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Due to the challenge of hundreds of potential mycotoxins that may be present in cereals, a rapid and reliable ultra-high performance liquid chromatography coupled with quadrupole-time-of-flight tandem mass spectrometry (UHPLC-Q-TOF MS) technique was developed for universal screening of 200 mycotoxins (prototype, emerging and related derivatives) in cereals. With satisfactory sensitivity of accurate mass full-spectrum acquisition, it is feasible to preliminarily identify tentative untargeted mycotoxins with a large range of polarity without reference materials. The current screening method was also validated by the determination of 33 typical mycotoxins, and the screening detection limits in the range of 0.5-100 μg kg

A reliable solid-phase extraction (SPE) procedure using reduced graphene oxide and gold nanoparticle (rGO/Au) composite as sorbent was proposed for purification and enrichment of aflatoxin B1 (AFB1), aflatoxin M1 (AFM1), ochratoxin A (OTA), zearalenone (ZEA), α-zearalenol (α-ZOL), β-zearalenol (β-ZOL), zearalanone (ZAN), α-zearalanol (α-ZAL) and β-zearalanol (β-ZAL) in milk. Main parameters affecting the performance of SPE procedure were thoroughly investigated. The optimized conditions included 2% acetonitrile in water as the loading solution, 5% methanol in water as the washing solution, and methanol/acetonitrile/formic acid (50/49/1, v/v/v) as the elution solvent. Satisfactory linearity (R2 ≥ 0.992), high sensitivity (limit of quantification in the range of 0.02–0.18 ng mL−1), adequate recovery (70.2–111.2%), and acceptable precision (RSD, 2.0–14.9%) were obtained when the optimal sample pretreatment protocol was combined with ultra-high-performance liquid chromatography-tandem mass spectrometry detection. The applicability of the validated method was further verified in real milk samples.

Yangxinshi Tablet (YXST) is a Chinese patent medicine commonly used to treat cardiovascular diseases. However, its detailed chemical basis and mechanisms of action have not been clarified. In this study, high performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight mass spectrometry (HPLC-ESI-Q-TOF-MS) was applied for comprehensive analysis of the chemical constituents in YXST. A total of 127 compounds, including 19 phenolic acids, 12 alkaloids, 51 flavanoids, 32 triterpenoids, 2 lignans, 2 phenylethanoid glycosides, 2 anthraquinones, 1 coumarin, and 6 other compounds, were identified or tentatively deduced by comparing their retention times and MS spectra with those of authentic standards or literature data. To further prove the antioxidant activity of YXST, its free radical scavenging capacity was assessed by 1,1-diphenyl-2-picrylhydrazyl (DPPH) spectrophotometric assay and the antioxidants in YXST were rapidly screened by DPPH-HPLC experiment. Especially, salvianolic acid A and salvianolic acid B showed excellent DPPH scavenging activities with the IC50 of 151.9 and 275.6μg/mL, respectively, which were stronger than that of l-ascorbic acid (positive control) with the IC50 of 297.1μg/mL. Additionally, these two most potent antioxidants were detectable in rat plasma after oral administration. In conclusion, this study reported important clues for the further pharmacological and clinical studies of YXST. Meanwhile, it provided a practical strategy for rapid screening and identifying of in vivo antioxidant in traditional Chinese medicine preparations.

Screening agonists of peroxisome proliferator-activated receptor-γ (PPARγ) from natural products is particularly motivated by the need for effective anti-diabetic agents. However, method for direct identification of PPARγ ligands from a complex sample is rarely reported. Here we propose a novel immobilized fusion protein affinity chromatography (IFPAC) to achieve rapid multicomponent screening. First, functional human PPARγ ligand binding domain (hPPARγLBD) was bacterially produced by fusion to glutathione S-transferase (GST). The unpurified GST-hPPARγLBD was directly applied to a 96-well filter plate prepacked with glutathione sepharose. Due to the strong affinity between GST and glutathione, the fusion protein could selectively attach to the glutathione matrix with an oriented immobilization, which was rapidly purified under non-denaturing conditions. Experimental results indicated that the prepared 96-affinity column array exhibited excellent selectivity and sensitivity for fishing novel interacting compounds. The proposed approach was applied in the high-throughput screening of PPARγ ligands from natural products, followed by rapid characterization of active compounds using HPLC-ESI-Q-TOF-MS/MS. Isochlorogenic acid A in Dendranthema indicum flowers was found to be a PPARγ ligand. Our findings suggested the IFPAC could be a powerful tool for drug discovery from natural products.

