HG/T 2535-1993 Determination of sensitizing dye content in photographic chemicals - Reversed-phase high performance liquid chromatography
Some standard content:
Chemical Industry Standard of the People's Republic of China
HG/T2535-93
Photographic Chemicals
Determination of Sensitizing Dye Content
Reversed Phase High Performance Liquid Chromatography
Published on September 6, 1993
Ministry of Chemical Industry of the People's Republic of China
Implementation on July 1, 1994
Chemical Industry Standard of the People's Republic of China
Photographic Chemicals
Determination of Sensitizing Dye Content Reversed Phase High Performance Liquid Chromatography 1 Subject Content and Scope of Application
This standard specifies the method for determining the content of sensitizing dyes by reversed phase high performance liquid chromatography. "This standard is applicable to alcohol-soluble sensitizing dyes with ultraviolet and visible absorption. 2 Method Summary
HG/T 2535 —93
Weigh the sample, dissolve it, inject the sample solution into the chromatographic column with a micro-syringe or quantitative injection valve, perform chromatographic separation, detect with a UV-visible wavelength detector of a specified wavelength, and calculate the content of the sensitizing dye by area normalization method or external standard method. 3 Reagents
3.1 Reagents for preparing mobile phase
. The mobile phase of reversed-phase high-performance liquid chromatography is usually prepared with reagents such as methanol, acetylene, and glacial acetic acid. The absorbance of the above reagents at 254nm should generally not exceed 0.03 (1.0cm absorption cell) and should not contain 0 .5μm or larger solid particles, it is best to use liquid chromatography special reagents when preparing mobile phase. When using analytical grade reagents, they should be redistilled or filtered with a 0.5m filter membrane. The mobile phase should be degassed before use. Common degassing methods include vacuum method, heating reflux method, ultrasonic method and ammonia method, which can be selected according to specific circumstances.
3.2 Distilled water
The water used for high performance liquid chromatography should be freshly distilled water, and then filtered through a 0.5μm filter membrane. 3.3 Standard sample
The main content of the standard sample shall not be less than 99 .0%, used for external standard method quantitative analysis of standard samples certified by the Photosensitive Materials and Photographic Chemicals Testing Center of the Ministry of Chemical Industry
4 Instruments
4.1 High-performance liquid chromatograph
High-performance liquid chromatograph is usually composed of infusion system. Injector, detector, microprocessor and other components, 4.1.1 Infusion pump
Generally, it is a reciprocating plunger pump, the flow rate is adjustable within the range of 0.10mL/mi, the flow rate stability is better than ±1%, and the maximum output pressure is generally not less than 250kg/cm,
4.1. 2 Injector
Generally a six-pass injector.
4.1.3 Viewer
Adjustable UV-visible wavelength detector, the wavelength setting accuracy [±2nm) and wavelength reproducibility (±1nm) of the variable wavelength detector should be good.
Approved by the Ministry of Chemical Industry of the People's Republic of China on September 6, 1993 and implemented on July 1, 1994
4.1.4 Microprocessor
Should have the function of recording and processing chromatograms. 4.2 Chromatographic column
HG/T 2535 -- 93
The instrument used for the analysis of the content of the sensitizing dye should be equipped with a carbon 18 chromatographic column or other suitable chromatographic column, and the material of the chromatographic column is generally stainless steel. The particle size of the filler is generally 5μm or 10 μm, the column length is generally 250mm or 150mm, the inner diameter is generally 3.9mm, 4.6mm or 5mm, and the column effect meets the needs of sensitizing dye analysis. 4.3 General laboratory instruments
4.3.1 Micro syringe: the penetration is 10μL, the minimum scale is 0.1.4.3.2. Analytical balance: the sensing plate is 0.01mg4.3.3 Recording bottle: the capacity is 25mL
5 Chromatographic operating conditions
Chromatographic operating conditions are mainly composed of eight factors such as chromatographic column, mobile phase, detection wavelength, detection sensitivity, flow rate, column temperature, recording paper speed and recorder sensitivity. The selected chromatographic conditions should make all components in the sensitizing dye sample completely separated, the chromatographic retention time of the main component is 520min, and the total elution time does not exceed 30min. On the basis of meeting the above requirements, try to select versatile chromatographic columns and reagents with relatively simple mobile phases that are easy to obtain and handle, and simplify the operation as much as possible. The chromatographic operating conditions of commonly used sensitizing dyes are shown in Table 1.
Operating conditions
Chromatographic column
Sample name
Mobile phase【Medium: water; V/W
Detection wavelength (nm)
Detection sensitivity (AUFS)
Flow rate (mL/min)
Column temperature (C℃)
Recording paper speed (mm/min)
Recorder sensitivity (mV full scale)
Chromatographic operating conditions of commonly used sensitizing dyes
For different liquid chromatographs and chromatographic columns from different manufacturers, the above parameters can be appropriately adjusted to obtain suitable spectral separation. The chromatograms of the control samples of commonly used sensitizing dyes are shown in Figures 1, 2, 3, and 4. Figure 1 Chromatogram of SG-03 sample separated by 254 nm detection wavelength www.bzxz.net
1, 2—impurity peaks (3.5, 4.0 min); 3—main dye peak (4.9 nm);
4—impurity peak (7.1 min)
HG/T 2535-93
Figure 2 Chromatogram of SB-16-2 main dye separated by 254 nm detection wavelength
1—main dye peak (1.2 nm);
2—impurity peak (3.4 min);
3—intermediate peak (4.3 nm); mia)
Figure 3 uses 254nm detection wavelength to separate the main dye and the previous step
two intermediates separation spectrum
1-main dye peak (1.9min);
2, 3-intermediate peaks (3.2, 4.3min) 6 Determination steps
HG/T 2535 93
Figure 4 uses 254mm detection wavelength to separate.SR-03-2 sample chromatogram
1, 2-impurity peaks (1.8, 3.5min)
3-main dye peak (4.7min)
The area normalization method is used. When the overall determination value is lower than 90%, or there is doubt about the determination result, the external standard method is used for arbitration analysis.
