2.43
1.65
1.98
0.73
1.88
1.81
2.43
2.2 用于相对分子质量分布校准曲线的标准物质：胰岛素、肌肽、甘氨酸-甘氨酸-酪氨酸-精氨酸、甘氨酸-甘氨酸-甘氨酸
3 仪器和设备
23.2
21.4
22.2
16.1
22.3
20.8
23.9
27.5
总体而言，Sustar 产品中的氨基酸比例高于 Zinpro 产品中的氨基酸比例。
第八部分 使用效果
不同来源的微量元素对产蛋后期蛋鸡生产性能和蛋品质的影响
生产过程
靶向螯合技术
剪切乳化技术
压力喷涂和干燥技术
制冷与除湿技术
先进的环境控制技术
附录A：肽相对分子质量分布的测定方法
采用标准：GB/T 22492-2008
1. 测试原则：
采用高效凝胶过滤色谱法测定。具体而言，以多孔填料为固定相，基于样品组分相对分子质量大小的差异进行分离，在220nm紫外吸收波长下检测肽键，利用凝胶过滤色谱法测定相对分子质量分布的专用数据处理软件（即GPC软件），对色谱图及其数据进行处理和计算，得到大豆肽的相对分子质量及其分布范围。
2. 试剂
实验用水应符合GB/T6682中二级水的规定，所用试剂除特殊规定外，应为分析纯。
2.1 试剂包括乙腈（色谱纯）、三氟乙酸（色谱纯），
2.2 用于相对分子质量分布校准曲线的标准物质：胰岛素、肌肽、甘氨酸-甘氨酸-酪氨酸-精氨酸、甘氨酸-甘氨酸-甘氨酸
3 仪器和设备
3.1 高效液相色谱仪（HPLC）：一种色谱工作站或积分仪，配有紫外检测器和 GPC 数据处理软件。
3.2 流动相真空过滤和脱气装置。
3.3 电子天平：刻度值为 0.000 1g。
4. 操作步骤
4.1 色谱条件和系统适应性实验（参考条件）
- 4.1.1 色谱柱：TSKgelG2000swxl300 mm×7.8 mm（内径）或其他同类型、性能相近的凝胶柱，适用于测定蛋白质和肽。
- 4.1.2 流动相：乙腈+水+三氟乙酸=20+80+0.1。
- 4.1.3 检测波长：220 nm。
- 4.1.4 流速：0.5 mL/min。
- 4.1.5 检测时间：30 分钟。
- 4.1.6 样品进样量：20μL。
- 4.1.7 色谱柱温度：室温。
- 4.1.8 为了使色谱系统满足检测要求，规定在上述色谱条件下，凝胶色谱柱效率，即理论塔板数（N），根据三肽标准品（甘氨酸-甘氨酸-甘氨酸）的峰计算，不小于10000。
- 4.2 相对分子质量标准曲线的制备
- 将上述不同相对分子质量的肽标准溶液（质量浓度均为1 mg/mL）通过流动相匹配配制，按一定比例混合，然后经孔径为0.2 μm~0.5 μm的有机相滤膜过滤后注入样品中，得到标准品的色谱图。通过绘制相对分子质量的对数与保留时间的关系图或进行线性回归，得到相对分子质量校准曲线及其方程。
4.3 样品处理
准确称取 10mg 样品于 10mL 容量瓶中，加入少量流动相，超声振荡 10min，使样品充分溶解混合，用流动相稀释至刻度，然后用孔径为 0.2μm~0.5μm 的有机相膜过滤，滤液按 A.4.1 中的色谱条件进行分析。
- 5. 相对分子质量分布的计算
- 在4.1的色谱条件下分析4.3中制备的样品溶液后，利用GPC数据处理软件，将样品色谱数据代入校准曲线4.2，即可得到样品的相对分子质量及其分布范围。不同肽段的相对分子质量分布可通过峰面积归一化法计算，公式为：X=A/Atotal×100
- 公式中：X - 样品中相对分子质量肽在总肽中的质量分数，%；
- A - 相对分子质量肽的峰面积；
- 总 A 值 - 各相对分子质量肽峰面积之和，计算至小数点后一位。
- 6. 重复性
- 在可重复性条件下获得的两次独立测定结果的绝对差值不得超过两次测定结果算术平均值的 15%。
- 附录B：游离氨基酸的测定方法
- 标准采纳：Q/320205 KAVN05-2016
- 1.2 试剂和材料
- 冰醋酸：分析纯
- 高氯酸：0.0500 mol/L
- 指示剂：0.1%结晶紫指示剂（冰醋酸）
- 2. 游离氨基酸的测定
样品在 80°C 下干燥 1 小时。
将样品放入干燥容器中，使其自然冷却至室温或冷却至可用温度。将约 0.1 克样品（精确到 0.001 克）称入 250 毫升干燥的锥形瓶中。尽快进行下一步，以免样品吸收环境水分。加入 25 毫升冰醋酸，搅拌均匀，搅拌时间不超过 5 分钟。加入2滴结晶紫指示剂用 0.0500 mol / L (±0.001) 高氯酸标准滴定溶液进行滴定，直至溶液颜色由紫色变为终点。
记录消耗的标准溶液的体积。
- 同时进行空白试验。
- 3. 计算和结果
- 试剂中游离氨基酸含量 X 以质量分数 (%) 表示，并根据以下公式计算：X = C × (V1-V0) × 0.1445/M × 100%，公式如下：
- C - 标准高氯酸溶液的浓度，单位为摩尔每升 (mol/L)
- V1 - 用标准高氯酸溶液滴定样品所用的体积，以毫升 (mL) 为单位。
- Vo - 用标准高氯酸溶液进行滴定空白所用的体积，单位为毫升（mL）；
M - 样品的质量，以克 (g) 为单位。
| 0.1445：相当于 1.00 mL 标准高氯酸溶液 [c (HClO4) = 1.000 mol / L] 的氨基酸的平均质量。 | 4.2.3 硫酸铈标准滴定溶液：浓度 c [Ce (SO4) 2] = 0.1 mol/L，按 GB/T601 制备。 | |