In this work, near infrared (NIR) spectroscopy was used in combination with chemometrics to determine the epimedin A, epimedin B, epimedin C, icariin, and moisture contents of Herba Epimedii. The variable selection method genetic algorithm (GA) and regression tool support vector machine (SVM) were used to improve the model performance. Four different calibration models, namely Full-PLS, GA-PLS, Full-SVM, and GA-SVM, were established, and their performances in terms of prediction accuracy and model robustness were systemically studied and compared. In conclusion, the performances of the models based on the efficient variables selected through GA were better than those based on full spectra, and the nonlinear models were superior over the linear models. In addition, the GA-SVM model demonstrated the optimal performance in predicting five quality parameters (viz. epimedin A, epimedin B, epimedin C, icariin, and moisture). For GA-SVM, the determination coefficient (Rp2), root-mean-square error (RMSEP), and residual predictive deviation (RPD) for the prediction set were 0.9015, 0.0268%, and 2.20 for epimedin A; 0.9089, 0.0656%, and 3.08 for epimedin B; 0.9056, 0.1787%, and 3.18 for epimedin C; 0.8192, 0.0657%, and 2.26 for icariin; and 0.9367, 0.2062%, and 4.12 for moisture, correspondingly. Results indicated that NIR spectroscopy coupled with GA-SVM calibration can be used as a reliable alternative strategy to measure the epimedin A, epimedin B, epimedin C, icariin, and moisture contents of Herba Epimedii because this technique is fast, economic, and nondestructive compared with traditional chemical methods.

For the first time, a reliable solid-phase extraction (SPE) procedure using iron (II, III) oxide (Fe3O4)/multi-walled carbon nanotube (MWCNT) composite as sorbents was proposed for purification and enrichment of zearalenone (ZEA) and four type A trichothecenes (T-2 toxin (T-2), HT-2 toxin (HT-2), neosolaniol (NEO) and diacetoxyscirpenol (DAS)) in Salviae miltiorrhiza Radix et Rhizoma (Danshen). The Fe3O4/MWCNT composite was synthesized by assembling Fe3O4 with MWCNT by sonication through an "aggregation wrap" mechanism and several key parameters affecting the performance of SPE procedure such as the composition of sample loading solutions, washing and elution solvents were thoroughly investigated. After optimization, 2% acetonitrile aqueous solution as the loading solution, 5% methanol aqueous solution as the washing solution and acetone containing 0.5% formic acid as the elution solvent presented an excellent purification efficiency for the five targets in Danshen. Under the optimal sample pretreatment conditions followed by analysis with ultra-high performance liquid chromatography-tandem mass spectrometry, satisfactory linearity (R2≥0.991), high sensitivity (limit of quantification in the range of 1.20-4.80μgkg-1), good recovery (73.7-91.9%) and acceptable precision (RSD, 2.1-13.3%) were obtained. The applicability of the validated method was further verified in real Salviae miltiorrhiza Radix et Rhizoma samples.

A simple and reliable analytical method for the simultaneous determination of alternariol (AOH), altenuene (ALT), tentoxin (TEN), altenusin (ALS), tenuazonic acid (TeA), and alternariol monomethyl ether (AME) in grapes was developed by ultra-high-performance liquid chromatography&ndash, tandem mass spectrometry (UHPLC-MS/MS). A modified QuEChERS (quick, easy, cheap, effective, rugged, and safe) procedure with the extraction by acetonitrile and purification by sodium chloride (0.5 g) and anhydrous magnesium sulfate (0.5 g) was established to recover the six Alternaria toxins. After validation by determining the linearity (R2 &gt, 0.99), recovery (77.8&ndash, 101.6%), sensitivity (limit of detection in the range of 0.03&ndash, 0.21 &mu, g kg&minus, 1, and limit of quantification in the range of 0.09&ndash, 0.48 &mu, 1), and precision (relative standard deviation (RSD) &le, 12.9%), the analytical method was successfully applied to reveal the contamination state of Alternaria toxins in grapes. Among 56 grape samples, 40 (incidence of 71.4%) were contaminated with Alternaria toxins. TEN was the most frequently found mycotoxin (37.5%), with a concentration range of 0.10&ndash, 1.64 &mu, 1, followed by TeA (28.6%) and AOH (26.8%). ALT (10.7%), AME (3.6%), and ALS (5.4%) were also detected in some samples. To the best of our knowledge, this is the first report about the Alternaria toxins contamination in grapes in China.

A variety of bioactive substances may account for the recognized efficacy and wide clinical application of Psoraleae Fructus in China. A high-performance liquid chromatography-diode array detector (HPLC-DAD) fingerprint method was developed to present the comprehensive phytochemical profile of the crude drug. Thirteen major compounds were separated and identified by HPLC coupled with time-of-flight mass spectrometry (HPLC/TOF-MS), namely psoralenoside (PO), isopsoralenoside (IPO), psoralen (PS), isopsoralen (IPS), neobavaisoflavone (NBF), bavachin (BC), corylin (CN), bavachromene (BCM), psoralidin (PD), isobavachalcone (IBC), bacachinin (BCN), corylifol A (CA) and bakuchiol (BK). Then quantitative analysis of multiple components by single marker (QAMS) was applied in content determination of PO, IPO, PS, IPS, BC, IBC, BCN, CA and BK, with NBF as the internal standard. The calculation results indicated no significant difference from the traditional external standard method (p > 0.05, RSD < 2.62%), suggesting that QAMS is a reliable and convenient method for content determination of multiple chemical compositions, especially when there is a shortage of reference substances. In conclusion, simultaneous qualitative and quantitative analysis of Psoraleae Fructus may be fulfilled through the newly proposed method of QAMS combined with HPLC-DAD/TOF-MS fingerprint.

