GB/T 26367-2020Hygienic requirements for biguanides disinfectants (English PDF)

4.1.3 Ethanol

It shall comply with the "Pharmacopeia of the People's Republic of China" (Part 2).

4.2 Production water

The production water shall meet the requirements of Article 30 of the "Sanitary specification for disinfection product manufacturers" (2009 edition).

5.1 Physical and chemical indicators

5.1.1 Active ingredient content

The content of the effective ingredients of the disinfectant shall meet the indicated content. Chlorhexidine disinfectants used in hand and skin disinfection shall meet the requirements of GB 27950, GB 27951, "Notice of the Ministry of Health on issuing the limit values of certain ingredients in skin and mucosal disinfectants"; the specified use concentration of chlorhexidine gluconate or chlorhexidine acetate shall be less than or equal to 45 g/L; the chlorhexidine disinfectant used for mucosal disinfection shall meet the requirements of GB 27954 and the "Notice of the Ministry of Health on issuing the limit values of certain ingredients in skin and mucosal disinfectants"; the use concentration of chlorhexidine or chlorhexidine acetate shall be less than or equal to 5 g/L; the use concentration of polyhexamethylenebiguanides disinfectant shall be less than or equal to 3 g/L.

5.1.2 Stability

The validity period is more than 12 months. The reduction rate of the effective ingredient content of the disinfectant shall be less than or equal to 10%, meanwhile the effective ingredient content after storage shall not be lower than the lower limit of the content specified by the product's corporate standard.

The validity period is more than 24 months. The reduction rate of the effective ingredient content of the disinfectant shall be less than or equal to 10%, meanwhile the effective ingredient content after storage shall not be lower than the lower limit of the content specified in the product's corporate standard.

5.1.3 Limits of lead, mercury, arsenic

Disinfectants used for hands, skin, mucous membranes shall have the following requirements: lead shall be less than or equal to 10 mg/kg, mercury shall be less than or equal to 1 mg/kg, arsenic shall be less than or equal to 2 mg/kg.

8.1 Active ingredient content

8.1.1 See Appendix A for the determination of chlorhexidine acetate, chlorhexidine gluconate, chlorhexidine hydrochloride.

8.1.2 See Appendix B for the determination of the content of polyhexamethylenebiguanide and polyhexamethylene mono-biguanides.

8.2 Stability test

It is stored in accordance with the method for determining the stability of disinfection products in the "Technical specifications for disinfection" (2002 edition); the content of active ingredients is determined in accordance with Appendix A or Appendix B.

8.3 Determination of lead, mercury, arsenic limits

Make determination according to the related methods of "Cosmetic safety technical specification" (2015 edition).

8.4 Test for killing microorganisms

It shall comply with the provisions of "Disinfection technical specifications" (2002 edition).

9.1 Transportation

It shall be airtight during transportation. Anticorrosion is required for shipping containers. It shall be handled with care and throwing is strictly prohibited. It shall be protected from sun, rain and moisture during transportation.

9.2 Storage

Store in a dry and dark place at room temperature. The packaging shall be tight and prevent moisture. The stack shall be more than 10 cm above the floor; the height of the stack shall not exceed 12 boxes; the distance from the wall shall be more than 20 cm.

A.1 Method 1: High-performance liquid chromatography

A.1.1 Overview

This method is suitable for single and compound disinfectants containing chlorhexidine acetate, chlorhexidine gluconate, chlorhexidine hydrochloride.

A.1.2 Principle

The samples are extracted by mobile phase ultrasonic wave, measured by high performance liquid chromatography-diode array detector, quantified by peak area external standard method.

A.1.3 Reagents or materials

Unless otherwise stated, the reagents used in this method are analytically pure; the experimental water is the first-grade water as specified in GB/T 6682.

A.1.3.1 Chlorhexidine acetate (C22H30Cl2N10·2C2H4O2, CAS number: 56-95-1), standard product.

A.1.3.2 Potassium dihydrogen phosphate (KH2PO4)

A.1.3.3 85% phosphoric acid (H3PO4).

A.1.3.4 Acetonitrile (CH3CN): chromatographically pure.

A.1.4 Instruments

A.1.4.1 High performance liquid chromatograph with diode array detector.