| 标准采纳：Q/70920556 71-2024 | 1. 测定原理（以铁为例） | 氨基酸铁络合物在无水乙醇中的溶解度很低，而游离金属离子可溶于无水乙醇，利用二者在无水乙醇中的溶解度差异来确定氨基酸铁络合物的螯合率。 |
| 公式中：V1 - 滴定试验溶液所消耗的硫酸铈标准溶液的体积，mL； | 无水乙醇；其余与GB/T 27983-2011第4.5.2条相同。 | 3. 分析步骤 |
| 同时进行两组试验。称取0.1g在103±2℃下干燥1小时的样品（精确至0.0001g），加入100mL无水乙醇溶解，过滤，滤渣用100mL无水乙醇洗涤至少三次，然后将滤渣转移至250mL锥形瓶中，按照GB/T27983-2011标准4.5.3条的规定加入10mL硫酸溶液，然后按照GB/T27983-2011标准4.5.3条的规定进行后续步骤“加热溶解，然后冷却”。同时进行空白试验。 | 4. 总铁含量的测定 | 4.1 确定原则与 GB/T 21996-2008 中的第 4.4.1 条相同。 |
4.2 试剂和溶液
| 4.2.1 混合酸：将 150 毫升硫酸和 150 毫升磷酸加入 700 毫升水中，充分混合。 | 4.2.2 二苯胺磺酸钠指示剂溶液：5g/L，按GB/T603制备。 | 4.2.3 硫酸铈标准滴定溶液：浓度 c [Ce (SO4) 2] = 0.1 mol/L，按 GB/T601 制备。 | |
| 4.3 分析步骤 | 同时进行两组平行试验。称取0.1g样品（精确至0.20001g），置于250mL锥形瓶中，加入10mL混合酸，溶解后，加入30mL水和4滴二苯胺磺酸钠指示剂溶液，然后按照GB/T21996-2008标准4.4.2条进行后续步骤。同时进行空白试验。 | 4.4 结果的表示 | 氨基酸铁复合物的总铁含量 X1（以铁的质量分数表示，%）按公式（1）计算： |
| X1=(V-V0)×C×M×10-3×100 | V0 - 用于滴定空白溶液的硫酸铈标准溶液的消耗量，mL； | V0 - 用于滴定空白溶液的硫酸铈标准溶液的消耗量，mL； | C - 硫酸铈标准溶液的实际浓度，mol/L5. 螯合物中铁含量的计算螯合物中铁含量 X2（以铁的质量分数表示，%）按以下公式计算：x2 = ((V1-V2) × C × 0.05585)/m1 × 100 |
| 公式中：V1 - 滴定试验溶液所消耗的硫酸铈标准溶液的体积，mL； | V2 - 用于滴定空白溶液的硫酸铈标准溶液消耗量，mL；nom1-样品的质量，g。取平行测定结果的算术平均值作为测定结果，平行测定结果的绝对差值不超过0.3%。 | 0.05585 - 亚铁离子的质量以克表示，相当于 1.00 mL 硫酸铈标准溶液 C[Ce(SO4)2.4H20] = 1.000 mol/L。nom1-样品的质量，g。取平行测定结果的算术平均值作为测定结果，平行测定结果的绝对差值不超过0.3%。 | 6. 螯合率的计算螯合率 X3，以百分比表示的值 X3 = X2/X1 × 100附录C：测定Zinpro螯合率的方法 |
标准采纳：Q/320205 KAVNO7-2016
1. 试剂和材料
a) 冰醋酸：分析纯；b) 高氯酸：0.0500mol/L；c) 指示剂：0.1% 结晶紫指示剂（冰醋酸）
2. 游离氨基酸的测定
2.1 将样品在 80°C 下干燥 1 小时。
2.2 将样品放入干燥的容器中，使其自然冷却至室温或冷却至可用温度。
2.3 称取约 0.1 g 样品（精确到 0.001 g）于 250 mL 干燥锥形瓶中。
2.4 迅速进行下一步，以避免样品吸收环境水分。
2.5 加入 25 毫升冰醋酸，搅拌均匀，搅拌时间不超过 5 分钟。
2.6 加入 2 滴结晶紫指示剂。
2.7 用 0.0500mol/L (±0.001) 高氯酸标准滴定溶液滴定，直至溶液颜色由紫色变为绿色，持续 15 秒，且不再变化，即为滴定终点。
2.8 记录消耗的标准溶液的体积。
2.9 同时进行空白试验。
- 3. 计算和结果
- 加泰罗尼亚语
- Physicochemical parameters
V1 - 用标准高氯酸溶液滴定样品所用的体积，以毫升 (mL) 为单位。
Vo - 用标准高氯酸溶液进行滴定空白所用的体积，单位为毫升（mL）；
c) Chelation rate: ≥ 95%
d) Arsenic: ≤ 2 mg/kg
e) Lead: ≤ 5 mg/kg
f) Cadmium: ≤ 5 mg/kg
g) Moisture content: ≤ 5.0%
h) Fineness: All particles pass through 20 mesh, with a main particle size of 60-80 mesh
地址：中国四川省成都市浦江县寿安镇青浦路147号
电话：86-18880477902
产品
无机微量元素
- 有机微量元素
- 斯瓦希里语
- 定制服务
- 快速链接
公司简介
