回收率分别为99.41%、101.80%、99.42%、98.47%、101.71%,102.27%和97.64%,RSD范围分别是2.32%、1.69%、2.38%、2.45%、1.24%、1.77%和1.91%。结论 建立的超高效液相色谱(UPLC)方法操作简便、稳定、准确且可行,可用于清热止咳颗粒的质量控制。     

Taste responses to neohesperidin dihydrochalcone in rats and baboon monkeys

Preference-aversion behavior to solutions containing neohesperidin dihydrochalcone (NHDHC) was studied rats and baboon monkeys. Electrophysiological responses evoked by application of NHDHC solutions to taste receptors innervated by the chorda tympani and the glossopharyngeal nerves were also measured. As a group, rats were indifferent to solutions containing up to 1.2×10^?3 M NHDHC in short and long-term preference tests. A solution containing the very high concentration of 8.2×10^?3 M NHDHC was consumed less than water by all rats. The aversive behavior of rats to the 8.2×10^?3 M NHDHC solution appeared to be due to taste quality rather than olfaction. When percent preferences were calculated on an individual basis for the long-term preference tests, 59% of the rats were indifferent to solutions containing up to 1.2×10^?3 M NHDHC, 33% of the animals found this solution aversive and less than 8% showed preference. Behavioral responses to a solution of 3.4×10^?4 M aspartame also varied considerably among rats. The electrophysiological data were in line with the behavioral responses suggesting weak taste responses for NHDHC in rats. More pronounced responses observed in the glossopharyngeal nerve as compared to the chorda tympani. Baboon monkeys showed a strong preference for solutions containing 1.6×10^?5 M–1.6×10^?3 M NHDHC. A solution of 1.6×10^?2 M was consumed to a lesser extent than water. It is concluded that baboon monkeys present a better experimental model than rats for investigating the sweetness of NHDHC.     

HPLC separation of naringin, neohesperidin and their C-2 epimers in commercial samples and herbal medicines

Flavanone glycosides, such as naringin and neohesperidin, are distributed in some Citrus species and have a chiral center in the C-2 position of the flavanone moiety. Naringin and neohesperidin (2S-form) were separated from the corresponding C-2 epimers (2R-epi-form) by normal-phase HPLC using a polysaccaride-derived chiral stationary phases (CSPs), CHIRALPAK^® IB. The analyses of commercial samples of naringin revealed that the relative ratios of naringin to the C-2 epimer were 29–89%. In the case of a commercial sample of neohesperidin, the relative ratio of the neohesperidin (2S-form) is 84%.

The HPLC application to Citrus species used as crude drugs in Japan (Kijitsu, Kikoku and Tohi) showed that the relative ratios of naringin to the C-2 epimer were 75–93% in Kijitsu, 74–79% in Kikoku and 54–64% in Tohi. However, there is a quite small ratio of the (2R)-epi-neohesperidin in Citrus. This result suggested that the averages of relative ratio of (2S)-naringin in Citrus species reduced according to the maturity of fruits (Kijitsu < Kikoku < Tohi). Since the relative ratios of (2S)-naringin of dry extracts of 5 Kampo formulations (including Kijitsu or Kikoku) decreased to 42–54%, the conversion from naringin to the (2R)-epimer might be enhanced during the decoction process of the formulations.     

高效液相色谱法测定枳壳中新橙皮苷的含量

目的:建立HPLC测定枳壳中新橙皮苷含量的方法。方法:选用色谱柱为Hypersil C18(250 mm×4.6 mm,5 μm); 流动相为乙腈0.5%磷酸水(pH=3)(19:81);检测波长283 nm;流速1.0 ml/min;柱温25℃。结果:新橙皮苷进样量在0.155-0.930μg范围内与峰面积呈良好线性关系(r=0.999 1,n=6);新橙皮苷的加样回收率103.12%(RSD= 1.22%)。结论:该法简便、准确、可靠,可用于枳壳药材的质量控制。     

The influence of compatibility of traditional Chinese medicine on the pharmacokinetic of main components in Fructus Aurantii

Ethnopharmacological relevance

Fructus Aurantii is one of the commonly used traditional Chinese medicines and the compatibility of Fructus Aurantii, Radix et Rhizoma Rhei and Semen Crotonis Pulveratum were the main components in a Chinese traditional patent medicine Weichang'an pill. Naringin and neohesperidin are two main flavonoid glycosides in the extract of Fructus Aurantii. The pharmacokinetics of naringin and neohesperidin, as the main marker constituents were analysis by a simple and accurate method.

