Senecio Species Research Articles

Chemical Diversity Investigation of Hepatotoxic Pyrrolizidine Alkaloids in Qianliguang (Senecio scandens)and Related Species by UHPLC-QTOF-MS1

Objective: Qianliguang (Senecio scandens) is a common Chinese medicinal herb. Qianliguang-containing herbal proprietary products are registered as over-the-counter remedies in China and exported to Western countries. The presence of hepatotoxic pyrrolizidine alkaloids (PAs) has raised concerns about the safety of using Qianliguang and its products. The present study aims at investigation of different types of PAs present in Qianliguang collected from representative locations in China. Methods: In this study, a simple but specific UHPLC-QTOF-MS method for the determination of toxic PAs was developed, based on the characteristic fragment ions specific to different types of PAs. It was successfully applied for the identification and distinguishing of PAs present in Qianliguang and related Senecio species growing in different locations of China. Results: Significant diversity of the PA types and quantities were revealed among the samples tested. The estimated total amounts of toxic PAs in three of the samples exceed the toxic limits of PA intake restricted by WHO, demonstrating the timely and highly demand for regulating both types and quantities of PAs present in Qianliguang. Conclusions: This study provides the methodology for simultaneous identification and quantification of PAs present in herbs without requiring corresponding standards, which could be further used for more systematic investigations of the PA distribution in Qianliguang and other PA-containing herbs.

Detection of dehydropyrrolizidine alklaoids in honey, pollen, eggs and the associated Senecio species

1,2-Dehydropyrrolizidine alkaloid (DHPAs) exposure in humans is most often via contamination of a primary food source such as in the co-harvesting of grains. More unknown is the potential for long term, low level exposure in foods, including pollen and honey, as well as eggs, milk and, perhaps, meat. Two such cases are presented here. First is the detection of DHPAs in honey and pollen from select experimental hives near fields infested with Senecio madagascariensis on the island of Hawaii. DHPAs concentration in honey from the hives ranged from 0.10 µg/g – 3.22 µg/g total alkaloid. No DHPAs were detected in honey from hives physically isolated from S. madagascariensis. Alkaloids were detected in two of four commercial samples at concentrations of 0.04 – 0.09 µg/g. Pollen contained much higher concentration of DHPAs in the range of 33.5 – 136.7 µg/g total alkaloids. Secondly, we report on an experiment to investigate the effect of dietary supplementation of chicken feed with Crotalaria pallida seeds during a 35 day feeding of commercial laying hens. Eggs collected during the 35 days of the treatment feedings contained usaramine at concentration range from 0.29 – 0.89 µg/g (dry egg matter). Seven days after treatment the concentration decreased to 0.009 – 0.043 µg/g usaramine.

Cypselar diversity in four species of <i>Senecio</i> L. (Asteraceae)

The genus Senecio L. is one of the largest genera of flowering plants and is an important member of the tribe Senecioneae (Asteraceae). Phenotypic information, including a broad range of morphological characters is very crucial for phylogenetic reconsideration of any family, tribe or genus. In the family Asteraceae, very little attention has been paid to cypselar diversity, though it is regarded as taxonomically valuable. A sincere attempt has been made to study detailed cypselas macro and micro-morphological features of four species of Senecio L. These diacritical features could be used to strengthen current inter-specific concept of Senecio L.DOI: http://dx.doi.org/10.3329/bjpt.v21i1.19251Bangladesh J. Plant Taxon. 21(1): 13-17, 2014 (June)

Three new caespitose species of Senecio (Asteraceae,Senecioneae) from South Peru.

Three new species of the genus Senecio (Asteraceae, Senecioneae) belonging to Senecio ser. Suffruticosi subser. Caespitosi were discovered in the tributaries of the upper Tambo River, Moquegua Department, South Peru. Descriptions, diagnoses and discussions about their distribution, a table with the morphological similarities with other species of Senecio, a distribution map, conservation status assessments, and a key to the caespitose Peruvian species of Senecio subser. Caespitosi are provided. The new species are Senecio moqueguensis Montesinos, sp. nov. (Critically Endangered) which most closely resembles Senecio pucapampaensis Beltrán, Senecio sykorae Montesinos, sp. nov. (Critically Endangered) which most closely resembles Senecio gamolepis Cabrera, and Senecio tassaensis Montesinos, sp. nov. (Critically Endangered) which most closely resembles Senecio moqueguensis Montesinos.

