OBJECTIVETo explore the genetic diversity and relationship of different Scrophularia ningpoensis cultivars.

METHODForty-eight germplasmic resources of S. ningpoensis cultivars were analyzed by Start Codon Targeted Polymorphism(SCOT) molecular markers. Genetic distance was calculated by TREECONW software and the systematic diagram of genetic relationship was clustered by UPGMA method.

RESULTA total of 279 bands were detected using 48 primers, among which 214 were polymorphic bands. The average percentage of polymorphic bands was 76.7%. Genetic distance was changed from 0.1507 to 0.4933. Clustering results showed that the genetic relationship of S. ningpoensis cultivars was more complex. There was significant correlation between some germplasm and its geographic origin while geographical distribution of some germplasm was not very obvious, but it was also showed that some of the S. ningpoensis from the same region were in the same group which presented the law of geographical distribution in the tested materials.

CONCLUSIONSignificant polymorphism and genetic diversity can be observed among S. ningpoensis germplasm resources which provided a wealth of genetic basis for cultivating fine varieties.

The morphological indexes of the Scuophularia ningpoensis seedlings including the longth, diameter and weight were measured, clustering analysis was used to set up the standard quality grading of seedlings of S. Ningpoensis by SPSS. Field experiment was carried out to measure the indicators of plants growth and development, the yield and the quality. The results showed that the growth and yield of class I seedlings were better than those of class II and III. The content of main active ingredients was affected barely by seedlings classification. To ensure the quality, class II seedlings or above should be used for plantation. The established quality classification standard of S. ningpoensis seedlings was scientific and feasible, and provides the basis for the standardized cultivation of S. ningpoensis.
Drugs, Chinese Herbal ; analysis ; standards ; Quality Control ; Scrophularia ; chemistry ; classification ; growth & development ; Seedlings ; chemistry ; growth & development

Modern drying technology was used to explore suitable drying process to provide scientific basis for improving drying processing methods of Scrophulariae Radix. Controlled temperature and humidity drying, vacuum drying apparatus, microwave vacuum drying apparatus, short infrared drying device were used to gain samples for analyzing. The character appearance, concentration of main components and power consumption indicators were chosen for preliminary judging. Six major components, including iridoids and phenylpropanoids were analyzed by UPLC-MS/MS method. The contents of polysaccharides were determined by UV-visible spectrophotometry. The character appearance with controlled temperature and humidity drying and short infrared drying meet the pharmacopoeia standard (Ch. p, edition 2015), while samples with vacuum and microwave vacuum drying apparatus didn't. Compared to fresh sample, concentrations of harpagide, harpagoside, aucubin and catalpol were lower in the dried samples. Angoroside-C showed no significant change before and after drying. Concentration of acteoside increased after drying. Samples with controlled temperature (70 degrees C) and humidity (15% - 10%) drying had high content and short drying time. The better drying process of Scrophulariae Radix was controlled temperature and humidity drying. The method will provide the reference for the drying technology standard of roots medicine.
Desiccation ; methods ; Drugs, Chinese Herbal ; chemistry ; Plant Roots ; chemistry ; Quality Control ; Scrophularia ; chemistry

The quality constant evaluation method was applied in the grade evaluation of Scrophulariae Radix pieces. Nineteen batches of Scrophulariae Radix were measured for the appearance index. Harpagide and harpagoside were taken as index components for the content determination. The traditional grading standard and the modern quality control index were combined to calculate the quality constant and grade Scrophulariae Radix pieces. The results showed that the quality constants ranged between 156. 75 and 491. 65; according to the percentage mass constants,more than 80% were graded as first-class pieces,50%-80% were graded as second-class pieces,and the rest were graded as third-class pieces. The quality constants of first-class Scrophulariae Radix were >393. 32,that of second-class Scrophulariae Radix was between 245. 83 and 393. 32,and that of third-class Scrophulariae Radix was < 245. 83. The study shows that this method can objectively,reasonably and effectively classify Scrophulariae Radix pieces,and further promote and apply the evaluation method of slice model quality constant to prove the rationality,scientificity and practicability.
Drugs, Chinese Herbal ; standards ; Plant Roots ; chemistry ; Quality Control ; Scrophularia ; chemistry

