ICS67.050
CCSX 04
中华人民共和国出入境检验检疫行业标准
SN/T5639—2024
出口畜禽肉中松香酸和脱氢松香酸的测定
Determination of abietic acid and dehydroabietic acid in livestock and poultry
muscles for export
2024-12-16发布2025-06-01实施
中华人民共和国海关总署发布

前 言
本文件按照GB/T 1.1—2020《标准化工作导则 第1部分:标准化文件的结构和起草规则》的规
定起草。
请注意本文件的某些内容可能涉及专利。本文件的发布机构不承担识别专利的责任。
本文件由中华人民共和国海关总署提出并归口。
本文件起草单位:中华人民共和国广州海关、中国肉类食品综合研究中心、中国海关科学技术研究
中心。
本文件主要起草人:李菊、谢建军、曾广丰、古瑾、王璐、韩深、席静、董洁、陈文锐、王志元、侯颖烨。
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SN/T5639—2024

出口畜禽肉中松香酸和脱氢松香酸的测定
1 范围
本文件规定了出口畜禽肉中松香酸和脱氢松香酸含量的高效液相色谱测定方法及液相色谱-质谱/
质谱确证方法。
本文件适用于鸡肉、鸭肉、鹅肉、猪肉、牛肉、羊肉中松香酸及脱氢松香酸残留量的测定和确证,其他
畜禽肉类食品可参照执行。
2 规范性引用文件
下列文件中的内容通过文中的规范性引用而构成本文件必不可少的条款。其中,注日期的引用文
件,仅该日期对应的版本适用于本文件;不注日期的引用文件,其最新版本(包括所有的修改单)适用于
本文件。
GB/T 6682 分析实验室用水规格和试验方法
3 术语和定义
本文件没有需要界定的术语和定义。
4 原理
试样中的松香酸和脱氢松香酸用乙腈提取,经硅胶键合C18 净化剂净化,用液相色谱二极管阵列或
紫外检测器测定,外标法定量,液相色谱-质谱/质谱法确证。
5 试剂和材料
除非另有规定,仅使用分析纯试剂,试验用水应符合GB/T 6682中规定的一级水。
5.1 乙腈:色谱纯。
5.2 甲醇:色谱纯。
5.3 甲酸:色谱纯
5.4 磷酸:优级纯。
5.5 无水硫酸镁。
5.6 氯化钠。
5.7 柠檬酸钠。
5.8 柠檬酸氢二钠。
5.9 0.1%磷酸水溶液:吸取1 mL磷酸(5.4),加水定容至1 000 mL,混匀。
5.10 松香酸 :纯度大于等于98%,松香酸标准物质的信息见附录A 中表A.1。
5.11 脱氢松香酸 :纯度大于等于98%,脱氢松香酸标准物质的信息见附录A 中表A.1。
5.12 松香酸标准储备溶液(1 000 mg/L):准确称取10 mg松香酸标准品(5.10)于小烧杯中,加少量
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甲醇(5.2)溶解,移入10 mL容量瓶中,用甲醇定容至刻度,摇匀,-18 ℃避光保存,有效期为6个月。
5.13 脱氢松香酸标准储备溶液(1 000 mg/L):精密称取10 mg脱氢松香酸标准品(5.11)于小烧杯
中,加少量甲醇(5.2)溶解,移入10 mL容量瓶中,用甲醇定容至刻度,摇匀,-18 ℃避光保存,有效期为
6个月。
5.14 标准中间溶液(100.0 mg/L):分别量取1 mL标准储备溶液(5.12和5.13),用乙腈(5.1)稀释并
定容至10 mL,摇匀,-18 ℃避光保存,有效期为1个月。
5.15 标准工作溶液:准确量取标准中间溶液(5.14),用乙腈提取液稀释成0.10 mg/L、0.20 mg/L、0.
