Caffeic acid

Examples of using this product（for reference only）

HPLC–MS

We used an HPLC system, which consisted of a binary solvent delivery system, an on-line degasser, an auto-sampler, a column temperature controller, and a diode array detector. Separation was conducted using a C18 column (5 μm, 150 mm × 4.6 mm), and the temperature was set to 40℃. The injection volume was 0.01 mL, and the flow rate was 1 mL/min. Gradient elution was conducted using a binary system consisting of 10 % formic acid in DW (A) and 10 % formic acid in acetonitrile (B). The utilized gradient was as follows: isocratic 1 % B for 6 min, 1% – 3% B for 4 min, 3 % – 15 % B for 20 min, 15 % – 40 % B for 20 min, 40 % – 90 % B for 5 min, 90 % B isocratic for 10 min, 90 % – 1 % B for 5 min, and 1 % B isocratic for 10 min. A re-equilibration operation followed. The UV detection wavelength was set to 280 nm.

The voltages of the sample cone and the capillary were set to 17 and 3000 V for the positive ion mode and 22 and 2000 V for the negative ion mode.

References：

[1]Zhang,Junhong,Tian,et al.Isolation and identification of phenolic compounds in Chinese purple yam and evaluation of antioxidant activity[J].LWT-Food Science & Technology, 2018, 96:161-165.

HPLC

HPLC was performed using an equipped with a DAD detector and a C18 (250 mm × 4.6 mm; 5 μm). The mobile phase included 0.2% phosphoric acid plus 0.4% sodium 1-heptanesulfonate (A) and methanol (B) at a flow speed of 1.0 ml/min in the condition of a column temperature of 30℃. The detection wavelength was set at 214 nm. The reference standard sample injection volume was 10 μl. The gradient elution was as follows: 0–5 min (100–100% A, 0%–0% B), 5–60 min (100–60% A, 0%–40% B), 60–120 min (60–30% A, 40–70% B), 120–125 min (30–30% A, 70–70% B), 125–125.1 min (30–100% A, 70–0% B), 125.1–132 min (100–100% A, 0–0% B). The chromatographic data and peak area scores were collected and analyzed using ChemStation software.

References：

Duan Q, Liu T, Huang C, Shao Q, Ma Y, Wang W, Liu T, Sun J, Fang J, Huang G, Chen Z. The Chinese Herbal Prescription JieZe-1 Inhibits Membrane Fusion and the Toll-like Receptor Signaling Pathway in a Genital Herpes Mouse Model. Front Pharmacol. 2021 Sep 24;12:707695. doi: 10.3389/fphar.2021.707695. PMID: 34630083; PMCID: PMC8497740.

HPLC

HPLC conditions: chromatographic column was SB-C18 column (4.6 × 250 mm, 5 μm) and the column temperature was maintained at 30℃. The wavelength of ultraviolet detector were 280 nm and 320 nm, and the gradient elution with a flow rate at 0.8 mL/min was performed by mobile phase A (2% acetic acid in deionized water) and solvent B (methanol; 0–10 min, 5–30%; 10–35 min, 30–50%; 35–40 min, 50–60%; 40–45 min, 60–70%; 45–50 min, 70–5%; 50–55 min, 5%).

References：

Xu J, Qi Y, Zhang J, Liu M, Wei X, Fan M. Effect of reduced glutathione on the quality characteristics of apple wine during alcoholic fermentation. Food Chem. 2019 Dec 1;300:125130. doi: 10.1016/j.foodchem.2019.125130. Epub 2019 Jul 4. PMID: 31325746.

