Test method of Chrysophanol(大黄酚检测方法) 标准溶液:精确称量5mg(美国sigma公司)标准 大黄酚,放入 25ml 的量杯, 加适量丙酮溶解,用超声波震动直至全溶。 然后将丙酮加至标度处,获得标准溶液。 测试溶液:精确称量5mg大黄酚粉末,放入25ml的量杯中,加入适当丙酮溶解,用超声波摇动直至溶解。然后加入丙酮至标度处,均匀摇晃,获得测试溶液
Test method of Chrysophanol(大黄酚检测方法)
1. Instrument 设备
a HPLC-9a (Shimadzu) 安捷伦色谱仪
b Sartorius electronic balance (Germany) 电子称
c. KQ-----model 100, Washing device of ultrasonic wave 100-KQ 超声波洗涤设备
2. Condition 条件
Chromatographic Column 色谱柱 ROM-150 ODS-c18(4.6mm×150mm, 5цm)
Mobile Phase: CH3OH: H₂O:CH3COOH =80∶20:1 ( V/V/V)
流动相:甲醇∶水∶冰醋酸=80:20:1 ( V/V/V)
Flow Rate: 1.0mL/min 流速
Wave Length: 254nm 波长
Injection volume:20μL进样量
Column: room temperature 测柱:25℃ 室温
3.Standard solution 标准溶液
Transfer about 5mg Chrysophanol standard(Sigma company U.S.A.), accurately weighed, to a 25mL volumetric flask, add the right amount CH3COCH3 to dissolve, Ultrasonic wave is shaken for 5 minutes till all dissolve in the CH3COCH3. Then add CH3COCH3 to scale mark, it is obtained.
精确称量5mg(美国sigma公司)标准 大黄酚,放入 25ml 的量杯, 加适量丙酮溶解,用超声波震动直至全溶。 然后将丙酮加至标度处,获得标准溶液。
4.Testing solution 测试溶液
Transfer 5mg Chrysophanol powder, accurately weighed, to a 25ml volumetric flask
, add the right amount CH3COCH3 to dissolve, Ultrasonic wave is shaken for 5 minutes till all dissolve in the water, Then add CH3COCH3 to scale mark, shake evenly, it is obtained.
精确称量5mg大黄酚粉末,放入25ml的量杯中,加入适当丙酮溶解,用超声波摇动直至溶解。然后加入丙酮至标度处,均匀摇晃,获得测试溶液。
5. Procedure 过程
Separately inject equal volumes (about 20 uL) of the Standard preparation and the Assay preparation into the chromatograph, record the chromatograms, and measure the responses for the major peaks.
A1 W2
X%=----- × ---- × 100%
A2 W1
In which is A1 the assay sample , A2 is the standard peak area, W2 is the concentration of standard, W1 is the concentration of assay sample.
将等量的(大约20 UL)标准溶液和测试溶液(大约20 UL)分别注入色谱仪,记录两个色相图谱 测量两个主峰的反应变化。
A1 W2
X%=----- × ---- × 100%
A2 W1
上式中,A1指测试样的峰面积,A2是标准溶液的峰面积, W2指标准溶液的浓度,W1指测试样的浓度。