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目的:建立同时测定3种不同大黄炮制品中没食子酸、桂皮酸、儿茶素含量的方法。方法:采用反相高效液相色谱法。色谱柱为Thermo ScientificTM Hypersil GOLD Dim,流动相为甲醇-0.1%磷酸水溶液(梯度洗脱),检测波长为278 nm,流速为1.0 mL/min,柱温为30 ℃,进样量为10 μL。结果:没食子酸、桂皮酸、儿茶素的检测进样量线性范围分别为0.126 2~1.262 0 μg(r=0.999 9)、0.036 2~0.362 0 μg(r=0.999 9)、0.177 9~1.779 4 μg(r=0.999 8);定量限分别为25.4、28.2、62.5 ng,检测限分别为6.2、3.6、11.8 ng;精密度、稳定性、重复性、耐用性试验的RSD均小于3%;加样回收率分别为94.64%~102.71%(RSD=2.74%,n=9)、95.35%~102.49%(RSD=2.44%,n=9)、93.65%~103.66%(RSD=3.27%,n=9)。含量测定结果显示,熟大黄中没食子酸、桂皮酸含量较高,两种成分含量大小顺序均依次为熟大黄>水蒸熟大黄>生大黄;生大黄中儿茶素含量较高,该成分含量大小顺序依次为生大黄>水蒸熟大黄>熟大黄。结论:本方法灵敏、可靠、重复性好,可用于同时测定3种不同大黄炮制品中没食子酸、桂皮酸和儿茶素的含量。
OBJECTIVE: To establish a method for simultaneous determination of gallic acid, cinnamic acid and catechin in 3 processed products of Rheum officinale. METHODS: RP-HPLC method was established. The determination was performed on Thermo ScientificTM Hypersil GOLD Dim column with mobile phase consisted of methanol-0.1% phosphoric acid solution (gradient elution) at the flow rate of 1.0 mL/min. The detection wavelength was set at 278 nm, and the column temperature was 30 ℃. The sample size was 10 μL. RESULTS: The linear range of gallic acid, cinnamic acid and catechin were 0.126 2-1.262 0 μg(r=0.999 9), 0.036 2-0.362 0 μg(r=0.999 9) and 0.177 9-1.779 4 μg(r=0.999 8), respectively. Quantitative limits were 25.4, 28.2, 62.5 ng, and detection limits were 6.2, 3.6, 11.8 ng, respectively. RSDs of precision, stability, repeatability and durability tests were all less than 3%. The recoveries ranged from 94.64%-102.71%(RSD=2.74%, n=9), 95.35%-102.49%(RSD=2.44%, n=9), 93.56%-103.66%(RSD=3.27%, n=9). The determination results showed that the contents of gallic acid and cinnamic acid in prepared R. officinale were higher, and the order of both were prepared R. officinale>steamed R. officinale>raw R. officinale. The content of catechin in raw R. officinale was higher, and the order of it was raw R. officinale> steamed R. officinale>prepared R. officinale. CONCLUSIONS: The method is sensitive, reliable and reproducible. It can be used to determine the contents of gallic acid, cinnamic acid and catechins in 3 processed products of R. officinale simultaneously.
大黄炮制品没食子酸桂皮酸儿茶素反相高效液相色谱法含量测定
Rheum officinaleProcessed productsGallic acidCinnamic acidCatechinRP-HPLCContent determi- nation
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