HG/T 2414-1993 Determination of coupler content in photographic chemicals - Reversed-phase high performance liquid chromatography
Some standard content:
Chemical Industry Standard of the People's Republic of China
HG / T 2414 93
Photographic chemicals
Determination of coupler content
Reversed phase high performance liquid chromatography
1993-(3-22 Issued
Ministry of Chemical Industry of the People's Republic of China
Implementation on 1993-10-01
Chemical Industry Standard of the People's Republic of China
Photographic chemicals
Determination of coupler content
Reversed phase high performance liquid chromatography
1 Content and scope of applicationwww.bzxz.net
HG / T 2414 — 93
This standard specifies the method for determining the content of colorants by reverse phase high performance liquid chromatography, including reagents and materials, instruments and devices, chromatographic operation procedures, determination steps, result calculation, etc. This standard is applicable to the determination of alcohol-soluble colorants with ultraviolet absorption. 2 Method Summary
The sample is weighed and dissolved, and the sample solution is injected into the ODS chromatographic column (or Ca, TMS chromatographic column) with a micro-syringe or quantitative injection, and chromatographic separation is performed. The detection is performed with a 2541m or other specified wavelength ultraviolet detector, and the content of the colorant is calculated by the area normalization method or the external standard method.
3 Reagents and Materials
3. 1 Reagents for preparing mobile phase
The mobile phase of reversed-phase HPLC is usually prepared with methanol, anhydrous ethanol, acetonitrile, glacial acetic acid and other reagents. The absorbance at 254m of the above reagents should generally not exceed 0.03 (1.0cm absorption cell), and should not contain solid particles larger than 0.5μm. It is best to use liquid chromatography-specific reagents when preparing mobile phase. When using analytically pure reagents, redistillation should be performed, and if necessary, precise fractionation should be performed to remove trace impurities. The mobile phase should be degassed before use. Common degassing methods include vacuum method, heating reflux method, ultrasonic method and nitrogen method, which can be selected according to specific circumstances.
3. 2 Water for chromatography
The water used for HPLC should be fresh distilled water to avoid the generation of microorganisms in the water and damage to the flow system of the HPLC. 3.3 Standard samples
High-purity colorant samples with a chromatographic main body content of not less than 99.0% are used for quantitative analysis by external standard method. Standard samples are tested and certified by the Photosensitive Material Quality Supervision and Inspection Center of the Ministry of Chemical Industry. 3.4 Control samples
are prepared from relevant colorants and their impurities, or relevant colorants and their difficult-to-separate intermediates, and are used to identify whether the chromatographic column for separating the colorants meets the separation requirements. Control samples are tested and certified by the Chemical Industry Ministry’s Photosensitive Material Quality Supervision and Inspection Center. Tested and certified by the Photosensitive Materials Quality Supervision and Inspection Center of the Ministry of Industry. 4 Instruments and devices