50 mg/L、1.00 mg/L、2.00 mg/L和5.00 mg/L的系列浓度标准工作溶液,现配现用。
5.16 硅胶键合C18 净化剂:粒径50 μm。
5.17 0.22 μm 微孔滤膜,有机系。
6 仪器和设备
6.1 高效液相色谱仪:配二极管阵列检测器(PDA)或紫外检测器。
6.2 高效液相色谱-质谱/质谱仪:配有电喷雾离子源(ESI)。
6.3 电子天平:感量为0.01 g和0.1 mg。
6.4 组织捣碎机。
6.5 涡旋混合器。
6.6 高速离心机:转速不低于4 000 r/min。
6.7 微型高速离心机:转速不低于10 000 r/min。
6.8 氮吹仪。
7 试样的制备与保存
7.1 在制样的操作过程中,应防止样品污染或发生残留量含量的变化。
7.2 从所取样品中取出有代表性样品(有皮畜禽肉类样品带皮)约500 g,用组织捣碎机捣碎,装入洁净
容器,一份作为试样,一份作为留样,密封并做好标识,于-18 ℃下保存。
8 测定步骤
8.1 提取
称取约5 g试样(精确到0.01 g)于50 mL离心管中,加入乙腈(5.1)10 mL、水10 mL,涡旋混匀
2 min后,加入无水硫酸镁(5.5)4.5 g、氯化钠(5.6)1 g、柠檬酸钠(5.7)0.3 g、柠檬酸氢二钠(5.8)
0.7 g,涡旋混合器中振摇25 min,4 000 r/min离心5 min,待净化。
8.2 净化
取1 mL上清液(见8.1)于装有100 mg C18(5.16)的2 mL离心管中,涡旋摇匀2 min,10 000 r/
min离心5 min,取上清液过滤膜(5.17),供高效液相色谱仪测定。
8.3 测定
8.3.1 液相色谱条件
由于测试结果取决于所使用的仪器,因此不可能给出液相色谱分析的通用参数。设定的参数应保
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证色谱分析时被测组分与其他组分能得到有效的分离,下列给出的参数可供参考。
液相色谱参考条件如下:
a) 色谱柱:C18 柱,100 mm ×2.1 mm(内径),粒度3.5 μm 或性能相当者;
b) 流动相:0.1%磷酸水溶液和乙腈,梯度洗脱,洗脱程序见表1;
c) 流速:0.5 mL/min;
d) 进样量:10 μL;
e) 柱温:35 ℃;
f) 检测波长:松香酸240 nm,脱氢松香酸210 nm。
表1 梯度洗脱程序
时间/min A/%(0.1%磷酸水溶液) B/%(乙腈)
0 70 30
4 70 30
15 10 90
17 10 90
17.5 70 30
20 70 30
8.3.2 标准工作曲线的绘制
按8.3.1的测定条件,将标准工作溶液浓度由低到高依次进样测定,以目标物色谱峰的峰面积为纵
坐标,与其对应的溶液浓度为横坐标作图,绘制标准工作曲线。高效液相色谱图见附录B中的图B.1
和图B.2。
8.3.3 试样测定
按8.3.1的条件测定样品和标准工作溶液,如果检测的松香酸和脱氢松香酸的色谱峰保留时间与
标准品保留时间相差不超过±2.5 %,则可初步确认样品中存在松香酸和脱氢松香酸。必要时,阳性样
品需用液相色谱-质谱/质谱法进行确认试验(见附录C),松香酸或脱氢松香酸定性离子对的相对丰度
(是用相对于最强离子丰度的强度百分比表示)与浓度相当基质标准工作溶液的相对丰度一致,相对丰
度允许偏差不超过表C.2规定的范围,且每个定性离子的信噪比均≥ 3,则可判断样品中存在松香酸和
脱氢松香酸。松香酸和脱氢松香酸的多反应监测色谱图见附录D 中的图D.1和图D.2。选取响应值
相近的基质混合标准工作液一起进行色谱分析,基质混合标准工作液和待测液中松香酸和脱氢松香酸
的响应值在仪器线性范围内,超过线性范围则应稀释后再进样分析。外标法定量。
8.4 空白试验
试剂空白试验:除不加试样外,均按上述步骤进行。
试样空白试验:取空白基质试样,除不加标准溶液外,均按上述步骤进行。
9 计算结果与表述
试样中松香酸或脱氢松香酸含量由色谱数据处理软件或按公式(1)计算获得,计算结果应扣除空
白值。
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X =C×V×1 000 m ×1 000 ………………………………( 1 )
式中:
X ———试样中松香酸或脱氢松香酸含量的数值,单位为毫克每千克(mg/kg);
C———从标准工作曲线得到的松香酸或脱氢松香酸浓度的数值,单位为微克每升(μg/L);
