The specific pigments of algae are chlorophyll, lutein and carotene. The three chlorophylls commonly found in planktonic algae are chlorophyll a, b and c. Chlorophyll a accounts for approximately 1-2% of the dry weight of organic matter in all planktonic algae. A good indicator of algae biomass. The chlorophyll content of a cell varies with the species or taxa, but also by age, growth rate, light and nutrient conditions. Pheophycolide a (a common degradation product of chlorophyll a) can Interfere with the determination of chlorophyll a, because if there is pheophytin a, it can absorb light and fluorescence in the same spectral region of chlorophyll a, resulting in an error in chlorophyll a value. When determining chlorophyll a also determine pheophytin a. Chlorophyll The ratio of a and pheophytin a can serve as a good indicator of the physiological conditions of phytoplankton. High-flux tissue grinders play a role in sample preparation and provide effective help for research.
The detection limit varies with the structure and flow rate of the fluorometer, but for most chlorophylls and their degradation products, the detection limit for each injection range is between 10 and 100 pg. The accuracy of the HPLC method depends mainly on the purity of the pigment standard. The more ideal method is to determine the absorption spectrum (350-750 nm) of the standard sample and compare it with the literature data. The purity of the pigment can also be determined by HPLC, provided that there is no overlap with the absorbance and fluorescence bands of the standard. Coelution of contaminants. If the data measured by spectrophotometry is corrected by demagnesium, the HPLC results are expressed as pigment equivalents (eg: chlorophyll a equivalent = chlorophyll a + chlorophyll a + chlorophyll ac, assuming correct molecular weight correction (Value), then HPLC and spectrophotometry provide pigment concentrations that are in good agreement with the EPA standards provided. Therefore, if pigment derivatives are clearly present, the data measured by spectrophotometry will be high. HPLC and fluorescence are required. The data measured by the method is consistent with the additional chlorophyll b, c and their derivatives.
Calibrate the working standard of the HPLC system with the standard prepared by the previous high-throughput tissue grinder. Mix 1 ml standard with 300 LL distilled water, shake and inject after equilibrating for 5 minutes. Rinse the syringe twice with 300 LL standard. Inhale the 500LL standard in the syringe. The liquid is used for injection, the syringe is inserted into the injection valve and filled with 200 LL sample loop. Mix 1 ml of 90% acetone with 300 LL of distilled water as the blank. Establish a standard curve of standard pigment concentration and fluorescence peak area (or height). The method is designed for the separation of chlorophyll and carotenoids and also allows the separation of the major chlorophyll breakdown products. The accuracy of the method is determined by the determination of phytoplankton colonies and plant extracts with three-fold injections. Use of appropriate internal standards is available Improve accuracy.
The detection limit varies with the structure and flow rate of the fluorometer, but for most chlorophylls and their degradation products, the detection limit for each injection range is between 10 and 100 pg. The accuracy of the HPLC method depends mainly on the purity of the pigment standard. The more ideal method is to determine the absorption spectrum (350-750 nm) of the standard sample and compare it with the literature data. The purity of the pigment can also be determined by HPLC, provided that there is no overlap with the absorbance and fluorescence bands of the standard. Coelution of contaminants. If the data measured by spectrophotometry is corrected by demagnesium, the HPLC results are expressed as pigment equivalents (eg: chlorophyll a equivalent = chlorophyll a + chlorophyll a + chlorophyll ac, assuming correct molecular weight correction (Value), then HPLC and spectrophotometry provide pigment concentrations that are in good agreement with the EPA standards provided. Therefore, if pigment derivatives are clearly present, the data measured by spectrophotometry will be high. HPLC and fluorescence are required. The data measured by the method is consistent with the additional chlorophyll b, c and their derivatives.
Calibrate the working standard of the HPLC system with the standard prepared by the previous high-throughput tissue grinder. Mix 1 ml standard with 300 LL distilled water, shake and inject after equilibrating for 5 minutes. Rinse the syringe twice with 300 LL standard. Inhale the 500LL standard in the syringe. The liquid is used for injection, the syringe is inserted into the injection valve and filled with 200 LL sample loop. Mix 1 ml of 90% acetone with 300 LL of distilled water as the blank. Establish a standard curve of standard pigment concentration and fluorescence peak area (or height). The method is designed for the separation of chlorophyll and carotenoids and also allows the separation of the major chlorophyll breakdown products. The accuracy of the method is determined by the determination of phytoplankton colonies and plant extracts with three-fold injections. Use of appropriate internal standards is available Improve accuracy.
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