Determination of 19 rhubarb constituents by high‐performance liquid chromatography–ultraviolet–mass spectrometry

Abstract

Rhubarb (Rhei rhizoma), a commonly used Chinese herb, contains anthraquinones, anthrones, galloylglucoses, stilbenes, and flavan-3-ols compounds, etc. as major constituents. Using 19 of these compounds as markers, an HPLC-UV-MS method was developed to estimate the quality of rhubarb samples within a period of 70 min. Extracts were analyzed with a Cosmosil 5C18-MS column and eluted with a gradient comprising an aqueous solution of acetic acid and methanol at a flow rate of 0.9 mL/min. Peaks were detected by absorbance measurements at 254 nm (6 and 8-19) and 280 nm (1-5 and 7), and the peaks of the marker substances were identified from their UV spectra and MS fragmentation patterns. The proposed method yielded a peak-area ratio RSD value with an intraday SD falling within 0.71-1.78% and an interday SD within 0.78-1.98% at a detection limit of 0.2-3.2 microg/mL. The ESI negative ion mode was used to collect data (molecular weight, CID fragments from MS and MS/MS spectra) for 19 compounds from four types of structure categories: anthraquinones, dianthrone glycosides, stilbenes, and galloylglucosides. The information gathered can be used to identify the structures of various peaks appearing in the LC chromatograms of rhubarb samples.

Ultrasonic cell grinder extraction of anthraquinones from Radix et Rhizoma Rhei and determination by ultra‐performance liquid chromatography

“…With the UPLC method, the usage of solvent decreased greatly, which is in favor of decreasing environmental pollution and financial expenses. Because of its advantages, such as speed and sensitivity, this technique is gaining considerable attention in recent years for pharmaceutical and biomedical analysis Guo-Ying Wang 1,2 Huan-Yang Qi 1 Yan-Ping Shi [22,24,25]. It is also suitable for the routine analysis of botanical drugs consisting of multiple components [26].…”

“…Radix et Rhizoma Rhei is an important Chinese herbal medicine with many pharmaceutical actions, such as liver protecting, choleretic, purgative, anti-inflammatory, antibacterial and anticarcinogenic effects [1,2]. From Radix et Rhizoma Rhei, some components belonging to types of anthraquinones, stilbenes, dianthrones, anthocyanins, flavonoids, galloyglucoses, phenylbutanones, organic acid and so on are extracted.…”

“…Some methods, such as TLC [5,6], HPLC [1,[7][8][9][10][11][12][13][14][15][16], CE [2,[17][18][19][20], high-speed counter-current chromatography (HSCCC) [21] and ultra-performance LC (UPLC) [22], have been reported for the determination of bioactive constituents in Radix et Rhizoma Rhei. With the objective of reducing analysis time and maintaining good efficiency, there has been substantial focus on high-speed chromatographic separations.…”

Simultaneous, Simple and Rapid Determination of Five Bioactive Free Anthraquinones in Radix et Rhizoma Rhei by Quantitative1H NMR

“…10 High-performance liquid chromatography (HPLC) for determination of five free anthraquinones is frequently used for quality control of Radix et Rhizoma Rhei according to the Chinese Pharmacopoeia and related literatures. 1,9,[11][12][13][14][15][16][17][18][19] Moreover, thin-layer chromatography (TLC), 20,21 capillary electrophoresis (CE), 22,23 ultra-performance liquid chromatography (UPLC), 6,24 UPLC-mass spectrometry (MS), 3 and high speed counter current chromatography (HSCCC) 25 also have been reported for determination of the main bioactive components in Radix et Rhizoma Rhei. However, all the analytical methods above for determining and quantifying Radix et Rhizoma Rhei suffer from the intrinsic limitations of chromatography-based methods, namely, the need of standard compounds for analyte identification or quantification and the relatively long analysis times.…”

Simultaneous determination of anthraquinone derivatives in Radix et Rhizoma Rhei-based medicines by ultra-performance LC-ESCI-MS/MS multiple reaction monitoring

“…1), which are the basis for the quality control of Radix et Rhizoma Rhei-based medicines [7][8][9][10]. The methods commonly used for the separation and determination of all or certain of the major anthraquinone derivatives in Radix et Rhizoma Rhei are thin-layer chromatography (TLC) with UV detection [11], highspeed counter current chromatography (HSCCC) with a UV monitor [12,13], capillary electrophoresis (CE) with a photodiode array detector [14], high-performance liquid chromatography (HPLC) with a UV detector [15,16], a photodiode array detector [17], and ultra-performance liquid chromatography (UPLC) with a photodiode array detector [10]. Among the developed methods, although HPLC/UPLC is well established and robust and is the most commonly used method for the determination of anthraquinone derivatives, it is not sufficiently sensitive to properly evaluate the Radix et Rhizoma Rhei-based medicines and the pharmacokinetics of anthraquinone derivatives because of the interference of matrices.…”