摘要
目的建立HPLC梯度洗脱法同时测定天麻首乌片(TSP)中12种活性成分芍药苷、2,3,5,4′-四羟基二苯乙烯-2-O-β-D-葡萄糖苷(二苯乙烯苷)、天麻素、阿魏酸、毛蕊花糖苷、甘草苷、丹参酮IIA、特女贞苷、欧前胡素、芦丁、蟛蜞菊内酯和大黄素的量。方法采用HPLC法,Hypersil GOLD C18(250 mm×4.6 mm,5μm)色谱柱;乙腈-0.1%磷酸水溶液为流动相,体积流量1.0 m L/min,梯度洗脱;进样量为10μL。结果 12种活性成分芍药苷、二苯乙烯苷、天麻素、阿魏酸、毛蕊花糖苷、甘草苷、丹参酮IIA、特女贞苷、欧前胡素、芦丁、蟛蜞菊内酯和大黄素分别在3.12~31.20μg/m L(r=0.999 5)、4.98~49.80μg/m L(r=0.999 4)、1.05~10.50μg/m L(r=0.999 5)、0.99~9.90μg/m L(r=0.999 2)、1.11~11.10μg/m L(r=0.999 2)、3.24~32.40μg/m L(r=0.999 2)、3.63~36.30μg/m L(r=0.999 2)、1.13~11.30μg/m L(r=0.999 3)、1.9~19.0μg/m L(r=0.999 5)、1.55~15.50μg/m L(r=0.999 5)、1.48~14.80μg/m L(r=0.999 5)、102.8~1028.0μg/m L(r=0.999 9)质量浓度与峰面积具有较好的线性关系;精密度、重复性良好,RSD均小于2.0%;平均加样回收率和相应的RSD分别为99.88%、1.55%,101.25%、0.98%,99.67%、1.29%,102.04%、1.17%,101.17%、1.67%,98.27%、1.51%,100.28%、1.20%,99.11%、0.95%,98.49%、1.67%,101.57%、0.94%,102.37%、0.58%,97.89%、0.69%。12批次供试品中12种指标成分质量分数分别为1.241~1.261、2.116~2.133、0.540~0.558、0.077~0.099、0.089~0.110、1.111~1.134、0.158~0.183、1.375~1.399、0.342~0.372、0.542~0.571、0.648~0.672、45.05~45.93 mg/g。结论建立的HPLC梯度洗脱法可同时测定TSP中12种活性成分,该方法操作简便、快速、准确,可作为TSP全面可靠的质量控制方法。
Objective To develop an HPLC combined with gradient elution method for the determination of the contents of paeoniflorin, 2,3,5,4′-tetrahydroxystilbene-2-O-β-D-glucosid, gastrodin, ferulic acid, acteoside, liquiritin, tanshinone IIA, specnuezhenide, imperatorin, rutinum, wedelolactone, and emod in Tianma Shouwu Pills（TSP） at the same time. Methods The chromatographic separation was achieved on a Hypersil GOLD C18（250 mm × 4.6 mm, 5 μm） with acetonitrile（A）-0.1% phosphoric acid solution（B） as mobile phase at the flow rate of 1.0 mL/min for gradient elution; sample quantity was 10 μL. Results The linear relation between peak area and concentration of the twelve active components were good, which were 3.12—31.20 μg/mL（r = 0.999 5） paeoniflorin, 4.98—49.80 μg/mL（r = 0.999 4） 2,3,5,4′-tetrahydroxystilbene-2-O-β-D-glucosid, 1.05—10.50 μg/mL（r = 0.999 5） gastrodin, 0.99—9.90 μg/mL（r = 0.999 2） ferulic acid, 1.11—11.10 μg/mL（r = 0.999 2） acteoside, 3.24—32.40 μg/mL（r = 0.999 2） liquiritin, 3.63—36.30 μg/mL（r = 0.999 2） tanshinone IIA, 1.13—11.30 μg/mL（r = 0.999 3） specnuezhenide, 1.9—19.0 μg/mL（r = 0.999 5） imperatorin, 1.55—15.50 μg/mL（r = 0.999 5） rutinum, 1.48—14.80 μg/mL（r = 0.999 5） wedelolactone, and 102.8—1028.0 μg/mL（r = 0.9999） emodin. The precision and repeatability were good, and RSD values were less than 2.0%. Moreover, the average recoveries and the corresponding RSD values were 99.88%（1.55%）, 101.25%（0.98%）, 99.67%（1.29%）, 102.04%（1.17%）, 101.17%（1.67%）, 98.27%（1.51%）, 100.28%（1.20%）, 99.11%（0.95%）, 98.49%（1.67%）, 101.57%（0.94%）, 102.37%（0.58%）, and 97.89%（0.69%）, respectively. The contents of 12 batches of the 12 active components were 1.241—1.261, 2.116—2.133, 0.540—0.558, 0.077—0.099, 0.089—0.110, 1.111—1.134, 0.158—0.183, 1.375—1.399, 0.342—0.372, 0.542—0.571, 0.648—0.672, and 45.05—45.93 mg/g. Conclusion An HPLC combined with gradient elution method has been successfully established for simultaneous determination of 12 components in TSP. The method is simple, quick, accurate, and helpful for the quality control of TSP.
出处
《中草药》
CAS
CSCD
北大核心
2017年第22期4682-4687,共6页
Chinese Traditional and Herbal Drugs
