Chromatography-diode Array Detector-tandem Mass Spectrometry Research Articles

Quantification of melittin and apamin in bee venom lyophilized powder from Apis mellifera by liquid chromatography–diode array detector–tandem mass spectrometry

A high-performance liquid chromatography–diode array detector–tandem mass spectrometry (HPLC–DAD–MS/MS) method was developed for simultaneous determination of melittin and apamin in crude bee venom lyophilized powder (CBVLP) as the traditional Chinese medicine possessing specific biological activity. Melittin and apamin were extracted with pure water from CBVLP samples followed by HPLC–DAD–MS/MS analysis. The method was validated to demonstrate its selectivity, linearity, limit of quantification (LOQ), intraday precision, interday precision, accuracy, recovery, matrix effect, and stability. The assay was linear over the concentration ranges of 1–100 and 0.2–25 μg/ml with limit of quantifications (LOQs) of 1.0 and 0.3 μg/ml for melittin and apamin, respectively. The precision results were expressed as coefficients of variation (CVs), ranging from 2.2% to 11.4% for intraday repeatability and from 3.2% to 13.1% for interday intermediary precision. The concentrations of endogenous melittin and apamin in CBVLP samples ranged from 46% to 53% and from 2.2% to 3.7% of dry weight, respectively. This rapid, simple, precise, and sensitive method allowed the simultaneous determination of melittin and apamin to evaluate authenticity and quality of CBVLP samples.

Inhibition of Three Selected Beverage Extracts on α-Glucosidase and Rapid Identification of Their Active Compounds Using HPLC-DAD-MS/MS and Biochemical Detection

Inhibition of alpha-glucosidase is a therapeutic approach for diabetes. In this study, a method based on online liquid chromatography-diode array detection-tandem mass spectrometry and biochemical detection (LC-DAD-MS/MS-BCD) was developed to screen and identify alpha-glucosidase inhibitors from selected beverage extracts, including pu-erh tea ( Camellia sinensis var. assamica), eagle tea ( Litsea coreana Levl.), and radix glycyrrhizae ( Glycyrrhiza uralensis Fisch.). As a result, two components, (-)-epigallocatechingallate (EGCG) and (-)-epicatechingallate (ECG), as potent alpha-glucosidase inhibitors, were found in pu-erh tea. The IC(50) values of EGCG and ECG on alpha-glucosidase (EC 3.2.1.20, from Saccharomyces cerevisiae ) were 175.1 and 246.9 microM, respectively, and both were lower than that of acarbose (IC(50) = 3553.0 microM), a commercial alpha-glucosidase inhibitor. Kinetic studies revealed that both EGCG and ECG inhibited alpha-glucosidase activity in a noncompetitive manner. The study suggests that the developed LC-DAD-MS/MS-BCD system is a powerful tool for rapid screening and identification of alpha-glucosidase inhibitors in complex samples and that EGCG and ECG may be good candidates as alpha-glucosidase inhibitors.

Analysis of the stable free radical scavenging capability of artificial polyphenol mixtures and plant extracts by capillary electrophoresis and liquid chromatography–diode array detection–tandem mass spectrometry

The oxidation process of phenolic compounds of an artificial mixture consisting of six polyphenols and the extract of eggplant ( Solanum melongena) skin was monitored by using capillary zone electrophoresis and liquid chromatography–diode array detection–tandem mass spectrometry. The methods developed enabled simultaneous evaluation of the antioxidative capability of each compound. The above oxidation process was carried out using two radicals, viz. the 2,2-diphenyl-1-picrylhydrazyl and hydroxyl radicals generated via the Fenton reaction. The radical scavenging effects of artificial and natural polyphenol mixtures were compared.

New C-Deoxyhexosyl Flavones and Antioxidant Properties of Passiflora edulis Leaf Extract

The flavonoids present in passion fruit (Passiflora edulis) leaves were identified by a high-performance liquid chromatography-diode array detection-tandem mass spectrometry (HPLC-DAD-MS/MS) method. Sixteen apigenin or luteolin derivatives were characterized, which included four mono-C-glycosyl, eight O-glycosyl- C-glycosyl, and four O-glycosyl derivatives. With the exceptions of C-hexosyl luteolin and C-hexosyl apigenin, all the compounds exhibited a deoxyhexose moiety. Moreover, the uncommon C-deoxyhexosyl derivatives of luteolin and apigenin have been identified for first time in P. edulis by HPLC-DAD-MS/MS. The antioxidative capacity of passion fruit leaves was checked against DPPH radical and several reactive oxygen species (superoxide radical, hydroxyl radical, and hypochlorous acid), revealing it to be concentration-dependent, although a pro-oxidant effect was noticed for hydroxyl radical.

Inhibition of lipid oxidation and dynamics of polyphenol content in mechanically deboned meat supplemented with sea buckthorn ( Hippophae rhamnoides) berry residues

Mechanically deboned meat (MDM) contains about 10 times more polyunsaturated fatty acids (PUFAs) and also more hemoproteins than hand deboned meat (HDM) and is essentially more susceptible to both chemical and biochemical oxidation. The oxidation, leading to the formation of potentially mutagenic and carcinogenic derivatives of PUFAs, can be inhibited by berry extracts rich in antioxidant polyphenols. Using the 2-thiobarbituric acid reactive substances (TBARS) method, we have established that the ethanol slurry of the juice-free solid residue of sea buckthorn ( Hippophae rhamnoides – SB) berries inhibits oxidation of unsaturated fatty acids, of both chicken and turkey MDM. The polyphenols, mainly flavonols, responsible for this inhibition, are comparatively stable during short-term cooking and 6-day storage of cooked SB-MDMs at +6 °C. About half of the polyphenols are lost, obviously oxidised, during the storage of the uncooked samples of turkey 2%SB-MDM at +6 °C. The loss of polyphenols is much smaller in the case of chicken MDM, which is characterised by an in situ lower content of fatty acids, including the polyunsaturated ones. The liquid chromatography–diode array detection–tandem mass spectrometry (LC–DAD–ESI-MS/MS) method was used for identification and ranking of the potent polyphenolic antioxidants in the berry residue.

Chemical fingerprint analysis of rhizomes of Gymnadenia conopsea by HPLC–DAD–MS n

A high-performance liquid chromatography–diode array detection–tandem mass spectrometry (HPLC–DAD–MS n ) method has been firstly developed for chemical fingerprint analysis of rhizomes of Gymnadenia conopsea R. Br. and rapid identification of major compounds in the fingerprints. Comparing the UV and MS spectra with those of reference compounds, seven main peaks in the fingerprints were identified as adenosine ( 1), 4-hydroxybenzyl alcohol ( 2), 4-hydroxybenzyl aldehyde ( 3), dactylorhin B ( 4), loroglossin ( 5), dactylorhin A ( 6) and militarine ( 7). Compounds 4– 7 were succinate derivative esters and firstly discovered from this species. The Computer Aided Similarity Evaluation System (CASES) for chromatographic fingerprint of traditional Chinese medicine was employed to evaluate the similarities of 10 samples of the rhizomes of G. conopsea collected from Sichuan, Qinghai and Hebei provinces, Tibet autonomous region of China, and Nepal. These samples from different sources had similar chemical fingerprints. This method is specific and may serve for quality identification and comprehensive evaluation of this traditional Tibetan remedy.