A simple, reliable method for simultaneous determination of the ten components in Danhong injection (DHI) in combination of chromatographic fingerprint analysis was developed by high performance liquid chromatography coupled with diode-array-detector (HPLC-DAD). The separation was performed on an Agilent Zorbax SB-C 18 column with a linear gradient elution of acetonitrile and 0.5% formic acid. The method was validated by linearity, precision, stability and recovery. The developed method was subsequently applied to evaluate 15 batches of DHI and testified to be suitable for its quality control.

In this study, a fast and effective high-performance liquid chromatography method was developed to obtain a fingerprint chromatogram and quantitative analysis simultaneously of four indexes including gallic acid, chlorogenic acid, albiflorin and paeoniflorin of the traditional Chinese medicine Moluodan Concentrated Pill. The method was performed by using a Waters X-bridge C

A generic procedure, which involved accelerated solvent extraction and homemade cleanup cartridges, has been developed for the extraction and purification of 35 mycotoxins in various traditional Chinese medicine (TCM) matrixes, i.e., rhizomes and roots, seeds, flowers, and grasses and leaves, for subsequent analysis by ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). All target analytes could be simultaneously quantitated in less than 17 min per run, showing narrow symmetrical peaks. The developed method was also successfully applied in routine monitoring programs, which implied a significant reduction of both effort and time, to investigate the contamination of TCMs. Among 60 commercial TCMs analyzed, 50 were positive. The achieved data underpin the practical application of the UHPLC-MS/MS method as a valuable tool for the trace analysis of multiple mycotoxins in TCMs.

An ultra-high pressure liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed for the quantification of three major ingredients in Chinese lantern preparations (CLP) in rat plasma. Following extraction by ethyl acetate, the analytes were separated on an Acquity UPLC BEH Shield RP C(18) column using a gradient mobile phase system of acetonitrile-water. Electrospray ionization (ESI) tandem interface was employed prior to mass spectrometric detection. The calibration curves were linear over the range of 5.0-500.0 ng/ml for physalin D, 2.3-230.0 ng/ml for physalin G and 0.71-71.0 ng/ml for 4,7-didehydroneophysalin B. The average extraction recoveries, examined at four concentration levels, carried from 57.1% to 76.9%, and the accuracies ranged from 94.0% to 113.3% with precision (RSD)

The application of near-infrared (NIR) spectroscopy for in-line monitoring of extraction process of scutellarein from Erigeron breviscapus (vant.) Hand-Mazz was investigated. For NIR measurements, two fiber optic probes designed to transmit NIR radiation through a 2 mm pathlength flow cell were utilized to collect spectra in real-time. High performance liquid chromatography (HPLC) was used as a reference method to determine scutellarein in extract solution. Partial least squares regression (PLSR) calibration model of Savitzky-Golay smoothing NIR spectra in the 5450-10,000 cm(-1) region gave satisfactory predictive results for scutellarein. The results showed that the correlation coefficients of calibration and cross validation were 0.9967 and 0.9811, respectively, and the root mean square error of calibration and cross validation were 0.044 and 0.105, respectively. Furthermore, both the moving block standard deviation (MBSD) method and conformity test were used to identify the end point of extraction process, providing real-time data and instant feedback about the extraction course. The results obtained in this study indicated that the NIR spectroscopy technique provides an efficient and environmentally friendly approach for fast determination of scutellarein and end point control of extraction process.

Antisolvent crystallization can be used as an alternative to cooling or evaporation for the separation and purification of solid product in the pharmaceutical industry. To improve the process understanding of antisolvent crystallization, the use of in-line tools is vital. In this study, the process analytical technology (PAT) tools including focused beam reflectance measurement (FBRM), particle video microscope (PVM), and near-infrared spectroscopy (NIRS) were utilized to monitor antisolvent crystallization of sodium scutellarein. FBRM was used to monitor chord count and chord length distribution of sodium scutellarein particles in the crystallizer, and PVM, as an in-line video camera, provided pictures imaging particle shape and dimension. In addition, a quantitative model of PLS was established by in-line NIRS to detect the concentration of sodium scutellarein in the solvent and good calibration statistics were obtained (r(2) = 0.976) with the residual predictive deviation value of 11.3. The discussion over sensitivities, strengths, and weaknesses of the PAT tools may be helpful in selection of suitable PAT techniques. These in-line techniques eliminate the need for sample preparation and offer a time-saving approach to understand and monitor antisolvent crystallization process.

