UPLC-DAD Method Research Articles

Transformation profiles of the isoflavones in germinated soybean based on UPLC–DAD quantification and LC–QTOF–MS/MS confirmation

Germinated soybean is one kind of food and a medicine. In the actual process of producing a large amount of naturally germinated soybean, it is difficult to strictly control the germination process conditions. However, sprout length may be more suitable as the terminal judgment indicator for naturally germinated soybean. An UPLC–DAD method was developed and validated to explore the transformation profiles of soybean isoflavones in germinated yellow or black soybean with different sprout lengths. Moreover, an LC − QTOF−MS/MS method was used to avoid false positive results. The contents of daidzein, glycitein, and genistein almost reached their corresponding maximum values when the sprout length ranged from 1.0 cm to 1.5 cm (P < 0.05). Therefore, yellow soybean is suggested to be the processing raw material with higher contents of those isoflavones, and the optimal sprout length for germinated soybean may be in the range of 1.0–1.5 cm.

Simultaneous assay of pyrimethamine and methylparaben in extemporaneous pediatric suspensions by an isocratic stability-indicating UPLC-DAD method

Pyrimethamine (PYM) is an antiparasitic drug used in the treatment of congenital toxoplasmosis, available only as tablets. Therefore, preparing extemporaneous formulations and their quality control is essential for safe pediatric treatment. This study aimed to develop and validate a stability-indicating method by ultra-performance liquid chromatography (UPLC) to quantify PYM and methylparaben (MP) in extemporaneous oral suspension and evaluate its applicability to the tablets assay. The analytical conditions included RP-C18 column, mobile phase composed of 0.05 M acetate buffer pH 4.5 and organic mixture (acetonitrile:methanol), at 60:40 (v/v), flow rate of 0.2 mL/min, injection volume of 2 µL and detection at 272 nm. PYM was detected at 3.9 min and MP at 4.7 min, without interference of the degradation products or other excipients. Linearity was observed in the range of 0.5 to 25 μg/mL. The method showed accuracy (mean recovery between 98.0 and 102.0%) and precision (RSD < 2%) for suspension and tablets; the method was robust regarding the variations in the analytical conditions. The developed method met the requirements of current guidelines, was indicative of stability, and was suitable for PYM determination in extemporaneous preparations and tablets.

Evaluation of antiplasmodial potential of Cissampelos pareira L.: Characterization and quantification of isoquinoline alkaloids responsible for the biological activity

Globally, based on traditional knowledge, different parts of Cissampelos pareira plant are being used individually or in combination in various forms to manage malaria. However, the scientific investigation for validating the most effective part of this plant against malaria parasite has not been done. Thus, the study aims to explore the antiplasmodial potential of different parts of the plant against malaria parasite by bio-assay guided isolation of compounds from different extracts and fractions. Antiplasmodial activities of the whole plant parent extract and its fractions along with decoctions (roots, stem, leaves and whole plant) and isolated molecules were evaluated against chloroquine-sensitive Pf3D7 and chloroquine-resistant PfINDO strains of Plasmodium falciparum. A UPLC-DAD based method was developed to quantify the marker compounds in all samples by utilizing isolated molecules as analytical standards. The chloroform fraction of the whole plant was found to be the most potent with IC50 (µg/mL) of 0.79 (Pf3D7) and 2.26 (PfINDO) followed by root decoction with IC50 (µg/mL) 10.22 (Pf3D7) and 7.7 (PfINDO). Further, phytochemical analysis of active chloroform fraction led to the isolation of two potent bisbenzylisoquinoline compounds, namely O,O-dimethylcurine (1) [IC50 (µM) 1.46 (Pf3D7) and 0.51 (PfINDO)], and curine (2) [IC50 (µM) 1.46 (Pf3D7) and 0.51 (PfINDO)]. Analysis of developed UPLC-DAD method showed the highest quantities of curine (2) (∼107 mg/g) and O,O-dimethylcurine (1) (∼15 mg/g) in chloroform fraction which might be responsible for its potent activity. This study showed that the root decoction was more effective than decoctions of each of the other parts of the plant and the whole plant hydroalcoholic extract.