A.1.4.2 Electronic balance with a sensitivity of 0.1 mg.

A.1.4.3 Ultrasonic cleaner.

A.1.5 Test procedure

A.1.5.1 Preparation of mobile phase and extract

Weigh 2.7 g of potassium dihydrogen phosphate. Put it into a 1000 mL graduated cylinder. Add about 950 mL of water to dissolve it. Add 1.5 mL of 85% phosphoric acid. Then add water to make the volume reach to 1000 mL. Mix […]

A.1.6 Test data processing

According to the standard curve, calculate the content of chlorhexidine acetate in the sample. The calculation is as shown in formula (A.1):

Where:

X - The content of chlorhexidine acetate in the sample, in grams per kilogram (g/kg) or grams per liter (g/L);

rho - The mass concentration of chlorhexidine acetate in the sample solution obtained from the standard curve, in milligrams per liter (mg/L);

V - Constant volume, in milliliters (mL);

m - The sample amount, in grams (g) or milliliters (mL).

Note: The determination of chlorhexidine gluconate content can use chlorhexidine acetate as the standard; the result is multiplied by 1.4352.

For the determination of chlorhexidine hydrochloride content, chlorhexidine acetate can be used as the standard; the result is multiplied by 0.9246.

A.1.7 Precision

The absolute difference between two independent test results obtained under repeatability conditions is not more than 10% of the arithmetic mean.

A.2 Method 2: Titration

A.2.1 Overview

This method is based on the determination method of chlorhexidine acetate in the Pharmacopoeia of the People's Republic of China (Part 2). It is suitable for simple-recipe disinfectants containing chlorhexidine acetate.

A.2.2 Principle

Use acetone and glacial acetic acid do dissolve the sample. Add methyl orange saturated acetone solution. Use perchloric acid titrant for titration. Stop the titration when the methyl orange indicator solution turns orange. Calculate the chlorhexidine acetate content by using the perchloric acid titrant.

A.2.3 Reagents or materials

Unless otherwise stated, the reagents used in this method are analytically pure; […]

A.2.7 Precision

The absolute difference between two independent determination results obtained under repeatability conditions is not more than 10% of the arithmetic mean.

A.3 Method 3: Ultraviolet spectrophotometry

A.3.1 Overview

This method is based on the determination method of chlorhexidine acetate ointment in the Pharmacopoeia of the People's Republic of China (Part 2). It is suitable for ointment or liquid preparation simple-recipe disinfectant containing chlorhexidine (including chlorhexidine acetate, chlorhexidine gluconate, chlorhexidine hydrochloride).

A.3.2 Principle

The matrix in chlorhexidine acetate in the sample is dissolved in chloroform and then extracted by 1.5 mol/L acetic acid solution. The absorbance value is determined at a wavelength of 260 nm to quantify it.

A.3.3 Reagents or materials

Unless otherwise stated, the reagents used in this method are analytically pure; the experimental water is the grade-1 water as specified in GB/T 6682.

A.3.3.1 Chlorhexidine acetate (C22H30Cl2N10·2C2H4O2, CAS number: 56-95-1), standard product.

A.3.3.2 Chloroform (CHCl3).

A.3.3.3 Glacial acetic acid (CH3COOH).

A.3.4 Instruments

A.3.4.1 Ultraviolet spectrophotometer.

A.3.4.2 Electronic balance with a sensitivity of 0.1 mg.

A.3.5 Test procedure

A.3.5.1 Standard solution determination

Accurately weigh about 10 mg of the standard chlorhexidine acetate. Put it in a 100 mL volumetric flask. Add 1.5 mol/L acetic acid solution to dissolve and dilute to the mark. Shake well. Accurately measure 5 mL. Place it in a 50 mL measuring flask. Use ethanol to dilute it to the mark. Measure the absorbance […]

B.1 Method 1: Visible spectrophotometry

B.1.1 Overview

This method is suitable for single-recipe disinfectants containing polyhexamethylene mono-biguanides and polyhexamethylenebiguanide.

B.1.2 Principle

Polyhexamethylene mono-biguanides (PHMG) and polyhexamethylenebiguanide (PHMB) can react with Eosin Y; the color changes from orange to pink; the absorbance value is measured at a wavelength of 545 nm. The absorbance value is directly proportional to PHMG, PHMB content.