Interspecific crossing and genetic mapping reveal intrinsic genomic incompatibility between two Senecio species that form a hybrid zone on Mount Etna, Sicily

Studies of hybridizing species can reveal much about the genetic basis and maintenance of species divergence in the face of gene flow. Here we report a genetic segregation and linkage analysis conducted on F2 progeny of a reciprocal cross between Senecio aethnensis and S. chrysanthemifolius that form a hybrid zone on Mount Etna, Sicily, aimed at determining the genetic basis of intrinsic hybrid barriers between them. Significant transmission ratio distortion (TRD) was detected at 34 (∼27%) of 127 marker loci located in nine distinct clusters across seven of the ten linkage groups detected, indicating genomic incompatibility between the species. TRD at these loci could not be attributed entirely to post-zygotic selective loss of F2 individuals that failed to germinate or flower (16.7%). At four loci tests indicated that pre-zygotic events, such as meiotic drive in F1 parents or gametophytic selection, contributed to TRD. Additional tests revealed that cytonuclear incompatibility contributed to TRD at five loci, Bateson-Dobzhansky-Muller (BDM) incompatibilities involving epistatic interactions between loci contributed to TRD at four loci, and underdominance (heterozygote disadvantage) was a possible cause of TRD at one locus. Major chromosomal rearrangements were probably not a cause of interspecific incompatibility at the scale that could be examined with current map marker density. Intrinsic genomic incompatibility between S. aethnensis and S. chrysanthemifolius revealed by TRD across multiple genomic regions in early-generation hybrids is likely to impact the genetic structure of the natural hybrid zone on Mount Etna by limiting introgression and promoting divergence across the genome.

Structural screening by multiple reaction monitoring as a new approach for tandem mass spectrometry: presented for the determination of pyrrolizidine alkaloids in plants

In tandem mass spectrometry the multiple reaction monitoring (MRM) mode is normally used for targeted analysis but this mode also has the potential to screen for structural similarities of analytes. On the basis of the fact that in general similar molecular structures result in similar fragments or losses of neutrals, this approach was used for pyrrolizidine alkaloid (PA) screening but could also be easily adapted to screen for other compound classes. PA are plant toxins of which several hundred individual compounds have been identified. Our MRM screening approach uses the structural relation and similar core structure of all PA which results in a common and thus predictable mass spectrometric fragmentation behaviour. On this basis a method was developed which screens for PA structures by MRM transitions and allows the detection of each individual PA down to a low microgram per kilogram concentration range. The approach was applied to investigate plants from the families of Asteraceae (several species of Senecio and Eupatorium), Boraginaceae (Echium, Cynoglossum, Borago and Anchusa officinalis as well as Heliotropium europaeum) and Fabaceae (Crotalaria incana) for a complete qualitative and quantitative PA characterisation. All analytes that were detected as possible PA by MRM screening were further investigated by recording product ion spectra. Analytes which exhibited a typical PA fragmentation pattern were either confirmed as PA or otherwise deleted as false positive signals (false positive rate was below 10%). Sum formulas of confirmed PA were determined by additional measurements applying high resolution mass spectrometry. In that way 121 unknown PA were identified and for the first time complete PA profiles of different PA plants were delivered.

Controle de Senecio spp. com pastoreio ovino de acordo com sua infestação e fenologia