Simultaneous and rapid quantification of multi-compounds in Nauclea officinalis based on UPLC-DAD

Nauclea officinalis is a Li-folk medicine in China, with antimalarial, antibacterial, and anti-inflammatory properties. However, the phytochemicals of Nauclea officinalis have not yet been comprehensively researched. In this paper, a reliable and efficient approach based on ultra-performance liquid chromatography (UPLC) with a diode array detector (DAD) was firstly established for simultaneous quantification of alkaloids, flavonoids, and phenolic acids in N. officinalis resources. The samples were extracted using ultrasound with 60 % ethanol for 20 min (1:50 g/mL), and analyzed by the developed UPLC-DAD method. Under the optimized extraction and chromatographic conditions, the proposed approach showed favorable performance in terms of the linearity (R2 > 0.9998), precision (RSD < 3.75 %), repeatability (RSD < 0.86 %), stability (RSD < 2.14 %), and accuracy (with the recovery rates between 95.15 % and 105.49 %) within an extremely short running time (12 min). Moreover, the developed approach was successfully employed in the quantification of ten phytochemicals in different parts (leaves, branches and trunks) of Nauclea officinalis. Based on analytical results, the content of ten phytochemicals in different organs of Nauclea officinalis was significantly different, and the superior contents of these phytochemicals were located in the leaves compared to the branches and trunks. In summary, the current work established an effective method for the assay of multi-compounds, which could supply a basis of the quality control and comprehensive utilization for Nauclea officinalis.

Development and validation of a stability indicating UPLC-DAD method coupled with MS-TQD for ramipril and thymoquinone in bioactive SNEDDS with in silico toxicity analysis of ramipril degradation products

Development and validation of a stability indicating UPLC-DAD method coupled with MS-TQD for ramipril and thymoquinone in bioactive SNEDDS with <i>in silico</i> toxicity analysis of ramipril degradation products

Chemical and biochemical characterization of Ipomoea aquatica: genoprotective potential and inhibitory mechanism of its phytochemicals against α-amylase and α-glucosidase.

Ipomea aquatica, also known as water spinach, is an aquatic non-conventional leafy vegetable and is considered a healthy and seasonal delicacy in ethnic food culture. The study revealed the presence of rich chemical and biochemical composition in I. aquatica and antioxidant activities. Moreover, the plant extracts demonstrated significant DNA damage prevention activity against UV/H2O2-induced oxidative damage. High-resolution mass spectrometric analysis by UPLC-qTOF-MS/MS resulted in the identification of over 65 different compounds and 36 important secondary metabolites. Most of the compounds identified represented polyphenolic compounds, viz. polyphenol glycosides and phenolic acids, followed by alkaloids and terpenoids. A UPLC-DAD method was developed and quantified for 10 different polyphenolic compounds. Out of all the metabolites examined, a significant number of compounds were reported to have various bioactive properties, including antibacterial, antiviral, antitumor, hepatoprotection, and anti-depressant effects. The plant extracts were found to contain various compounds, including euphornin, lucidenic acid, and myricitin glycosides, which possess significant medicinal value. Metabolite analysis utilizing GC-MS revealed the presence of various fatty acids, amino acids, sugars, and organic acids. The analysis revealed the presence of essential unsaturated fatty acids such as α-linolenic acid as well as beneficial substances such as squalene., The evaluation of glycemic control activity was carried out by comprehending the inhibitory potential of α-amylase and α-glucosidase, outlining the kinetics of the inhibition process. The inhibitory activities were compared to those of acarbose and revealed stronger inhibition of α-glucosidase as compared to α-amylase. Furthermore, the mechanism of inhibition was determined using in silico analysis, which involved molecular docking and molecular dynamic simulation of the identified IA phytochemicals complexed with the hydrolase enzymes. The study generates convincing evidence that dietary intake of I. aquatica provides a positive influence on glycemic control along with various health-protective and health-promoting benefits.

