Method for measuring weight-average molecular weight and content of polyguluronate sulfate
A technology of guluronic acid sulfate and weight-average molecular weight, which is applied in the field of natural medicinal chemistry and its quality control, can solve the problems of differences in structure and color rendering efficiency, the inability to accurately quantify acidic sugars, etc., and solve the problem of large measurement deviations Effect
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Embodiment 1
[0083] Example 1 SEC and ICP-MS reveal the separation mechanism of acidic sugars in size exclusion chromatography, and determine the weight average molecular weight and content of polyguluronic acid sulfate sodium salt
[0084] Step 1: SEC-ICP-MS to determine the peak position and content of metal ions
[0085] (1) Use the column Waters ACQUITY SEC 1.7um 4.6×150mm column, 0.08mol / L NH4Ac as mobile phase, flow rate 0.1mL / min.
[0086] (2) Combine SEC with ICP-MS: dissolve the sample in water, the concentration is 0.02mg / ml, inject 20ul, collect 23 Na, 43 Ca, 44 Ca signaling. Concentrations of 0.1ppm, 0.5ppm, 1.0ppm, 5.0ppm and 10.0ppm Na ion standard solutions were prepared respectively, and the content of metal ions in the sample (A value) was calculated by the external standard method. The results of A values are shown in Table 1.
[0087] Step 2: Determination of weight average molecular weight and content by SEC-MALS
[0088] (1) Use the column Waters ACQUITY SE...
Embodiment 2
[0099] Embodiment 2 SEC-MALS measures the weight-average molecular weight and content of polyguluronic acid sulfate potassium salt
[0100] Step 1: Determination of the ratio of metal ions by ion chromatography
[0101] K ion standard solution is configured as 25, 10, 12.5, 6.25, 3.125ppm. The sample concentration was 2 mg / ml.
[0102] Ion chromatography conditions: pre-column: Metrosep C4Guard / 4.0; separation column: Metrosep
[0103] C5-150 / 4.0; mobile phase: ultrapure aqueous solution of 5mM nitric acid; flow rate: 0.9ml / min; injection volume 20ul. The content of metal ions was determined by external standard method.
[0104] Step 2: Determine dn / dc
[0105] (1) Weigh the sample with constant weight, and prepare a series of solutions with concentrations of 0.2mg / ml, 0.4mg / ml, 0.8mg / ml, 1.2mg / ml, 1.8mg / ml, and 2mg / ml with mobile phase.
[0106] (2) Manually inject mobile phase, 0.2mg / ml, 0.4mg / ml, 0.8mg / ml, 1.2mg / ml, 1.8mg respectively with a differential refractive ind...
Embodiment 3
[0115] Embodiment 3 SEC-MALS measures the weight-average molecular weight and content of polyguluronic acid sulfate calcium salt
[0116] Step 1: Determination of metal ion content by flame atomic absorption method:
[0117] Collect the spectral lines of Ca and prepare Ca standard solutions of 1mg / L, 10mg / L, 50mg / L, 75mg / L and 100mg / L; the concentration of the sample solution is prepared to be 200mg / L.
[0118] Step 2: Determine dn / dc
[0119] With embodiment 2 step 2. See Table 3 for the measured values.
[0120] Step 3: Determination of weight average molecular weight and content by SEC-MALS
[0121] (1) Use a chromatographic column Shodex OHpak SB-803HQ 7.8×300mm (Shodex), 0.15mol / L NaCl as the mobile phase, and a flow rate of 0.5mL / min.
[0122] (2) Combine SEC with MALS, in the mobile phase of the sample solution, the concentration is 10mg / ml, and the injection volume is 20ul, that is, the Y value in the formula is 200ug. The experimental results are shown in Table 3...
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