HPLC-DAD变波长法同时测定博尔宁胶囊中12种指标成分

Determination of 12 kinds of index components in Boerning Capsules by HPLC-DAD variable wavelength method

目的建立HPLC-DAD波长切换联合梯度洗脱法同时测定博尔宁胶囊中12种指标成分毛蕊异黄酮葡萄糖苷、澳洲茄碱、澳洲茄边碱、重楼皂苷VII、重楼皂苷VI、重楼皂苷II、重楼皂苷I、特女贞苷、迷迭香酸、大黄酸、大黄酚和大黄素的量。方法采用HPLC-DAD法,Atlantis T3 C_(18)(250 mm×4.6 mm,5μm)色谱柱;乙腈-甲醇-0.1%甲酸水溶液为流动相,体积流量0.8 m L/min,梯度洗脱;变波长扫描;进样量为20μL。结果 12种指标成分毛蕊异黄酮葡萄糖苷、澳洲茄碱、澳洲茄边碱、重楼皂苷VII、重楼皂苷VI、重楼皂苷II、重楼皂苷I、特女贞苷、迷迭香酸、大黄酸、大黄酚和大黄素分别在1.97~19.70μg/m L(r=0.999 2)、1.022~10.220μg/m L(r=0.999 3)、0.982~9.820μg/m L(r=0.999 1)、1.1~11.0μg/m L(r=0.999 6)、1.154~11.540μg/m L(r=0.999 8)、1.114~11.140μg/m L(r=0.999 5)、1.102~11.020μg/m L(r=0.999 3)、2.768~27.680μg/m L(r=0.999 3)、3.04~30.40μg/m L(r=0.999 6)、3.379~33.790μg/m L(r=0.999 5)、3.286~32.860μg/m L(r=0.999 4)、3.507~35.070μg/m L(r=0.999 7)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100.08%、1.27%;98.11%、1.15%;99.68%、1.13%;101.38%、0.87%;101.87%、0.95%;100.53%、0.74%;98.52%、0.83%;99.52%、0.88%;97.84%、1.33%;98.31%、0.71%;99.66%、0.57%;101.73%、1.41%。12批次供试品中12种指标成分质量分数分别为0.085~0.1_(18) mg/g、0.065~0.085 mg/g、0.051~0.075 mg/g、1.822~1.888 mg/g、1.532~1.599 mg/g、1.027~1.148 mg/g、2.420~2.621 mg/g、6.428~6.937 mg/g、0.258~0.289 mg/g、0.122~0.143mg/g、0.159~0._(18)4 mg/g、0.222~0.273 mg/g。结论建立的HPLC-DAD波长切换联合梯度洗脱法同时测定博尔宁胶囊中的12种成分,方法操作简便、快速、准确,可作为博尔宁胶囊全面可靠的质量控制方法。

Objective To develop an HPLC-DAD wavelength switching combined with gradient elution method for the determination of the contents of calycosin 7-O-β-D-glucopyranoside, solasonine, solamargine, chonglou saponin I, chonglou saponin II, chonglou saponin VI, chonglou saponin VII, specnuezhenide, rosmarinic acid, rhein, chrysophanol, and emod in Boerning Capsules（BC） simultaneously. Methods The chromatographic separation was achieved on an Atlantis T3 C_（18）（250 mm × 4.6 mm, 5 μm） with acetonitrile-methanol（A）-0.1% formic acid solution（B） as mobile phase at the flow rate of 0.8 m L/min for gradient elution; variable wavelength method; sample quantity was 20 μL. Results The 12 active components were well separated and showed good linearity, such as calycosin 7-O-β-D-glucopyranoside, solasonine, solamargine, polyphyllin I, polyphyllin II, polyphyllin VI, polyphyllin VII, specnuezhenide, rosmarinic acid, rhein, chrysophanol, and emod 1.97—19.70 μg/m L（r = 0.999 2）, 1.022—10.220 μg/m L（r = 0.999 3）, 0.982—9.820 μg/m L（r = 0.999 1）, 1.102—11.020 μg/m L（r = 0.999 3）, 1.114—11.140 μg/m L（r = 0.999 5）, 1.154—11.540 μg/m L（r = 0.999 8）, 1.114—11.140 μg/m L（r = 0.999 5）, 2.768—27.680 μg/m L（r = 0.999 3）, 3.04—30.40 μg/m L（r = 0.999 6）, 3.379—33.790 μg/m L（r = 0.999 5）, 3.286—32.860 μg/m L（r = 0.999 4）, and 3.507—35.070 μg/m L（r = 0.999 7）. The precision and repeatability were good, and RSD values were less than 2.0%. The average recoveries and the corresponding RSD values were 100.08%（1.27%）, 98.11%（1.15%）, 99.68%（1.13%）, 101.38%（0.87%）, 101.87%（0.95%）, 100.53%（0.74%）, 98.52%（0.83%）, 99.52%（0.88%）, 97.84%（1.33%）, 98.31%（0.71%）, 99.66%（0.57%）, and 101.73%（1.41%）, respectively. The contents of 12 batches of the tevelve active components were 0.085—0.1_（18）, 0.065—0.085, 0.051—0.075, 1.822—1.888, 1.532—1.599, 1.027—1.148, 2.420 — 2.621, 6.428 — 6.937, 0.258 — 0.289, 0.122 — 0.143, 0.159 — 0._（18）4, and 0.222 — 0.273 mg/g. Conclusion An HPLC wavelength switching combined with gradient elution method has been successfully established for simultaneous determination of 12 components in BC. The method is simple, quick, accurate, and helpful for the quality control of BC.