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目的:建立同时测定牛黄清胃丸中绿原酸、栀子苷、连翘酯苷A、柚皮苷、黄芩苷、甘草酸铵、大黄酚7个成分含量的方法。方法:采用高效液相色谱(HPLC)-波长切换法。色谱柱为Agilent ZORBAX SB-C18,流动相为乙腈(A)-0.1%磷酸溶液(B),梯度洗脱;流速为1.0 mL/min;检测波长分别为348 nm(绿原酸)、238 nm(栀子苷)、330 nm(连翘酯苷A)、280 nm(柚皮苷和黄芩苷)、237 nm(甘草酸铵)、254 nm(大黄酚);柱温为30 ℃;进样量为10 μL。结果:绿原酸、栀子苷、连翘酯苷A、柚皮苷、黄芩苷、甘草酸铵和大黄酚的进样量线性范围分别为0.011 67~0.233 4 μg(r=0.999 4)、0.042 91~0.858 1 μg(r=0.999 4)、0.125 0~2.500 μg(r=0.999 9)、0.118 0~2.360 μg(r=0.999 9)、0.119 6~2.392 μg(r=0.999 7)、0.030 57~0.611 4 μg(r=0.999 6)和0.006 201~0.124 0 μg(r=0.999 4),定量限分别为1.167、0.858、1.250、1.180、1.196、0.611、0.620 μg/mL;精密度试验的RSD分别为0.98%、1.04%、0.59%、1.50%、0.83%、1.24%和1.32%(n=6);稳定性试验的RSD分别为1.21%、0.97%、1.42%、0.71%、0.98%、1.87%和1.63%(n=6,12 h);平均回收率分别为98.32%、98.11%、98.81%、98.50%、98.30%、98.16%和97.83%,RSD分别为1.37%、1.41%、0.64%、1.01%、1.18%、1.16%和1.16%(n=6)。结论:建立的含量测定方法操作简单、重复性好,可用于牛黄清胃丸的质量控制。
OBJECTIVE: To develop a method for simultaneous determination of 7 components of Niuhuang qingwei pills as chlorogenic acid, geniposide, forsythoside A, narirutin, baicalin, ammonium glycyrrhetate, chrysophanol. METHODS: HPLC-wavelength switching method was adopted. The determination was performed on Agilent ZORBAX SB-C18 column with mobile phase consisted of acetonitrile (A)-0.1% phosphoric acid (B) gradient elution at the flow rate of 1.0 mL/min. The detection wavelengths were set at 348 nm (chlorogenic acid), 238 nm (geniposide), 330 nm (forsythoside A), 280 nm (narirutin and baicalin), 237 nm (ammonium glycyrrhetate), 254 nm (chrysophanol). The column temperature was 30 ℃, and sample size was 10 μL. RESULTS: The linear ranges of chlorogenic acid, geniposide, forsythoside A, narirutin, baicalin, ammonium glycyrrhetate, chrysophanol were 0.011 67-0.233 4 μg (r=0.999 4), 0.042 91-0.858 1 μg (r=0.999 4), 0.125 0-2.500 μg (r=0.999 9), 0.118 0- 2.360 μg (r=0.999 9), 0.119 6-2.392 μg (r=0.999 7), 0.030 57-0.611 4 μg (r=0.999 6), 0.006 201-0.124 0 μg(r=0.999 4), respectively; the limits of quantitation were 1.167, 0.858, 1.250, 1.180, 1.196, 0.611, 0.620 μg/mL, respectively; RSDs of precision tests were 0.98%, 1.04%, 0.59%, 1.50%, 0.83%, 1.24% and 1.32% (n=6), respectively. RSDs of stability tests were 1.21%, 0.97%, 1.42%, 0.71%, 0.98%, 1.87% and 1.63% (n=6, 12 h), respectively. Average recoveries were 98.32%, 98.11%, 98.81%, 98.50%, 98.30%, 98.16% and 97.83%, and the RSDs were 1.37%, 1.41%, 0.64%, 1.01%, 1.18%, 1.16% and 1.16% (n=6), respectively. CONCLUSIONS: Established method is easy and reproducible. It can be used for the quality control of Niuhuang qingwei pills.
牛黄清胃丸高效液相色谱-波长切换法绿原酸栀子苷连翘酯苷A柚皮苷黄芩苷甘草酸铵大黄酚含量测定
Niuhuang qingwei pillsHPLC-wavelength switching methodChlorogenic acidGeniposideForsythoside ANarirutinBaicalinAmmonium glycyrrhetateChrysophanolContent determination
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