V———试样溶液最终定容体积的数值,单位为毫升(mL);
m ———最终定容体积试样溶液所代表试样质量的数值,单位为克(g)。
计算结果保留3位有效数字。
10 检出限、定量限、回收率和精密度
10.1 检出限和定量限
本方法松香酸和脱氢松香酸的检出限均为0.20 mg/kg,定量限均为 0.50 mg/kg。
10.2 回收率和精密度
本方法分别以鸡肉、鸭肉、鹅肉、羊肉、猪肉、牛肉为空白样品基质,进行三个浓度水平的添加回收试
验,每个浓度水平进行6次重复实验,测得各种基质中松香酸和脱氢松香酸的回收率范围与精密度,见
附录E中的表E.1。
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附 录 A
(资料性)
松香酸和脱氢松香酸标准物质的信息
松香酸和脱氢松香酸的中文名称、英文名称、CAS号、分子式、结构式和相对分子质量见表A.1。
表A.1 松香酸和脱氢松香酸标准物质的信息
中文名称英文名称CAS号分子式结构式相对分子质量
松香酸Abietic acid 514-10-3 C20H30O2 302.46
脱氢松香酸Dehydroabietic acid 1740-19-8 C20H28O2 300.44
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附 录 B
(资料性)
松香酸和脱氢松香酸标准溶液的高效液相色谱图
松香酸和脱氢松香酸标准溶液的高效液相色谱图见图B.1与图B.2。
图B.1 松香酸标准溶液的高效液相色谱图(2.50 mg/L)
图B.2 脱氢松香酸标准溶液的高效液相色谱图(2.50 mg/L)
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附 录 C
(资料性)
液相色谱-质谱/质谱法确证条件1)
1) 非商业性声明:附录C所列参考质谱条件是在 AB 4000Q trap型液质联用仪上完成的,此处列出试验用仪器型
号仅为提供参考,并不涉及商业目的,鼓励标准使用者尝试不同厂家或型号的仪器。
C.1 液相色谱条件
液相色谱条件如下:
a) 色谱柱:C18 柱,100 mm ×2.1 mm(内径),3.5 μm,或性能相当者;
b) 流动相:0.1%甲酸-乙腈(等度洗脱8 min,15∶85,体积比);
c) 流速:0.5 mL/min;
d) 柱温:35 ℃;
e) 进样量:10 μL。
C.2 质谱参考条件
质谱参考条件如下:
a) 电喷雾电压:5 000 V;
b) 离子源温度:550 ℃;
c) 气帘气:25 psi;
d) 雾化气:50 psi ;
e) 加热辅助气:50 psi ;
f) 松香酸和脱氢松香酸的主要质谱参考参数见表C.1,定性确证时相对离子丰度的最大允许偏
差见表C.2。
表C.1 松香酸和脱氢松香酸的主要质谱参考参数
化合物电离模式
母离子
m/z
子离子
m/z
去簇电压
V
碰撞能量
eV
驻留时间
s
松香酸ESI+ 303.2 257.3 70.0 19.3 0.2
123.1* 70.0 21.7 0.2
脱氢松香酸ESI+ 301.0 173.2* 89.4 19.0 0.2
159.2 89.4 24.1 0.2
注: 带“*”的离子为定量离子。
表C.2 定性确证时相对离子丰度的最大允许偏差
相对离子丰度(基峰)/% >50 >20~50(含) >10~20(含) ≤10
允许的相对偏差/% ±20 ±25 ±30 ±50
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附 录 D
(资料性)
松香酸和脱氢松香酸标准溶液的监测色谱图
松香酸和脱氢松香酸标准溶液的监测色谱图见图D.1和图D.2。
a) 松香酸标准溶液定性离子色谱图
b) 松香酸标准溶液定量离子色谱图
图D.1 松香酸标准溶液的监测色谱图(0.50 mg/L)
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a) 脱氢松香酸标准溶液定量离子色谱图
b) 脱氢松香酸标准溶液定性离子色谱图
图D.2 脱氢松香酸标准溶液的监测色谱图(0.50 mg/L)
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附 录 E
(资料性)
畜禽肉中松香酸和脱氢松香酸的添加回收率和精密度(n=6)
畜禽肉中松香酸和脱氢松香酸的添加回收率和精密度(n=6)见表E.1.
