Quantitative Analysis Of Major Constituents Research Articles

Qualitative Profiling and Quantitative Analysis of Major Constituents in Jinmu-tang by UHPLC-Q-Orbitrap-MS and UPLC-TQ-MS/MS.

Jinmu-tang (JMT) is a traditional herbal medicine consisting of five herbal medicines: Poria cocos Wolf, Paeonia lactiflora Pallas, Zingiber officinale Roscoe, Atractylodes japonica Koidzumi, and Aconitum carmichaeli Debeaux. In this study, the JMT components were profiled using UHPLC-Q-Orbitrap-MS, and 23 compounds were identified and characterized. In addition, UPLC-TQ-MS/MS analysis was performed in the positive and negative ion modes of an electrospray ionization source for the simultaneous quantification of the identified compounds. The multiple reaction monitoring (MRM) method was established to increase the sensitivity of the quantitative analysis, and the method was verified through linearity, recovery, and precision. All analytes showed good linearity (R2 ≤ 0.9990). Moreover, the recovery and the relative standard deviation of precision were 86.19-114.62% and 0.20-8.00%, respectively. Using the established MRM analysis method, paeoniflorin was found to be the most abundant compound in JMT. In conclusion, these results provide information on the constituents of JMT and can be applied to quality control and evaluation.

The Effects of Aronia melanocarpa Extract on Testosterone-Induced Benign Prostatic Hyperplasia in Rats, and Quantitative Analysis of Major Constituents Depending on Extract Conditions

This study aimed to investigate the beneficial effects of A. melanocarpa on testosterone propionate (TP)-induced benign prostatic hyperplasia (BPH) in Wistar rats. Moreover, the bioactive constituents in the extract were determined using LC/MS and HPLC analyses. The dried fruits of A. melanocarpa were extracted using accelerated solvent extraction (ASE) under different extract conditions (temperature, 30 °C or 100 °C; extract solvent, 60% or 100% ethanol) to yield four extracts (T1~T4). Of the four A. melanocarpa extracts, T1 extracted under the condition of 100% ethanol/low temperature (30 °C) exhibited the greatest inhibitory activity on TP-induced prostatic hyperplasia in rats. The administration of T1 (100 mg/kg body weight, p.o.) for six weeks attenuated TP-induced prostate enlargement and reduced the levels of dihydrotestosterone (DHT) and 5α-reductase in both serum and prostate tissue. The suppression of PCNA mRNA expression in prostate tissue was remarkable in T1-treated rats. In LC/MS analysis, the levels of main anthocyanins and phenolics were significantly higher in T1 than in the other extracts. Furthermore, the quantitative study showed that the contents of cyanidin-3-glucose and cyanidin-3-xylose in T1 exhibited 1.27~1.67 and 1.10~1.26 folds higher compared to those in the other extracts. These findings demonstrated that A. melanocarpa extract containing anthocyanins as bioactive constituents attenuated the development of testosterone-induced prostatic hyperplasia, and suggested that this extract has therapeutic potential to treat prostate enlargement and BPH.

UPLC–MS/MS quantitative analysis and structural fragmentation study of five Parmotrema lichens from the Eastern Ghats

Comparative phytochemical analysis of five lichen species [Parmotrema tinctorum (Delise ex Nyl.) Hale, P. andinum (Mull. Arg.) Hale, P. praesorediosum (Nyl.) Hale, P. grayanum (Hue) Hale, P. austrosinense (Zahlbr.) Hale] of Parmotrema genus were performed using two complementary UPLC–MS systems. The first system consists of high resolution UPLC-QToF-MS/MS spectrometer and the second system consisted of UPLC–MS/MS in Multiple Reaction Monitoring (MRM) mode for quantitative analysis of major constituents in the selected lichen species. The individual compounds (47 compounds) were identified using Q-ToF-MS/MS, via comparison of the exact molecular masses from their MS/MS spectra, the comparison of literature data and retention times to those of standard compounds which were isolated from crude extract of abundant lichen, P. tinctorum. The analysis also allowed us to identify unknown peaks/compounds, which were further characterized by their mass fragmentation studies. The quantitative MRM analysis was useful to have a better discrimination of species according to their chemical profile. Moreover, the determination of antioxidant activities (ABTS+ inhibition) and Advance Glycation Endproducts (AGEs) inhibition carried out for the crude extracts revealed a potential antiglycaemic activity to be confirmed for P. austrosinense.