Identification and quantitation of NF-κB inhibitory components in weichang'an pill based on UHPLC-QE-MS and spectrum-effect relationship

Weichang'an pill (WCAP) is a traditional Chinese patent medicine, which is clinically used for the treatment of bowel syndrome and functional dyspepsia such as diarrhea, abdominal distension, and enteritis. So far, quality control studies of WCAP have mainly focused on the determination of chemical composition content, which has little relevance to biological activity and clinical effects. With the aim of identifying the multi-index ingredients with NF-κB inhibitory activities related to WCAP clinical effect, this present work described the chemical profile of WCAP by UHPLC-QE-MS, established the correlated relationship between chromatographic fingerprints and the NF-κB inhibitory activities based on multivariate statistical analysis, including hierarchical clustering analysis (HCA), Pearson correlation analysis, and Partial least squares regression analysis (PLSR). The spectrum-effect relationship analysis indicated 10 compounds, which were ferulic acid, naringin, narirutin, hesperidin, neohesperidin, aloe emodin, emodin, honokiol, magnolol, and physcion, might be the potential NF-κB inhibitory constituents in the pill. The NF-κB inhibitory effects of the ten compounds were verified by in vitro dual luciferase reporting detection system. Considering that the detection index should be representative of more medicinal materials, a rapid and efficient UPLC-DAD method was eventually developed to determine the content of the 13 components. Our findings will provide data support for WCAP quality control and advance the understanding of the quality assessment of traditional Chinese patent medicines.

Development and Validation of a Stability-indicating UPLC-DAD Method for the Simultaneous Determination of Ivermectin and Praziquantel in Pharmaceutical Tablets and Dissolution Media.

Currently, there is no single rapid and accurate stability-indicating quantitative method that can simultaneously determine both ivermectin and praziquantel and their related compounds. Thus, the goal of this research is to develop and validate a new rapid, accurate, and stability-indicating ultra-performance liquid chromatography (UPLC) method. The method uses a water, acetonitrile, and methanol gradient. The chromatographic separation was achieved on a C18 (1.7 μm, 2.1 × 50 mm) column with a flow rate of 0.7 mL/min, and the column temperature was maintained at 40°C. Analytes are detected at 245 nm. The method was validated in accordance with ICH Q2R1 guidelines. The linearity (R2) was >0.9987 and 0.9997 for praziquantel and ivermectin, respectively. The corresponding accuracy ranged between 98.0 and 102.0%. Intermediate precision (assessed as inter-day precision) was determined by calculating the cumulative %CV of eighteen assay preparations and was less than 2.0% for both praziquantel and ivermectin. The specificity of the method was shown by the resolution of the two active pharmaceutical ingredients (APIs) from any interfering excipients, impurities, or degradation products. The limit of detection and quantitation for ivermectin was 26.80 ng/mL and 81.22 ng/mL, respectively. The limit of detection and quantitation for praziquantel was 1.39 μg/mL and 4.22 μg/mL, respectively. The robustness study proved that method performance is stable against small variations in sample processing parameters (shaking, sonication time, and acetonitrile % in solvent solution) and also against small variations in the initial % of mobile phase components and gradient slope. Using ICH Q2R2 criteria, the method was demonstrated to be specific, accurate, stability indicating, and robust to small variations of chromatographic variables.

Design of Experiment-Based Green UPLC-DAD Method for the Simultaneous Determination of Indacaterol, Glycopyrronium and Mometasone in their Combined Dosage Form and Spiked Human Plasma.