| Application object | Suggested dosage (g/t full-value material) | Content in full-value feed (mg/kg) | Efficacy |
| 古吉拉特语 | 点击查询 | © 版权所有 - 2010-2025：保留所有权利。 | 网站地图 热门搜索 电话 |
| 电话 | 86-18880477902 | 爪哇语 | 电子邮件 |
| 8618880477902 | 中国人 | 法语 | |
| Bird | 中国人 | 法语 | 德语 西班牙语 |
| Aquatic animals | 日本人 | 韩国人 | 阿拉伯 希腊语 |
| 土耳其 | 意大利语 | ||
| Ruminant animal g/head day | January 0.75 | 印度尼西亚 南非荷兰语 瑞典 |
抛光
- 巴斯克
- 加泰罗尼亚语
- Physicochemical parameters
印地语
老挝
c) Chelation rate: ≥ 95%
d) Arsenic: ≤ 2 mg/kg
e) Lead: ≤ 5 mg/kg
f) Cadmium: ≤ 5 mg/kg
g) Moisture content: ≤ 5.0%
h) Fineness: All particles pass through 20 mesh, with a main particle size of 60-80 mesh
绍纳语
保加利亚语
- 宿务语
- This product is chemically stable and can significantly reduce its damage to vitamins and fats, etc. The use of this product is conducive to improving feed quality;
- The product is absorbed through small peptide and amino acid pathways, reducing the competition and antagonism with other trace elements, and has the best bio-absorption and utilization rate;
- 克罗地亚语
荷兰语
| Application object | 乌尔都语 越南语 | Content in full-value feed (mg/kg) | Efficacy |
| 古吉拉特语 | 海地 | 豪萨语 | 卢旺达语 苗族 匈牙利 |
| Piglets and fattening pigs | 伊博语 | 爪哇语 | 卡纳达语 高棉语 库尔德 |
| 吉尔吉斯 | 拉丁 | ||
| Bird | 300～400 | 45～60 | 马其顿 马来语 马拉雅拉姆语 |
| Aquatic animals | 200～300 | 30～45 | 1. Promote growth, improve feed conversion; 2. Improve anti-stress abolity, reduce morbidity and mortality. |
挪威
- 普什图语
- Appearance: brownish-yellow granules
- Physicochemical parameters
塞尔维亚
塞索托语
c) Chelation rate: ≥ 95%
d) Arsenic: ≤ 2 mg/kg
e) Lead: ≤ 5 mg/kg
f) Cadmium: ≤ 5 mg/kg
g) Moisture content: ≤ 5.0%
h) Fineness: All particles pass through 20 mesh, with a main particle size of 60-80 mesh
绍纳语
信德语
This product is an all-organic trace mineral chelated by a special chelating proces with pure plant enzymatic small molecule peptides as chelating substrates and trace elements;
斯瓦希里语
塔吉克
泰米尔语
泰卢固语
泰国
| Application object | 乌尔都语 越南语 | Content in full-value feed (mg/kg) | Efficacy |
| 意第绪语 | 约鲁巴 | 祖鲁语 | 卢旺达语 奥里亚语 土库曼 |
| 维吾尔族 | 250～400 | 37.5～60 | 1. Improving the immunity of piglets, reducing diarrhea and mortality; 2. Improving palatability, increasing feed intake, increasing growth rate and improving feed conversion; 3. Make the pig coat bright and improve the carcass quality and meat quality. |