The sample weight (including standard sample) specified in this experiment is determined by the sample solution injection oxygen, chromatographic column load, impurity peak orange detection limit, detector sensitivity, microprocessor attenuation and other factors. The sample weight can be appropriately adjusted according to different types of chromatographs. 5.1 Area normalization method
6.1.1 Sample solution preparation
Weigh the specified plate sample with an accuracy of 0.1m, place it in a sealed glass container, and dissolve and mix it with 10mL methanol. 4
The sample weight of commonly used sensitizing dye samples is shown in Table 2. HG/T 2535 — 93
Table 2 Sample weight of commonly used sensitizing dye samples
Sample name
Sample weight, mg
6 1.2. Setting
Debug the high performance liquid chromatograph according to the chromatographic operating conditions required by the sample to be tested, and make it stable. Note: Draw the sample solution to be tested for chromatographic separation to obtain a chromatogram. The injection volume of commonly used sensitizing dye sample solutions is shown in Table 3. Table 3 Injection volume of commonly used sensitizing dye sample solutions Sample solvent name
Injection volume, μ
6.1.3 Calculation of results
The sensitizing dye content is calculated according to formula (1):
Wu Zhong: X
The percentage of sensitizing dye in the sample to be tested, %: - The peak area of sensitizing dye in the sample to be tested: A
EA——The sum of the peak areas of each component in the sample to be tested, 6.1.4 Allowable difference
This method is measured twice in parallel, and the results are taken as the average of the two measurements. The difference between the results of the two measurements should not be greater than 1.0%: 6.2 External standard method
6 2. 1 Preparation of standard solution
Weigh the specified sensitizing dye standard sample to an accuracy of 0.01 mg, place it in a 25 mL volumetric flask, dissolve it with methanol, dilute it to the mark, and mix it. The standard solution should be prepared before use. The sample weights of commonly used sensitizing dye standards are shown in Table 4. Table 4 Sample weights of commonly used sensitizing dye samples
Name of standard mixture
Sample weight, mg
6.2.2 Preparation of sample solution
Prepare the sample solution according to the sample weight and preparation method specified in 6.2.1. 6.2.3 Determination
Adjust the HPLC according to the chromatographic operating conditions required by the sample to be tested and make it stable. Inject the specified amount of the standard solution of the sensitizing dye, the sample solution, the sample solution and the standard solution respectively for determination to obtain their respective chromatograms. The injection volume of the commonly used sensitizing dye sample solution (including the corresponding standard solution) is shown in Table 5.5
Sample name
Injection volume, L
6.2.4 Calculation
HG/T 2535—93
Table 5 Injection amount of commonly used sensitizing dye sample solutions (including corresponding standard solutions) SG-03
The sensitizing dye content is calculated according to formula (2),
Wherein: X
The percentage of sensitizing dye in the sample to be tested, %; A: The average of the main peak areas of two tested sensitizing dye standard sample solutions: A,
The average of the main peak areas of two tested sensitizing dye sample solutions; M, The mass of the tested sensitizing dye standard sample, mg: AS
The mass of one tested sensitizing dye sample, ng; The total amount of the tested sensitizing dye standard sample, %, 6.2.5 Allowable difference
This method is measured twice in parallel, and the result is the average of the two measurements. The difference between the two results should not be greater than 1.5%. The test report should include the following: All information about the sample: batch number, date, time, instrument model used in the test: 8. Analysis results; c. Abnormal phenomena observed during the determination; Any operation or freely selected test conditions not included in this standard. Additional notes: This standard was proposed by the Technical Supervision Department of the Ministry of Chemical Industry and is under the jurisdiction of the Photosensitive Material Technology Development Center of the Ministry of Chemical Industry. SR-03
This standard was jointly drafted by the Photosensitive Materials Technology Development Center of the Ministry of Chemical Industry and Shanghai Photosensitive Film Factory; Xiamen Fuda Company participated in the drafting.
The main drafters of this standard are Ding Peiling, Chen Yangfang, E Chunhui, Cai Ting, and Tong Yu.
Chemical Industry Standards of the People's Republic of China
Xixiang Chemicals
Determination of Sensitizing Dye Content Reversed-Phase High Performance Liquid Chromatography Method HG/T2535 -93
Sales Department of Chemical Industry Standards
(Standardization Research Institute of Ministry of Chemical Industry)
Postal Code: 100013
Standardization Research Institute of Ministry of Chemical Industry
Copyright reserved. No reproduction allowed
Book size 880×[2301/16 Number of printed sheets 12000 First edition in May 1994
First printing in June 1994
Number of prints【—500
Cost 2.90 Yuan
HG/T 2535--93
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