Fmoc-Ser(tBu)-OH CAS 71989-33-8 Fmoc-O-tert-Butyl-L-Serine Purity >98.5% (HPLC)

Inspection procedures
1. Appearance
-- Visual inspection
2. Purity (HPLC)
2.1 Instrument
High performance liquid chromatography, PDA detector.
Electronic analytical balance
2.2 Reagent
Acetonitrile (chromatographic grade), trifluoroacetic acid (chromatographic grade)
2.3 Chromatographic conditions
2.3.1 Column: YMC-ODS-AM, 5 μl, 250x4.6 mm
2.3.2 Detection wavelength: INC220nm
Flow rate: 1.0mL/min
Sample size:10 μl (reference)
Diluent: acetonitrile
Data collection time: 25.00min
2.4 Mobile phase preparation
Mobile phase A (0.1% trifluoroacetic acid water): precision absorption 2. Dilute Oml trifluoroacetic acid with water to 2000m1, mix well, and degassing;
Mobile phase B (0.1% acetonitrile trifluoroacetic acid): precise absorption 2.0ml trifluoroacetic acid was diluted to 2000m1 with acetonitrile, mixed and degassing;
2.5 Gradient program of mobile phase
Time (min) A% B%
0.00 90 10
13.00 10 90
18.00 10 90
18.01 90 10
23.00 90 10
2.6 Preparation of sample solution
Weigh and dissolve 0.1g sample with acetonitrile and dilute to 100ml, shake well for use, or the same concentration. Prepare two samples in parallel.
2.7 Sample Determination
Analyze the sample according to the following sampling procedure:
More than 1 injection of blank solution
1 stitch sample solution 1#
1 stitch sample solution 2#
2.8 Result calculation
2.8.1 The peak area normalization method was used to calculate the HPLC purity by deducting blank space.
2.8.2 The relative mean deviation of purity of two needles shall not be greater than 1%
2.8.3 If the results of both injections meet the acceptance criteria, the average purity is taken as the final result.
3, Melting point -- RY-1 melting point instrument
4. Loss on drying detection method
4.1 Instruments:
Electric thermostatic drying oven, one in ten thousand balance.
4.2 Procedure:
In a flat weighing bottle with a ground mouth cover with constant weight and over-drying, weigh 1 gram of sample (accurate to 0.0001 gram). The sample should be evenly spread at the bottom of the weighing bottle with a thickness of no more than 10mm, put in a constant temperature electric drying oven, dry at 105-110 ℃ for 3 hours, and then move into the drying room, cool to room temperature and weigh.
Calculation: Dry weight loss %= (M1-M2) ÷M×100
Where: M1: mass of sample and weighing bottle before drying, g
M2: Mass of sample and weighing bottle after drying, g
M: Sample mass, gram
5. Specific rotation
-- Specific rotation is measured in accordance with GB/T613-1988
Sample preparation: 0.5000g of sample was accurately weighed and transferred to a clean and dry 50ml volumetric bottle with 20ml Ethyl acetate. The bottle was corked and shaken to dissolve the Ethyl acetate.
Test: Adjust the zero of the gyroscope before the test, then fill the test tube with sample solution, record the deflection Angle of rotation, and calculate the specific rotation of the sample using the following formula.
[α]20/D= (r×50) ÷(L×W)
Where:
[α]20/D: Specific rotation of sample solution at 25℃
r: Rotation observed at 25℃ for sample solution
50: Prepare sample solution volume (ml)
w: Sample weight (g)
L: Optical tube length (dm)
6. TCL Analysis
Sample preparation: 0.5000g sample was accurately weighed and moved to a clean and dry 50ml volumetric bottle, 40ml EtoH was added, the bottle was capped and shaken to dissolve, and then diluted to the scale with EtoH.
Preparation of control sample: 0.5000g sample was accurately weighed and moved to a clean and dry 50ml volumetric bottle, 40ml water and 2 drops of 6N hydrochloric acid were added, the bottle was capped and shaken to dissolve, and then diluted to the scale with water. Then take 0.5 of the above liquid and dilute it with ethanol in a 100ml volume bottle to a reference sample with 1.0% amino acid residue scale.
10 microliter samples were taken with a microsampling syringe, and the samples were placed on the GF254 silicone plate. 5 microliter, 10 microliter and 15 microliter point plates were taken for comparison points, respectively.
Developing agent: n-butanol: glacial acetic acid: water =4:1:1.
Color developing agent: 5% ninhydrin methanol solution.
Color development operation: first, use a hair dryer to dry the developed silica gel plate, then dip it into 5% ninhydrin methanol solution, then blow dry, and bake at high temperature in an electric furnace for color development.
Judgment: Look at the product color time, spot size, color concentration and comparison with the control product, judge that it is in that range.