ABSTRACT
OBJECTIVE：To establish a method for the simultaneous determination of eight constituents ，such as scopolin ， scopoletin，chlorogenic acid ，cryptochlorogenic acid ，neochlorogenic acid ，isochlorogenic acid A ，isochlorogenic acid B ，and isochlorogenic acid C ，in different medicinal parts （stems，twigs and leaves ）of Porana racemosa ，and to compare the contents of eight constituents . METHODS ：HPLC method was adopted. The determination was performed on Agilent TC-C 18 column with mobile phase consisted of acetonitrile- 0.1％ phosphate acid （gradient elution ） at the flow rate of 1.0 mL/min. The column temperature was 30 ℃，and the detection wavelength was set at 345 nm. The sample size was 10 μL. The contents of above constituents in stems ，twigs and leaves of P. racemosa were compared ，and the principal component analysis was carried out by Markerlynx XS software. RESULTS ：The linear range of scopolin ，scopoletin，chlorogenic acid ，cryptochlorogenic acid ， neochlorogenic acid ，isochlorogenic acid A ，isochlorogenic acid B ，and isochlorogenic acid C were 0.076 4- 7.64，0.062 8-6.28， 0.090 8-9.08，0.080 0-8.00，0.057 6-5.76，0.094 4-9.44，0.086 0-8.60，0.078 8-7.88 mg/L，respectively（all r＞0.999）. RSDs of precision，stability（24 h）and reproducibility tests were all lower than 2.0％. The average recoveries were 99.71％（RSD＝1.36％， n＝6），100.39％（RSD＝1.76％，n＝6），99.20％（RSD＝1.75％，n＝6），100.04％（RSD＝2.63％，n＝6），98.57％（RSD＝1.99％， n＝6），99.68％（RSD＝1.84％，n＝6），99.90％（RSD＝1.88％，n＝6），99.76％（RSD＝1.47％，n＝6），respectively. The average contents of above eight constituents were 9.725 3，1.286 5，7.271 3，1.347 6，0.997 7，0.710 9，0.656 3，0.364 7 mg/g in stems ； those were 0.690 3，0.411 7，4.394 3，0.639 6，0.531 3，1.392 7，0.989 1，1.129 2 mg/g in twigs ；those were 1.195 1，0.691 1， 27.952 9，6.173 4，1.405 1，0.549 7，0.288 8，0.794 2 mg/g in leaves ，respectively. The results of principal component analysis showed that different parts of P. racemosa could be divided into 3 categories. Among them ，most of the stems ofP. racemosa gathered in the first quadrant of score plot ，all the twigs gathered in the third quadrant ， and all the leaves 分m gathered in the fourth quadrant. CONCLUSIONS ：Established method is simple and reproducible ，and can be used for the determination of 8 constituents in different medicinal parts of P. racemosa. The average contents of neochlorogenic acid ，chlorogenic acid and cryptochlorogenic acid in the leaves of P. racemosa are relatively high ；the contents of isochlorogenic acid B ，isochlorogenic acid A and isochlorogenic acid C in the twigs are relatively high；the average contents of scopolin and scopoletin in the stems are also relatively high.