Intoxicação por Senecio spp. é a principal causa de morte de bovinos no Rio Grande do Sul (RS) e não há medidas terapêuticas eficazes, mas os ovinos são mais resistentes e têm sido usados como controladores naturais da planta. O objetivo desse trabalho foi avaliar a eficiência do pastoreio com ovinos em diferentes estações do ano (primavera-inverno) determinando a melhor época pela interferência desse pastoreio na fenologia da planta, especialmente nas fases reprodutivas, e em relação à infestação na pastagem. O experimento foi desenvolvido em duas propriedades localizadas na região da Campanha do RS, em módulos de 50m X 100m, com água disponível e com diferentes graus de infestação por Senecio spp. O grau de infestação, independentemente da espécie, foi determinado pela média do número de plantas em três pontos aleatórios com diâmetro de 2m cada, dentro da área de estudo. Em 12 pontos de observação fenológica definidos em cada módulo, uma a três plantas representou infestação baixa (IB) e quatro a seis plantas infestação média (IM), ambos localizados na propriedade A; sete a nove plantas infestação alta (IA) e 10 ou mais plantas foi considerada infestação muito alta (IMA), localizados na propriedade B. Os módulos localizaram-se em propriedades distintas porque não foram encontrados todos os níveis de infestação numa única propriedade. Nesses pontos foram avaliados os seguintes parâmetros no dia 0 (antes da entrada dos ovinos) e a cada 15 dias durante a primavera de 2009 (primeiro ano) e inverno e primavera de 2010 (segundo ano): número de exemplares das espécies de Senecio presentes, fenofases reprodutivas, vigor e consumo. O controle foi feito no segundo ano do experimento, em seis pontos de observação fenológica, estabelecidos de forma idêntica ao módulo, em área contígua a cada um dos módulos, sem pastoreio ovino, com pastoreio periódico de bovinos e equinos, seguindo o manejo de rodízio aplicado nas propriedades. No momento da instalação, cada controle tinha o mesmo grau de infestação de Senecio spp. correspondente à área experimental. No intervalo de tempo experimental, entre a primavera de 2009 e o inverno de 2010, os ovinos foram retirados e os módulos abertos ao pastoreio de bovinos e equinos. A análise estatística foi feita através de regressão simples e a comparação das médias por covariância, sendo que as variáveis fenofase reprodutiva, vigor e consumo, que não apresentaram distribuição normal, foram comparadas pelo teste não paramétrico de Wilcoxon. Os resultados demonstraram que o pastoreio ovino interferiu negativamente no desenvolvimento das espécies de Senecio acompanhadas e que, no inverno, inibe as futuras fenofases reprodutivas das plantas. Considerando as condições fenológicas, ambientais e de maior risco para os bovinos, o controle de Senecio spp. com ovinos, no inverno, é mais eficaz do que na primavera. Pela interferência negativa na planta, especialmente em relação às fenofases reprodutivas, diminuindo a produção de sementes, o pastoreio ovino é uma alternativa de controle biológico de Senecio spp. que reduzirá a infestação a médio e longo prazo no RS, consequentemente, o prejuízo à bovinocultura.

The fate of <i>Robinsonia</i> (Asteraceae): sunk in <i>Senecio</i>, but still monophyletic?

Robinsonia is a genus of eight species and is endemic to the Juan Fernández Islands. Previous studies based on ITS phylogenies place Robinsonia deeply nested within Senecio, however its monophyly remains uncertain. In this paper, we use phylogenies reconstructed from plastid, ITS-ETS, and combined data to test its monophyly. Plastid phylogenies support Robinsonia as monophyletic, whereas ITS-ETS trees suggest that Robinsonia berteroi may be more closely related to a South American clade of Senecio species rather than to the remaining Robinsonia species. Maximum parsimony and Bayesian inference analyses of the combined data are congruent with the plastid trees, whereas maximum likelihood analyses are congruent with the ITS-ETS data. Nodal support for either hypothesis is generally low, and Shimodaira-Hasegawa tests in which Robinsonia was either constrained to be monophyletic or to be non-monophyletic showed that these trees do not have significantly lower likelihood scores than trees from unconstrained analyses. Thus the monophyly of Robinsonia remains inconclusive despite additional data and analyses. The results of the present paper further corroborate the results of previous studies that Robinsonia is deeply nested within Senecio. We therefore propose to reduce Robinsonia to synonymy and present new names and combinations of the Robinsonia species under Senecio.

A New Genus and Species of Asteraceae-Inhabiting Aphid (Hemiptera: Aphididae) from Costa Rica and Mexico

Ucrimyzus villalobosi Mier Durante & Perez Hidalgo gen. n., sp. n. (Hemiptera: Aphididae: Macrosiphini) are described from apterous and alate viviparous females collected on species of genera Bidens, Schkuhria, Senecio and Stevia (Asteraceae: Asteroideae) in Costa Rica and Mexico. Principal components analysis (PCA) was done to verify that the studied aphids belong to a single species regardless of their geographical origin or host plant. Molecular analyses were carried out on the sequences of a fragment of the mitochondrial gene encoding for cytochrome c oxidase subunit 1 (COI) and of a fragment of the nuclear gene encoding elongation factor 1α (EF1α). The taxonomic discussion takes into account the conclusions of the molecular analyses and the morphologic study compared with other genera of Macrosiphini. The identification keys by Blackman & Eastop (2006) to aphids living on each mentioned plant genus are modified to include the new species.

