Determination of Hu Huanglian content, referring to the Chinese Pharmacopoeia (2025 edition), Part I

Coptis chinensis is the dried root and rhizome of the plant Coptis chinensis from the Scrophulariaceae family. It is harvested in autumn, with the fine roots and soil removed, and then sun-dried. Its functions and main uses: reducing deficient heat, eliminating pediatric nutritional heat, and clearing damp-heat. It is used for steaming bone fever, nutritional heat in children, damp-heat diarrhea, jaundice with red urine, and painful hemorrhoids.

According to the analysis method under the determination of Coptis chinensis content in the Chinese Pharmacopoeia (2025 edition, Volume I), the theoretical plate number calculated using the peak of Coptis chinensis glycoside II should not be less than 3,000.

Following the pharmacopoeia method, we tested with both the DAISOPAK ODS-RPS and DAISOPAK ODS-P columns, and both meet the pharmacopoeia requirements.

ODS-RPS分析结果

DAISOPAK ODS-RPS

The test results of the sample solution on the column are shown in Figure 1 and Table 1. The theoretical plate number of berberine glycoside II in the sample solution is 6,469, which is higher than the 3,000 specified in the pharmacopoeia, and the separation between the main peak and the impurity peaks meets the pharmacopoeia requirements.

图1.png

Figure 1 Test Results of the ODS-RPS Chromatography Column

Table 1 Peak Integration Results (Test Sample Solution)

表1.png

【Chromatography Conditions】

Column：DP ODS-RPS 5μm; 4.6×250mm

Mobile phase: methanol / water / phosphoric acid = 35 / 65 / 0.1

flow rate：1.0mL/min

Column Temperature：25 ℃

Detector：PDA 275nm

Injection volumn：10μL

Preparation of the test solution: Take about 0.1 g of the powder (sifted through a No. 3 sieve), accurately weigh it, place it in a stoppered conical flask, add 50 mL of methanol precisely, seal it, weigh it, treat it with ultrasound (power 250W, frequency 33kHz) for 30 minutes, let it cool, weigh it again, add methanol to make up for any lost weight, shake well, filter it, accurately measure 1 mL of the filtrate, place it in a 5 mL volumetric flask, add methanol up to the mark, shake well, and it's ready.

ODS-P Analysis Results

The test results of the DAISOPAK ODS-P column with the sample solution are shown in Figure 2 and Table 2. The theoretical plate number of berberine glucoside II in the sample solution is 6,567, which is higher than the 3,000 required by the pharmacopeia, and the separation between the main peak and impurity peaks meets pharmacopeia standards.

图2.png

Figure 2 Test Results of the ODS-P Chromatography Column

Table 2 Peak Integration Results (Test Sample Solution)

表2.png

【Chromatography Conditions】

Column：DP ODS-P 5 μm; 4.6×250 mm

Mobile phase: methanol / water / phosphoric acid

Flow rate：1.0 mL/min

Column Temperature：25 ℃

Detector：PDA 275 nm

Injection Volumn：10 μL

Preparation of the test solution: Take about 0.1 g of the powder (sifted through a No. 3 sieve), accurately weigh it, place it in a stoppered conical flask, precisely add 50 mL of methanol, tightly seal, weigh, sonicate (power 250 W, frequency 33 kHz) for 30 minutes, let it cool, then weigh again. Add methanol to make up for any lost weight, shake well, filter, accurately measure 1 mL of the filtrate, put it into a 5 mL volumetric flask, add methanol up to the mark, shake well, and it's ready.

In summary, using the DAISOPAK SP-120-5-ODS-RPS 4.6 mm i.d. × 250 mm and DAISOPAK SP-100-5-ODS-P 4.6 mm i.d. × 250 mm columns, the test results all meet the requirements of the Chinese Pharmacopoeia (2025 edition). Because the sample matrix is quite complex, we can't be sure if the target peak still contains unresolved impurities. The above experimental results are provided for users' reference.

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