表E.1 畜禽肉中松香酸和脱氢松香酸的添加回收率和精密度(n=6)
基质添加浓度/(mg/kg)
回收率范围/% RSD/%
松香酸脱氢松香酸松香酸脱氢松香酸
鸡肉
0.500 84.4~99.6 104~107 6.61 1.34
1.00 94.0~106 96.2~108 4.06 4.55
5.00 89.4~95.2 87.6~90.6 2.21 1.56
鸭肉
0.500 98.0~101 96.0~102 1.16 2.01
1.00 86.2~107 86.0~97.6 7.47 5.21
5.00 83.6~91.8 86.9~89.4 3.81 1.20
鹅肉
0.500 90.8~109 91.2~104 8.00 5.66
1.00 94.0~106 88.6~97.0 5.14 3.40
5.00 88.8~93.0 89.0~95.2 2.23 2.48
猪肉
0.500 93.2~102 87.2~98.0 3.71 4.38
1.00 93.6~106 92.6~106 5.12 5.01
5.00 90.0~94.4 85.4~90.6 1.70 2.28
牛肉
0.500 96.4~100 88.0~98.8 1.37 4.26
1.00 84.6~92.4 88.8~90.8 3.52 0.73
5.00 85.0~93.6 90.2~98.8 3.24 4.17
羊肉
0.500 90.0~106 100~104 7.90 1.78
1.00 96.4~110 99.6~104 5.73 1.83
5.00 86.6~94.2 89.6~94.0 3.05 1.95
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Foreword
This standard was drafted according to GB/T 1.1—2020.
Please note that some of the contents of this document may involve patents. The publisher of this
document does not assume the responsibility of identifying these patents.
This standard was proposed by and under the charge of General Administration of Customs of the
Pepople’s Republic of china.
This standard was drafted by GuangZhou Customs District P. R. China, China Meat Research Center,
Science and Technology Research Center of China Customs.
This standard was mainly drafted by Li Ju, Xie Jian Jun, Zeng Guang Feng,Gu Jin, Wang Lu,Han
Shen,Xi Jing, Dong Jie, Chen Wen Rui, Wang Zhi Yuan, Hou Ying Ye.
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Determination of abietic acid and dehydroabietic acid
in livestock and poultry muscles for export
1 Scope
This standard specifies the method of determination of abietic acid and dehydroabietic acid in livestock
and poultry muscles for export by HPLC, and confirmation by LC-MS/MS.
This standard is applicable to the determination and confirmation of abietic acid and dehydroabietic
acid in animal origin foodstuffs shuch as chicken, duck, goose meat, pork, beef, mutton etc.
2 Normative reference
The following referenced documents are indispensable for the application of this document.
For dated references, only the edition cited applies.For undated references, the latest edition of the
referenced document (including any amendments) applies.
GB/T 6682 water for analytical laboratory use—Specification and test method
3 Terms and definitions
There is no term or definition to be defined in this document.
4 Principle
The abietic acid and dehydroabietic acid in the test sample is extracted with acetonitrile , thereafter
being cleaned up by C18 purifying agent bonded silica. the contents are determined by HPLC with PDA
(photo-diode array) or UV detector, quantifired by external standard method, and the positive sample
is qualitatively confirmed by LC-MS/MS.
5 Reagent and materials
Unless otherwise specified, all regents used should be of analytical reagent(AR), “water” is the first
grade water prescribed by GB/T 6682.
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5.1 Acetonitrile: Chromatogram grade.
5.2 Methanol: Chromatogram grade.
5.3 Formic acid:Chromatogram grade.
5.4 Phosphoric acid: Guaranteed reagent.
5.5 Magnesium sulfate anhydrous.
5.6 Sodium chloride.
5.7 Trisodium citrate.
5.8 Disodium hydrogen citrate sesquihydrate.
5.9 0.1% Phosphoric acid solution: accurately pipette 1 mL phosphoric acid(5.4) dilute to 1000
mL with water,mix to homogenous.
5.10 Abietic acid: purity(ca 98.0%),the information of abietic acid is listed in Table A.1 in
Annex A.
5.11 Dehydroabietic acid: purity(ca 98.0%),the information of dehydroabietic acid is listed in Table
A.1 in Annex A.
