SPDS Aloe Vera Method Book

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Standard Operating Procedures

Subject: Aloe Polysaccharides by O-Acetyl Department: Laboratory Original Date: 06/20/2016 Revision #: 1

Procedures #: CA17011

Approval Date:

1. Purpose To determine amount of aloe polysaccharides in raw materials. 2. Reference Aloe Products for Food Raw Material, Chinese National Standard, QB/T 2489 2007 The acetyl groups on polysaccharides are reacted with hydroxylamine to form acetohydroxamic acid. The resulting acetohydroxamic acid is reacted with ferric trichloride to form a ferric ‐ acethydroxamic acid complex and measured 540 nm. 3. Scope This method is intended for raw materials and finished product. Products may contain acetic acid that would react to acetylcholine resulting in false positive results. 4. Materials and Equipment 4.1. UV-VIS spectrophotometer set to 540 nm and 1 cm path length cell

4.2. Ultrasonic bath 4.3. Deionized water 4.4. Hydroxylamine hydrochloride [5470-11-1] 4.5. Acetylcholine chloride [60-31-1] 4.6. Ferric chloride hexahydrate [10025-77-1] fw 270 4.7. Hydrochloric acid, concentrated [7467-01-0] 4.8. Sodium hydroxide [1310-73-2] fw 40 4.9. Sodium acetate trihydrate [6131-90-4] fw 136 5. Preparation of Solutions 5.1. Hydrochloric acid (4 M)

5.1.1.Dilute 33.3 ml of concentrated HCl to 100 ml with DI water. 5.2. Hydrochloric acid (0.1 M) 5.2.1.Dilute 2.5 ml of 4 M HCl to 100 ml with DI water. 5.3. Sodium Hydroxide (3.5 M) 5.3.1.Dissolve 14.0 g in 100 ml of DI water. 5.4. Ferric chloride – hydrochloric acid (0.37 M) 5.4.1.Dissolve 10.0 g in 100 ml of 0.1 M HCl 5.5. Sodium acetate (0.001 M, pH 4.5) 5.5.1.Dissolve 136 mg in 1000 ml of DI water 5.6. Hydroxylamine hydrochloride (2M) 5.6.1.Dissolve 13.9 g in 100 ml of DI water (keep refrigerated for up to one week)