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目的:建立测定不同贮存年限(1~19年)广陈皮药材中主要活性成分含量的方法。方法:采用高效液相色谱法(HPLC)测定药材中橙皮苷含量:色谱柱为Diamonsil C18,流动相为甲醇-醋酸-水(35 ∶ 4 ∶ 61,V/V/V),流速为1.0 ml/min,检测波长为283 nm,柱温为25 ℃,进样量为5 μl。采用HPLC法测定药材中川陈皮素、桔皮素含量:色谱柱为Diamonsil C18,流动相为水-甲醇(梯度洗脱),流速为1.0 ml/min,检测波长为326 nm,柱温为25 ℃,进样量为10 μl。采用HPLC法测定药材中辛弗林含量:色谱柱为Diamonsil C18,流动相为甲醇 - 磷酸二氢钾溶液(取磷酸二氢钾0.6 g、十二烷基磺酸钠1.0 g、冰醋酸1 ml,用水溶解至1 000 ml)(65 ∶ 35,V/V);流速为1.0 ml/min,检测波长为275 nm,柱温为25 ℃,进样量20 μl。结果:橙皮苷、川陈皮素、桔皮素、辛弗林检测进样量线性范围分别为500~4 500 ng(r=0.999 8)、38.816~388.16 ng(r=0.999 6)、19.936~199.36 ng(r=0.999 5)、640~2 560 ng(r=0.999 9);精密度、稳定性、重复性试验的RSD<3%;加样回收率分别为96.42%~102.75%(RSD=2.54%,n=6)、97.42%~99.95%(RSD=2.46%,n=6)、99.26%~106.19%(RSD=2.31%,n=6)、97.47%~99.76%(RSD=1.95%,n=6)。结论:该方法操作简便、稳定、重复性好,可用于不同贮存年限广陈皮药材中主要活性成分含量的测定。陈皮“陈久者良”可能与上述4种活性成分含量变化无关,推测与其挥发油含量减少而缓和燥性有关。
OBJECTIVE:To establish a method for the main active components in Citrus chacheiensis with different storage time (1-19 years). METHODS:HPLC was conducted to determine the content of hesperidin: the column was Diamonsil C18 with mobile phase of methanol-acetic acid-water (35 ∶ 4 ∶ 61,V/V/V) at a flow rate of 1.0 ml/min, detection wavelength was 283 nm, column temperature was 25 ℃, and the injection volume was 5 μl; the contents of nobiletin and tangeretin: the column was Diamonsil C18 with mobile phase of water-methanol(gradient elution) at a flow rate of 1.0 ml/min, detection wavelength was 326 nm, column temperature was 25 ℃, and the injection volume was 10 μl; and the content of synephrine: the column was Diamonsil C18 with mobile phase of methanol-potassium dihydrogen phosphate solution(taking 0.6 g potassium dihydrogen phosphate, 1.0 g sodium dodecyl sulfate,1 ml glacial acetic acid dissolved to 1 000 ml) (65 ∶ 35,V/V) at a flow rate of 1.0 ml/min, detection wavelength was 275 nm,column temperature was 25 ℃, and the injection volume 20 μl. RESULTS: The linear range was 500-4 500 ng for hesperidin(r=0.999 8), 38.816-388.16 ng for nobiletin (r=0.999 6), 19.936-199.36 ng for tangeretin(r=0.999 5) and 640-2 560 ng for synephrine(r=0.999 9); RSDs of precision, stability and reproducibility tests were lower than 3%; recoveries were 96.42%-102.75%(RSD=2.54%,n=6), 97.42%-99.95% (RSD=2.46%,n=6),99.26%-106.19%(RSD=2.31%,n=6) and 97.47%-99.76%(RSD=1.95%,n=6), respectively. CONCLUSIONS: The method is simple and stable with good reproducibility, and can be used for the contents determination of main active components in C. chacheiensis with different storage time. Pericarpium citri “the older the better” may be irrelevant to the change of the contents of the above-mentioned 4 active components, and it is speculated related to the release of volatile oil content to ease dryness.
广陈皮贮存年限活性成分高效液相色谱法含量测定
Citrus chacheiensisStorage timeactive componentsHPLCContent determination
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