Abstract Indacaterol, is an ultra-long-acting β2 agonist, glycopyrronium is a long-acting muscarinic-antagonist and mometasone is a synthetic corticosteroid. They were used recently in combination for the treatment of severe asthma symptoms and chronic obstructive pulmonary disease. In this work, it was the first time to develop a green and environment friendly ultra-performance liquid chromatographic method using design expert program for the analysis of the three drugs in their combined dosage form. Also, the method was bioanalytically validated for the analysis of the three drugs in spiked human plasma samples. The method was linear in range from 0.50 to 100.0 μg mL−1 for indacaterol and mometasone and from 1.0 to 150.0 μg mL−1 for glycopyrronium. It showed high accuracy where, the % recovery for indacaterol, glycopyrronium and mometasone in plasma were ranged from 94.27 to 97.86%, 96.43 to 98.75% and 96.86 to 98.43%, respectively. Also, it was precise where, the % relative standard deviation for the inter-day precision was ranged from 2.571 to 3.484%, 3.180 to 4.123% and 3.150 to 3.984% and the intra-day precision was ranged from 2.351 to 3.125%, 2.512 to 3.544% and 2.961 to 3.983% for indacaterol, glycopyrronium and mometasone, respectively. The limit of detection and the limit of quantification for indacaterol and mometasone were 0.03 and 0.10 μg mL−1 while for glycopyrronium, they were 0.16 and 0.50 μg mL−1. Highlights

Eco-Efficient Quantification of Glucosinolates in Camelina Seed, Oil, and Defatted Meal: Optimization, Development, and Validation of a UPLC-DAD Method.

Camelina sativa (camelina) seed, oil, and defatted meal are widely used for food, animal feed, and other purposes. The accurate quantification of camelina glucosinolates is critical as their functionalities are highly dose-dependent. The classic quantification of glucosinolates in camelina products involves tedious desulfation steps, toxic reagents, and a lengthy instrument time because glucosinolates are easy to degrade and subject to interference in the liquid chromatography. Thus, we developed and validated an eco-efficient UPLC-DAD method for determining glucoarabin (GS9), glucocamelinin (GS10), and homoglucocamelinin (GS11) in camelina seed, oil, and defatted meal. Glucosinolates were extracted using 80% cold methanol to denature myrosinase, and were separated by an HSS T3 column without desulfation. Glucotropaeolin was used as an internal standard to track analyte degradation and loss during sample preparation. The method has shown high precision (relative standard deviations ranging from 4.12% to 6.54%) and accuracy (&gt;94.4% spike recovery) for GS9-11, and all validation parameters passed the industry-consensus AOAC Appendix F criteria. To our best knowledge, this is the first eco-efficient and low-cost analytical method that is validated against strict AOAC criteria for the quantification of intact camelina glucosinolates. The method is suitable to be adopted as a new industrial testing standard to assist in the quality control of camelina products.

Main components in different varieties of Xihuangcao by UPLC-Q-TOF-MS and UPLC-DAD

This study established an ultra-high performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS) method to analyze the main components in different varieties of Xihuangcao and established a UPLC-DAD method to simultaneously determine the five active components(caffeic acid, rosmarinic acid, schaftoside, isoschaftoside, and oridonin).The chromatographic separation was performed on a Waters ACQUITY UPLC BEH C_(18) column(2.1 mm×100 mm, 1.7 μm) with a gradient elution of methanol(B)-water containing 0.1% formic acid(A) at a flow rate of 0.3 mL·min~(-1).The column temperature was 30 ℃.The Q-TOF-MS discriminant analysis was performed under positive electrospray ion mode and the split ratio was 1∶1. Quantitative analysis was carried out by UPLC-DAD.The determination wavelength was set at 245 nm.Thirty-two main components of Xihuangcao were separated and identified by UPLC-Q-TOF-MS, where 19 were identified in Rabdosia serra, nine in R.nervosa, 10 in R.lophanthoides, 15 in R.lophanthoides var.graciliflora, 10 in R.lophanthoides var.gerardianus, and seven in R.stracheyi.The UPLC-DVD method was developed for simultaneously determining five active components in different varieties of Xihuangcao.The standard curves for five compounds showed good linearity with correlation coefficients higher than 0.999 0.The precision, repeatability, and stability were good.The average recoveries(n=6) were between 97.01% and 102.7% with RSD&lt;3.0%.The results of UPLC-Q-TOF-MS analysis provided a scientific basis for the use of R.stracheyi as a medicinal material of Xihuangcao and the equivalent use of R.lophanthoides var.gerardianus with R.lophanthoides var.graciliflora to some extent.The UPLC-DAD method for simultaneously determining five active components is simple, rapid, and accurate.This study can provide the basis for the quality control of different varieties of Xihuangcao.

