Li Yao-Li, Shang Ming-Ying, Geng Chang-An, Zhang Xue-Mei, Chen Ji-Jun, Cai Shao-Qing
State Key Laboratory of Phytochemistry and Plant Resources in West China, Kunmning Institute of Botany, Chinese Academy of Sciences, Kunming 650204, China.
Zhongguo Zhong Yao Za Zhi. 2013 May;38(9):1394-400.
To develop an HPLC method for simultaneous determination of swertiamarin, gentiopicroside, sweroside, mangiferin, erythrocentaurin, and to detect these five constituents in eight Qingyedans derived from Swertia mileensis, S. cincta, S. patens, S. punicea, S. delavayi, S. nervosa, S. macrosperma and S. yunnanensis.
The separation was carried out on a Thermo BDS Hypersil C18 (4. 6 mm x 250 mm, 5 microm) column eluted with mobile phase of water containing 0. 1% phosphoric acid and methanol (B) in gradient program (0-10 min, 18%-20% B; 10-30 min, 20%-35% B; 30-35 min, 35%-60% B). The column temperature was 32 degrees C , and the detection wavelength was set at 250, 260, 225 nm. The flow rate was 0. 7 mL . min-1 from 0 to 30 min, and be increased to 1. 0 mL . min-1 in 35 min.
The five compounds were well separated. The linear response ranges of swertiamarin, gentiopicroside, sweroside, mangiferin, erythrocentaurin were 0. 072-13. 39, 0. 1204. 518, 0. 060-5. 050, 0. 025-1. 518, and 0. 031-0. 210 microg, respectively. The mean recoveries of five compounds were 97.03% -102. 7% (RSD 1. 8% -6.2% ). There are swertiamarin, gentiopicroside and sweroside in most samples, and mangiferin in half samples. But erythrocentaurin was only detected in a few samples. The contents of five compounds were different in different samples. The contents of swertiamarin in S. mileensis, S. patens, S. yunnanensis and S. delavayi are up to 34. 47-118.05 mg . g-1, the contents of gentiopicroside are up to 25. 91 mg . g-1 in S. cincta. In S. puncea all contents of swertiamarin, gentiopicroside, sweroside and mangiferin are higher, especially the content of sweroside. There are Xiao-Qingyedans and Da-Qingyedans called in markets, and they can be identified by the contents of swertiamarin, gentiopicroside and sweroside. S. punicea can be identified by the content of sweroside, and the ratio gentiopicroside/total content can be used for identification of S. cincta from other seven Qingyedan species.
The method was certified to be accurate and reliable and can be used for identification and quality evaluation of traditional Chinese medicine Qingyedan derived from Swertia species.
建立高效液相色谱法同时测定獐牙菜苦苷、龙胆苦苷、獐牙菜苷、芒果苷、红百金花素，并检测来自青叶胆、显脉獐牙菜、湿生獐牙菜、紫红獐牙菜、川东獐牙菜、抱茎獐牙菜、大籽獐牙菜和云南獐牙菜的8种青叶胆中这5种成分的含量。
采用Thermo BDS Hypersil C18（4.6 mm×250 mm，5μm）色谱柱，以含0.1%磷酸的水和甲醇（B）为流动相进行梯度洗脱（0 - 10 min，18% - 20%B；10 - 30 min，20% - 35%B；30 - 35 min，35% - 60%B）。柱温32℃，检测波长设定为250、260、225 nm。0至30 min流速为0.7 mL·min⁻¹，35 min时增至1.0 mL·min⁻¹。
5种化合物分离良好。獐牙菜苦苷、龙胆苦苷、獐牙菜苷、芒果苷、红百金花素的线性响应范围分别为0.072 - 13.39、0.120 - 4.518、0.060 - 5.050、0.025 - 1.518、0.031 - 0.210μg。5种化合物的平均回收率为97.03% - 102.7%（RSD 1.8% - 6.2%）。多数样品中含有獐牙菜苦苷、龙胆苦苷和獐牙菜苷，半数样品中含有芒果苷。但红百金花素仅在少数样品中检测到。不同样品中5种化合物的含量不同。青叶胆、湿生獐牙菜、云南獐牙菜和川东獐牙菜中獐牙菜苦苷含量高达34.47 - 118.05 mg·g⁻¹，显脉獐牙菜中龙胆苦苷含量高达25.91 mg·g⁻¹。在紫红獐牙菜中，獐牙菜苦苷、龙胆苦苷、獐牙菜苷和芒果苷的含量均较高，尤其是獐牙菜苷。市场上有小青叶胆和大青叶胆之分，可通过獐牙菜苦苷、龙胆苦苷和獐牙菜苷的含量进行鉴别。紫红獐牙菜可通过獐牙菜苷的含量进行鉴别，龙胆苦苷与总含量的比值可用于鉴别显脉獐牙菜与其他7种青叶胆。
该方法准确可靠，可用于獐牙菜属中药青叶胆的鉴别和质量评价。