5.12 Abietic acid standard stock solution(1 000 mg/L): Accurately weigh 10 mg of abietic acid
standard (5.10) in a small beaker, dissolve with a small amount methanol(5.2), to scale in the
10 mL volumetric flask then make up to graduation with Methanol, shake. the standard stock solution
should be kept away from light and stored at -18 ℃. The term of validity is 6 months.
5.13 Dehydroabietic acid standard stock solution(1 000 mg/L): Accurately weigh 10 mg of dehydroabietic
acid standard (5.11) in a small beaker, dissolve with a small amount Methanol, to scale
in the 10 mL volumetric flask then make up to graduation with Methanol, shake. the standard stock
solution should be kept away from light and stored at -18 ℃. The term of validity is 6 months.
5.14 Mixed standard intermediates solution(100 μg/mL): Accurately transfer 1.00 mL standard
stock solution(5.12, 5.13)into 10 mL volumetric flask, dilute to the scale with acetonitrile(5.1).
should be kept away from light and stored at -18 ℃. The term of validity is 1 month.
5.15 Mixed matrix standard working solution: Dilute the mixed standard intermediate solution
(5.14) with acetonitrile(5.1), a series of concentration of the standard solution : 0.10 mg/L 、0.20
mg/L 、0.50 mg/L、1.00 mg/L、2.00 mg/L and 5.00 mg/L, Prepare just before use.
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5.16 Silica Gel bonded C18 purifying agent:particle size 50 μm.
5.17 Membrane filter: 0.22 μm, organic type.
6 Apparatus and equipment
6.1 High performance liquid chromatograph : equipped with PDA or UV detector.
6.2 High performance liquid chromatography tandem mass spectrometry: equipped with electrospray
ionization source (ESI).
6.3 Analytical balance: capable of accurately weighing to 0.01 g and 0.1 mg.
6.4 Tissue blender.
6.5 Vortex mixer.
6.6 Centrifuge: not less than 4 000 r/min.
6.7 High speed centrifuge: not less than 10 000 r/min.
6.8 Nitrogen evaporator.
7 Preparation and storage of test sample
7.1 In the course of sample preparation, precaution must be taken avoid the contamination or any
factors which may cause the charge of residue content.
7.2 Take a representative sample(livestock meat samples with skin) of about 500 g , grind and and
homogenize, put into a clean container,divide into two groups, with one part as the sample and the
other as the retention sample. Seal and make labeling. stored at -18 ℃.
8 Procedure
8.1 Extraction
Weigh 5.0 g (accurate 0.01 g) of the test sample into 50 mL centrifuge tube. Add 10 mL acetonitrile
(5.1) and 10 mL water, blend with cap, Vortex for 2 min, then add 4.5 g Magnesium sulfate anhydrous(
5.5), 1 g Sodium chloride(5.6), 0.3 g Trisodium citrate(5.7), 0.7 g Disodium hydrogen citrate
sesquihydrate(5.8), oscillate for 25 min, centrifuge at 4 000 r/min for 5 min after well blend,
for clean up.
8.2 Clean up
Transfer 1 mL the supernatant(8.1) into a 2 mL centrifuge tube which had 100 mg C18(5.16). Vortex
for 2 min, centrifuge at 10 000 r/min for 5 min, then filter the supernatant through 0.22 μm Membrane
filter(5.17) and ready for analysis by HPLC.
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8.3 Determination
8.3.1 Liquid chromatography conditions
As the test result depends on test condition of instrument, so it is not possible to take the general
parameters for the LC chromatography , the parameters should be able to identify the components of
the test group and the other components can be separated effectively, the following parameters can
be used for reference:
HPLC operation conditions are as follows:
a) LC column: C18 LC column (100 mm ×2.1 mm id .), 3.5 μm or equivalent;
b) Mobile phase:A is Phosphoric acid (5.3),B is acetonitrile(5.1),the grade of mobile phase is listed
in table 1.
c) Flow rate: 0.5 mL/min.
d) Injection volumn: 10 μL.
e) Column temperature: 35 ℃.
f) Wave length: abietic acid is 240 nm, dehydroabietic acid is 210 nm.