In this paper, to evaluate the effect of the region of origin on the quality consistency of Shaoyao-Gancao Decoction (SGD), the SGD fingerprint was developed for the first time. Chemometric methods including similarity analysis, hierarchical clustering analysis and principal component analysis were employed to study the quality consistency of SGD. Meanwhile, high-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight mass spectrometry was applied for comprehensive analysis of SGD and 93 compounds were tentatively characterized. Furthermore, a high-performance liquid chromatography method with multi-wavelength switching for simultaneous determination of 16 characteristic ingredients comprising gallic acid, oxypaeniflorin, albiflorin, paeoniflorin, liquiritin apioside, liquiritin, isoliquiritin apioside, galloylpaeoniflorin, 1,2,3,4,6-penta-O-galloyl-d-galactopyranose (PGG), ononin, isoliquiritin, liquiritigenin, benzoylpaeoniflorin, glycyrrhizic acid, isoliquiritigenin and formononetin, was established. All 16 analytes show excellent linearity (R2 ≥ 0.9990) with recoveries ranging from 96.58 to 104.61% and limits of detection and quantification of 0.022-0.291 and 0.037-0.635 μg/mL, respectively. Finally, it was successfully applied to determine 15 batches of SGD. The results of our research indicate that different regions of origin have a significant effect on the quality consistency of SGD, and its fingerprint combined with chemometrics and multi-ingredient determination comprise an efficient and reliable approach for quality consistency evaluation.

An ultra-pressure LC (UPLC) method has been developed and validated for the quality evaluation of a traditional Chinese medicine (Radix Linderae) by chemical fingerprint analysis with chromatograms collected at two wavelengths (260 and 320 nm). Eleven characteristic peaks in the fingerprints were identified by comparing their retention times, UV spectra and ESI-MS/MS data with those of the reference substances or the data in the literatures. Both correlation coefficient of similarities in chromatograms and relative peak areas of common peaks were calculated for quality expression of the Radix Linderae samples collected from different areas in China. The results showed high variation of relative peak area and correlation coefficients among the samples collected from various habitats, which indicated that the quality consistency of Radix Linderae is still a problem worthy of serious concern.

A reliable isotope dilution method for simultaneous determination of fumonisin B1, fumonisin B2 and fumonisin B3 in traditional Chinese medicines by ultra-high-performance LC-MS/MS was developed, and a special focus was placed on the optimization of extraction, cleanup, ultra-high-performance LC separation and MS/MS conditions. Homogenized samples were extracted by 50% acetonitrile aqueous solution and purified with MultiSep 211 Fum columns. A linear gradient mobile phase, consisting of water containing 0.2% formic acid and acetonitrile/methanol (50:50 v/v) and an Acquity UPLC HSS T3 column (100 mm×2.1 mm, 1.8 μm) were employed to obtain the best resolution of the analytes for the positive ESI(+) analysis. The established method was validated by determining the linearity (R(2)≥0.9991), sensitivity (LOQ, 0.08-0.16 ng/mL), recovery (88.2-113.3%) and precision (RSD≤12.3%). Finally, the validated method was successfully applied to the determination of fumonisins in different traditional Chinese medicines. Of 35 samples, 18 were contaminated with fumonisins. The mean levels (incidence) of fumonisin B1, fumonisin B1 and fumonisin B1 in positive samples were 8.55 (94.4%), 4.88 (77.8%) and 0.52 μg/kg (33.3%), respectively. Based on the contaminant situations, a possible association between the contamination levels and the selected herbs was clarified in this study.

A reliable ultra-high-performance liquid chromatography–tandem mass spectrometry (UHPLC–MS/MS) method for simultaneous determination of aflatoxins B1, B2, G1, G2, M1 and M2 in peanuts and their derivative products was developed. The sample was extracted by 84% of acetonitrile aqueous solution and the extract was purified by a reliable solid phase extraction-based clean-up method. Then, the analytes were separated on Acquity UPLC HSS T3 column (100 mm × 2.1 mm, 1.8 μm particle size), and eluted with a mobile phase consisting of (A) water containing 0.1% formic acid and (B) acetonitrile/methanol (50/50, v/v). The separated compounds were detected with a Waters Micromass Quattro Ultima Pt tandem quadrupole mass spectrometer operating in positive electro-spray ionization using multiple reaction monitoring mode. The established method was extensively validated by determining the linearity ( R 2 ≥ 0.9990), average recovery (74.7–86.8%) and precision (relative standard deviation ≤ 10.9%). It was shown to be a suitable method for simultaneous determination of the six aflatoxins in peanuts and their derivative products. Finally, a total of 73 samples randomly collected from different areas in Zhejiang province were screened for aflatoxins with the proposed method. The results showed that 31 samples of peanut butter, 14 samples of fresh peanut and 5 samples of musty peanut were contaminated with aflatoxins. Meanwhile, this was the first report on aflatoxins M1 and M2, which were found in unprocessed peanuts and their derivative products.