Qualitative and quantitative analysis of major constituents from Dazhu Hongjingtian capsule by UPLC/Q-TOF-MS/MS combined with UPLC/QQQ-MS/MS.

In this work, a sensitive and efficient method was established and validated for qualitative and quantitative analysis of major bioactive constituents in Dazhu Hongjingtian capsule by liquid chromatography tandem mass spectrometry. A total of 32 compounds were tentatively identified using ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry. Furthermore, 12 constituents, namely gallic acid, 3,4-dihydroxybenzoic acid, salidroside, p-coumaric acid-4-O-β-d-glucopyranoside, bergeninum, 4-hydroxybenzoic acid, 4-hydroxyphenylacetic acid, syringate, 6''-O-galloylsalidroside, rhodiosin, rhodionin and kaempferol-7-O-α-l-rhamnoside, were simultaneously quantified by the developed ultra-performance liquid chromatography coupled with a triple quadrupole mass spectrometry method in 9 min. All of them were analyzed on an Agilent ZorBax SB-C18 column (3.0 × 100 mm, 1.8 μm) with linear gradient elution of methanol-0.1% formic acid water. The proposed method was applied to analyze three batches of samples with acceptable linearity (R, 0.9979-0.9997), precision (RSD, 1.3-4.7%), repeatability (RSD, 1.7-4.9%), stability (RSD, 2.2-4.9%) and recovery (RSD, 0.6-4.4%) of the 12 compounds. As a result, the analytical method possessing high throughput and sensitivity is suitable for the quality control of Dazhu Hongjingtian capsule.

Qualitative and quantitative analysis of major constituents of Paeoniae Radix Alba and Paeoniae Radix Rubra by HPLC-DAD-Q-TOF-MS/MS

In order to explore the differences of chemical constituents of Paeoniae Radix Alba and Paeoniae Radix Rubra, a qualitative analytical method of liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (HPLC-Q-TOF-MS/MS) was developed for identification of multi-constituents and an HPLC-DAD analytical method was developed for simultaneously determining 14 major compounds (gallic acid, protocatechuic acid, paeoniflorin sulfonate, protocatechuic aldehyde, methyl gallate, oxypaeoniflorin, catechin, albiflorin, and paeoniflorin, ethyl gallate, benzoic acid, pentagaloylglucose, benzoyl-paeoniflorin, and paeonol) in Paeoniae Radix Alba and Paeoniae Radix Rubra. Q-TOF/MS qualitative analysis was performed under negative ion mode and inferred 38 components of Paeoniae Radix Alba and 30 components of Paeoniae Radix Rubra. HPLC-DAD quantitative method result showed the contents of 8 ingredients were different between Paeoniae Radix Alba and Paeoniae Radix Rubra. The results indicated that the new approach was applicable in qualitative and quantitative quality control of Paeoniae Radix Alba and Paeoniae Radix Rubra.

Qualitative and quantitative analysis of major constituents in Tetrastigma hemsleyanum by HPLC-Q-TOF-MS and UPLC-QqQ-MS

A qualitative analytical method of liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (HPLC-Q-TOF-MS) was developed for identification of multi-constituents and an analytical method was developed for simultaneously determining 4 major compounds (rutin, isoquercitrin, kaempferol-3-0-rutinoside, and astragalin) in Tetrastigma hemsleyanum Diels et Gilg. The HPLC-Q-TOF-MS assay was performed on a Welch Ultimate XB-C18 column (4.6 mm x 150 mm, 5 microm) with the mobile phase consisting of acetonitrile (A) and water containing 0.1% Formic acid (B) in gradient mode at a flow rate of 0.8 mL x min(-1). The column temperature was at 30 degrees C, and negative ion mode was used for TOF-MS. The UPLC-QqQ-MS assay was performed on a Waters CORTECS C18 (2.1 mm x 100 mm, 1.6 microm) with the mobile phase consisting of acetonitrile (A) and water containing 0.1% formic acid (B) in gradient mode at a flow rate of 0.25 mL x min(-1). The column temperature was at 45 degrees C, and MRM mode was used for QqQ-MS. Based on the retention time and MS spectra, 24 compounds were identified or tentatively characterized by comparing with reference substances or literatures. For quantitative the linear range of 4 detected compounds were good (r > 0.9966), and the overall recoveries ranged from 98.27% to 101.58%, with the RSD ranging from 3.15% to 5.88%. The results indicated that new approach conbined HPLC-Q-TOF-MS and UPLC-QqQ-MS was applicable in qualitative and quantitative quality control of Tetrastigma hemsleyanum.