The standard decoction of Chinese herbal decoction pieces is a standard reference substance to measure whether different dosage forms of Chinese medicine are basically consistent with those of clinical decoction,and provides new ideas and methods for effectively solving the problems of uneven quality in Chinese medicine dispensing granules. In this study,a systematic method for evaluating the quality of Scrophulariae Radix decoction was established from the perspective of " standard decoction",providing reference for the quality control of the Scrophulariae Radix dispensing granules. 15 batches of Scrophulariae Radix decoction pieces from different origins were collected,and 15 batches of standard decoctions were prepared according to the standardized process with water as solvent.Harpagide and harpagoside were used as quantitative detection indicators to determine the content,calculate the transfer rates and determine the extraction rate. The high performance liquid chromatography( HPLC) was used to establish a standard decoction fingerprint analysis method. The results showed that the transfer rates of harpagide and harpagoside in 15 batches of Scrophulariae Radix pieces standard decoction were( 70. 84±13. 39) % and( 48. 56±6. 40) % respectively; the extraction rate was( 57. 47±5. 89) %. Nine peaks were identified in the HPLC fingerprint,and the similarity was higher than 0. 97 between the fingerprints of 15 batches of standard decoction and the control fingerprint. In this study,the preparation process of standard decoction of Scrophulariae Radix pieces conformed to the traditional decoction preparation method. The sources of the samples were representative,and the established fingerprint method was stable and feasible,which can provide reference for the preparation and quality control of Scrophulariae Radix dispensing granules.
Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; standards ; Plant Roots ; chemistry ; Quality Control ; Scrophularia ; chemistry

Phytochemical investigation of the roots of Scrophularia buergeriana Miq. (Scrophulariaceae), resulted in the isolation of a new iridoid derivative named as buergerinin (1). Its structure was elucidated as rel-(1R, 5R, 6R)-(2-oxa-bicyclo[3.3.0]oct-7-en-6, 7-diyl)dimethoxypropane based mainly on MS and 1D and 2D NMR spectroscopic analyses.
Iridoids ; chemistry ; isolation & purification ; Magnetic Resonance Spectroscopy ; Mass Spectrometry ; Plant Roots ; chemistry ; Scrophularia ; chemistry

By establishing the preparation process of Scrophulariaceae Radix reference extract(SRRE) and calibrating it, we discussed its feasibility as a substitute for single reference substance in the quality control of Scrophulariae Radix. The SRREs were prepared by solvent extraction method and chromatographic separation technology, and then calibrated with the reference substances of harpagide, angoroside C and harpagoside. The HPLC content determination method of Scrophulariae Radixl was established with SRREs of the known content and the reference substances of harpagide, angoroside C and harpagoside respectively as the control ones. Then the content of three components in Scrophulariae Radix was determined, and the t-test method was used to compare the results of the two methods. With SRRE as references, harpagide, angoroside C and harpagoside were in a good linear relationship(r≥0.999 8) within each range, and the average recovery rate was 98.55% to 100.6%. The t-test results showed that the P values of two determination methods were 0.493, 0.155 and 0.171 for harpagide, angoroside C and harpagoside respectively, indicating no significant diffe-rence between the two methods of content determination. The SRRE can be used as a substitute for the reference in the quality control of Scrophulariaceae Radix. The SRRE can replace the corresponding reference substance for the quality control of Scrophulariae Radix. The results of this study provide new methods and new ideas for the quality evaluation of Scrophulariae Radix, and provide a scientific basis for the application of reference extracts in the quality research of traditional Chinese medicine.
Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; Quality Control ; Scrophularia ; Scrophulariaceae

