Qualitative and quantitative evaluation of melittin in honeybee venom and drug products containing honeybee venom

Abstract

a b s t r a c t a reverse-phase high-performance liquid chromatographic (rp-hplc) method was developed and validated for the analysis of honeybee venom samples and drug products containing honeybee venom. the validation parameters were linearity, sensitivity, precision, and recovery. melittin is the main component of honeybee venom was extracted with pure water, and then evaluated by rp-hplc with a photodiode array (pda) detector. separation of the samples was achieved on a europa protein c 18 column with linear gradient elution of acetonitrile and 0.4% phosphoric acid at 25°c. there was a flow rate of 1 ml/min. detection was set at 220 nm. limits of detection (lod) and quantification (loQ) for melittin were 1.1 and 3.2 µg/ml, respectively. the amount of melittin in honeybee venom samples ranged from 21.9 to 66.4 %.

Quantitative determination of seven chemical constituents and chemo-type differentiation of chamomiles using high-performance thin-layer chromatography

“…Another method described the use of chemometrics analysis in quality assessment of commercial chamomile oils. Moricz et al., reported a GC–MS method , chemotaxonomic authentication of herbal drug chamomile , and UHPLC–UV method was reported for the chamomile. These reported methods suffer with one or other disadvantages such as long run times, sophisticated and expensive equipment, no quantification, no differentiation studies on different types of chamomiles, and applicability for commercial products.…”

Effects of Bee Venom on Activity and Expression of 15-Lipoxygenase-1 in Human HT29 Colon Cancer

“…In the same study, the area under the curve of the melittin fraction was about 50% (49.85%) and its retention time was 42 min (27). In another research, the melittin of bee venom was separated by a gradient of 5 ñ 52% acetonitrile in reverse-phase high-performance liquid chromatography with a retention time less than 15 min (28). Because of the impurity of our bee venom and different working methods in the HPLC analysis, in comparison to other studies, the retention time of our bee venom was high.…”

The Use of Chicken Igy in a Double Antibody Sandwich Elisa for the Quantification of Melittin in Bee Venom and Bee Venom Melittin Content in Cosmetics

“…However, there is no sensitive method to detect melittin in bee venom and melittin in those cosmetic products which contain bee venom. The reliable, accurate technique, HPLC, quantifies a protein, but the previously reported LOQ for melittin (3.2 μg/mL) is inadequate for a concentration measurement in cosmetic samples (Haghi et al, 2013). Such samples typically keep melittin levels below 0.05% (unpubl.…”

“…1). The Limit of Quantitation (LOQ) of this developed DAS ELISA was approximately 1000 times and 400 times more sensitive than those obtained from reverse-phase HPLC (3.2. μg/mL) and HPLC-DAD-MS/MS (1.0 μg/mL), respectively (Zhou et al, 2010;Haghi et al, 2013). The cream sample dilutions 1:2000, 1:4000, and 1:8000, were appropriate for the detection system.…”

“…A recently used method of melittin quantification is High Performance Liquid Chromatography (HPLC). This method requires a laborious sample preparation, costly reagents, and great expertise (Haghi et al, 2013). Although it can determine a concentration with satisfactory accuracy, HPLC is less practical when quantifying a bulk mixture of proteins.…”