A Uplc-Dad Method to Separate and Quantify Carotenoids and Chlorophylls, Including Geometric Xanthophyll Isomers, from Diverse Plant Matrices

A Uplc-Dad Method to Separate and Quantify Carotenoids and Chlorophylls, Including Geometric Xanthophyll Isomers, from Diverse Plant Matrices

Rapid Detection of Chloramphenicol Residues in Fermented Foods Based on UPLC-DAD

in the detection of chloramphenicol residues in fermented food, there are often problems of slow detection speed. Using UPLC-DAD method, a rapid detection method of chloramphenicol residues in fermented food based on UPLC-DAD method is designed. According to the characteristics of chloramphenicol, set up the detection reagent, select the detection equipment, and form the detection laboratory. It is usingUPLC-DAD method to design the test paper, using the set test reagent to deal with the sample to be tested, according to the design results of the test process, combining the reagent with the sample, to determine its specificity. Chloramphenicol residue was detected by test paper. So far, the rapid detection method of chloramphenicol residues in fermented food based on UPLC-DAD method has been designed. Compared with the original detection method, the detection speed of the detection method designed in this paper is significantly higher than the original method. In conclusion, the rapid detection method of chloramphenicol residues in fermented food based on UPLC-DAD method is effective.

Stability Study of Admixtures Combining Ziconotide With Morphine or Sufentanil in Polypropylene Syringes

The association of morphine ziconotide or sufentanil ziconotide was used to manage cancer pain. Moving these patients is sometimes difficult. In order to transport these syringes for pump refilling, it could be interesting to demonstrate the stability of the mixture and so to be able to ensure the best transport conditions of syringes. A stability indicating UPLC-DAD method was developed and validated according to the ICH guidelines. Fur mixtures of each association have been stored in 5 ± 3°C and 25 ± 2°C and were evaluated for seven days and compared to the initial observed concentrations. The stability of these associations was demonstrated at 5°C for seven days thanks to relative concentrations (95% confidence intervals of the mean of three samples) systematically positioned between 95% and 105%. No degradation product was observed during the stability study. This study shows the stability of these association morphine ziconotide or sufentanil ziconotide at 5°C for seven days in polypropylen syringes. This result will allow the transport of the preparation under optimal conditions. Advance preparations for intrathecal pump refills could also be feasible.

Evaluation of Anti-Obesity Activity of an Herbal Formulation (F2) in DIO Mice Model and Validation of UPLC-DAD Method for Quality Control

Obesity is considered a chronic metabolic disorder that can be associated with multiple medical complications. Currently, there is no or limited curative therapy for obesity. This study focused on the assessment of anti-obesity activity and UPLC standardization of a polyherbal formulation (F2). An anti-obesity activity was investigated using the diet-induced obese (DIO) mice model, where obesity was developed in C57BL/6J mice by providing a high-fat diet (HFD) for five weeks without treating drugs. After the successful development of obesity, the obese mice were treated with F2 for seven weeks with continuing HFD feeding. The major obesity-related parameters such as body weight gain, food efficiency ratio, serum lipid profile, and white adipose tissue (WAT) mass were found to be significantly reduced in F2 treated obese mice. These results were supported by the down-regulation of specific adipogenic transcription factors (PPARγ, SREBP-1c, and ap2) in epididymal WAT. Histological evaluation of liver and WAT also revealed reduced fat deposition in the tissues by F2 compared to the HFD control group. The overall observations indicated that the F2 exhibited pronounced obesity-controlling activity through the inhibition of adipocyte differentiation and triglyceride accumulation in the tissues, and serum lipid depletion. In addition, F2 ameliorated obesity-induced insulin resistance. Furthermore, the UPLC-DAD method for quality control of F2 was validated and standardized using five reference compounds: astragalin, ellagic acid, fisetin, fustin, and sulfuretin.

