Technology Transfer and Scale Up of a Potential Cancer‐Preventive Plant Dynamic Extraction of Glucoraphanin

Fisher, Derek; Garrard, Ian; Heuvel, Remco van den; Sutherland, Ian; Chou, F. Edward; Fahey, Jed W.

doi:10.1081/jlc-200063563

Cited by 31 publications

(12 citation statements)

References 5 publications

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“…The GR content of the broccoli sprout preparations used in this study was determined using paired-ion high performance liquid chromatography (HPLC) and hydrophilic interaction HPLC [ 21 , 22 ]. GR was quantitated by comparing peak areas with the peak areas of an authentic GR standard prepared as previously described [ 23 ].…”

Section: Methodsmentioning

confidence: 99%

Sulforaphane Bioavailability from Glucoraphanin-Rich Broccoli: Control by Active Endogenous Myrosinase

et al. 2015

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Glucoraphanin from broccoli and its sprouts and seeds is a water soluble and relatively inert precursor of sulforaphane, the reactive isothiocyanate that potently inhibits neoplastic cellular processes and prevents a number of disease states. Sulforaphane is difficult to deliver in an enriched and stable form for purposes of direct human consumption. We have focused upon evaluating the bioavailability of sulforaphane, either by direct administration of glucoraphanin (a glucosinolate, or β-thioglucoside-N-hydroxysulfate), or by co-administering glucoraphanin and the enzyme myrosinase to catalyze its conversion to sulforaphane at economic, reproducible and sustainable yields. We show that following administration of glucoraphanin in a commercially prepared dietary supplement to a small number of human volunteers, the volunteers had equivalent output of sulforaphane metabolites in their urine to that which they produced when given an equimolar dose of glucoraphanin in a simple boiled and lyophilized extract of broccoli sprouts. Furthermore, when either broccoli sprouts or seeds are administered directly to subjects without prior extraction and consequent inactivation of endogenous myrosinase, regardless of the delivery matrix or dose, the sulforaphane in those preparations is 3- to 4-fold more bioavailable than sulforaphane from glucoraphanin delivered without active plant myrosinase. These data expand upon earlier reports of inter- and intra-individual variability, when glucoraphanin was delivered in either teas, juices, or gelatin capsules, and they confirm that a variety of delivery matrices may be equally suitable for glucoraphanin supplementation (e.g. fruit juices, water, or various types of capsules and tablets).

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Section: Methodsmentioning

confidence: 99%

Sulforaphane Bioavailability from Glucoraphanin-Rich Broccoli: Control by Active Endogenous Myrosinase

et al. 2015

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“…Branded countercurrent chromatography (CCC) systems, DE-Mini, DE-Midi and DE-Maxi Centrifuges, can now provide manufacturers with a range of scales from mg-quantity of products to kg-quantity. For instance, 50 g glucoraphanin has been prepared in multiple runs within 3 days using a DE-Midi Centrifuge with a 1-L column volume [2] and, in association with Dynamic Extractions Ltd., this process was subsequently scaled up for making 1 kg glucoraphanin * Corresponding author. Tel.…”

Section: Introductionmentioning

confidence: 99%

The importance of column design for protein separation using aqueous two-phase systems on J-type countercurrent chromatography

Guan¹,

Bourton²,

Hewitson³

et al. 2009

Separation and Purification Technology

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“…It permits enhanced mixing of the ATPS in a manner similar to the much larger multilayered toroidal coils that have been pioneered for use with ATPS on commercially available and highly scalable J‐type CCC configurations (Guan et al , ; Sutherland, ; Sutherland et al , ). This work and that of others suggests that scale‐up of this method should be facile and predictable (Fisher et al, ; Guan et al, ; Ito et al, ; Wood et al, ; Sutherland et al , ), and that the use of CCCs with greater column capacity will result in a concomitant increase in the yield of catalytically active myrosinase, if they can be adapted to this or other tubing design that permits greater retention of ATPS phases. Thus, based only upon first principles, 80 g of dried moringa leaves might be used to charge one of the largest available CCCs today (18 L column capacity), for a per‐run yield of about 740 Units in a gram.…”

Section: Resultsmentioning

confidence: 99%

Purification of Active Myrosinase from Plants by Aqueous Two‐Phase Counter‐Current Chromatography

Wade

Ito

Ramarathnam

et al. 2014

Phytochemical Analysis

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Introduction Myrosinase (thioglucoside glucohydrolase; E.C. 3.2.1.147), is a plant enzyme of increasing interest and importance to the biomedical community. Myrosinase catalyses the formation of isothiocyanates such as sulforaphane (frombroccoli) and 4-(α-l-rhamnopyranosyloxy)benzyl isothiocyanate (from moringa), which are potent inducers of the cytoprotective phase-2 response in humans, by hydrolysis of their abundant glucosinolate (β-thioglucoside N-hydroxysulphate) precursors. Objective To develop an aqueous two-phase counter-current chromatography (CCC) system for the rapid, three-step purification of catalytically active myrosinase. Methods A high-concentration potassium phosphate and polyethylene glycol biphasic aqueous two-phase system (ATPS) is used with a newly developed CCC configuration that utilises spiral-wound, flat-twisted tubing (with an ovoid cross-section). Results Making the initial crude plant extract directly in the ATPS and injecting only the lower phase permitted highly selective partitioning of the myrosinase complex before a short chromatography on a spiral disk CCC. Optimum phase retention and separation of myrosinase from other plant proteins afforded a 60-fold purification. Conclusion Catalytically active myrosinase is purified from 3-day broccoli sprouts, 7-day daikon sprouts, mustard seeds and the leaves of field-grown moringa trees, in a CCC system that is predictably scalable.

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Technology Transfer and Scale Up of a Potential Cancer‐Preventive Plant Dynamic Extraction of Glucoraphanin

Cited by 31 publications

References 5 publications

Sulforaphane Bioavailability from Glucoraphanin-Rich Broccoli: Control by Active Endogenous Myrosinase

Sulforaphane Bioavailability from Glucoraphanin-Rich Broccoli: Control by Active Endogenous Myrosinase

The importance of column design for protein separation using aqueous two-phase systems on J-type countercurrent chromatography

Purification of Active Myrosinase from Plants by Aqueous Two‐Phase Counter‐Current Chromatography

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