Method for measuring contents of protocatechuic acid and protocatechualdehyde in Yi Xue'an particles

Abstract

The invention discloses a method for detecting protocatechuic acid and protocatechualdehyde in Yi Xue'an particles. The method comprises the following steps: (1) adding Yi Xue'an particles into methanol, and carrying out ultrasonic extraction to prepare a sample solution; (2) subjecting the sample solution to high performance liquid chromatography (HPLC) detection under the following chromatographic conditions: gradient elution, and UV detection wavelength: 256 nm (0 to 20 minutes), and 279 nm (20 to 30 minutes). The measurement conditions such as chromatographic column, detection wavelength, chromatographic mobile phase, column temperature, sample pretreatment method, and the like are all optimized to ensure good repeatability and stability of the detection method, and thus the quality of finished product of Yi Xue'an particles is effectively controlled.

Description

Technical field

[0001] The invention relates to the field of drug detection, in particular to a method for determining the content of protocatechuic acid and protocatechualdehyde in Yixuean granules. Background technique

[0002] Yixuean granules is a Chinese patent medicine for the treatment of gynecological aemia. The main ingredients are Yunnan Gui Aina, Motherwort, Corydalis, Licorice, etc. It has the effects of promoting blood circulation and stopping bleeding, promoting Qi and relieving pain. Clinically used for postpartum lochia, uterine bleeding after artificial abortion is not clean, TCM dialectics is blood stasis syndrome.

[0003] Among them, the monarch medicine Diangui Ai Naxiang is a folk herbal medicine of the Yao nationality. It is derived from the dried whole plant of Diangui Ai Naxiang [Blumeariparia(BL)DC.] of the Compositae family. Naxiang was discovered and used by modern people, so there is no record in ancient literature. It is mainly produced in southeaster...

Examples

Embodiment 1

[0055] Example 1: Optimization of chromatograph conditions

[0056] The preparation method of the test solution: Take 5g of Ixanol granules, place a stoppered Erlenmeyer flask, add 15mL of a volume fraction of 80% methanol aqueous solution, ultrasonically extract for 45 minutes, let cool, and use a volume fraction of 80% methanol aqueous solution to make up for loss reduction The weight, filter, take the filtrate, centrifuge, take the supernatant, and get the test solution.

[0057] 1-1 Selection of Column

[0058] The other measurement conditions of the high performance liquid chromatograph are the same. Welchrom C18 column (4.6mm×250mm, 5μm), Hypersil C18 column (4.6mm×250mm, 5μm) and Phenomenex Gemini C18 column (4.6mm×250mm, 5μm) were investigated. Chromatographic column, the chromatogram obtained is divided into figure 1 , figure 2 , image 3 Shown by image 3 It can be seen that the Phenomenex Gemini C18 column (4.6mm×250mm, 5μm) has the best resolution, symmetrical and sharp...

Embodiment 2

[0067] Example 2: Optimization of sample pretreatment method

[0068] The chromatographic conditions of the high performance liquid chromatograph are: Phenomenex Gemini C18 column (4.6mm×250mm, 5μm) is used as the chromatographic column, Phenomenex C18 (4×3.0mm) is used as the guard column, gradient elution, and the mobile phase is methanol and 0.1% Chromatography uses a gradient eluent composed of glacial acetic acid, column temperature 25℃, flow rate 1.0mL·min -1 ; The gradient elution program is set to time gradient 0min→9min→19min→20min→30min, and the corresponding gradient eluent is composed of mobile phase A methanol and mobile phase B 0.1% glacial acetic acid according to the following volume ratio, mobile phase The volume percentage gradient of A is 8%→9%→10%→14%→15%, the volume percentage gradient of mobile phase B is 92%→91%→90%→86%→85%; UV detection wavelength: 0~20min 256nm, 20-30min is 279nm.

[0069] 2-1 Selection of extraction solvent

[0070] Three extraction solven...

Embodiment 3

[0089] Example 3 Determination of the content of protocatechuic acid and protocatechualdehyde in Yixuean granules

[0090] 3-1 Preparation of test solution: Test 10 batch numbers (120203, 120302, 120303, 120602, 120603, 120701, 120702, 120801, 120802, 120803) Ixan granules, sample 4 for each batch number; respectively Weigh 5 g of Ixan granules, place a stoppered Erlenmeyer flask, precisely add 15 mL of 80% methanol aqueous solution, ultrasonically extract for 45 min, let cool, 80% methanol aqueous solution to make up the lost weight, filter, take the additional filtrate, centrifuge, and take it The clear solution is the test solution.