Genomic evidence for the parallel evolution of coastal forms in the Senecio lautus complex

Instances of parallel ecotypic divergence where adaptation to similar conditions repeatedly cause similar phenotypic changes in closely related organisms are useful for studying the role of ecological selection in speciation. Here we used a combination of traditional and next generation genotyping techniques to test for the parallel divergence of plants from the Senecio lautus complex, a phenotypically variable groundsel that has adapted to disparate environments in the South Pacific. Phylogenetic analysis of a broad selection of Senecio species showed that members of the S.lautus complex form a distinct lineage that has diversified recently in Australasia. An inspection of thousands of polymorphisms in the genome of 27 natural populations from the S.lautus complex in Australia revealed a signal of strong genetic structure independent of habitat and phenotype. Additionally, genetic differentiation between populations was correlated with the geographical distance separating them, and the genetic diversity of populations strongly depended on geographical location. Importantly, coastal forms appeared in several independent phylogenetic clades, a pattern that is consistent with the parallel evolution of these forms. Analyses of the patterns of genomic differentiation between populations further revealed that adjacent populations displayed greater genomic heterogeneity than allopatric populations and are differentiated according to variation in soil composition. These results are consistent with a process of parallel ecotypic divergence in face of gene flow.

Analysis of Pyrrolizidine Alkaloids and Evaluation of Some Biological Activities of Algerian Senecio Delphinifolius (Asteraceae)

Although Senecio species are known as sources of potentially toxic pyrrolizidine alkaloids (PAs), some species of this genus are traditionally used as remedies, notably in Algeria. In this paper, the evaluation of biological activities and the analysis of PAs of Algerian specimens of Senecio delphinifolius Vahl are reported. The n-butanolic extract of the herb showed a weak antibacterial effect against Escherichia coli with a MIC of 1 mg/mL, but was inactive against Staphylococcus aureus and Pseudomonas aeruginosa. The n-butanolic extracts of the roots, stems and herb showed a modest antioxidant activity, scavenging the free radical DPPH with respective IC50 values of 55.3, 50.2 and 13.3 microg/mL. A cytotoxic effect against a series of human tumor cell lines was observed with the n-butanolic extract from stems (IC50 ranging between 34 and 88 microg/mL). The herb of the evaluated sample contains 140 ppm of PAs (senecionine, seneciphylline, integerrimine, senkirkine) and PA-related alkaloids (dehydrosenkirkine and neosenkirkine). As the major PAs belong to the toxic series (1,2-unsaturation in the pyrrolizidine cycle and macrocyclic diester), the use of S. delphinifolius should be discouraged in traditional medicine.

A phylogenetic analysis and new delimitation of Senecio sect. Crociseris (Compositae: Senecioneae), with evidence of intergeneric hybridization

AbstractSenecio sect. Crociseris is one of the three main sections of western Eurasian/northern African Senecio species. Previous studies indicated that species commonly assigned to this section are closely related to those of sect. Doria. The aim of the research presented here was to test the morphology–based delimitation of sect. Crociseris using phylogenetic analyses of plastid and nuclear DNA sequence data. The results of these studies are used to propose a new delimitation of sect. Crociseris. Species of sect. Doria were nested within a clade formed by species of sect. Crociseris. Section Doria is therefore best considered a synonym of sect. Crociseris. In addition, S. racemosus, traditionally included in sect. Crociseris, is transferred to the genus Jacobaea. One of the two J. kirghisica specimens that were sequenced contains ITS copies that are most closely related to those of sect. Crociseris as well as copies that nest among ITS accessions of Jacobaea. This suggests intergeneric hybridization between Senecio and Jacobaea.