| Bird | 300～400 | 45～60 | 1. Improve feather glossiness; 2. improve the laying rate, fertilization rate and hatching rate of breeding eggs, and strengthen the coloring ability of egg yolk; 3. Improve anti-stress ability and reduce mortality; 4. Improve feed conversion and increase growth rate. |
| Aquatic animals | January 300 | 45 | 1. Promote growth, improve feed conversion; 2. Improve anti-stress abolity, reduce morbidity and mortality. |
| Ruminant animal g/head day | 2.4 | 1. Improve milk yield, prevent mastitis and foof rot, and reduce somatic cell content in milk; 2. Promote growth, improve feed conversion and improve meat quality. |
4. Manganese Amino Acid Chelate Feed Grade
- Product Name: Manganese Amino Acid Chelate Feed Grade
- Appearance: brownish-yellow granules
- Physicochemical parameters
a) Mn: ≥ 10.0%
b) Total amino acids: ≥ 19.5%
c) Chelation rate: ≥ 95%
d) Arsenic: ≤ 2 mg/kg
e) Lead: ≤ 5 mg/kg
f) Cadmium: ≤ 5 mg/kg
g) Moisture content: ≤ 5.0%
h) Fineness: All particles pass through 20 mesh, with a main particle size of 60-80 mesh
n=0, 1,2,...indicates chelated manganese for dipeptides, tripeptides, and tetrapeptides
Characteristics of Manganese Amino Acid Chelate Feed Grade
This product is an all-organic trace mineral chelated by a special chelating proces with pure plant enzymatic small molecule peptides as chelating substrates and trace elements;
This product is chemically stable and can significantly reduce its damage to vitamins and fats, etc. The use of this product is conducive to improving feed quality;
The product is absorbed through small peptide and amino acid pathways, reducing the competition and antagonism with other trace elements, and has the best bio-absorption and utilization rate;
The product can improve the growth rate, improve feed conversion and health status significantly; and improve the laying rate, hatching rate and healthy chick rate of breeding poultry obviously;
Manganese is necessary for bone growth and connective tissue maintenance. It is closely related to many enzymes; and participates in carbohydrate, fat and protein metabolism, reproduction and immune response.