Materials and Methods

In the present study, pharmacokinetic differences of two flavonoid glycosides were detected after oral administration of the extract of single herb and different compatibility of Fructus Aurantii, Radix et Rhizoma Rhei and Semen Crotonis Pulveratum which at doses of approximately 700 mg/kg naringin and 300 mg/kg neohesperidin.

Results

It indicated that the pharmacokinetic process of naringin and neohesperidin in four groups were demonstrated bimodal phenomenon after oral administration. Comparing to pharmacokinetic parameters of the single herbs administration, there were no significant difference after the compatibility of Fructus Aurantii and Radix et Rhizoma Rhei. However, the absorption of two flavonoid glycosides was increased dramatically after the compatibility of Fructus Aurantii or Semen Crotonis Pulveratum. The same results were attained after co-administration of the extract of three herbs.

Conclusion

Some components in the extract of Semen Crotonis Pulveratum promoted the absorption of naringin and neohesperidin.     

正交试验优选枳实中新橙皮苷的提取工艺

目的 优选枳实中新橙皮苷的最佳提取工艺.方法 采用正交试验,以新橙皮苷含量为指标,考察溶剂浓度、溶剂用量、回流时间、回流次数对枳实中新橙皮苷提取工艺的影响.结果 提取次数为影响枳实中新橙皮苷提取效果的主要因素.确定提取工艺为6倍量的70%乙醇提取3次,每次1 h.结论 优选的提取工艺稳定,提取率高.     

HPLC法测定枳术丸中4种有效成分的含量

目的建立高效液相色谱法同时测定枳术丸中白术内酯Ⅰ等4种成分的含量。方法采用反相高效液相方法分离检测柚皮苷、橙皮苷及新橙皮苷和白术内酯Ⅰ。色谱柱:symmtryC18(250mm×4.6mm,5μm);以甲醇-0.1%磷酸溶液(梯度洗脱)为流动相;测定波长:283nm和224nm;流速:1.0mL·min-1;柱温:30℃;进样量:10μL。结果测定3批枳术丸中,柚皮苷、橙皮苷、新橙皮苷及白术内酯Ⅰ含量分别为10.77±0.61、1.16±0.13、9.41±0.09、0.38±0.02mg·g-1,并且4种成分得到完全分离,柚皮苷平均回收率为98.81%(RSD=1.11%,n=6);橙皮苷平均回收率为100.63%(RSD=1.90%,n=6);新橙皮苷平均回收率为99.32%(RSD=1.44%,n=6);白术内酯Ⅰ平均回收率为99.65%(RSD=1.53%,n=6)。结论该方法准确、可靠、重复性好,可作为枳术丸质量控制方法。     

HPLC法同时测定胃苏颗粒中柚皮苷、橙皮苷和新橙皮苷的含量

目的:建立同时测定胃苏颗粒中柚皮苷、橙皮苷和新橙皮苷含量的方法。方法:采用高效液相色谱法。色谱柱为Agilent Zorbax SB-C18柱,流动相为甲醇-醋酸-水(35∶4∶61,V/V/V),流速为1.0 ml/min,检测波长为283 nm,进样量为5μl,柱温为35℃。结果:柚皮苷、橙皮苷、新橙皮苷检测质量浓度分别在9.413～188.3、4.929～98.59、5.112～102.2μg/ml范围内与峰面积积分值呈良好的线性关系(r=0.999 9);精密度、稳定性、重复性试验的RSD≤1.3%;平均加样回收率分别为101.08%、99.89%、101.46%,RSD分别为0.62%、1.35%、0.45%(n=6)。结论:该方法简便、准确、重复性好,适用于测定胃苏颗粒中3种黄酮苷类成分的含量。