Table 1—Grade of mobile phase
Time/min A/%(0.1% Phosphoric acid solution) B/%(Acetonitrile)
0 70 30
4 70 30
15 10 90
17 10 90
17.5 70 30
20 70 30
8.3.2 Draw curve of the working solution curve
Base on the determination conditions of 8.3.1, the standard working solution was injected from low
to high, and the chromatographic peak area is taken as the ordinate, and the corresponding solution
concentration is taken as abscissa, draw curve of the standard working solution. The liquid chromatography
of them are shown in figures B.1 and figures B.2 in Annex B.
8.3.3 Qualitation and quantification determination of sample
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Determine the solution of test sample and the standard working solution according to the determination
conditions in 8.3.1.If the retention time of the chromatographic peak of abietic acid and dehydroabietic
acid from the test sample is less than ±2.5% compared with the standard solution, the
presence of abietic acid and dehydroabietic acid in the test sample can be preliminarily confirmed. If
necessary, the positive sample should be qualitatively confirmed by LC-MS/MS (referenced Annex
C).The relative intensities (Expressed as a percentage of intensity relative to the strongest ion
abundance)of abietic acid and dehydroabietic acid from sample transitions shall correspond to those
of mixed matrix standard solution transitions for confirmation, the concentration of mixed matrix
standard solution should be same with those of sample solution. The deviation of the relative abundance
does not exceed the provisions in Table C.2 in Annex C, and the S/N ≥3,it can be judged that
there are abietic acid and dehydroabietic acid in the sample. The multiple reaction monitoring (MRM)
chromatograms of abietic acid and dehydroabietic acid are shown in figures D.1 and D.2 in Annex
D. Select the mixed matrix standard working solution with similar response value for chromatographic
analysis ,the responses of abietic acid and dehydroabietic acid in the mixed matrix standard
working solution should be in the linear range of the instrumental detection, if the concentration of
the test sample exceed the line range, the test sample should be diluted before injection analysis again.
And quantifired by external standard method.
8.4 Blank test
Solvent blank: The operation of the blank test is the same as that described in the method of determination,
but with the omission of sample addition.
Sample blank: The operation of the sample blank test is the same as that described in the method of
determination, but with the omission of abietic acid and dehydroabietic acid addition.
9 Calculation and expression of the result
Calculate the content of abietic acid and dehydroabietic acid in the test sample by LC data processor
or according to the followed formula(1), the blank values should be deducted from the calculation
result:
X=C×V×1 000
m ×1 000
…………………………( 1 )
Where:
X———the content of abietic acid or dehydroabietic acid in the test sample, mg/kg;
C———the concentration of abietic acid or dehydroabietic acid of test sample in the standard
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working solution,
g/L;
V———the final volume of the test sample solution, mL;
m ———the mass of the test sample in final sample solution, g.
three significant figure should be retained.
10 Limit of detection(LOD),limit of quantitation(LOQ) ,recoveries and accuracy
10.1 LOD and LOQ
The LOD of this method for abietic acid or dehydroabietic acid is 0.20 mg/kg, the LOQ for abietic
acid or dehydroabietic acid is 0.50 mg/kg.
10.2 Recoveries and accuracy
This method was carried out in an indoor recovery experiment with different samples (chicken,
duck, goose meat, pork, beef, mutton) as a sample blank for three concentration levels of additive
recovery experiments. Each concentration level conducting 6 times repeated experiments. The
measured range of abietic acid and dehydroabietic acid recovery in Various matrix are shown in table
E.1 in Annex E.
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Annex A
(informative)
The compound information of abietic acid and dehydroabietic acid
The compound name, CAS No., Molecular formula, Structural formula and Molecular
weight of abietic acid and dehydroabietic acid are shown in table A.1.
Table A.1—The compound information of abietic acid and dehydroabietic acid
compound name CAS No. Molecular formula Structural formula Molecular weight
Abietic acid 514-10-3 C20H30O2 302.46
Dehydroabietic acid 1740-19-8 C20H28O2 300.44
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Annex B
(informative)
The liquid chromatograms of abietic acid and dehydroabietic acid standard solution
The liquid chromatography of abietic acid and dehydroabietic acid starndard solution are shown in
figures B.1 and B.2 in Appendix B.
Figure B.1—Liquid chromatograms of abietic acid standard solution(2.50 mg/L)
Figure B.2—Liquid chromatograms of dehydroabietic acid standard solution(2.50 mg/L)
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Annex C
(informative)
LC-MS/MS operating condition1)
1) Non-commercial statement: Referenced conditions in Annex C were obtained from AB 4000Q trap LC-MS/MS.