Usage and Efficacy of Manganese Amino Acid Chelate Feed Grade
| Application object | Suggested dosage (g/t full-value material) | Content in full-value feed (mg/kg) | Efficacy |
| Breeding pig | 200～300 | 30～45 | 1. Promote the normal development of sexual organs and improve sperm motility; 2. Improve the reproductive capacity of breeding pigs and reduce reproductive obstacles. |
| Piglets and fattening pigs | 100～250 | 15～37.5 | 1. It is beneficial to improve immune functions, and improve anti-stress ability and disease resistance; 2. Promote growth and improve feed conversion significantly; 3. Improve meat color and quality, and improve lean meat percentage. |
| Bird | 250～350 | 37.5～52.5 | 1. Improve anti-stress ability and reduce mortality; 2. Improve laying rate, fertilization rate and hatching rate of breeding eggs, improve eggshell quality and reduce shell breaking rate; 3. Promote bone growth and reduce the incidence of leg diseases. |
| Aquatic animals | 100～200 | 15～30 | 1. Promote growth and improve its anti-stress ability and disease resistance; 2. Improve sperm motility and hatching rate of fertilized eggs. |
| Ruminant animal g/head day | Cattle 1.25 | 1. Prevent fatty acid synthesis disorder and bone tissue damage; 2. Improve reproductive capacity, prevent abortion and postpartum paralysis of female animals, reduce the mortality of calves and lambs, and increase the newborn weight of young animals. | |
| Goat 0.25 |
Part 6 FAB of Small Peptide-mineral Chelates
| S/N | F: Functional attributes | A: Competitive differences | B: Benefits brought by competitive differences to users |
| 1.52 | Selectivity control of raw materials | Select pure plant enzymatic hydrolysis of small peptides | High biological safety, avoiding cannibalism |
| 2 | Directional digestion technology for double protein biological enzyme | High proportion of small molecular peptides | More "targets", which are not easy to saturation, with high biological activity and better stability |
| 3 | Advanced pressure spray & drying technology | Granular product, with uniform particle size, better fluidity, not easy to absorb moisture | Ensure easy to use, more uniform mixing in complete feed |
| Low water content (≤ 5%), which greatly reduces the influence caused by vitamins and enzyme preparations | Improve the stability of feed products | ||
| 4 | Advanced production control technology | Totally enclosed process, high degree of automatic control | Safe and stable quality |
| 5 | Advanced quality control technology | Establish and improve scientific and advanced analytical methods and control means for detecting factors affecting product quality, such as acid-soluble protein, molecular weight distribution, amino acids and chelating rate | Ensure quality, ensure efficiency and improve efficiency |
Part 7 Competitor Comparison
Standard VS Standard
Comparison of peptide distribution and chelation rate of products
| Sustar's products | Proportion of small peptides（180-500） | Zinpro's products | Proportion of small peptides（180-500） |
| AA-Cu | ≥74% | AVAILA-Cu | 78% |
| AA-Fe | ≥48% | AVAILA-Fe | 59% |
| AA-Mn | ≥33% | AVAILA-Mn | 53% |
| AA-Zn | ≥37% | AVAILA-Zn | 56% |
| Sustar's products | Chelation rate | Zinpro's products | Chelation rate |
| AA-Cu | 94.8% | AVAILA-Cu | 94.8% |
| AA-Fe | 95.3% | AVAILA-Fe | 93.5% |
| AA-Mn | 94.6% | AVAILA-Mn | 94.6% |
| AA-Zn | 97.7% | AVAILA-Zn | 90.6% |
The ratio of small peptides of Sustar is slightly lower than that of Zinpro, and the chelation rate of Sustar's products is slightly higher than that of Zinpro's products.
