Method development for the determination of elements in Hypericum perforatum L. (St John's wort) herb and preparations using inductively coupled plasma-optical emission spectroscopy and microwave digestion.

OBJECTIVES: A method was developed to analyze St John's wort (Hypericum perforatum L.) herb and preparations using inductively coupled plasma-optical emission spectroscopy (ICP-OES) to determine the quantity of 11 elements (Al, B, Ba, Ca, Cu, Fe, Mg, Mn, Ni, Sr and Zn). METHODS: This study includes the evaluation of digestion acids and calibration methods, as well as instrumental parameters such as choice of nebulizer and emission wavelength. KEY FINDINGS: Two nebulizers (Conikal and SeaSpray) performed similarly for most elements, and two optimum wavelengths were determined for each element. Five acids were evaluated for the digestion of the Polish Certified Reference Material Tea Leaves (INCT-TL-1), while three were taken forward to use for the different St John's wort formulations (i.e. herb, capsule and tablet). A simple protocol using 5 ml HNO3 was sufficient in most cases; however, variability was observed for elements often bound in silicates (e.g. Al, Fe and Zn). An external weighted calibration was also found to be preferential over unweighted, and the use of standard addition affected some concentration values up to 20%. CONCLUSIONS: Therefore, this paper presents the development and optimized method parameters to be used with ICP-OES that will allow the analysis of 11 key elements present in St John's wort herb and preparations.

Elemental fingerprinting of Hypericum perforatum (St John's Wort) herb and preparations using ICP-OES and chemometrics.

St. John's wort (SJW) (Hypericum perforatum) is a herbal remedy commonly used to treat mild depression. The elemental profiles of 54 samples (i.e., dry herbs, tablets and capsules) were evaluated by monitoring 25 elements using Inductively Coupled Plasma Optical Emission Spectroscopy (ICP-OES). The major elemental constituents in the SJW samples were Ca (300-199,000µg/g), Mg (410-3,530µg/g), Al (4.4-900µg/g), Fe (1.154-760µg/g), Mn (2.4-261µg/g), Sr (0.88-83.6µg/g), and Zn (7-64µg/g). For the sixteen elements that could be reliably quantified, principal component analysis (PCA) was used to investigate underlying patterns in the data. PCA models identified 7 key elements (i.e., Ba, Ca, Cd, Mg, Mo, Ni and Y), which described 85% of the variance in the dataset in the first three principal components. The PCA approach resulted in a general delineation between the three different formulations and provides a basis for monitoring product quality in this manner.

Native CB1 receptor affinity, intrinsic activity and accumbens shell dopamine stimulant properties of third generation SPICE/K2 cannabinoids: BB-22, 5F-PB-22, 5F-AKB-48 and STS-135.

In order to investigate the in vivo dopamine (DA) stimulant properties of selected 3rd generation Spice/K2 cannabinoids, BB-22, 5F-PB-22, 5F-AKB-48 and STS-135, their in vitro affinity and agonist potency at native rat and mice CB1 receptors was studied. The compounds bind with high affinity to CB1 receptors in rat cerebral cortex homogenates and stimulate CB1-induced [(35)S]GTPγS binding with high potency and efficacy. BB-22 and 5F-PB-22 showed the lowest Ki of binding to CB1 receptors (0.11 and 0.13 nM), i.e., 30 and 26 times lower respectively than that of JWH-018 (3.38 nM), and a potency (EC50, 2.9 and 3.7 nM, respectively) and efficacy (Emax, 217% and 203%, respectively) as CB1 agonists higher than JWH-018 (EC50, 20.2 nM; Emax, 163%). 5F-AKB-48 and STS-135 had higher Ki for CB1 binding, higher EC50 and lower Emax as CB1 agonists than BB-22 and 5F-PB-22 but still comparatively more favourable than JWH-018. The agonist properties of all the compounds were abolished or drastically reduced by the CB1 antagonist/inverse agonist AM251 (0.1 µM). No activation of G-protein was observed in CB1-KO mice. BB-22 (0.003-0.01 mg/kg i.v.) increased dialysate DA in the accumbens shell but not in the core or in the medial prefrontal cortex, with a bell shaped dose-response curve and an effect at 0.01 mg/kg and a biphasic time-course. Systemic AM251 (1.0 mg/kg i.p.) completely prevented the stimulant effect of BB-22 on dialysate DA in the NAc shell. All the other compounds increased dialysate DA in the NAc shell at doses consistent with their in vitro affinity for CB1 receptors (5F-PB-22, 0.01 mg/kg; 5F-AKB-48, 0.1 mg/kg; STS-135, 0.15 mg/kg i.v.). 3rd generation cannabinoids can be even more potent and super-high CB1 receptor agonists compared to JWH-018. Future research will try to establish if these properties can explain the high toxicity and lethality associated with these compounds.