A new separation and quantification method using liquid chromatography under ultra-high-pressure in combination with tandem mass spectrometry (MS/MS) was developed for simultaneous determination of four alkaloids in Lindera aggregata . The analysis was performed on an Acquity UPLC BEH C 18 column (50 mm × 2.1 mm, 1.7 μm particle size; Waters, Milford, MA, USA) utilizing a gradient elution profile and a mobile phase consisting of (A) water containing 10 mM ammonium acetate adjusted to pH 3 with acetic acid and (B) acetonitrile. An electrospray ionization (ESI)–tandem interface in the positive mode was employed prior to mass spectrometric detection. The calibration curve was linear over the range of 17.1–856 ng for boldine, 42.4–2652 ng for norboldine, 6.1–304 ng for reticuline and 0.5–50 ng for linderegatine, respectively. The average recoveries ranged from 99.2 to 101.4% with RSDs ≤ 2.7%. Then, four L. aggregata samples from different batches were analyzed using the established method. The results indicated that ultra-high-pressure liquid chromatography–tandem mass spectrometry provided improved chromatographic parameters resulting in significantly increased sample throughput including lower solvent consumption and lower limits of quantitation (LOQs) for most of target analytes compared to previous method employing conventional high-performance liquid chromatography (HPLC) separation. So, the established method was validated, sensitive and reliable for the determination of four alkaloids in L. aggregata .

A solid-phase extraction (SPE) procedure using multi-walled carbon nanotubes (MWCNTs) as sorbents coupled with ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) was developed for simultaneous determination of four type A trichothecenes in maize, wheat and rice for the first time. Several key parameters including the composition of sample loading solutions, washing and elution solvents were thoroughly investigated to achieve optimal SPE recoveries and efficiency. Performance of the MWCNTs materials was significantly affected by pH, and after optimization, n-hexane and 5% methanol aqueous solution as the washing solutions and methanol containing 1% formic acid as the elution solvent presented an excellent purification efficiency for the four targets in the different matrices. The method was validated by determining the linearity (R(2)≥0.992), recovery (73.4-113.7%), precision (1.2-17.1%) and sensitivity (limit of quantification in the range of 0.02-0.10μg/kg), and was further applied for simultaneous determination of type A trichothecenes in 30 samples. Although low contamination levels of type A trichothecenes in wheat, maize and rice were observed revealing mitigated risks to consumers in Shanghai, China, the developed method has proven to be a valuable tool for type A trichothecenes monitoring in complex crop matrices.

A rapid and efficient method for the identification of antioxidants in the traditional Chinese medicine Polygalae Radix (PR) by HPLC-ESI-Q-TOF-MS/MS is described. The method is based on the hypothesis that upon reaction of antioxidants with 1,1-diphenyl-2-picrylhydrazyl (DPPH), the peak areas of compounds with potential antioxidant activities in the HPLC chromatogram will be significantly reduced in comparison to the untreated sample. The identity confirmation was achieved by Q-TOF-MS/MS technique. With this method, eight components were proposed possessing potent antioxidant activity. They were identified as sibiricose A5, sibiricose A6, sucrose monoester, polygalaxanthone III, tenuifoliside B, 3',6-disinapoylsucrose (DISS), sucrose diester, tenuifoliside C, respectively. DISS was proposed to be the most potent one. The antioxidant activity of DISS was evaluated by DPPH, ABTS radical scavenging assay and ferric-reducing antioxidant power (FRAP) assay in vitro. Vitamin C (Vc) was used as positive control substance. DISS showed moderate DPPH (DISS's IC50 value was 1024.17 μg/mL, Vc's was 294.68 μg/mL) and ABTS (IC50 324.13 μg/mL, Vc's was 117.50 μg/mL) free radical scavenging capacity and ferric-reducing antioxidant power. DISS can be used as a new source of natural antioxidant in foods and cosmetics.

A simple and reliable method of high-performance liquid chromatography with diode array detection method was developed for fingerprint analysis and simultaneous determination of six compounds including puerarin, salvianolic acid B, berberine hydrochloride, palmatine chloride, dehydrocorydaline, and icariin in the Chinese medicine preparation Yangxinshi Pian. The separation was performed on an Agilent Eclipse XDB-C18 reserved-phase column (250 mm × 4.6mm I.D., 5 μm) using gradient elution with 50 mmol/L monopotassium phosphate aqueous solution and methanol as mobile phase at a flow rate of 1.0 mL/min. The column operating temperature was set at 30°C, and the detection wavelength was 280 nm. The method was validated by linearity, precision, accuracy, stability, and recovery. For fingerprint analysis, 25 peaks were selected as the common peaks, and four kinds of similarities including cosine similarity (S), ratio of similarity (S'), projection content similarity (C), and content similarity (P) were applied to evaluate the quality consistency of different batches of Yangxinshi Pian. The results showed that the developed method was an efficient tool for quality evaluation of Yangxinshi Pian.