Quantitative analysis of major constituents in green tea with different plucking periods and their antioxidant activity.

The objective of this study was to determine the relationship between the plucking periods and the major constituents and the antioxidant activity in green tea. Green tea was prepared from leaves plucked from the end of April 2013 to the end of May 2013 at intervals of one week or longer. The contents of theanine, theobromine, caffeine, catechin (C), and gallocatechin gallate (GCg) were significantly decreased, whereas those of epicatechin (EC), epigallocatechin gallate (EGCg) and epigallocatechin (EGC) were significantly increased along with the period of tea leaf plucking. In addition, antioxidant activity of green tea and standard catechins was investigated using ABTS, FRAP and DPPH assays. The highest antioxidant activity was observed in relatively the oldest leaf, regardless of the assay methods used. Additionally, the order of antioxidant activity of standard catechins was as follows: EGCg ≥ GCg ≥ ECg > EGC ≥ GC ≥ EC ≥ C. Moreover, the cis-catechins contents were the key factor affecting the antioxidant activity of green tea in all assays employed (ABTS, r = 0.731, p < 0.01; FRAP, r = 0.886, p < 0.01; DPPH, r = 0.778, p < 0.01).

A Rapid Method for Qualitative and Quantitative Analysis of Major Constituents in Dengzhanxixin Injection by LC-DAD-ESI–MSn

For the first time a rapid method for qualitative and quantitative analysis of constituents in Dengzhanxixin injection was established by liquid chromatography with electrospray ionization–tandem mass spectrometry. Eighteen compounds including flavonoids and phenolic acids were characterized or tentatively identified. Ten of these compounds, including 5-O-caffeoylquinic acid, caffeic acid, 1,3-O-dicaffeoylquinic acid, 6′-O-caffeoylerigeroside, scutellarin, 3,4-O-dicaffeoylquinic acid, 3,5-O-dicaffeoylquinic acid, erigoster B, apigenin-7-O-glucuronide and 4,5-O-dicaffeoylquinic acid, were further quantified as marker substances by LC-DAD using a C18 column at 0.4 mL min−1 within 37 min. The quantitative method was validated for ten interesting compounds, including linearity, accuracy, precisions, LOQ and LOD, which was proved to be sensitive, reproducible and accurate. The study might provide a comprehensive method for the quality assessment of dengzhanxixin injection.

Modulation of cytochrome P450 1A1 expression and activity in intestinal Caco-2 cells by components of Ginkgo biloba-based dietary supplements

Ginkgo biloba is largely consumed as food supplement, because it could contribute to memory improvement by acting on cerebrovascular insufficiency. Among the large variety of phytochemicals contained in Ginkgo biloba leaves and extracts, flavonols (mainly kaempferol, quercetin and isorhamnetin) represent the largest part. Several unique terpenoids (ginkgolides A, B, C, J and bilobalide) are also present. In the present study, we investigated the modulation of cytochrome P450 (CYP) 1A1 enzymatic activity and expression in the intestinal Caco-2 cell line by either Ginkgo biloba extracts, by major individual secondary metabolites or by mixtures of these constituents in realistic proportions. Plausible used intestinal concentrations were calculated on the basis of recommended daily intake and quantitative analysis of major constituents in dietary supplements. Preliminary results suggest that (−)-bilobalide, but not the other terpenoids, induces a dose-dependent increase of CYP1A1 activity (measured through 7-ethoxyresorufin O-deethylation, EROD) after 6 h treatment of the Caco-2 cells. RT-PCR highlighted an increase of mRNA after 3 h treatment with (−)-bilobalide, suggesting the involvement of a transcriptional mechanism. Furthermore, a strong dose-dependent inhibition of benzo[a]pyrene (B[a]P)-induced EROD activity was shown after 6 h co-treatment with kaempferol, quercetin or isorhamnetin. This antagonistic effect was conserved with mixtures of the three flavonols and of the eight pre-cited major secondary metabolites. These preliminary results suggest that frequent consumption of Ginkgo biloba-based dietary supplements may influence CYP1A1 expression and activity in intestinal cells, leading to possible toxicological interactions.

