SPDS Aloe Vera Method Book

Date Issued: Supersedes:

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Procedure Title: Determination of Polysaccharides Molecular Weight in Aloe vera Fresh Leaf and Concentrated Juice Powder by SEC-RI

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Effective Date:

QCL

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Prepared By: Yuehong Zhang, Zhichao Bao Department: R&D Applies To: Herbalife International

CONTENTS Procedures Definitions References Forms/Chromatograms Ownership/Management Sign-off Revision History

SECTION 1.0 2.0 3.0 4.0 5.0 6.0

PURPOSE The purpose of the test method is to determine the polysaccharides molecular weight in Aloe vera fresh leaf and concentrated juice powder by Size Exclusion Chromatography Coupling with Refractive Index Detector (SEC-RI).

REAGENTS 

1 : 0.342, 2 :1.32, 3 : 6.2, 4 : 10.0, 5 : 21.7, 6 : 48.8,

Pullulan Standards and Molecular Weight (MW) (M w

7 :113, 8 : 200, 9 : 366, and 10 : 805 kDa), Sigma-Aldrich, Product No.96351 or equivalent  Sodium chloride, Sinopharm Reagents, Catalog# 10019318 or equivalent  Methanol, HPLC grade  Ultrapure Water or equivalent EQUIPMENT  Analytical Balance  Nitrocellulose Membrane, 0.22μm GSWP or equivalent  Disposable Syringe Filters, 25mm, 0.45 μm Hydrophilic PTFE membrane filter or equivalent  Disposable Syringes (1mL)  Disposable Plastic Pipette  Vortex Mixer  SCIENTZ-12N Lyophilizer  1-L Volumetric Flask  50-mL Volumetric Flask  5-mL Volumetric Flask  1.5-mL Glass HPLC Vials HPLC  HPLC System: Waters 2695 Alliance Separations Module consisting of pump and auto sampler and Empower 3 GPC/SEC Software or equivalent  Waters 2414 Refractive Index Detector or equivalent  Size Exclusion Chromatographic (SEC) Column: Phenomenex PolySep™-SEC GFC-P Linear, LC Column 300 x 7.8 mm, Product No. CHO-9232 or equivalent  Guard Column: Phenomenex PolySep™-SEC GFC-P Guard, LC Guard Column 35 x 7.8 mm, Product No. CHO-9225 or equivalent  HPLC chromatographic conditions: - Mobile phase: 100mM Sodium chloride solution - Flow rate: 0.7 mL/min

- Column temperature: 35 o C - Sample temperature: 20 o C - Refractive index detector temperature: 35 o C - Injection volume: 100 μL - Run time: 30 minutes

1.0

PROCEDURES

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