THE POPULATION GENETICS OF SPOROPHYTIC SELF-INCOMPATIBILITY IN THREE HYBRIDIZINGSENECIO(ASTERACEAE) SPECIES WITH CONTRASTING POPULATION HISTORIES

Hybridization generates evolutionary novelty and spreads adaptive variation. By promoting outcrossing, plant self-incompatibility (SI) systems also favor interspecific hybridization because the S locus is under strong negative frequency-dependent balancing selection. This study investigates the SI mating systems of three hybridizing Senecio species with contrasting population histories. Senecio aethnensis and S. chrysanthemifolius native to Sicily, form a hybrid zone at intermediate altitudes on Mount Etna, and their neo-homoploid hybrid species, S. squalidus, has colonized disturbed urban habitats in the UK during the last 150 years. We show that all three species express sporophytic SI (SSI), where pollen incompatibility is controlled by the diploid parental genome, and that SSI is inherited and functions normally in hybrids. Large-scale crossing studies of wild sampled populations allowed direct comparison of SSI between species and found that the main impacts of colonization in S. squalidus compared to Sicilian Senecio was a reduced number of S alleles, increased S allele frequencies, and increased interpopulation S allele sharing. In general, many S alleles were shared between species and the S locus showed reduced intra- and interspecific population genetic structure compared to molecular genetic markers, indicative of enhanced effective gene flow due to balancing selection.

Plant-defensive sesquiterpenoids from Senecio species with biopesticide potential

Senecio species have been used in folk medicine for treatment of wounds, as antiemetic, anti-inflammatory and their crude extract or dry powder as crop protection agents. The toxicity exhibited to livestock by these plants has been attributed to their content in pyrrolizidine alkaloids and furanoeremophilane type sesquiterpenes. Sesquiterpenoids with eremophilane, cacalol, bisabolane, silphinene, caryophillane, humulane, germacrane and benzofurane skeletons have been isolated from this genus. Here we focus on bioactive sesquiterpenoids with plant defensive properties isolated from Senecio.

Gastroprotective effect of Senecio candicans DC on experimental ulcer models

Ethnopharmacological relevanceSenecio candicans DC (Asteraceae) is used as a remedy for gastric ulcer and stomach pain in the Nilgiris district, Tamil Nadu for which no scientific evidence exists. Aim of the studyThe present study was performed to evaluate the gastroprotective effects and acute oral toxicity of aqueous leaf extract of Senecio candicans (AESC) in experimental models. Materials and methodsThe antiulcerogenic activity of AESC was performed in two different ulcer models viz., pylorus-ligated model and ethanol-induced model using Wistar albino rats. Acute toxicity study was also performed to get information on the admissible dose for treatment of ulcer. Preliminary phytochemical screening of AESC was performed to find the active principles present, which are thus responsible for the antiulcerogenic activity. DPPH assay was performed to confirm the antioxidant activity of AESC. ResultsThe acute toxicity study did not show any mortality up to 2500mg/kg b.w. of AESC. Both the ulcer models showed gastroprotective effect comparable to that of the standard Omeprazole. The results of antioxidant enzymes, histopathology sections, ATPase and mucus content of gastric secretion showed that several mechanisms are involved in the gastroprotective effect. The preliminary phytochemical screening revealed the presence of alkaloids, flavonoids and steroids in AESC. The DPPH assay confirmed the antioxidant activity of AESC. ConclusionThe traditional consumption of AESC for the treatment of gastric ulcer is thus true, the antioxidant constituents present in the extract plays a major role in the gastroprotective activity, but since Senecio species are known for the presence of pyrrolizidine alkaloids, a detailed study in future is required to describe the safe dose for a prolonged period.

Chromosome numbers on the Korean species of Senecio L. and two related genera (Asteraceae)

The somatic chromosome numbers of the Korean species of Senecio L. and two related genera are investigated here. Three different chromosome numbers were found: 2n = 40 in Senecio argunensis Turcz., S. nemorensis L., S. vulgaris L., Tephroseris flammea (DC.) Holub and 2n = 44 in T. phaeantha (Nakai) C. Jeffrey & Y.L. Chen. and 2n = 48 in Sinosenecio koreanus (Kom.) B. Nord., T. kirilowii (Turcz. ex DC.) Holub and T. pierotii (Miq.) Holub. The chromosome numbers of Sinosenecio koreanus (Kom.) B. Nord., Tephroseris flammea (DC.) Holub and T. phaeantha (Nakai) C. Jeffrey & Y.L. Chen. were different from those in previous reports.

