Hangzhou King Techina Feed Co.

, LTD
Analysis method of CS PLUS
1. Appearance: white to offwhite granule;

2. Particle Size: Through 2.00mm (10 mesh) sieve, not less than 90%;

3. Identification: The retention time of major peak of test solution corresponds to that of standard

solution in Assay test.

4. Assay:

4.1. Reagent

4.1.1 Acetonitrile: chromatographic pure;

4.1.2 Sodium 1-heptanesulfonate: analytical reagent;

4.1.3 Phosphoric acid: analytical reagent;

4.1.4 Methanol: chromatographic pure;

4.1.5 Water: ultra-pure water;

4.1.6 50% Methanol water solution: volume ratio of methanol and water is 1:1;

4.1.7 0.017 mol/L Sodium 1-heptanesulfonate solution (contain 0.02% phosphoric acid):

weight 1.7 g of sodium 1-heptanesulfonate, and transfer it to 500 mL volumetric flask, add 0.1 mL

H3PO4, dilute it to volume with ultra-pure water, and mix;

4.1.8 Mobile phase: prepare a mixture of acetonitrile and 0.017 mol/L sodium 1-heptanesulfonate

solution (15: 85, volume ratio). Pass through 0.45 μm PTFE filter and degas.

4.1.9 Standard solution: Dissolve an accurately weighed quantity of cysteamine hydrochloride

RS (≥98%) in 50% methanol water solution. The concentration of cysteamine hydrochloride in

the solution should between 150 mg/100 mL and 200 mg/100 mL. Prepare it when needed.

4.2 Chromatographic system

HPLC column C18 column (250mm×4.6mm i.d, 5μm.)

Flow rate 1.0 mL/min

Detection wavelength 214 nm

Injection volume 20 μL

4.3 Assay preparation

Transfer about 0.5 g of sample, accurately weighed, to a 100-mL volumetric flask. Add about 40

mL of 50% methanol water solution, heat in boiling water bath for 20 minutes. After cooling,
 Hangzhou King Techina Feed Co., LTD
dilute with 50% methanol water solution to volume, and mix. Place it for 15 minutes, filter it with

ordinary filter paper first, and then filter it with 0.45 μm PTFE filter member again. The filtrate is

for HPLC injection.

4.4 Procedure

Separately inject equal volumes (about 20µL) of the Standard solution and the Assay preparation

into the chromatograph, record the chromatograms, and measure the responses for the major peaks.

Calculate the percentage of cysteamine hydrochloride taken by the formula:

A1×C1×100
X= —————— ×100%
A2×m1
Where,

X—The content of cysteamine hydrochloride in the sample;

A1—Peak area of cysteamine hydrochloride in sample test;

C1—The concentration of standard solution, mg/mL;

A2—Peak area of cysteamine hydrochloride in standard solution test;

m1—Mass of the test sample, mg.

5. Moisture: Dry it in vacuum at 60℃ for 4 hours. It loses not more than 5.0% of its weight.

6. Pb: it should be not more than 2mg/kg;

7. Arsenic: it should be not more than 2mg/kg.