UHPLC

Chromatographic separation of SPLFA and SPLF was done on an Agilent 1290 UHPLC system coupled to an auto sampler, a diode array detector (DAD) and an ACQUITY UHPLC? HSS T3 column (2.1 ×100 mm, 1.8 μm; Waters, Milford, MA, USA)maintained at 30 ℃. Eluent A was 0.05% formic acid in water and eluent B was 0.05% formic acid in acetonitrile. Elution of the SPLPA compounds was achieved using the following linear gradient elution (in %B): 0 min, 5%; 20 min, 60%; 20.1 min, 5%; 30 min, 5%, and for the SPLF compounds: 0 min, 20%, 25 min, 50%, 25.1 min, 20%, 35 min, 20%. The injection volume was 5 μL, with a flow rate of 0.3 mL/ min. All sample solutions were filtered through a 0.22 μm PTFE filter (Agilent Technologies) before analysis. The detection wavelength was 320 nm for SPLPA and 254 nm for SPLF.

References：

[1] Luo D , Mu T , Sun H .Profiling of phenolic acids and flavonoids in sweet potato (Ipomoea batatas L.) leaves and evaluation of their anti-oxidant and hypoglycemic activities[J].Food Bioscience, 2020, 39(44):100801.DOI:10.1016/j.fbio.2020.100801.

HPLC

HPLC analysis employed an Agilent 1260 HPLC instrument and a diode array detector (Agilent, Santa Clara, CA, USA). The chromatographic separation was achieved by a Zobax Stablebond Analytical SB-C18 column (250 × 4.6 mm, 5 μm). The mobile phases, containing a gradient of solvent A (acetic/water, 2:98, v/v) and solvent B (acetic acid/acetonitrile/water, 5:50:45, v/v) were used to elute the column. The solvent gradient was programmed from 90% to 55%A and 10% to 45%B for 50 min, 55% to 0%A and 45% to 100%B for 10 min, and 0% to 90%A and 100% to 10%B for 5 min. The flow velocity was 1.0 mL·min-1 , the column temperature was 30℃, and the injection volume was 20 μL

References：

Liang D, Deng H, Deng Q, Lin L, Lv X, Wang J, Wang Z, Xiong B, Zhao X, Xia H. Dynamic Changes of Phenolic Compounds and Their Associated Gene Expression Profiles Occurring during Fruit Development and Ripening of the Donghong Kiwifruit. J Agric Food Chem. 2020 Oct 14;68(41):11421-11433. doi: 10.1021/acs.jafc.0c04438. Epub 2020 Sep 30. PMID: 32936614.

HPLC-DAD

The identification and quantification of phenolics in the S. miltiorrhiza leaves extract before and after enrichment were analyzed using an Agilent 1260 Infinity LC system (Agilent Co., Palo Alto, CA, USA).10 μL of sample dissolved in 80 % methanol was injected into the HPLC system, and phenolics were chromatographed over a YMC-Pack ODS-A (250 × 4.6 mm, 5 μm) column at room temperature with a combination of 0.1 % trifluoroacetic acid (A) and acetonitrile (B) at a flow rate of 1 mL/min. The procedure for elution was as follows: 0? 5 min, 10 % B; 5? 10 min, 10 %–12 % B; 10? 20 min, 12 %–15 % B; 20? 25 min, 15 %–20 % B; 25? 60 min, 20 %–30 % B; 60? 65 min, 30 %–100 % B; 65? 70 min, 100 % B; 70? 80 min, 100 %-10 % B. Phenolics were detected by a diode array detector (DAD) at 280 nm.

References：

[1] Hou M , Hu W , Hao K ,et al.Enhancing the potential exploitation of Salvia miltiorrhiza Bunge: Extraction, enrichment and HPLC-DAD analysis of bioactive phenolics from its leaves[J].Industrial Crops and Products, 2020, 158:113019.DOI:10.1016/j.indcrop.2020.113019.

Remark：These protocols are for reference only. Solarbio does not independently validate these methods.

Note：

1. The products are all for scientific research use only. Do not use it for medical, clinical diagnosis or treatment, food and cosmetics, etc. Do not store them in ordinary residential areas.

2. For your safety and health, please wear laboratory clothes, disposable gloves and masks.

3. The experimental results may be affected by many factors, after-sale service is limited to the product itself and does not involve other compensation.