Direct and indirect effects of grazing constrain shrub encroachment in semi‐arid Patagonian steppes

Abstract Question: What are the long‐term effects of grazing exclusion on the population structure and dynamics of, and interactions among, three dominant shrub species? Location: Grass‐shrub Patagonian steppe, Chubut, Argentina. Methods: Permanent plots were established in grazed paddocks and paddocks excluded from grazing in representative Patagonian rangelands. Shrub abundance, population size‐structure, short‐term (two 3‐yr periods) and long‐term (matrix models) population dynamics, and neighborhood interactions of three native and codominant shrub species (Mulinum spinosum, Senecio filaginoides and Adesmia volckmanni) were measured and analysed using different statistical approaches. Results: The total density of shrubs was 74% higher in paddocks excluded from grazing, owing mainly to increases in Mulinum (80%) and Senecio (68%) species. However, differences in size structure between ungrazed and grazed paddocks were only detected in Mulinum. Demographic rates differed between shrub species, time‐periods and grazing conditions. In particular, recruitment in the short term (especially in wet years) and population growth rate in the long term (λ) were higher in paddocks excluded from grazing only in Mulinum populations. Senecio populations showed a marginal increase in recruitment and mortality independent of the grazing condition in the wet and dry period. Grazing exclusion modified the balance of neighborhood interactions among the three shrub species. In grazing‐exclusion paddocks, there was a balance between positive and negative interspecific interactions, while in grazed paddocks there were more negative intraspecific and interspecific interactions, resulting in a net negative balance of neighborhood interactions. Conclusions: Our understanding of woody encroachment in arid rangelands can be informed through evaluation of direct and indirect effects of grazing exclusion on the abundance and demography of dominant woody species. In Patagonian arid steppes, the occurrence of woody encroachment in rangelands excluded from grazing can be explained by altered responses in plant‐animal and plant‐plant interactions among shrub species.

Pyrrolizidine alkaloids of senecio sp from Peru

Six pyrrolizidine alkaloids (PAs) (two saturated macrocyclic, three unsaturated macrocyclic and one unsaturated seco-macrocyclic) were isolated from native Peruvian Senecio species. The structures of these alkaloids were established by a complete NMR spectroscopic analysis, chemical transformations and comparison of their NMR data with those published for similar alkaloids. Three PAs were then tested for antifungal activity against Fusarium moniliforme, F. (Sheldon), F. oxysporum fs. lycopersici (Scheldt) and F. solani (Mart), no significant activity being observed.

Semiquinol and phenol compounds from seven Senecio species

AbstractIn this paper, examination of the aerial parts of seven Senecio species from Serbia and Montenegro is reported. Phytochemical investigation of Senecio erucifolius led to the isolation and characterization of semiquinol butyl 2-(1-hydroxy-4-oxocyclohexa-2,5-dienyl)acetate (I), along with methyl 2-(1-hydroxy-4-oxocyclohexa-2,5-dienyl)acetate (jacaranone, II), and methyl 2-(4-hidroxyphenyl)acetate (III). The structure of I was established based on spectroscopic studies (1H- and 13C-NMR, IR, and CI-MS). Compound II was also isolated from S. carpathicus and S. subalpinus. The presence of jacaranone in the methanol extracts of S. wagneri, S. othonnae, and S. paludosus was confirmed by LC/ESI-TOF MS.

A new approach for simultaneous screening and quantification of toxic pyrrolizidine alkaloids in some potential pyrrolizidine alkaloid-containing plants by using ultra performance liquid chromatography–tandem quadrupole mass spectrometry

A rapid, but sensitive and selective method for simultaneous screening and quantification of toxic pyrrolizidine alkaloids (PAs) by ultra performance liquid-chromatography (UPLC) coupled with tandem mass spectrometry (MS/MS) on a tandem quadrupole mass spectrometer (TQ-MS) is described. This was accomplished by incorporating the precursor ion scan (PIS) acquisition and multiple reaction monitoring (MRM) acquisition in the same UPLC–MS/MS run. Notably, the developed PIS approach for detecting two pairs of characteristic product ions at m/ z 120/138 or 168/150, allowed specific identification of toxic retronecine and otonecine types PAs. This PIS method is highly sensitive with over 10-fold sensitivity improvement upon previously published LC–MS method. Moreover, this new approach is suitable for high sample throughput and was applied to the screening and quantifying toxic PAs in 22 samples collected from seven Parasenecio species and four Senecio species. In addition, coupling the MRM with PIS approach generated quantitative results equivalent to those obtained by conventional MRM-only approach. This coupled MRM with PIS approach could provide both qualitative and quantitative results without the need of repetitive analyses.