Development and Validation of a UPLC-DAD Method for the Simultaneous Quantification of Eight Antihypertensive Drugs in the Pharmaceutical Matrix.

Hypertension is a critical health problem; it is a prevalent risk factor for cardiovascular disease. Many treatments to combat hypertension are available, however many patients are resistant to the standard therapeutic approaches. The association of two or more substances in a fixed-dose combination is effective and tolerated as a substitute for the standard therapeutic approach. The new ultra performance liquide chromatography method was developed and validated to assay a combination of eight antihypertensive drugs including a diuretic: hydrochlorothiazide, dihydropyridine calcium channel blocker: Amlodipine and angiotensin II type 1 receptor blockers (sartans): valsartan, candesartan, eprosartan, olmesartan, losartan, and irbesartan in the pharmaceutical matrix. Chromatographic separation was performed on an Acquity® UPLC C18 1.7 µm 2.1 × 100 mm column, with a gradient of buffer solution and acetonitrile, in the proportion of (80:20 v/v). Good resolution was obtained, and an optimal analysis time of less than 5 min was achieved. The method was validated according to the International Conference on Harmonization guidelines following the classical approach and accuracy profile, and it is shown to be suitable for intended applications. The method was successfully used for quality control laboratories and the determination of these drugs combinations in pharmaceutical dosage forms.

Bryophyllum pinnatum markers: CPC isolation, simultaneous quantification by a validated UPLC-DAD method and biological evaluations

Bryophyllum pinnatum (Lam.) Oken (Crassulaceae) is widely used as leaf juice or extracts in traditional medicine all over tropical areas, especially in Brazil, to relieve inflammation-associated symptoms. Flavonol glycosides with unusual sugar moiety are among the major metabolites. Nevertheless, there are not enough quality control studies that can contribute to authentication of B. pinnatum and determination of their markers. As it is also used as medicinal plant in several countries, it is necessary to provide data related to safety, efficacy and quality. In this context, this work aims to isolate the major flavonoids from B. pinnatum hydroethanolic extract, to validate a method to quantify the content of chemical markers and to evaluate their xanthine oxidase inhibition and antioxidant activity. The extract was submitted to centrifugal partition chromatography (CPC). The solvents system CyHex-EtOAc-EtOH-H2O, 0.5:9:3:5.5, v/v/v/v was selected by shake-flask method. Four flavonoids (quercetin 3-O-α-L-arabinopyranosyl-(1→2)-O-α-L-rhamnopyranoside (1), kaempferol 3-O-α-L-arabinopyranosyl-(1→2)-O-α-L-rhamnopyranoside (2), quercetin 3-O-α-L-rhamnopyranoside (3) and kaempferol 3-O-α-L-rhamnopyranoside (4)) were isolated in a single and fast CPC run and their structures were confirmed by NMR analysis. An UPLC-DAD quantification method was established for the first time with validation of required parameters, according to RDC 166/2017. The calibration curves were linear with correlation coefficient ranging from 0.9996 to 0.9997 while the values of LOD (0.0077–1.984 ng.mL−1), LOQ (0.0263–6.012 ng.mL−1), recovery (≥ 80.7 %) and inter-day (%RSD ≤ 3.581) and intra-day precision (%RSD ≤ 2.628) were satisfactory. Quantitative analysis of these compounds showed that the proportion of 1, 2 and 3 were 2.43, 0.25 and 0.33 % (24.3 mg.g−1, 0.25 mg.g−1 and 0.33 mg.g−1 of extract), respectively. Moreover, in vitro xanthine oxidase (XO), DPPH and ABTS inhibition were evaluated for the extract and the major flavonoids. Compounds 2 (168 μM) and 3 (124 μM) moderately inhibited XO, while compounds 1 and 3 displayed average radical scavenging activity. In conclusion, our results suggest the flavonoid 1 as a specific marker which may be used for quality control of B. pinnatum hydroethanolic leaves extract.