Near Infrared Analysis of Whole Blood and Plasma in Blood-Collecting Tubes

An easy-to-use measurement method requiring no cleaning between samples, was developed and applied for the determination of cows' plasma constituents. Whole blood and plasma samples were measured with blood-collecting tubes as sample cells, providing easy sample handling and presentation. For whole blood, near infrared (NIR) spectra in the short wavelength region from 600 to 1000 nm were recorded with a portable NIR instrument (Fruit Tester 20: FT20). Plasma samples were measured in both the short wavelength region with the FT20 and in the long wavelength region from 1100 to 2500 nm with an NIRSystems 6500. Both instruments were equipped with specially designed sample holders for blood-collecting tubes and fibre optics. Partial least squares (PLS) regression was applied to derive calibrations for total protein, total cholesterol and glucose. Whole blood showed inferior performance to plasma because of scattering interference and absorption by red blood cells. Plasma results heavily depended on the constituent analysed. Total protein was determined accurately in both wavelength regions due to its high concentration and high absorptivity. For total cholesterol, the long wavelength region provided much better results because of its higher absorptivity. Results for glucose were only mediocre in both regions as its concentration and absorptivity were low. Overall, results indicate that blood-collecting tubes can be easily and effectively used for quantitative analysis of major constituents in blood plasma.

Quantification of germanium and boron in heterostructures Si/Si 1− xGe x/Si by SIMS

The use of secondary ions mass spectrometry (SIMS) generally leads to very high sensitivity and depth resolution for silicon-based components. Low concentration elements such as dopants, are easy to quantify in silicon, whereas high concentration elements are subject to “matrix effects”, i.e. the intensity does not vary linearly with the concentration. Matrix effects are presently studied in SiGe alloys using two primary beams O 2 + and Cs + and a CAMECA IMS4F instrument. Rapid thermal chemical vapor deposition grown Si/Si 1− x Ge x /Si multilayer structures with x varying from 0 to 23.5% are preliminarily characterized by Rutherford backscattering spectrometry (RBS) in thickness and atomic composition. The linear variation of the SIMS intensity ratio I Ge +/ I Si + with the RBS concentration ratio x/(1− x) is confirmed for both O 2 + and Cs + beams. However, the linearity does not imply the absence of matrix effects. A variation of the ionization yields with Ge concentration and a similar behavior for τ Ge+ and τ Si+ actually lead to an exact compensation of the intensity ratio variation. At any energy, O 2 + experiments systematically over-estimate the real Ge concentration, whereas matrix effects under Cs + beam are weak. For polyatomic MCs + ions, strong matrix effects are observed in Si 1− x Ge x alloys. A quantitative analysis of major constituents and a dopant (boron) is used to determine the depth profiles of a thin base of a SiGe HBT heterostructure. Two different procedures are used depending on the primary beam. Consistent results are obtained for the concentrations of the major elements. The fully autonomous SIMS analysis with the Cs + beam is still penalized by the lack of sensitivity for boron in the secondary positive mode. © 1997 Elsevier Science S.A. All rights reserved.

Use of indirect multivariate calibration for quality control of agricultural products by near infrared spectroscopy

In the near infrared (NIR) region, (λ=800–2500 nm) of the spectrum, most organic molecules have weak but reproducibly measurable absorption bands. This phenomenon has been increasingly exploited for the rapid, quantitative analysis of major constituents of agricultural products. As there is strong spectral overlap and an interaction of constituents in NIR, the calibration of instrumentation has to be performed indirectly with a training set of samples applying multivariate methods. A software package for IBM compatible personal computers includes programs to select samples to be used for calibration, to compute multiple linear regression equations, to cross-validate regression equations and to detect unusual samples. These methods are applied to rapidly determine the oil content of over a thousand rapeseed samples of the harvest of 1986. A considerable saving of time, labour, and costs can be achieved, the agreement with the standard laboratory method is very satisfactory.