The equipments and their types involved in the standard method are not related to commercial aims, and it is
encouraged to use equipments of different corporation of different type.
C.1 LC operating conditions
LC operating conditions is flowing:
a) LC column: C18 LC column (100 mm ×2.1 mm id.),3.5 μm, or equivalent.
b) Mobile phase: 0.1% formic acid-acetonitrile(isogradient elution 8 min, 15∶85,V/V).
c) Flow rate: 0.5 mL/min.
d) Column temperature: 35 ℃.
e) Injection volumn: 10 μL.
C.2 MS/MS operating conditions
MS/MS operating conditions is flowing:
a) Capillary voltages :5 000 V.
b) Source temperature :550 ℃.
c) Curtain Gas pressure: 25 psi.
d) Nebulizer Gas pressure: 50 psi.
e) Aux Gas pressure: 50 psi.
f) Mass spectrometry parameters of abietic acid and dehydroabietic acid are shown in table C.1,
the maximum allowable deviation of relative ion abundance are shown in table C.2.
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Table C.1—Mass spectrometry parameters of abietic acid and dehydroabietic acid
compound name
Scanning
model
Parent ion
(m/z)
Danghter ion
(m/z)
de clustering
voltage/V
collision
energy/eV
dwell time/
s
abietic acid ESI+ 303.2
257.3 70.0 19.3 0.2
123.1* 70.0 21.7 0.2
dehydroabietic acid ESI+ 301.0
173.2* 89.4 19.0 0.2
159.2 89.4 24.1 0.2
Note: “*”represents the quantitative ion.
Table C.2—The maximum allowable deviation of relative ion abundance
Relative ion abundance(base peak)/% >50 >20~50 >10~20 ≤10
Allowable relative deviation/% ±20 ±25 ±30 ±50
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Annex D
(informative)
The MRM chromatograms of abietic acid and dehydroabietic acid standard solution
The MRM chromatograms of abietic acid and dehydroabietic acid standard solution are shown in figures
D.1 and figures D.2
a) Qualitative ion chromatogram of abietic acid standard solution
b) Quantitative ion chromatogram of abietic acid standard solution
Table D.1—The MRM chromatograms of abietic acid standard solution(0.50 mg/L)
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a) Quantitative ion chromatogram of dehydroabietic acid standard solution
b) Qualitative ion chromatogram of dehydroabietic acid standard solution
Table D.2—The MRM chromatograms of dehydroabietic acid standard solution(0.50 mg/L)
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Annex E
(informative)
Range of recovery and RSDs for abietic acid and dehydroabietic
acid in livestock and poultry muscles(n=6)
Range of recovery and RSDs for abietic acid and dehydroabietic acid in livestock and poultry muscles
are shown in table E.1.
Table E.1—Range of recovery and RSDs for abietic acid and dehydroabietic acid
in livestock and poultry muscles(n=6)
Matrix Spiked level/(mg/kg)
Range of recovery/% RSD/%
abietic acid dehydroabietic acid abietic acid dehydroabietic acid
chicken
0.500 84.4~99.6 104~107 6.61 1.34
1.00 94.0~106 96.2~108 4.06 4.55
5.00 89.4~95.2 87.6~90.6 2.21 1.56
duck
0.500 98.0~101 96.0~102 1.16 2.01
1.00 86.2~107 86.0~97.6 7.47 5.21
5.00 83.6~91.8 86.9~89.4 3.81 1.20
goose meat
0.500 90.8~109 91.2~104 8.00 5.66
1.00 94.0~106 88.6~97.0 5.14 3.40
5.00 88.8~93.0 89.0~95.2 2.23 2.48
pork
0.500 93.2~102 87.2~98.0 3.71 4.38
1.00 93.6~106 92.6~106 5.12 5.01
5.00 90.0~94.4 85.4~90.6 1.70 2.28
beef
0.500 96.4~100 88.0~98.8 1.37 4.26
1.00 84.6~92.4 88.8~90.8 3.52 0.73
5.00 85.0~93.6 90.2~98.8 3.24 4.17
mutton
0.500 90.0~106 100~104 7.90 1.78
1.00 96.4~110 99.6~104 5.73 1.83
5.00 86.6~94.2 89.6~94.0 3.05 1.95
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