SPDS Aloe Vera Method Book

OFFICIAL DOCUMENT

Date Issued: Supersedes:

Page:

3 of 15

Procedure Title: Identification and quantitation of Aloe Vera Using 1 H NMR

N/A

Effective Date:

TBD

SOP #:

Revision #:

00

Prepared By: Department: Applies To:

Isaac Lee Quality Herbalife QC Laboratories

1.3.1 Not less than 300 MHz NMR spectrometer (e.g. Bruker, Ascend 400 with Topspin software, or equivalent) 1.3.2 Lyophilizer (e.g. Free Zone Plus 4.5L Cascade Dry System or equivalent) 1.3.3 Vortex mixer 1.3.4 Sonicator 1.3.5 5 mm NMR tube (e.g. Norell, part# 507-HP-7, or equivalent) 1.3.6 Test tubes 1.3.7 Glass bottle 1.3.8 Glass vial with Teflon lined cap 1.4 NMR Conditions ( The following parameters can be modified for optimization ) 1.4.1 Acquisition parameters 1.4.1.1 Relaxation delay (d1) 2-6 sec 1.4.1.2 Number of scans (NS) Not less than 16 depending on the concentration 1.4.1.3 Sweep width (SW) 15 ppm 1.4.1.4 Pulse angle 30 o to 90 o depending on the sample (it is recommended to use water suppression pulse sequence with 90 o angle on the sample containing water) 1.4.1.5 Temperature 20 o C – 25 o C 1.4.1.6 Date points: Not less than (NLT) 64K 1.4.1.7 Dummy scans: 4

1.5

Blank preparation 1.5.1

Dissolve 3mg of DSS with 0.7mL of D 2

O in a test tube. Transfer the solution to a NMR

tube.

1.6

Reference Standard Preparation 1.6.1

Weigh 5 mg of Dextrose and Malic acid reference standards in the separate 12x75 mm test tubes and add 3 mg of DSS. Add 0.7 mL of deuterium oxide and vortex to dissolve. Transfer the solution to a NMR tube. 1.6.2 Filter each dilution through a 0.45 μm filter prior to NMR analysis, if required. 1.7 Nicotinamide solution 1.7.1 Weigh 357.14mg of Nicotinamide reference standard and 214.29mg of DSS in a glass bottle, and add 50mL of deuterium oxide. Mix the solution until it observes the clear dissolution.

1.8

System suitability solution 1.8.1

Transfer 0.7mL of Nicotinamide solution to a NMR tube.

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