Comparison of the content of 17 amino acids in different products
| Name of amino acids | Sustar's Copper Amino Acid Chelate Feed Grade | Zinpro's AVAILA copper | Sustar's Ferrous Amino Acid C helate Feed Grade | Zinpro's AVAILA iron | Sustar's Manganese Amino Acid Chelate Feed Grade | Zinpro's AVAILA manganese | Sustar's Zinc Amino Acid Chelate Feed Grade | Zinpro's AVAILA zinc |
| aspartic acid (%) | 1.88 | 0.72 | 1.50 | 0.56 | 1.78 | 1.47 | 1.80 | 2.09 |
| glutamic acid (%) | 4.08 | 6.03 | 4.23 | 5.52 | 4.22 | 5.01 | 4.35 | 3.19 |
| Serine (%) | 0.86 | 0.41 | 1.08 | 0.19 | 1.05 | 0.91 | 1.03 | 2.81 |
| Histidine (%) | 0.56 | 0.00 | 0.68 | 0.13 | 0.64 | 0.42 | 0.61 | 0.00 |
| Glycine (%) | 1.96 | 4.07 | 1.34 | 2.49 | 1.21 | 0.55 | 1.32 | 2.69 |
| Threonine (%) | 0.81 | 0.00 | 1.16 | 0.00 | 0.88 | 0.59 | 1.24 | 1.11 |
| Arginine (%) | 1.05 | 0.78 | 1.05 | 0.29 | 1.43 | 0.54 | 1.20 | 1.89 |
| Alanine (%) | 2.85 | 1.52 | 2.33 | 0.93 | 2.40 | 1.74 | 2.42 | 1.68 |
| Tyrosinase (%) | 0.45 | 0.29 | 0.47 | 0.28 | 0.58 | 0.65 | 0.60 | 0.66 |
| Cystinol (%) | 0.00 | 0.00 | 0.09 | 0.00 | 0.11 | 0.00 | 0.09 | 0.00 |
| Valine (%) | 1.45 | 1.14 | 1.31 | 0.42 | 1.20 | 1.03 | 1.32 | 2.62 |
| Methionine (%) | 0.35 | 0.27 | 0.72 | 0.65 | 0.67 | 0.43 | January 0.75 | 0.44 |
| Phenylalanine (%) | 0.79 | 0.41 | 0.82 | 0.56 | 0.70 | 1.22 | 0.86 | 1.37 |
| Isoleucine (%) | 0.87 | 0.55 | 0.83 | 0.33 | 0.86 | 0.83 | 0.87 | 1.32 |
| Leucine (%) | 2.16 | 0.90 | 2.00 | 1.43 | 1.84 | 3.29 | 2.19 | 2.20 |
| Lysine (%) | 0.67 | 2.67 | 0.62 | 1.65 | 0.81 | 0.29 | 0.79 | 0.62 |
| Proline (%) | 2.43 | 1.65 | 1.98 | 0.73 | 1.88 | 1.81 | 2.43 | 2.78 |
| Total amino acids (%) | 23.2 | 21.4 | 22.2 | 16.1 | 22.3 | 20.8 | 23.9 | 27.5 |
Overall, the proportion of amino acids in Sustar's products is higher than that in Zinpro's products.
Part 8 Effects of use
Effects of different sources of trace minerals on the production performance and egg quality of laying hens in the late laying period
Production Process
- Targeted chelation technology
- Shear emulsification technology
- Pressure spray & drying technology
- Refrigeration & dehumidification technology
- Advanced environmental control technology
Appendix A: Methods for the Determination of relative molecular mass distribution of peptides
Adoption of standard: GB/T 22492-2008
1 Test Principle:
It was determined by high performance gel filtration chromatography. That is to say, using porous filler as stationary phase, based on the difference in the relative molecular mass size of the sample components for separation, detected at the peptide bond of the ultraviolet absorption wavelength of 220nm, using the dedicated data processing software for the determination of relative molecular mass distribution by gel filtration chromatography (i.e., the GPC software), the chromatograms and their data were processed, calculated to get the size of the relative molecular mass of the soybean peptide and the distribution range.
2. Reagents
The experimental water should meet the specification of secondary water in GB/T6682, the use of reagents, except for special provisions, are analytically pure.
2.1 Reagents include acetonitrile (chromatographically pure), trifluoroacetic acid (chromatographically pure),
2.2 Standard substances used in the calibration curve of relative molecular mass distribution: insulin, mycopeptides, glycine-glycine-tyrosine-arginine, glycine-glycine-glycine
3 Instrument and equipment
3.1 High Performance Liquid Chromatograph (HPLC): a chromatographic workstation or integrator with a UV detector and GPC data processing software.
3.2 Mobile phase vacuum filtration and degassing unit.
3.3 Electronic balance: graduated value 0.000 1g.
4 Operating steps
4.1 Chromatographic conditions and system adaptation experiments (reference conditions)
4.1.1 Chromatographic column: TSKgelG2000swxl300 mm×7.8 mm (inner diameter) or other gel columns of the same type with similar performance suitable for the determination of proteins and peptides.
4.1.2 Mobile phase: Acetonitrile + water + trifluoroacetic acid = 20 + 80 + 0.1.
4.1.3 Detection wavelength: 220 nm.
4.1.4 Flow rate: 0.5 mL/min.
4.1.5 Detection time: 30 min.
4.1.6 Sample injection volume: 20μL.
4.1.7 Column temperature: room temperature.
4.1.8 In order to make the chromatographic system meet the detection requirements, it was stipulated that under the above chromatographic conditions, the gel chromatographic column efficiency, i.e., the theoretical number of plates (N), was not less than 10000 calculated on the basis of the peaks of the tripeptide standard (Glycine-Glycine-Glycine).