The present study investigated the chemical constituents of Scrophulariae Radix and their antitumor activities in vitro. The compounds in the ethyl acetate extract were separated and purified by conventional column chromatographies(such as silica gel, Sephadex LH-20, and ODS column) and semi-preparative high-performance liquid chromatography(HPLC), and their structures were identified by various spectral techniques such as nuclear magnetic resonance(NMR) and mass spectrometry(MS). Twenty-three compounds were isolated and identified as benzyl-β-D-(3',6'-di-O-acetyl) glucoside(1), 5-O-p-methoxybenzoyl kojic acid(2), 5-O-methoxybenzoyl kojic acid(3), 7-O-methylbenzoyl kojic acid(4), 5-O-benzoyl kojic acid(5), methyl ferulate ethyl ether(6), trans-ferulic acid(7), trans-isoferulic acid(8), trans-caffeic acid(9), trans-caffeic acid methyl ester(10), caffeic acid ethyl ester(11), trans-cinnamic acid(12), trans-p-methoxycinnamic acid(13), trans-p-hydroxycinnamic acid(14), trans-p-hydroxycinnamic acid methyl ester(15), 2-(3,4-dihydroxyphenethyl) alcohol(16),(p-hydroxyphenyl) propanoic acid(17), coniferaldehyde(18), sinapaldehyde(19), benzyl β-primeveroside(20), 5-(hydroxymethyl) furfural(21), furan-2-carboxylic acid(22), and decanedioic acid(23). Among them, compound 1 is a new benzyl glucoside, compounds 2-4 are new pyranone compounds, compound 5 is a new natural product of pyranone. The NMR data of compounds 5 and 6 are reported for the first time. Compounds 6 and 20 were isolated from the Scrophularia plant for the first time. Compounds 8, 11, 14, 16, 18, 19, 22, and 23 were isolated from this plant for the first time. The in vitro cytotoxic activities of these compounds against three tumor cell lines(HepG2, A549, and 4 T1) were evaluated. The results showed that compounds 10 and 15 showed cytotoxic activities against HepG2 cells with IC_(50) values of(19.46±0.48) μmol·L~(-1) and(46.10±1.21) μmol·L~(-1).
Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal/chemistry* ; Mass Spectrometry ; Plant Roots/chemistry* ; Scrophularia/chemistry*

Traditional Chinese medicine Scrophulariae Radix, which is also called Yuan Shen, black Shen, is the dried root of Scrophularia ningpoensis of the Scrophulariaceae family. Research has indicated that the chemical constituents of Scrophulariae Radix mainly include terpenoids, phenylpropanoids, organic acids, volatile oils, steroids, sugars, flavonoids, alkaloids and phenols, among which iridoids and phenylpropanoids were the main active constituents. It has been reported that extracts of Scrophulariae Radix or its active substances have anti-inflammatory, antioxidant, hepatoprotective, anti-tumor, anti-fatigue, uric acid-lowering, anti-depression, myocardial cell-protective and other pharmacological activities, and can regulate cardiovascular system, central nervous system and immune system. This paper reviewed the present research achievements of Scrophulariae Radix in chemical constituents, pharmacological activities, processing methods, toxicity and other aspects, and the clinical application of Scrophulariae Radix in ancient and modern times was illustrated. This paper aimed to provide reference for further research of Scrophulariae Radix and facilitated its clinical application.
Medicine, Chinese Traditional ; Drugs, Chinese Herbal/chemistry* ; Chromatography, High Pressure Liquid ; Plant Roots/chemistry* ; Scrophularia/chemistry*

The genetic diversity of three Tibetan herbs, i. e., Sang-Di, E-Dewa and Ye-Xingba (Tibetan names), was studied based on the field collection, specimen identification and DNA sequence analysis. Swertia hispidicalyx, Gentiana lhassica and Scrophularia dentata, as the original plants of the three Tibetan herbs, were collected and identified. The regions of ITS, matK, rbcL, rpoC1, trnL(UAA), psbA-trnH, atpB-rbcL, trnS (GCU)-trnG(UCC), rpl20-rps12, trnL(UAA)-trnF(GAA) and nadl 2nd intron were amplified and sequenced. The ITS regions of S. hispidicalyx and S. dentata were cloned and sequenced, and the sequences were classified into different genotypes. All the sequences were analyzed and compared with those of closely related species. Our studies may provide reference for the genetic diversity analysis and molecular identification of the three Tibetan herbs.
Genetic Variation ; Gentiana ; classification ; genetics ; Phylogeny ; Plant Proteins ; genetics ; Plants, Medicinal ; classification ; genetics ; Scrophularia ; classification ; genetics ; Swertia ; classification ; genetics ; Tibet