Study of Physicochemical Stability of Ziconotide in Medication Cassette Reservoir

To determine the physicochemical stability of ziconotide solutions for intrathecal administration in the Medication Cassette Reservoir (MCR). A stability indicating UPLC-DAD method was developed and validated according to the ICH guidelines. Two mixtures of ziconotide (0.40 μg/mL and 0.60 μg/mL) stored in MCR stored at 25 ± 2°C were evaluated for 14 days and compared to the initial observed concentrations. The physicochemical stability of the two solutions was demonstrated for two days thanks to relative concentrations, pH measurement, visual inspections, and turbidity assays. A degradation product was observed and increased during the study. This study showed a very low physicochemical stability of diluted ziconotide stored at 25 ± 2°C in the MCR. The intrathecal administration of ziconotide does not seem appropriate with this device for outpatients.

Content determination and principal component analysis of nine active components in Paeonia lactiflora flowers from different cultivars and drying methods

This study aims to establish an UPLC-DAD method for the simultaneous determination of nine active components in phenolic acids, anthocyanins, monoterpene glucosides and flavonoids in the petals of Paeonia lactiflora flowers from different cultivars and processing methods. UPLC analysis was performed on a Waters ACQUITY UPLC HSS T3 C_(18) column(2.1 mm × 100 mm, 1.8 μm)for gradient elution with a mobile phase consisting of 0.5% formic acid solution(A)-acetonitrile(B)at a flow rate of 0.3 mL·min~(-1).The detection wavelength was determined with a switched wavelength method and the column temperature maintained at 20 ℃, with an injection volume of 2 μL.The contents of the above components in the samples with different sources and treatment methods were obtained. The principal component analysis(PCA)of P. lactiflora flowers was conducted by using SIMCA 14.0 software, and the results showed that the P. lactiflora flowers could be classified into two categories according to cultivars, including P. lactiflora 'Bozhou-shaoyao' as one category with high contents of the above components, P. lactiflora 'Baihuachuanshaoyao' and P. lactiflora 'Honghuachuanshaoyao' as the other category. Dried P. lactiflora flowers could be classified into three categories, including baking-drying, sun-drying and shade-drying, with baking-drying method as the optimal one.The P. lactiflora flowers were not greatly affected by origins, growing altitudes, growing years, or blooming stages.The results of contents determination can be used to guide the cultivar selection, harvest and processing of P. lactiflora flowers.

Determination of Phenolic Compounds in the Methanolic Extract of Pourouma guianensis Leaves by UPLC-DAD-MS

Background: The presence of phenolic compounds is common in Urticaceae. However, only one report of the isolation of an apigenin in the Pourouma genus is available. Pourouma guianensis occurs widely throughout Brazil, mainly in the Southeastern region, and is popularly known as “pitinga”. The chemical profile of P. guianensis is marked by the presence of steroids and triterpenes. Objective: This study carried out the characterization of phenolic substances using UPLC-DAD-MS separation, in order to increase the chemical knowledge on the Pourouma genus. Methods: The extract was analyzed by UPLC-DAD-MS using a C-18 column, DAD detector at 190- 400nm and ESI-Q-TOF mass spectrometer. The solvent system consisted of water with 0.1% formic acid (phase A) and acetonitrile with 0.1% formic acid (phase B). Results: The extract analysis identified the presence of ten compounds: two aryl propanoids (5-Ocaffeoylquinic acid and 3-O-feruloylquinic acid) and eight glycosylated flavones derived from apigenin, luteonin, and chrysoeryol (vicenin 2, saponarin, orientin, isoorientin, isoschaftoside, schaftoside, isovitexin, and isoscoparin). Conclusion: The developed UPLC-DAD method was proven to be an efficient tool for the separation of phenolic mixtures and the UPLC-DAD-MS analysis enabled the identification of these compounds, reported for the first time in Pourouma genus.