4.2 Production of relative molecular mass standard curves
The above different relative molecular mass peptide standard solutions with a mass concentration of 1 mg / mL were prepared by mobile phase matching, mixed in a certain proportion, and then filtered through an organic phase membrane with the pore size of 0.2 μm~0.5 μm and injected into the sample, and then the chromatograms of the standards were obtained. Relative molecular mass calibration curves and their equations were obtained by plotting the logarithm of relative molecular mass against retention time or by linear regression.
4.3 Sample treatment
Accurately weigh 10mg of sample in a 10mL volumetric flask, add a little mobile phase, ultrasonic shaking for 10min, so that the sample is fully dissolved and mixed, diluted with mobile phase to the scale, and then filtered through an organic phase membrane with a pore size of 0.2μm~0.5μm, and the filtrate was analyzed according to the chromatographic conditions in A.4.1.
5. Calculation of relative molecular mass distribution
After analyzing the sample solution prepared in 4.3 under the chromatographic conditions of 4.1, the relative molecular mass of the sample and its distribution range can be obtained by substituting the chromatographic data of the sample into the calibration curve 4.2 with GPC data processing software. The distribution of the relative molecular masses of the different peptides can be calculated by the peak area normalization method, according to the formula: X=A/A total×100
In the formula: X - The mass fraction of a relative molecular mass peptide in the total peptide in the sample, %;
A - Peak area of a relative molecular mass peptide;
Total A - the sum of the peak areas of each relative molecular mass peptide, calculated to one decimal place.
6 Repeatability
The absolute difference between two independent determinations obtained under conditions of repeatability shall not exceed 15% of the arithmetic mean of the two determinations.
Appendix B: Methods for the Determination of Free Amino Acids
Adoption of standard: Q/320205 KAVN05-2016
1.2 Reagents and materials
Glacial acetic acid: analytically pure
Perchloric acid: 0.0500 mol/L
Indicator: 0.1% crystal violet indicator (glacial acetic acid)
2. Determination of free amino acids
The samples were dried at 80°C for 1 hour.
Place the sample in a dry container to cool naturally to room temperature or cool down to a usable temperature.
Weigh approximately 0.1 g of sample (accurate to 0.001 g) into a 250 mL dry conical flask.
Quickly proceed to the next step to avoid the sample from absorbing ambient moisture
Add 25 mL of glacial acetic acid and mix well for no more than 5 min.
Add 2 drops of crystal violet indicator
Titrate with 0.0500 mol / L (±0.001) standard titration solution of perchloric acid until the solution changes from purple to the end point.
Record the volume of standard solution consumed.
Carry out the blank test at the same time.
3. Calculation and results
The free amino acid content X in the reagent is expressed as a mass fraction (%) and is calculated according to the formula: X = C × (V1-V0) × 0.1445/M × 100%, in tne formula:
C - Concentration of standard perchloric acid solution in moles per liter (mol/L)
V1 - Volume used for titration of samples with standard perchloric acid solution, in milliliters (mL).
Vo - Volume used for titration blank with standard perchloric acid solution, in milliliters (mL);
M - Mass of the sample, in grams (g ).
0.1445: Average mass of amino acids equivalent to 1.00 mL of standard perchloric acid solution [c (HClO4) = 1.000 mol / L].
Appendix C: Methods for the Determination of Sustar's chelation rate
Adoption of standards: Q/70920556 71-2024
1. Determination principle (Fe as an example)
Amino acid iron complexes have very low solubility in anhydrous ethanol and free metal ions are soluble in anhydrous ethanol, the difference in solubility between the two in anhydrous ethanol was utilized to determine the chelation rate of amino acid iron complexes.
2. Reagents & Solutions
Anhydrous ethanol; the rest is the same as clause 4.5.2 in GB/T 27983-2011.
3. Steps of analysis
Do two trials in parallel. Weigh 0.1g of the sample dried at 103±2℃ for 1 hour, accurate to 0.0001g, add 100mL of anhydrous ethanol to dissolve, filter, filter residue washed with 100mL of anhydrous ethanol for at least three times, then transfer the residue into a 250mL conical flask, add 10mL of sulfuric acid solution according to clause 4.5.3 in GB/T27983-2011, and then perform the following steps according to clause 4.5.3 “Heat to dissolve and then let cool” in GB/T27983-2011. Carry out the blank test at the same time.