A sensitive and specific method was developed for the determination of sophoridine (SRI), sophocarpine (SC) and matrine (MT) in rabbit plasma by HPLC-MS. After an administration of Kuhuang by injection, blood samples were collected and extracted with methanol. The extract solutions were analysed by HPLC-MS method. The separation was performed on a ZORBAX Extend-C18 column using methanol/water/diethylamine (50:50:0.07, v/v/v) as mobile phase. The quinolizidine alkaloids were detected by using mass spectrometry in the SIM mode. There was a good linear relationship between peak area and concentration of analytes over the concentration range of 13.2-995.0 ng mL(-1) for SRI, 7.0-530.0 ng mL(-1) for SC and 8.8-655.0 ng mL(-1) for MT, respectively. The absolute recovery of this method was more than 57% for SRI, 87% for SC and 91% for MT. The accuracy of assay was more than 90%. The limits of detection (LODs) were 6.8 ng mL(-1) for SRI, 3.5 ng mL(-1) for SC and 4.2 ng mL(-1) for MT, respectively. The limits of quantitation (LOQs) were 13.2 ng mL(-1) for SRI, 7.0 ng mL(-1) for SC and 8.8 ng mL(-1) for MT, respectively. The intra-day and inter-day coefficients of variation (RSDs) were less than 10.1, 6.3 and 5.8% for SRI, SC and MT, respectively. The developed method was applied to determine the concentration-time profiles of SRI, SC and MT in rabbit plasma after injection of Kuhuang.

An on-line detection method combining near infrared (NIR) spectroscopy with local partial least squares regression (PLSR) was investigated for the elution process of sodium aescinate. A 2 mm pathlength flow cell which transmitted NIR radiation through two fiber optic probes was placed in bypass of the macroporous resin column to collect real-time spectra of the sodium aescinate eluate. To compare the predictive accuracy, both local and global PLSR were employed to build mathematical models between NIR spectra and reference values. Meanwhile, Mahalanobis distance was introduced to select the appropriate local model for the prediction of unknown samples. Experimental results demonstrated that local PLSR was superior to global PLSR in both calibration performance and predictive accuracy. Moreover, the on-line detection method was proven to be feasible in real application and thereby would be of great value for monitoring the elution process of sodium aescinate in real time as well as determining the start and end points of eluate collection.

Physalins are important bioactive compounds from genus Physalis. They often occur as isomers, which makes the structural elucidation difficult. In the present study, the fragmentation behavior and UV characteristics of seven physalins from genus Physalis were firstly investigated using electrospray ionization tandem mass spectrometry (ESI-MS/MS) and diode array detection (DAD). Combined with ultra-performance liquid chromatography (UPLC) and DAD, the established approach to the structural identification of physalins by ESI-MS/MS was then applied to the analysis of Physalis alkekengi L. According to the UPLC retention behavior, the diagnostic UV spectra and the molecular structural information provided by MS/MS spectra, about 19 fingerprint peaks were identified, including 14 physalins and 5 other compounds. Finally, the established fingerprint method was applied to the analysis of 31 P. alkekengi L. samples collected from different locations, which reflected their similar chemical constituent properties. The proposed method provides a scientific and technical platform to the herbal industry for quality control and safety assurance of herbal preparations that contain this class of physalins.

A rapid and reliable ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method for simultaneous determination of zearalenone (ZEN), α-zearalenol (α-ZOL), β-zearalenol (β-ZOL), zearalanone (ZAN), α-zearalanol (α-ZAL) and β-zearalanol (β-ZAL) in traditional Chinese medicines (TCMs) was developed. The development of the method and investigations of the matrix influence were described in particular. After evaluation of the matrix effects of different TCMs, i.e., rhizomes and roots, seeds, flowers, grasses and leaves, by the post-extraction spiked method, a reliable sample clean-up method based on home-made clean-up cartridges, a suitable internal standard and the matrix calibration were combined using to minimize the matrix effects to ensure the accuracy of the method. The established method was further validated by determining the linearity (R(2)≥0.9990), sensitivity (limit of quantitation 0.11-0.99 ng mL(-1)), average recovery (86.6-113.5%) and precision (relative standard deviation ≤13.5%). It was shown to be a suitable method for simultaneous determination of ZEN, α-ZOL, β-ZOL, ZAN, α-ZAL and β-ZAL in different TCMs. Finally, the established method was successfully applied to the determination of the six mycotoxins in various TCMs and the results were presented to provide relevant insights to researchers in TCM analysis.