4.1 General laboratory instruments.
4.2 Analytical balance: sensitivity is 0.01mg.
4.3 Microdisc syringe: capacity 10uL, minimum engraving is 0.1μL, 4.4 High-performance filtration chromatograph:
High-performance liquid chromatograph is usually composed of infusion pump, injector, detector, microprocessor and other components, approved by the Ministry of Chemical Industry of the People's Republic of China on March 22, 1993
Implementation on October 1, 1993
HG / T 2414 93
4.4.1 Infusion pump: generally a reciprocating plunger pump, the flow rate should be adjustable within the range of 0-10mL/min, the flow rate stability should be better than ±1%, and the maximum output pressure should not be less than 25MPa. 4.4.2 Sample injector: generally a six-way sample injector, or a dual-flow sample injector. The injection setting error should not be greater than 0.1.JuL. 4.4.3 UV detector: a 2541m fixed wavelength detector is selected, or a variable wavelength (200-390nm) detector can be used. The wavelength setting error should not be greater than 2nm, and the wavelength fluctuation range should not be greater than 1nm. 4.4.4 Microprocessor: It should have the function of recording and processing chromatograms. 4.5 Chromatographic column: The chromatographic instrument used for the analysis of the coloring agent content should be equipped with at least two ODS chromatographic columns. If conditions permit, it should also be equipped with C (octane) chromatographic column and methyl chromatographic column. The material of the chromatographic column is generally stainless steel; the particle size of the filler is generally 5μm or 10um: the column length is generally 250mm or 150mm; the inner diameter is generally 3.9mm or 4.6mm or 5.0mm. The column effect meets the needs of colorant analysis. If necessary, the colorant control sample should be used for calibration. 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 recording sensitivity: the selected chromatographic operating conditions should make all components in the colorant sample completely separated, the chromatographic retention time of the main component is 5 to 20min, and the total elution time is about 30min. On the basis of meeting the above requirements, try to use a chromatographic column with strong versatility and a mobile phase with relatively simple reagent composition and easy to obtain and handle, so as to simplify the operation as much as possible. The chromatographic analysis operating conditions of commonly used couplers are shown in Tables 1 and 2. Table 1 Mobile phase used for chromatographic analysis of commonly used couplers Name of product
Sample name
Operating conditions
Chromatographic column
Detection wavelength, nm
Detection sensitivity, AUFS
Flow rate, mL/min
Column source ℃
Recording paper speed, mm/min
Recording sensitivity, mV/Fs
Methanol: water = 94:6
Composition of mobile phase (VV)
Anhydrous ethanol: water = 88:12
Methanol: water: ethyl acetate 87:4:
Methanol: water 90:10
Methanol (containing 0. [% glacial acetic acid): water (containing 0. 1% glacial acetic acid) = 95:5Methanol (containing 0.1% glacial acetic acid): water (containing 0.1% glacial acetic acid) = 92:8Methanol! Water = 93:7
Table 2 Other chromatographic analysis operating conditions of commonly used colorantsCOC-1
HG / T 2414 -- 93
For different HPLCs and ODS columns from different manufacturers, the above parameters can be adjusted appropriately to obtain appropriate chromatographic separation and meet the requirements of the chromatographic separation diagram of the reference sample. The chromatograms of the reference samples of commonly used colorants are shown in Figures 1, 2, 3, 4, 5, 6 and 7.
10 1 12 :13 [4 (min)
Figure 1 Chromatographic separation diagram of COC-1 reference sample
1—Impurity 1 (B.2 min);
2—CoC-1 (9. 4 min);
3--Impurity II (10.9min)
Figure 2: Chromatographic separation diagram of coc-2 control sample-Intermediate phenyl ester (5.6min):
2-COC-2 (7. 0 min)
HG / T 2414—93
891011 12 13 14.1516 171819
Figure 3 Chromatographic separation diagram of COM-1 control sample
1-Impurity 1 (11.1 min);2—COM-1(12.5min):3—Impurity II (14.1min);4 Impurity III (15.0min)HG / T 2414: 93
Figure 4 Chromatographic separation diagram of COM-2 control sample
1 Impurity 1 (7. 2 min);
2-Impurity II (8, 7 min);
3—com-2 (10.5min);:
4—Impurity II (12.7min)
Figure 5: Chromatographic separation diagram of COM-4 control sample 1—Intermediate amino compound (4. 4 min);
2-COM-4 (6. 2 mia):
3—Intermediate ethyl ester (6. 8 min)
HG / T 2414 -
345678910111213(min)
Figure 6 Chromatographic separation diagram of COY-2 control sample 1—Intermediate amino compound (6.1min):
2-COY-2 (6.8 min);
3-Intermediate hydride (8.0mim)
Determination steps
012345
6789011213141516(
Figure 7 Chromatographic separation diagram of COY-5 control sample
1-Intermediate hydrogen base (6.5min);
2-Intermediate chloride (8.7mn);
3--COY-5 (10.1min)
Generally, the area normalization method is used. When the content determination value is lower than 90%, or when 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 test is determined by the injection volume of the sample solution, the chromatographic column load, the impurity bee detection limit, the detection sensitivity, and the microprocessor attenuation. The sample weight specified in this test is determined under the condition of high sensitivity and low attenuation. The sample weight can be appropriately adjusted according to different models of chromatographs. 6.1 Area normalization method
6.1.1. Preparation of sample solution
Weigh the specified sample accurately to 0.1 mg, place it in a stoppered glass container, dissolve it with 10 mL of methanol (for COC-2, use 10 mL of ethanol instead, for COM-I, add about 8 drops of ethyl acetate to assist dissolution), mix well. The sample weight of commonly used color-forming agents is shown in Table 3.