4. Determination of total iron content
4.1 The principle of determination is the same as clause 4.4.1 in GB/T 21996-2008.
4.2. Reagents & Solutions
4.2.1 Mixed acid: Add 150mL of sulfuric acid and 150mL of phosphoric acid to 700mL of water and mix well.
4.2.2 Sodium diphenylamine sulfonate indicator solution: 5g/L, prepared according to GB/T603.
4.2.3 Cerium sulfate standard titration solution: concentration c [Ce (SO4) 2] = 0.1 mol/L, prepared according to GB/T601.
4.3 Steps of analysis
Do two trials in parallel. Weigh 0.1g of sample, accurate to 020001g, place in a 250mL conical flask, add 10mL of mixed acid, after dissolution, add 30ml of water and 4 drops of sodium dianiline sulfonate indicator solution, and then perform the following steps according to clause 4.4.2 in GB/T21996-2008. Carry out the blank test at the same time.
4.4 Representation of results
The total iron content X1 of the amino acid iron complexes in terms of mass fraction of iron, the value expressed in %, was calculated according to formula (1):
X1=(V-V0)×C×M×10-3×100
In the formula: V - volume of cerium sulfate standard solution consumed for titration of test solution, mL;
V0 - cerium sulfate standard solution consumed for titration of blank solution, mL;
C - Actual concentration of cerium sulfate standard solution, mol/L
5. Calculation of iron content in chelates
The iron content X2 in the chelate in terms of the mass fraction of iron, the value expressed in %, was calculated according to the formula: x2 = ((V1-V2) × C × 0.05585)/m1 × 100
In the formula: V1 - volume of cerium sulfate standard solution consumed for titration of test solution, mL;
V2 - cerium sulfate standard solution consumed for titration of blank solution, mL;
C - Actual concentration of cerium sulfate standard solution, mol/L;
0.05585 - mass of ferrous iron expressed in grams equivalent to 1.00 mL of cerium sulfate standard solution C[Ce(SO4)2.4H20] = 1.000 mol/L.
m1-Mass of the sample, g. Take the arithmetic mean of the parallel determination results as the determination results, and the absolute difference of the parallel determination results is not more than 0.3%.
6. Calculation of chelation rate
Chelation rate X3, the value expressed in %, X3 = X2/X1 × 100
Appendix C: Methods for the Determination of Zinpro's chelation rate
Adoption of standard: Q/320205 KAVNO7-2016
1. Reagents and materials
a) Glacial acetic acid: analytically pure; b) Perchloric acid: 0.0500mol/L; c) Indicator: 0.1% crystal violet indicator (glacial acetic acid)
2. Determination of free amino acids
2.1 The samples were dried at 80°C for 1 hour.
2.2 Place the sample in a dry container to cool naturally to room temperature or cool down to a usable temperature.
2.3 Weigh approximately 0.1 g of sample (accurate to 0.001 g) into a 250 mL dry conical flask
2.4 Quickly proceed to the next step to avoid the sample from absorbing ambient moisture.
2.5 Add 25mL of glacial acetic acid and mix well for no more than 5min.
2.6 Add 2 drops of crystal violet indicator.
2.7 Titrate with 0.0500mol/L (±0.001) standard titration solution of perchloric acid until the solution changes from purple to green for 15s without changing color as the end point.
2.8 Record the volume of standard solution consumed.
2.9 Carry out the blank test at the same time.
3. Calculation and results
The free amino acid content X in the reagent is expressed as a mass fraction (%), calculated according to formula (1): X=C×(V1-V0) ×0.1445/M×100%...... .......(1)
In the formula: C - concentration of standard perchloric acid solution in moles per liter (mol/L)
V1 - Volume used for titration of samples with standard perchloric acid solution, in milliliters (mL).
Vo - Volume used for titration blank with standard perchloric acid solution, in milliliters (mL);
M - Mass of the sample, in grams (g ).
0.1445 - Average mass of amino acids equivalent to 1.00 mL of standard perchloric acid solution [c (HClO4) = 1.000 mol / L].
4. Calculation of chelation rate
The chelation rate of the sample is expressed as mass fraction (%), calculated according to formula (2): chelation rate = (total amino acid content - free amino acid content)/total amino acid content×100%.
Post time: Sep-17-2025