Physalin D is an important constituent of some traditional Chinese medicines, and has several known bioactivities. An UPLC-MS/MS method for the determination of physalin D in rat plasma and tissues was developed and the pharmacokinetic and tissue distribution characteristics of physalin D after intravenous administrations were investigated. The bio-samples were prepared by a simple protein precipitation, and the separation of physalin D was achieved on a UPLC HSS T3 column with a mobile phase consisting of methanol/acetonitrile (70:30, v/v) and water (containing 0.1% formic acid and 10 mM ammonium acetate) at a flow rate of 0.3 mL/min. The MS/MS detection was carried out by monitoring the fragmentation of m/z 544.9→508.8 for physalin D and m/z 286.7→152.8 for luteolin (internal standard; IS) on a triple-quadrupole mass spectrometer. The total run time was only 3.6 min. The analyte showed good linearity over a wide concentration range (R(2)0.995) and its lower limit of quantification was 2 ng/mL. The pharmacokinetic study found that physalin D was distributed and eliminated rapidly in rats (t(1/2)10 min). Tissue distribution showed the highest level was observed in kidney, then in liver, but no physalin D was detected in brain, which indicated that kidney was the major distribution tissue for physalin D in rats and that physalin D does not cross the blood-brain barrier.

A speedy and selective ultra-HPLC-MS/MS method for simultaneous determination of deoxynivalenol (DON), 3-acetyldeoxynivalenol (3-ADON), 15-ADON, nivalenol and fusarenon X in traditional Chinese medicines (TCMs) was developed. The method was based on one-step sample cleanup using reliable homemade cleanup cartridges. A linear gradient mobile-phase system, consisting of water containing 0.2% aqueous ammonia and acetonitrile/methanol (90:10, v/v) at a flow rate of 0.4 mL/min, and an Acquity UPLC HSS T3 column (100 mm x 2.1 mm, 1.8 microm) were employed to obtain the best resolution of the target analytes. [(13)C(15)]-DON was used as the internal standard to accomplish as accurate as possible quantitation. The established method was further validated by determining the linearity (R(2) > or = 0.9990), sensitivity (LOQ, 0.29-0.99 microg/kg), recovery (88.5-119.5%) and precision (RSD < or = 15.8%). It was shown to be a suitable method for simultaneous determination of DON, 3-ADON, 15-ADON, nivalenol and fusarenon X in various TCM matrices. The utility and practical impact of the method was demonstrated using different TCM samples.

OBJECTIVE To develop a HPLC method for simultaneous determination of caffeic acid, rosmarinic acid, oleanolic acid and ursolic acid in Spica Prunellae. METHOD After ultrasonic extraction with 75% ethanol solution containing 1% formic acid, the ethanol-extract of Spica Prunellae was analyzed on an Elite SinoChrom ODS-AP column using gradient elution of 0.01% phosphoric acid (A) and acetonitrile (B) at a flow rate of 0.9-1.0 mL x min(-1). A wavelength switch program was used for detection at 330 nm (0-33 min) and 203 nm (33-40 min). The column temperature was set at 20 degrees C and injection volume was 50 microL. RESULT The calibration curves of all analytes were linear. The average recoveries were 93.7%-105.2% with RSDs not more than 4.5%. The contents of caffeic acid, rosmarinic acid, ursolic acid and oleanolic acid in Spica Prunellae were 0.0401-0.0968, 0.99-2.57, 0.243-0.556, 4.06-8.13 mg x g(-1), respectively. CONCLUSION The described method is sensitive, convenient and accurate, and is suitable for the simultaneous determination of caffeic acid, rosmarinic acid, oleanolic acid and ursolic acid in Spica Prunellae.

To develop an HPLC method for simultaneous determination of three major sesquiterpene lactones in Radix Linderae.The chromatographic separation was achieved on a Diamonsil C18 column (4.6 mm x 250 mm, 5 microm) using isocratic elution of acetonitrile-water (containing 0.1% H3 PO4) (45 : 55) at a flow rate of 1.0 mL x min(-1). Detection was carried out using a photodiode array detector at 220 nm.The calibration curves were linear in the range of 0.001 8-0.036 0 g x L(-1) for hydroxylinderstrenolide (R2 = 0.999 8), 0.016 2-0.323 2 g x L(-1) for neolinderalactone (R2 = 0.999 9), 0.010 5-0.209 9 g x L(-1) for linderane (R2 = 0.999 9), respectively. The average recoveries were 100.0% for hydroxylinderstrenolide, 98.8% for neolinderalactone and 98.9% for linderane with RSD not more than 3.3%.The established method was proved to be simple, sensitive and credible, and can be applied to quality control of Radix Linderae.