Product name
Sample weight, mg
Sample weight of commonly used color-forming agents
6.1.2 Determination
HG/ T 24i4 ~- 93
Debug the high performance liquid chromatograph according to the chromatographic operating conditions required for the sample to be tested and make it stable. Inject the specified amount of the sample solution to be tested for chromatographic separation to obtain the chromatogram.
The injection amount of the commonly used color former sample solution is shown in Table 4. Table 4 Injection amount of commonly used color former sample solution Sample solution name
Injection, uL
6. 1.3 Result calculation
COoC-2
Color former content (X,) is calculated according to formula (1): X,
Where: X,——Percentage content of color former in the sample to be tested, %; A,——Peak area of color former in the sample to be tested; ZA
Sum of peak areas of each component in the sample to be tested. 6. 1. 4 Repeatability
× 100
This method is measured twice in parallel, and the result is the average of the two measurements. The difference between the two determination results should not be greater than 1.0%. 6.2 External standard method
6.2.1 Preparation of standard solution
Weigh the specified amount of color former standard sample, accurate to 0.01 mg, place it in a 25 mL volumetric flask, and dissolve it in methanol (for COC-2, use 10 mL of anhydrous alcohol instead). For COM-1 standard sample, add 10 drops of ethyl acetate to assist dissolution, dilute to the scale, and mix. The standard solution should be prepared as needed
The sample weight of commonly used color former standard samples is shown in Table 5. Table 5 The sample weight of commonly used color former standard samples Standard sample name
Sample weight, mg
6.2.2 Preparation of sample solution
According to 6.2. 1 Specified sample weight and preparation method Prepare sample solution 6.2.3 Determination
According to the chromatographic operating conditions required by the sample to be tested, debug the high performance liquid chromatograph and make it stable. Sequentially inject the specified amount of the colorant standard solution, sample solution, sample solution and standard solution to be tested and obtain the respective chromatograms. The injection amount of the commonly used colorant sample solution (including the corresponding standard solution) is shown in Table 6. Table 6 Injection amount of commonly used colorant sample solution (including the corresponding standard solution) Liquid name
Injection amount, uL
Standard 2. 4 Result calculation
Colorant content (X) is calculated according to formula (2): 7
HG / T 2414— 93
A,-m,p
Where: X2—weight percentage of colorant in the sample to be tested, %; A,—average of the main peak area of the two tested colorant standard solutions: A2—average of the main peak area of the two tested sample solutions;,
Mass of the tested colorant standard sample, mg: Mass of the tested sample, mg;
Content of the tested colorant standard sample, %: 6. 2. 5 Repeatability
This method is measured twice in parallel, and the result is the average of the two measurements. The difference between the two measurement results should not be greater than 1.5%. 7 Test report
The test report should include the following:
All information about the sample: batch number, date, time and location, instrument type used in the test; a
h. Analysis results and expression methods;
Abnormal phenomena observed in the measurement;
Any operation or freely selected test conditions not included in this standard. Additional Notes:
This standard was proposed by the Science and Technology Department of the Ministry of Chemical Industry of the People's Republic of China. This standard is under the jurisdiction of the Photosensitive Material Technology Development Center of the Ministry of Chemical Industry. This standard was drafted by the Photosensitive Material Technology Development Center of the Ministry of Chemical Industry, and participated in by the Shenyang Chemical Research Institute of the Ministry of Chemical Industry. The main drafters of this standard are Ding Peiling, Pan Liren, Cai Ting, Guan Ruibao, and Zhuang Youqin.