[0091] 3-2 Preparation of reference solution

[0092] Accurately weigh 10.04mg of protocatechuic acid reference substance and 2.58mg of protocatechualdehyde reference substance into a 25mL volumetric flask, add methanol to dissolve and dilute to the mark, shake well to obtain a concentration of 0.4016mg·mL -1 Protocatechin reference substance soluti...

Technical field

[0001] The invention relates to the field of drug detection, in particular to a method for determining the content of protocatechuic acid and protocatechualdehyde in Yixuean granules. Background technique

[0002] Yixuean granules is a Chinese patent medicine for the treatment of gynecological aemia. The main ingredients are Yunnan Gui Aina, Motherwort, Corydalis, Licorice, etc. It has the effects of promoting blood circulation and stopping bleeding, promoting Qi and relieving pain. Clinically used for postpartum lochia, uterine bleeding after artificial abortion is not clean, TCM dialectics is blood stasis syndrome.

[0003] Among them, the monarch medicine Diangui Ai Naxiang is a folk herbal medicine of the Yao nationality. It is derived from the dried whole plant of Diangui Ai Naxiang [Blumeariparia(BL)DC.] of the Compositae family. Naxiang was discovered and used by modern people, so there is no record in ancient literature. It is mainly produced in southeaster...

Examples

Embodiment 1

[0055] Example 1: Optimization of chromatograph conditions

[0056] The preparation method of the test solution: Take 5g of Ixanol granules, place a stoppered Erlenmeyer flask, add 15mL of a volume fraction of 80% methanol aqueous solution, ultrasonically extract for 45 minutes, let cool, and use a volume fraction of 80% methanol aqueous solution to make up for loss reduction The weight, filter, take the filtrate, centrifuge, take the supernatant, and get the test solution.

[0057] 1-1 Selection of Column

[0058] The other measurement conditions of the high performance liquid chromatograph are the same. Welchrom C18 column (4.6mm×250mm, 5μm), Hypersil C18 column (4.6mm×250mm, 5μm) and Phenomenex Gemini C18 column (4.6mm×250mm, 5μm) were investigated. Chromatographic column, the chromatogram obtained is divided into figure 1 , figure 2 , image 3 Shown by image 3 It can be seen that the Phenomenex Gemini C18 column (4.6mm×250mm, 5μm) has the best resolution, symmetrical and sharp...

Embodiment 2

[0067] Example 2: Optimization of sample pretreatment method

[0068] The chromatographic conditions of the high performance liquid chromatograph are: Phenomenex Gemini C18 column (4.6mm×250mm, 5μm) is used as the chromatographic column, Phenomenex C18 (4×3.0mm) is used as the guard column, gradient elution, and the mobile phase is methanol and 0.1% Chromatography uses a gradient eluent composed of glacial acetic acid, column temperature 25℃, flow rate 1.0mL·min -1 ; The gradient elution program is set to time gradient 0min→9min→19min→20min→30min, and the corresponding gradient eluent is composed of mobile phase A methanol and mobile phase B 0.1% glacial acetic acid according to the following volume ratio, mobile phase The volume percentage gradient of A is 8%→9%→10%→14%→15%, the volume percentage gradient of mobile phase B is 92%→91%→90%→86%→85%; UV detection wavelength: 0~20min 256nm, 20-30min is 279nm.

[0069] 2-1 Selection of extraction solvent

[0070] Three extraction solven...

Embodiment 3

[0089] Example 3 Determination of the content of protocatechuic acid and protocatechualdehyde in Yixuean granules

[0090] 3-1 Preparation of test solution: Test 10 batch numbers (120203, 120302, 120303, 120602, 120603, 120701, 120702, 120801, 120802, 120803) Ixan granules, sample 4 for each batch number; respectively Weigh 5 g of Ixan granules, place a stoppered Erlenmeyer flask, precisely add 15 mL of 80% methanol aqueous solution, ultrasonically extract for 45 min, let cool, 80% methanol aqueous solution to make up the lost weight, filter, take the additional filtrate, centrifuge, and take it The clear solution is the test solution.

[0091] 3-2 Preparation of reference solution

[0092] Accurately weigh 10.04mg of protocatechuic acid reference substance and 2.58mg of protocatechualdehyde reference substance into a 25mL volumetric flask, add methanol to dissolve and dilute to the mark, shake well to obtain a concentration of 0.4016mg·mL -1 Protocatechin reference substance soluti...

Images