B.1.3 Reagents or materials

Unless otherwise stated, the reagents used in this method are analytically pure; the experimental water is the grade-1 water as specified in GB/T 6682.

B.1.3.1 Polyhexamethylene biguanide hydrochloride [PHMB, (C8H17N5)n ·xHCl], standard product, polyhexamethylene mono-biguanides hydrochloride standard product [PHMG, (C7H16N3)n ·xHCl, which may be raw materials with a purity greater than 95%].

B.1.3.2 Eosin Y (C20H6Br4Na2O5).

B.1.3.3 Sodium acetate trihydrate (CH3COONa·3H2O).

B.1.4 Instruments

B.1.4.1 Visible light spectrophotometer, 5 cm cuvette.

B.1.4.2 Electronic balance with a sensitivity of 0.1 mg.

B.1.5 Test procedure

B.1.5.1 Preparation of indicator solution

B.1.7 Precision

The absolute difference between two independent determination results obtained under repeatability conditions is not more than 10% of the arithmetic mean.

B.2 Method 2: Ultraviolet spectrophotometry

B.2.1 Overview

This method is suitable for single-recipe disinfectants containing polyhexamethylenebiguanide.

B.2.2 Principle

Polyhexamethylene biguanide has ultraviolet absorption at 234 nm; the absorbance value is proportional to the PHMB content within a certain concentration range.

B.2.3 Reagents or materials

Unless otherwise specified, the experimental water is the grade-1 water as specified in GB/T 6682. Polyhexamethylenebiguanide hydrochloride [PHMB, (C8H17N5)n·xHCl] standard product.

B.2.4 Instruments

B.2.4.1 UV spectrophotometer, 1 cm quartz cuvette.

B.2.4.2 Electronic balance with a sensitivity of 0.1 mg.

B.2.5 Test procedure

B.2.5.1 Preparation of PHMB standard solution

Accurately weigh 10 mg of PHMB standard substance and use water to dilute to 100 mL, to obtain a 100 mg/L standard solution.

B.2.5.2 Preparation of standard curve

Use water to dilute the standard solution into a standard series of 2 mg/L, 4 mg/L, 6 mg/L, 8 mg/L, 10 mg/L, 12 mg/L, 16 mg/L. Use a 1 cm quartz cuvette to determine the absorbance value at 234 nm of the UV spectrophotometer; meanwhile draw a standard curve. […]

B.3.3 Reagents or materials

Unless otherwise stated, the reagents used in this method are analytically pure; the experimental water is the grade-1 water as specified in GB/T 6682.

B.3.3.1 Polyhexamethylenebiguanide hydrochloride [PHMB, (C8H17N5)n · xHCl], standard product, polyhexamethylene mono-biguanides hydrochloride standard product [PHMG, (C7H16N3)n · xHCl, which can be raw materials with purity greater than 95%], chlorhexidine acetate (CHA, C22H30Cl2N10 · 2C2H4O2) standard.

B.3.3.2 Sodium tetraborate (Na2B4O7·10H2O, > 99.5%).

B.3.3.3 Sodium hydroxide (NaOH): superior pure.

B.3.3.4 Sodium dodecyl sulfate (SDS, 99%).

B.3.3.5 Sodium deoxycholate (SD, 98%).

B.3.3.6 Polyethylene glycol 20000 (PEG20000).

B.3.4 Instruments

B.3.4.1 Capillary electrophoresis instrument with diode array detector (PDA).

B.3.4.2 Electronic balance with a sensitivity of 0.1 mg.

B.3.5 Test procedure

B.3.5.1 Preparation of standard solution

Accurately weigh 50 mg each of PHMG, CHA, PHMB; place them in a 15 mL plastic centrifuge tube. Use a pipette to add 10 mL of water and vortex to mix to obtain a standard stock solution with a mass concentration of 5 g/L. Store in refrigerator at 4 °C.

B.3.5.2 Preparation of standard curve

Use sample solution to dilute the standard stock solutions of PHMG, PHMB, CHA step by step, to respectively prepare them into 7.5 mg/L, 15 mg/L, 30 mg/L, 60 mg/L, 120 mg/L PHMG working solutions, PHMB and CHA mixed working solution. […]