Chemical Industry Standards of the People's Republic of China
Photographic Chemicals
Determination of blue content in couplers Reversed high performance liquid chromatography HG/T 2414 ---93
Editor: Chemical Industry Standards Editorial Department
(Standardization Research Institute of the Ministry of Chemical Industry)
Postal Code: 100013
Printer: Standardization Research Institute of the Ministry of Chemical Industry
Ownership: No reproduction allowed
Format: 880 × 1230 1 / 16 [
Print Sheet%Number of Words: 16000
First Edition: June 1993
First Printing: June 1993
Print Quantity: 1—500
Cost: RMB 4.10
Mass of the color former standard sample to be tested, mg: Mass of the sample to be tested, mg;
Content of the color former standard sample to be tested, %: 6. 2. 5 Repeatability
This method is measured twice in parallel, and the result is the average value of the two measurements. The difference between the two measurement results should not be greater than 1.5%. 7 Test report
The test report should include the following contents:
All information about the sample: batch number, date, time and location, instrument type used in the test; a
h. Analysis results and expression methods;
Abnormal phenomena observed in the measurement;
Any operation or freely selected test conditions not included in this standard. Additional notes:
This standard was proposed by the Science and Technology Department of the Ministry of Chemical Industry of the People's Republic of China, and this standard is under the jurisdiction of the Photosensitive Material Technology Development Center of the Ministry of Chemical Industry. This standard was drafted by the Photosensitive Material Technology Development Center of the Ministry of Chemical Industry, and the Shenyang Chemical Industry Research Institute of the Ministry of Chemical Industry participated in the drafting. This standard is mainly drafted by Ding Peiling, Pan Liren, Cai Ting, Guan Ruibao, Zhuang Youqin, and is published by the People's Republic of China. Chemical Industry Standards Photographic Chemicals Determination of blue content in couplers Reverse high performance liquid chromatography HG/T 2414---93 Editor Chemical Industry Standards Editorial Department (Standardization Research Institute, Ministry of Chemical Industry) Postal Code: 100013 Printing Standardization Research Institute, Ministry of Chemical Industry Copyright reserved Reproduction prohibited Format 880 × 1230 1 / 16 [
Printing Sheet % Number of words 16000 First edition in June 1993 First printing in June 1993 Number of copies 1-500 Cost 4.10 yuan
Mass of the color former standard sample to be tested, mg: Mass of the sample to be tested, mg;
Content of the color former standard sample to be tested, %: 6. 2. 5 Repeatability
This method is measured twice in parallel, and the result is the average value of the two measurements. The difference between the two measurement results should not be greater than 1.5%. 7 Test report
The test report should include the following contents:
All information about the sample: batch number, date, time and location, instrument type used in the test; a
h. Analysis results and expression methods;
Abnormal phenomena observed in the measurement;
Any operation or freely selected test conditions not included in this standard. Additional notes:
This standard was proposed by the Science and Technology Department of the Ministry of Chemical Industry of the People's Republic of China, and this standard is under the jurisdiction of the Photosensitive Material Technology Development Center of the Ministry of Chemical Industry. This standard was drafted by the Photosensitive Material Technology Development Center of the Ministry of Chemical Industry, and the Shenyang Chemical Industry Research Institute of the Ministry of Chemical Industry participated in the drafting. This standard is mainly drafted by Ding Peiling, Pan Liren, Cai Ting, Guan Ruibao, Zhuang Youqin, and is published by the People's Republic of China. Chemical Industry Standards Photographic Chemicals Determination of blue content in couplers Reverse high performance liquid chromatography HG/T 2414---93 Editor Chemical Industry Standards Editorial Department (Standardization Research Institute, Ministry of Chemical Industry) Postal Code: 100013 Printing Standardization Research Institute, Ministry of Chemical Industry Copyright reserved Reproduction prohibited Format 880 × 1230 1 / 16 [
Printing Sheet % Number of words 16000 First edition in June 1993 First printing in June 1993 Number of copies 1-500 Cost 4.10 yuan
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