Analysis of Flavonoids Chemical Components in Waste Material during Industrialization of Pueraria thomsonii Resources / 中国药房

OBJECTIVE：To study the composition and contents of flavonoids chemical components in waste material during industrialization of Pueraria thomsonii resources，and to provide reference for comprehensive development and reasonable utilization of the variety. METHODS ：Using“No. 2 Gange”of P. thomsonii from Jiangxi as objects ，UPLC-Q-TOF-MS and HPLC method were adopted to detect the components and contents of flavonoids in the root （with or without cortex ），cortex，flower， fibrous root ，stem，head and dregs （with or without cortex ）of P. thomsonii as well as dry matter of industrial wastewater （with or without cortex ）after precipitation of pueraria powder. RESULTS ：The linearity ，precision，repeatability，stability and recovery of the established method for content determination of 7 flavonoids（puerarin，daidzin，iridoxine-7-O-xylose glucoside ，genistin， iridin，daidzein and kakkalide ）were all in line with the requirements. Totally 12 kinds of flavonoids were identified ，among which the flavonoids in the root ，cortex，stem，fibrous root ，head and dregs of P. thomsonii as well as dry matter of industrial wastewater were the same ，mainly were puerarin ，daidzin，genistein，daidzein and malonyl-daidzein. The flower of P. thomsonii mainly included iridoxine- 7-O-xylose glucoside ，genistin，iridin，kakkalide，6″-O-xylosyldaidzein，but the components as puerarin ， daidzin and its aglycone were not be detected. The content of puerarin in the head of P. thomsonii was the highest （5.765％）. The contents of puerarin in root and dregs of P. thomsonii as well as dry matter of industrial waste-water in samples with cortex were all higher than in corresponding peeled sample. CONCLUSIONS ：The waste material from the industrialization of P. thomsonii resources contains a lot of flavonoids with rich species and high content ，and can be used as an important raw material for obtaining flavonoids such as puerarin.

Analysis of genomic SSRs in Pueraria lobata and P. thomsonii and establishment of DNA identity card for different germplasms of P. thomsonii of Jiangxi province / 中国中药杂志

The type and frequency of simple sequence repeats( SSRs) in the genomes was investigated using the DNA sequence data of Pueraria lobata and P. thomsonii. Based on these SSRs,20 pairs of SSR primers were designed and 5 high polymorphism primer pairs were selected to analyze genetic diversity of 9 cultivars of P. thomsonii in Jiangxi province. The results showed that the 5 pairs of primers could generate 16 polymorphic alleles bands. The average polymorphism information content( PIC) of each SSR primer pair was 0. 600 7.According to the genetic similarity coefficients,the 9 cultivars of P. thomsonii can be classified into 6 germplasms. This study established DNA identity cards with 5 pairs of SSR primers for different germplasm resources of P. thomsonii in Jiangxi province,which provided reference information for the selection of fine germplasms of P. thomsonii and the theoretical basis for the study of Dao-di herbs.

Simultaneous Determination of 14 Kinds of Isoflavones in Pueraria radix by LC-MS/MS / 中国药房

OBJECTIVE:To establish the method for simultaneous determination of 14 kinds of isoflavones in Pueraria radix. METHODS:LC-MS/MS was adopted to detect 14 kinds of isaflavones in 14 batches of P. radix(P. radix:PL-1 to PL-7,P. thomsonii:PT-1 to PT-7). The determination was performed on Extend C18 with mobile phase consisted of methanol-water(gradient elution)at the flow rate of 0.8 mL/min. The column temperature was set at 22℃. The sample size was 5μL. Ion source was ESI source. The detection mode was negative ion detection. Scanning mode was MRM with jet voltage of -4500 V;ion source temperature was set at 600 ℃, and atomization gas was 60 psi. The auxiliary gas was 60 psi,collision gas was 7 psi,air curtain gas was 30 psi. SIMCA 13.0 software wasused for cluster analysis of above batches. RESULTS:The linear range of 14 kinds of isoflavones ranged 10-1000 ng/mL(puerarin of 10-5000 ng/mL,r>0.9900). RSDs of precision,stability and reproducibility tests were all lower than 5％. The recoveries were 95％-105％(RSD were 0.8％-4.5％,n=6). The total content of isoflavones were different significantly between P. radix and P. thomsonii. The contents of isoflavones in P. radix from different origins were different significantly. Among 14 kinds of isoflavones, the content of puerarin was the highest. Results of cluster analysis showed that 14 batches of P. radix could be clustered into 4 categories,including PL-2 as Ⅰ category,PL-3 and PL-4 as Ⅱ category,PL-5,PL-6 and PL-7 as Ⅲ category,other as Ⅳcategory. CONCLUSIONS:The method is simple and reproducible. It can be used for content determination of 14 kinds of isoflavones in P. radix and quality control.

Tissue culture and induction of autotetraploid of Pueraria thomsonii / 中草药

Objective To establish a tissue culture system of Pueraria thomsonii and to cultivate a new breed autotetraploid.Methods Using the MS medium as the basic medium,the explants shoot apexes and stem segments were cultured to differentiate the regenerations.The autotetraloid with genetic material doubled was achieved by treating shoot tips with colchicine of different concentrations for different long times.Results The proper media MS+6-BA 1.0 mg/L + 2,4-D 1.0 mg/L+NAA 0.2 mg/L is for pro-(tocorn) inducing,MS+6-BA 1.0 mg/L+2,4-D 0.2 mg/L for diffrentiation and propagation,and 1/2(MS+) IBA 0.2 mg/L for rooting.The best effect of the autotetraploid induction can be achieved by treating the 0.3-0.5 cm shoot tips cut from the 1 cm plantlets with 0.4%-0.5% colchicine for 48 h,and it is unlikely to succeed by treating shoot tips with colchicine of very low or high concentration or for very long or short time.Conclusion The autotetraploid gives a chance to improve the content of medical materials in the root of P.thomsonii.