Analysis of small samples of limonoids of neem (Azadirachta indica) using solid phase extraction from tissue culture

Abstract

The limonoids (triterpenoids), including the natural insecticide azadirachtin, from the seeds of the neem tree (Azadirachta indica) can be partially purified and separated into groups for final analysis very quickly by the use of solid phase extraction procedures. The individual compounds can then be accurately quantified in the separated fractions by supercritical fluid chromatography or high pressure liquid chromatography. The method is particularly useful for the small samples available from tissue culture experiments, but is equally applicable to analysis of seed extracts. Two alternative procedures are described, one that gives six fractions and more accurate quantification, and a second shorter, faster procedure for smaller samples that gives two fractions. Both procedures include the quantification of azadirachtin. Copyright © 2000 John Wiley & Sons, Ltd.

A comprehensive review of phytochemical profile, bioactives for pharmaceuticals, and pharmacological attributes of Azadirachta indica

“…Azadirachtin has been isolated and quantified in several reports from the seeds of the tree (Deota et al, ; Jarvis & Morgan, ; Kumar et al, ; S. M. Lee, Olsen, Schweizer, & Klocke, ). Nimbidinin and nimbidic acid were also isolated from extract of seed kernels (Mitra, Garg, & Pandey, ).…”

“…Azadirachtin has been isolated and quantified in several reports from the seeds of the tree (Deota et al, 2000;Jarvis & Morgan, 2000;Kumar et al, 1996; S. M. Lee, Olsen, Schweizer, & Klocke, 1988).…”

Limonoids: Overview of Significant Bioactive Triterpenes Distributed in Plants Kingdom

“…Tests have shown azadirachtin to be effective on at least 200 different insect species. 3,23,24) Azadirachtin and related limonoids from neem tree have shown strong antifeedant and growth regulating activity and at the same time these compounds are highly bio-degradable, are very slightly toxic towards non-target organisms, while they are non-toxic towards humans and mammals in general, with very low persistence in systemic action, 21,25) this is the reason why an array of insecticides and pesticides comprising of azadirachtin as principal constituent and based on neem extract are available today in the market. Koul et al 26,27) have further shown that azadirachtin being the most active compound in neem is not synergized or influenced by any other limonoid, but other non-azadirachtin limonoids show synergism in specific combination, which may be due to their different modes of action.…”

Neem: Today and in the New Millennium

“…Standardization and quality control of the product. There is as yet no standard method for the determination of azadirachtin (although HPLC is much used) and certainly no standards for the lesser compounds in azadirex (but see Jarvis and Morgan, 2000), and no agreed standards of quality control. An important consideration for azadirex is the control of aflatoxin in the product.…”

“…This was also achieved during the AZTEC project and the results are described in Jarvis and Morgan (2000). A rapid and sensitive analysis of azadirachtin and related triterpenoids by high-performance liquid chromatography-atmospheric pressure chemical ionisation mass spectrometry from tissue culture samples has also been developed (Schaaf et al, 2000).…”