RP-HPLC荧光法同时检测喜树组织CPT和HCPT的含量

Simultaneous Determination of CPT and HCPT Content of Camptotheca acumianta Tissues by Means of RP-HPLC with Fluorescence Detector

建立高效液相色谱荧光法同时测定喜树组织中喜树碱(CPT)和10-羟基喜树碱(HCPT)含量的方法。使用XTerra RP18柱(5μm,4.6×250 mm)色谱柱,以水∶乙腈=25∶75(V/V)为流动相,流速为1.0 ml/min,30℃下检测。检测器激发波长为363 nm,发射波长为550 nm。分别检测喜树苗1月、6月、12月不同生长期的根、茎、叶和不同培养基诱导的愈伤组织中的CPT和HCPT的含量。结果表明:喜树苗中CPT含量叶>茎>根,1月期喜树苗中HCPT含量叶>根或茎,6月期和12月期喜树苗中HCPT含量根>茎>叶;培养基SH+NAA1.0 mg/L+6-BA 5.0 mg/L上诱导的喜树愈伤组织有较高的CPT和HCPT含量。此方法简单、可靠,分离效果好,专属性强,灵敏度高。

Determination the CPT and HCPT content of different tissues of Camptotheca acumianta by means of RP-HPLC with fluorescence detector.The CPT and HCPT were separated on a XTerra RP18 column（5 μm,4.6 mm × 250 mm） at 30 ℃,using water ∶ acetonitrile = 25 ∶ 75（V/V） system with 1.0 ml/min as the mobile phase,the excitation and emission wavelengths were 363 nm and 550 nm respectively.The leaves,stems and roots of one month,six months and twelve months Camptotheca acumianta seedling and calli induced on different medium were determined. The results show that different dates of Camptotheca acumianta,the CPT content leavesstemsroots,and for HCPT leavesroots or stems when one month,rootsstemsleaves when six months and twelve months;On medium SH＋NAA1.0 mg/L＋6-BA 5.0 mg/L,calli had more content of CPT and HCPT.This method is simple,reliable,sensitive,and suitable for the quantitive determination of CPT and HCPT.