An ultra-performance LC in combination with MS/MS method was established to evaluate the quality of wu-yao (the dried root of Lindera aggregata (Sims) Kosterm. (Lauraceae)) through simultaneous quantitation of five sesquiterpene lactones, linderagalactone D (1), linderagalactone C (2), hydroylindestenolide (3), neolinderalactone (4) and linderane (5). All compounds were separated on a Waters Acquity ultra-performance LC HSS T3 column (2.1 mm x 100 mm, 1.8 microm particle size) with linear gradient elution of acetonitrile and 0.1% formic acid. An ESI interface in the positive mode was employed prior to MS detection. All the compounds showed good linearity (R(2)or = 0.9992). The recoveries, measured at three concentration levels, varied from 97.3 to 103.4% with RSDs4.8%. The simple, rapid and reliable method was successfully applied to quantify the five components in 13 samples of wu-yao from different areas.

A reliable ultra-high-performance liquid chromatography-mass spectrometry method for simultaneous determination of bovine alpha-lactalbumin (alpha-La) and beta-lactoglobulin (beta-Lg) was developed. Compared to the previous methods, the developed approach with mass spectrometer operated in selected area monitoring mode offered increased speed and enhanced lower detection limit. A linear gradient mobile phase, consisting of (A) water containing 0.1% trifluoroacetic acid (TFA) and (B) acetonitrile containing 0.1% TFA, and an Acquity UPLC BEH300 C18 column (150mmx2.1mm, 1.7microm) were employed to obtain the best resolution of the target analytes. The accurate quantitation was achieved by employing human alpha-lactalbumin as the internal standard. The established method was extensively validated by determining the linearity (R(2)or=0.9991), sensitivity (limit of quantitation, 0.15-0.19microgmL(-1)), recovery (94.0-98.7%), precision (relative standard deviationor=11.1%) and repeatability (relative standard deviationor=5.7%). It was shown to be a suitable method for simultaneous determination of the major whey proteins in biological samples. Current validated method was successfully applied to the nutrient investigation of infant formulae.

The paper reported a reliable analytical method for simultaneous determination of ochratoxin A (OTA) and ochratoxin B (OTB) in traditional Chinese medicines (TCMs) by ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). The development of the method and investigations on the matrix influence were described in particular. The matrix effects were thereby minimized by using a reliable internal standard and a simple sample pretreatment. The established method was further validated by determining the linearity (R(2) > or = 0.9990), average recovery (86.3-114.2%), sensitivity (limit of quantitation 0.03-0.19 ng mL(-1)) and precision (relative standard deviation < or = 13.1%). It was shown to be a suitable method for simultaneous determination of OTA and OTB in various TCMs. Finally, a total of 51 TCMs widely used in China were screened for OTA and OTB with the proposed method. The results showed that only 4 samples were contaminated with ochratoxins at low levels, indicating that it was low risk of ochratoxins to consumers who occasionally used TCMs.

An ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method for simultaneous determination of aflatoxins B1, B2, G1, G2, M1 and M2 in traditional Chinese medicines (TCMs) was developed. The approach was characterized in details and a special focus was placed on the recovery rates of isolation procedure in different TCM matrices, i.e. rhizomes and roots, seeds, flowers, grasses and leaves. For this purpose, [(13)C(17)]-aflatoxinB1 was employed as the internal standard and a reliable solid phase extraction-based clean-up method was developed. The observed recovery rates of the six aflatoxins ranged from 85.6% to 117.6% in different matrices. Then, the established method was successfully applied to the determination of the six aflatoxins in various TCMs. For 30 commercial samples analyzed, 16 were contaminated with aflatoxins. The mean levels (incidence) of aflatoxins B1, B2, G1 and G2 in positive samples were 1.40 (68.8%), 1.27 (50.0%), 0.50 (43.8%) and 0.94 (43.8%) microg kg(-1), respectively. Interestingly, aflatoxin M1 was detected in two samples with the maximal content of 0.70 microg kg(-1). No sample was contaminated with aflatoxin M2. Meanwhile, a possible association between the contamination levels and the selected herbs was clarified in the present study.

To develop an HPLC method for simultaneous determination of four major alkaloids in Lindera aggregate.The analysis was carried out on an Agilent ZORBAX SB-C18 column (4.6 mm x 250 mm, 5 microm) with gradient elution using acetonitrile-water (containing 0. 15% diethylamine, adjusted to pH = 3.0 with acetic acid) as mobile phase. Flow rate was 1.0 mL x min(-1) and the detection wavelength was at 289 nm.The calibration curves were linear over the range of 0.428-8.560 microg for boldine, 2.122-31.83 microg for norboldine, 0.760-15.20 microg for reticuline and 0.020 4-0.400 8 microg for linderegatine, respectively. The average recoveries were 99.18% for boldine, 101.0% for norboldine, 100.3% for reticuline and 99.17% for linderegatine, respectively. with RSD not more than 3.0%.The described method is reliable and convenient and could be used for the quality control of Lindera aggregate.