A method for separation and determination of spin-labeled anti-tumor podophyllotox in derivatives by HPLC coupled with electron paramagnetic resonance(EPR) has been developed.The analytical conditions of reverse phase HPLC have been discussed and an separation of GP-3,GP-5,GP-7 was obtained by using MeOH-H2O system.The detection limits were 10-8g/mL.EPR detector was then used to determine HPLC.This new technique has been proved to be useful for pharmacokinetics study of spin-labeled anti-tumor podophyllotoxin derivatives.This method was assayed.It is simple,rapid and accurate.

以决明发根为材料, 在培养基中附加0.001～10 mg·L-1 Eu3+(EuCl3), 测量发根的鲜重和干重, 用高效液相色谱法测定游离蒽醌化合物含量. 发现极低剂量(0.001, 0.01, 1.0 mg·L-1)Eu3+抑制生长, 0.001 mg·L-1 Eu3+抑制作用最强, 干重比对照降低22%;较低剂量(10 mg·L-1)Eu3+促进生长, 干重比对照增加27%;0.001, 0.01, 0.1, 1.0 mg·L-1 Eu3+各个处理的游离蒽醌化合物总含量均低于对照, 而10 mg·L-1 Eu3+处理的含量高于对照26%, 高于Eu3+ 1.0 mg·L-1处理97%. 10 mg·L-1 Eu3+能有效增强决明发根的生长、提高游离蒽醌化合物总含量和产量.

Effect of EuCl3 on the callus growth and cell ultrastructure of Rheum was studied. The effect of EuCl3 on the growth of the callus was a biphasic one, with stimulation at low concentrations, and inhibition at higher concentrations. The explants would die when the concentration of EuCl3 was 100 mg. L-1. The stimulative effect of EuCl3 could be inhibited by calmodulin antagonist-chlorpromazine. 10 mg.L-1 of EuCl3 would cause variations of the cell ultrastructure.

目的：研究稀土元素对药用植物细胞组织培养的作用和意义。方法：以药用植物黄连为材料，研究不同浓度的稀土化合物对愈伤组织鲜重增加和生物碱含量的影响。结果：０．１ｍｇ·Ｌ－１Ｙｂ３＋和１ｍｇ·Ｌ－１Ｅｕ３＋促进愈伤组织的生长，高浓度稀土化合物抑制生长。在不含稀土化合物的培养基上的继代培养中，来源于稀土化合物处理的愈伤组织鲜重增加值均比对照低。稀土化合物影响愈伤组织小檗碱、药根碱和巴马汀含量。结论：低浓度的稀土化合物对黄连愈伤组织的生长具有促进作用，低浓度和高浓度的Ｙｂ３＋和Ｅｕ３＋对黄连愈伤组织后续的继代培养均有不利的影响。Ｙｂ３＋抑制愈伤组织中三种生物碱的积累，０．１ｍｇ·Ｌ－１Ｅｕ３＋有利于药根碱和巴马汀的累积

不同生长季节的黄连外植体在一系列培养基上诱导产生愈伤组织.经高效液相色谱法测定愈伤组织内小檗碱和药根碱的含量.结果表明,幼叶柄外植体在67-V 0.5mg/L 2,4-D上产生的愈伤组织的小檗碱和药根碱含量明显高于对照组.花苔外植体在67-V 0.5mg/L 2,4-D上诱导的愈伤组织小檗碱含量高于对照组.

Objective: To explore the approach to producing plant secondary metabolites using hairy root culture method. Methods: The medicinal plant Rheum wittrochii Lundstr., was genetically with Ri plasmid harbouring in Agrobacterium rhizogenes 9402 strain to induce hairy roots; transformed the opines from the hairy roots were assayed by paper electrophoresis; the contents of free anthroquinones such as aloe emodin, rhein, chrysophanol, emodin, physcion, 8 O methylchrysophanol from the hairy roots were quantitatively determined by HPLC technique. Results: The hairy root was successfully induced and the hairy root culture system was set up; The opines assay was positive for mannopine and agropine; The result of determination showed that there were a lot of anthroquinone constituents in the rhubarb hairy root and the contents of rhein, aloe emodin, emodin or 8 O methylchrysophanol were respectively higher than those in wild plant root. Conclusion: The hairy root cultures of Rheum wittrochii Lundstr. produced similar constitutents as the medicinal site of the wild plant.