Targeted trace ingredients coupled with chemometric analysis for consistency evaluation of Panax notoginseng saponins injectable formulations / 中国天然药物

Evaluating the consistency of herb injectable formulations could improve their product quality and clinical safety, particularly concerning the composition and content levels of trace ingredients. Panax notoginseng Saponins Injection (PNSI), widely used in China for treating acute cardiovascular diseases, contains low-abundance (10%-25%) and trace saponins in addition to its five main constituents (notoginsenoside R1, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, and ginsenoside Rd). This study aimed to establish a robust analytical method and assess the variability in trace saponin levels within PNSI from different vendors and formulation types. To achieve this, a liquid chromatography-triple quadrupole mass spectrometry (LC-MS/MS) method employing multiple ions monitoring (MIM) was developed. A "post-column valve switching" strategy was implemented to eliminate highly abundant peaks (NR1, Rg1, and Re) at 26 min. A total of 51 saponins in PNSI were quantified or relatively quantified using 18 saponin standards, with digoxin as the internal standard. This study evaluated 119 batches of PNSI from seven vendors, revealing significant variability in trace saponin levels among different vendors and formulation types. These findings highlight the importance of consistent content in low-abundance and trace saponins to ensure product control and clinical safety. Standardization of these ingredients is crucial for maintaining the quality and effectiveness of PNSI in treating acute cardiovascular diseases.

A new comprehensive procedure for the quality control of Semen Cassiae and its application in evaluating commercially available material in China / 中国天然药物

AIM@#To establish a more comprehensive and suitable procedure for the quality control of Semen Cassiae which can be used to supplement the evaluation procedure adopted by the Pharmacopoeia of the Peoples Republic of China.@*METHODS@#A HPLC assay-based comprehensive quality evaluation procedure for Semen Cassiae using three bioactive compounds including anthraquinones and naphthopyrones, i.e., chrysophanol-1-O-β-D-glucopyranosyl-(1→3)-β-D-glucopyranosyl-(1→6)-β-D-glucopyranoside (1), rubrofusarin-6-O-β-D-gentiobioside (2) and toralactone-9-O-β-D-gentiobioside (3) as the index components was established. The resultant data were further analyzed by principal component analysis (PCA) and data distribution methods using software SPSS 16.0.@*RESULTS@#Sixty-six batches of Semen Cassiae obtained from various regions of China were analyzed with the procedure. Based on the test results of these batches, the content limits of the three bioactive compounds in Semen Cassiae were proposed.@*CONCLUSION@#The procedure established herein is more comprehensive and appropriate for the quality evaluation of Semen Cassiae commercially available in China, and can be used as a useful supplement to the current official method in China for the quality evaluation of Semen Cassiae.

Genetic variability of Brazilian isolates of Alternaria alternata detected by AFLP and RAPD techniques / Variabilidade genética de isolados Brasileiros de Alternaria alternata por meio de marcadores moleculares de AFLP e RAPD

The Alternaria brown spot (ABS) is a disease caused in tangerine plants and its hybrids by the fungus Alternaria alternata f. sp. citri which has been found in Brazil since 2001. Due to the recent occurrence in Brazilian orchards, the epidemiology and genetic variability of this pathogen is still an issue to be addressed. Here it is presented a survey about the genetic variability of this fungus by the characterization of twenty four pathogenic isolates of A. alternata f. sp. citri from citrus plants and four endophytic isolates from mango (one Alternaria tenuissima and three Alternaria arborescens). The application of two molecular markers Random Amplified Polymorphic DNA (RAPD) and Amplified Fragment Length Polymorphism (AFLP) had revealed the isolates clustering in distinct groups when fingerprintings were analyzed by Principal Components Analysis (PCA). Despite the better assessment of the genetic variability through the AFLP, significant modifications in clusters components were not observed, and only slight shifts in the positioning of isolates LRS 39/3 and 25M were observed in PCA plots. Furthermore, in both analyses, only the isolates from lemon plants revealed to be clustered, differently from the absence of clustering for other hosts or plant tissues. Summarizing, both RAPD and AFLP analyses were both efficient to detect the genetic variability within the population of the pathogenic fungus Alternaria spp., supplying information on the genetic variability of this species as a basis for further studies aiming the disease control.

A mancha marrom ou mancha de Alternaria é uma doença causada pelo fungo Alternaria alternata f. sp. citri, encontrada no Brasil desde 2001 em plantas de tangerina e seus híbridos. Por se tratar de uma doença recente no Brasil, a epidemiologia e variabilidade genética deste patógeno compõem importantes pontos a serem estudados. Este trabalho teve como objetivo avaliar a variabilidade genética deste patógeno por meio da caracterização de vinte e quatro isolados de A. alternata f. sp. citri de plantas de citros juntamente com quatro isolados endofíticos de manga, sendo um deles identificado como Alternaria tenuissima e outros três como Alternaria arborescens. A análise de componentes principais (PCA) do perfil de bandas obtidos pela aplicação de duas técnicas de marcadores moleculares, Amplificação Aleatória de Polimorfismos de DNA (RAPD) e Polimorfismo de Comprimento de Fragmentos Amplificados (AFLP), mostrou a formação de quatro grupos distintos. Apesar do mais amplo perfil de análise por meio da técnica de AFLP, não foi observado nenhuma modificação significativa dentro dos grandes grupos obtidos quando comparado ao RAPD, exceto no posicionamento dos isolados LRS 39/3 e 25M. Além disso, em ambas as análises, somente os isolados de plantas de limão agruparam entre si. Considerando outros hospedeiros ou tecidos de planta não foi possível encontrar grupos específicos. Concluindo, ambas as análises (RAPD e AFLP) são eficientes no estudo de variabilidade genética de Alternaria spp., fornecendo informações sobre a diversidade genética desta espécie, servindo como base para futuramente correlacionar este estudo com estudos adicionais objetivando o controle da doença.