Optimization and Validation of a Multiresidue Method for Screening of 126 Pesticide Residues in Herbal Raw Materials and Extracts Used as Ingredients in Ayurvedic Medicines and Dietary Supplements.

BACKGROUND: Testing for pesticide levels in herbal products is an important aspect in determining product safety. Plants and their extracts are widely used as ingredients in botanical dietary supplements and traditional medicines. The extracts of plants, especially those prepared out of organic solvents, are rich in secondary metabolites and pigments, and adequate clean-up is required since the extracts completely dissolve in organic solvents. OBJECTIVE: The study aims at reporting a multiresidue analytical method for 126 different pesticides in raw material biomass as well as extracts of plants, which are widely used as ingredients in ayurvedic medicines as well as dietary supplements using LC-MS/MS and GC-MS/MS with a rugged sample preparation technique for accurate results. METHOD: QuEChERS (quick, easy, cheap,effective, rugged, and safe) procedure, gel permeation chromatography (GPC), GPC coupled with solid phase extraction (SPE), and liquid-liquid extraction (LLE) coupled with SPE sample preparation methods were compared against each other for suitability to test pesticides in selected herbal raw materials and their alcoholic and aqueous extracts. The standard addition method was used for quantifying the level of pesticides below 10 µg/kg. RESULTS: Single laboratory validation for sample preparation involving GPC and SPE resulted linearity in the range of 2.5-500 ng/mL, average intraday and interday precision of 6.6% RSD, and average recovery (spiked at 10 µg/kg) of 92% for all analytes tested. The method was repeatable with different analysts and days. CONCLUSIONS: The sample preparation technique combining GPC and SPE as well as LLE and SPE was the most suitable for the selected herbal alcoholic extracts, whereas any of the regular techniques involving LLE, SPE, and QuEChERS were suitable for raw material biomass as well as aqueous extracts. HIGHLIGHTS: The method was found to be capable of determining selected pesticides in the selected matrixes at 10 µg/kg concentration. Provision of recycling solvents used in the GPC+SPE method was adopted to make the method environmentally friendly.

Ethanolic extract of Moringa oleifera Lam. leaves protect the pre-pubertal spermatogonial cells from cyclophosphamide-induced damage.

ETHNOPHARMACOLOGICAL RELEVANCE: Moringa oleifera Lam. is widely cultivated in Asian and African countries for its medicinal and dietary significance. The leaves are highly nutritious and are known to possess various biological activities. MATERIALS AND METHODS: Pre-pubertal Swiss albino male mice were injected with single dose of cyclophosphamide (CP, 200mg/kg body weight) or ethanolic extract of Moringa oleifera leaves (MOE, 100mg/kg body weight) intraperitoneally. In combination group, MOE was administered 24h prior to CP injection. RESULTS: CP induced a significant decrease in testicular weight (p<0.01) and depletion of germ cells (p<0.001) and higher level of DNA damage (p<0.001) compared to control. The expression of P53, Bax, Cytochrome C (Cyt C) was increased while there was a decrease in the expression of Bcl2, c-Kit and Oct4. Administration of MOE 24h prior to CP treatment ameliorated the depletion (p<0.001), DNA damage (p<0.001) and apoptosis (p<0.01) of germ cells induced by CP. The mitigating effect of MOE appears to be mediated by up-regulating the expression of c-Kit and Oct4 transcripts in P53-independent manner. CONCLUSION: MOE protects the spermatogonial cells from CP-induced damage by modulating the apoptotic response elicited by CP and therefore can be considered as an efficient method of male fertility preservation.